Pepper plant having multiple flowers per node

Abstract

The present invention relates to a Capsicum annuum plant comprising QTL MF2.1 and QTL MF2.2 conferring multiple flowers per node. The present invention further relates to a seed produced by the plant according to the present invention, a seed from which a plant according to present invention can be grown, a fruit produced by a plant according to the present invention and a part of a plant according to the present invention. The present invention further relates to a method of identifying and / or selecting a plant or plant part according to the present invention. The present invention further relates to a method for producing a Capsicum annuum plant having the multiple flowers per node phenotype according to the present invention. The present invention further relates to a method for increasing the number of flowers per node in a Capsicum annuum plant and the use of QTL MF2.1 and QTL MF2.2 for increasing the number of flowers per node in a Capsicum annuum plant. The present invention further relates to genetic markers specific for the QTLs according to the present invention and the use thereof for selecting a Capsicum annuum plant having an increased number of flowers per node.

Description

[0001] Nunhems Netherlands B.V. 230055WO011 PEPPER PLANT HAVING MULTIPLE FLOWERS PER NODEFIELD OF THE INVENTION[1] This invention relates to the field of plant breeding. Provided is a Capsicum annuum plantcomprising QTL MF2.1 and QTL MF2.2 conferring multiple flowers per node relative to a plant lacking said QTL MF2.1 and QTL MF2.2. Further provided is a seed produced by the plant ac- cording to the present invention, a seed from which a plant according to present invention can be grown, a fruit produced by a plant according to the present invention and a part of a plant accord- ing to the present invention. The present invention further provides a method of identifying and / or selecting a plant or plant part according to the present invention. Further provided is a method forproducing a Capsicum annuum plant having the multiple flowers per node phenotype accordingto the present invention. Further provides is a method for increasing the number of flowers pernode in a Capsicum annuum plant and the use of QTL MF2.1 and QTL MF2.2 for increasing thenumber of flowers per node in a Capsicum annuum plant. Further provided are genetic markersspecific for the QTLs according to the present invention and the use thereof for selecting a Cap-sicum annuum plant having an increased number of flowers per node.BACKGROUND[2] Capsicum annuum plants are herbaceous plants of the family Solanaceae. The plantreaches about 0.5–1.5 meters (20–60 in). Single white flowers bear the pepper fruit which is green when unripe, changing principally to red, although some varieties may ripen to brown or purple. While the species can tolerate most climates, they are especially productive in warm and dryclimates. Pepper plants of the species Capsicum annuum include different types of peppers, suchas bell peppers, cayenne peppers, paprika, and jalapeños.[3] The yield of pepper fruits is one of the most important characteristic pepper plants and ismainly influenced by the number of flowers per node. Capsicum annuum plants produce a singleflower per node, whereas other species of the genus Capsicum, such as Capsicum chinense,produce several flowers per node. The genetic mechanism underlying the control of the floweringtraits in Capsicum remains largely unknown. Zhu et al. (2019) Scientific Reports 9:1054 doi:10.1038 / s41598-018-38370-0 describe the construction of a high-density genetic map of an in-terspecific cross of Capsicum chinense and Capsicum annuum and QTL analysis of floraltraits.Kim et al (2022) Frontiers in Plant Science 13:884338 doi: 10.3389 / fpls.2022.884338 de-scribe the identification of genetic factors controlling the formation of multiple flowers per node inpepper. Although these studies succeeded in identifying QTLs controlling floral traits in Capsicumchinense, such as flowering time and flower number per node, the genetic incompatibility betweenCapsicum chinense and Capsicum annuum so far precluded the development of a Capsicumannuum plant having multiple flowers per node.[4] There is a constant need to improve the yield of Capsicum annuum cultivars. It is thereforean object of the invention to provide new flowering trait QTLs that confer multiple flowers pernode, relative to a Capsicum annuum plant lacking said QTLs and / or having an improved pepperfruit yield. It is a further object of the invention to provide a Capsicum annuum plant, and cells,tissues, fruits and other parts of such plant comprising in their genome QTLs capable of conferringmultiple flowers per node and / or having an improved pepper fruit yield. It is a further object of theinvention to provide a method of identifying and / or selecting a plant or plant part comprising inNunhems Netherlands B.V. 230055WO012their genome QTLs capable of conferring multiple flowers per node and / or an improved pepperfruit yield. It is a further object of the invention to provide a method for producing a Capsicumannuum plants comprising in their genome QTLs capable of conferring multiple flowers per nodeand / or an improved pepper fruit yield. It is a further object of the invention to provide a method forincreasing the number of flowers per node and / or to improve pepper fruit yield. It is a further objectof the invention to provide genetic markers specific for QTLs capable of conferring multiple flowersper node, relative to a Capsicum annuum plant lacking said QTLs and / or conferring an improvedpepper fruit yield. SUMMARY OF INVENTION[5] The present invention provides a Capsicum annuum plant comprising in its genome QTLMF2.1 located between 158,316,753 bp and 163,630,465 bp on chromosome 2 of C. annuumDempsey V1.1 reference genome; and QTL MF2.2 located between 137,347,441 bp and151,147,982 bp on chromosome 2 of C. annuum Dempsey V1.1 reference genome, wherein saidQTL MF2.1 and QTL MF2.2 confer multiple flowers per node relative to a plant lacking said QTLMF2.1 and QTL MF2.2.[6] The present invention further provides a seed produced by the Capsicum annuum plantaccording to the present invention, wherein the seed comprises QTL MF2.1 and QTL MF2.2 as described herein. The present invention further provides a seed from which the Capsicum an-nuum plant according to the present invention can be grown. The present invention further pro-vides a plant cell, tissue or plant part of the Capsicum annuum plant according to the presentinvention or of the seed according to the present invention, comprising QTL MF2.1 and QTLMF2.2 as described herein. The present invention further provides a haploid plant or dihaploidplant derived from the Capsicum annuum plant as provided by the present invention.[7] In addition, the present invention provides a method for identifying and / or selecting a Cap-sicum annuum plant or plant part comprising determining whether said plant or plant part com-prises in its genome QTL MF2.1 and QTL MF2.2 according to the present invention or the recom- binant introgression on chromosome 2 according to the present invention and optionally deter-mining whether said plant or plant part comprises in its genome the QTL ft12.1 on chromosome12 according to the present invention.[8] In addition, the present invention provides a method for producing a Capsicum annuumplant having a multiple flowers per node phenotype, said method comprising the step(s) of: (i)crossing a first Capsicum annuum plant and a second Capsicum plant, wherein the first Capsicumannuum plant is a plant according to the present invention comprising in its genome QTL MF2.1and QTL MF2.2 as described herein or the recombinant introgression on chromosome 2 compris-ing QTL MF2.1 and QTL MF2.2 as described herein; (ii) optionally harvesting seed from the cross-ing of (i) and selecting seed comprising said QTL MF2.1 and QTL MF2.2 or said recombinantintrogression on chromosome 2 comprising QTL MF2.1 and QTL MF2.2 in its genome.[9] In addition, the present invention provides a method for increasing the number of flowersper node in a Capsicum annuum plant, said method comprising introgressing QTL MF2.1 andQTL MF2.2 according to the present invention or the recombinant introgression on chromosome2 according to the present invention into said Capsicum annuum plant.Nunhems Netherlands B.V. 230055WO013

[0010] In addition, the present invention provides the use of QTL MF2.1 and QTL MF2.2 accordingto the present invention or the recombinant introgression on chromosome 2 according to the pre-sent invention for increasing the number of flowers per node in a Capsicum annuum plant.

[0011] In addition, the present invention provides the use of a genetic marker specific for QTLMF2.1 and QTL MF2.2 according to the present invention or the recombinant introgression onchromosome 2 according to the present invention for selecting a Capsicum annuum plant havingan increased number of flowers per node phenotype.BRIEF DESCRIPTION OF THE FIGURES

[0012] Figure 1: Capsicum annuum plant showing the multiple flowers per node phenotype of thepresent invention. DETAILED DESCRIPTION OF THE INVENTION

[0013] The present invention provides Capsicum annuum plants and plant parts thereof, whereinsaid plants have a multiple flowers per node phenotype.

[0014] The present invention provides Capsicum annuum plant comprising in its genome QTLMF2.1 located between 158,316,753 bp and 163,630,465 bp on chromosome 2 of C. annuumDempsey V1.1 reference genome; and QTL MF2.2 located between 137,347,441 bp and151,147,982 bp on chromosome 2 of C. annuum Dempsey V1.1 reference genome, wherein saidQTL MF2.1 and QTL MF2.2 confer multiple flowers per node relative to a plant lacking said QTLMF2.1 and QTL MF2.2.

[0015] The inventors found that Capsicum annuum plant comprising in its genome QTL MF2.1located between 158,316,753 bp and 163,630,465 bp on chromosome 2 of C. annuum DempseyV1.1 reference genome; and QTL MF2.2 located between 137,347,441 bp and 151,147,982 bpon chromosome 2 of C. annuum Dempsey V1.1 reference genome, show multiple flowers pernode relative to a plant lacking said QTL MF2.1 and QTL MF2.2. It was surprisingly found thatboth QTL MF2.1 and QTL MF2.2 as described herein are required to obtain the multiple flowerstrait according to the present invention. A Capsicum annuum plant comprising either QTL MF2.1or QTL MF2.2 do not show the desired increased number of flowers per node phenotype.

[0016] QTLs controlling floral traits, such as flowering time and flower number per node, in Capsi-cum chinense and Capsicum annuum have been described in the prior art. Kim (2021) http: / / dcol-lection.snu.ac.kr / common / orgView / 000000157331 describes the generation of 85 recombinantinbred lines between C. annuum “TF68” and C. chinense “Habanero”, which allowed the identifi-cation of four new QTLs on chromosome 1, 2, 7 and 11 for multiple-flower per node trait. Of saidfour QTLs, the QTL on chromosome 2, named as “TH-mf-2”, is described to be located at position128.6-139.6 Mbp using the C. annuum “Dempsey” reference genome. Zhu et al. (2019) Loc cit.describe the construction of a molecular genetic linkage map from 150 F2 populations from aninterspecific cross between the inbred lines 740 (C. chinense) and CA1 (C. annuum). Amongothers, three flower number per node QTLs were identified on chromosome 2, 7 and 10, respec-tively. Of said three QTLs, the QTL on chromosome 2, named as “Mf2.1I” was mapped to0.382 cM, and the interval physically represents approximately 1400 kb in the Zunla-1 referencegenome. The prior art fails to describe a Capsicum annuum plant comprising in its genomeNunhems Netherlands B.V. 230055WO014specifically QTL MF2.1 and QTL MF2.2 as further described herein, wherein said QTL MF2.1 islocated between 158,316,753 bp and 163,630,465 bp on chromosome 2 of C. annuum DempseyV1.1 reference genome and said QTL MF2.2 is located between 137,347,441 bp and 151,147,982bp on chromosome 2 of C. annuum Dempsey V1.1 reference genome. The prior art also fails tosuggest that specifically QTL MF2.1 and QTL MF2.2 as further described herein must be selectedfrom the numerous QTLs described in the prior art to be associated to flower number per node inpepper to provide a Capsicum annuum plant showing the multiple flowers per node phenotype ofthe present invention. It was previously not possible to elucidate the underlying mechanisms thatcontrol multiple flowers in pepper and to develop reliable markers for maker-assisted selection topyramid the genes that control the multiple flower trait into Capsicum annuum plants due to com-plex underlying genetics and the segregation distortion that is common in interspecific cross pop-ulations between C. chinense and C. annuum.

[0017] The term “number of flowers per node” refers to the average number of flowers which areproduced by a given plant in a segment of said plant which lies between two adjacent nodes. Theterm “increased number of flowers per node phenotype” or “multiple flowers per node phenotype”or “multiple flowers phenotype” refers to a plant phenotype which comprises a (statistically signif-icant) increased number of flowers per node when compared to an appropriate control, e.g. a wildtype Capsicum annuum plant, not comprising QTL MF2.1 and QTL MF2.2 as described herein.In such a wild type Capsicum annuum plant the number of flowers per node is about 1. It isunderstood that comparisons between different plant lines involves growing a number of plants of a line (or variety) (e.g. at least 5 plants, preferably at least 10 plants per line) under the same conditions as the plants of one or more control plant lines (preferably wild type plants) and the determination of differences, preferably statistically significant differences, between the plant lines when grown under the same environmental conditions. Preferably the plants are of the same line or variety, such as isogenic plants.

[0018] The term "isogenic plant" refers to two plants which are genetically identical except for theallele to be investigated, such as the multiple flowers allele of the present invention. In order to investigate the impact of a flowering trait, accordingly, one can cross a plant line (or variety) of interest with a plant comprising the multiple flowers allele and select for progeny expressing the desired trait. Optionally one may have to self the progeny one or more times to be able to deter- mine the genetic determinants for the multiple flowers phenotype. Said progeny can then be back- crossed (at least 2 times, e.g.3, 4, or preferably 5 or 6 times) with the plant line (or variety) of interest while selecting for progeny having the same phenotype as the plant line (or variety) of interest and expressing the genetic determinants for the multiple flowers phenotype. The impact of the multiple flowers allele causing the multiple flowers phenotype can then be compared be- tween the plant line (variety) of interest and its isogenic line not comprising the genetic determi- nants for the multiple flowers phenotype.

[0019] The term “multiple flowers allele” refers to the alleles of QTL MF2.1 and QTL MF2.2 asdescribed in more detail herein, which in combination cause the multiple flowers phenotype ac- cording to the present invention.

[0020] The term "genome" relates to the genetic material of an organism. It consists of DNA. Thegenome includes both the genes and the non-coding sequences of the DNA.Nunhems Netherlands B.V. 230055WO015

[0021] The term "gene" means a (genomic) DNA sequence comprising a region (transcribed re-gion), which is transcribed into a messenger RNA molecule (mRNA) in a cell, and an operably linked regulatory region (also described herein as regulatory sequence, e.g. a promoter). A gene may thus comprise several operably linked sequences, such as a promoter, a 5' leader sequence comprising e.g. sequences involved in translation initiation, a (protein) coding region (cDNA or genomic DNA) and a 3' non-translated sequence comprising e.g. transcription termination sites. Different alleles of a gene are thus different alternative forms of the gene, which may be in the form of e.g. differences in one or more nucleotides of the genomic DNA sequence (e.g. in the promoter sequence, the exon sequences, intron sequences, etc.), mRNA and / or amino acid se- quence of the encoded protein. A gene may be an endogenous gene (in the species of origin) or a chimeric gene (e.g. a transgene or cis-gene). The "promoter" of a gene sequence is defined as a region of DNA that initiates transcription of a particular gene. Promoters are located near the genes they transcribe, on the same strand and upstream on the DNA. Promoters can be about 100-1000 base pairs long. In one aspect the promoter is defined as the region of about 1000 base pairs or more e.g. about 1500 or 2000, upstream of the start codon (i.e. ATG) of the protein encoded by the gene.

[0022] "Expression of a gene" refers to a process wherein a DNA region, which is operably linkedto appropriate regulatory regions, particularly a promoter, is transcribed into an RNA, which isbiologically active, i.e. which is capable of being translated into a biologically active protein orpeptide or which is active itself (e.g. in posttranscriptional gene silencing or RNAi). The coding sequence may be in sense-orientation and encodes a desired, biologically active protein or pep- tide.

[0023] The terms "protein" and "polypeptide" are used interchangeably and refer to molecules con-sisting of a chain of amino acids, without reference to a specific mode of action, size, 3 -dimen-sional structure or origin. A "fragment" or "portion" of a protein may thus still be referred to as a"protein". An "isolated protein" is used to refer to a protein which is no longer in its natural envi- ronment, for example in vitro or in a recombinant bacterial or plant host cell.

[0024] The terms “peptide sequence” and “amino acid sequence” refer to the primary amino acidsequence of a protein or polypeptide.

[0025] The term "locus" (plural loci) means a specific place or places or a site on a chromosomewhere for example a gene or genetic marker is found.

[0026] As is used herein, a QTL (quantitative trait locus) is a hereditary unit (often indicated by oneor more molecular markers) that occupies a specific location on a chromosome and that contains the genetic instruction for a particular phenotypic characteristics or trait in a plant. In contrast to a gene, the exact boundaries of a QTL are not known, but can be found without undue burden by a person skilled in the art by using fine mapping techniques well known in the art of genetic map- ping and subsequent DNA sequencing routines. The QTL encodes at least one gene of which the expression, alone or in combination with other genes, results in the phenotypic trait being ex- pressed, or that encodes at least one regulatory region that controls the expression of at least one gene the expression of which, alone or in combination with other genes, results in the phe- notypic trait being expressed. A QTL may be defined by indicating its genetic location in theNunhems Netherlands B.V. 230055WO016 genome of the donor of the introgression that contains the QTL using one or more molecular markers. These one or more markers, in turn, indicate a specific locus.

[0027] The term "allele(s)" means any of one or more alternative forms of a gene at a particularlocus, all of which alleles relate to one trait or characteristic at a specific locus. In a diploid cell of an organism, alleles of a given gene are located at a specific location, or locus (loci plural) on a chromosome. One allele is present on each chromosome of the pair of homologous chromo- somes. A diploid plant species may comprise a large number of different alleles at a particular locus. These may be identical alleles of the gene (homozygous) or two different alleles (hetero- zygous).

[0028] An allelism test is a test known in the art that can be used to identify whether two genesconferring the same trait are located at the same locus.

[0029] The word "trait" in the context of this application refers to the phenotype of the plant. Whena plant shows the traits of the invention, its genome comprises the genetic determinant causing the trait of the invention. The plant, thus, comprises a genetic determinant according to the inven- tion, wherein said genetic determinant may be one or more of the specific QTLs as described herein, the recombinant introgression as described herein, the mutant allele as described herein, or the mutant gene as described herein. It is understood that when referring to a plant comprisingthe trait of the plant of the invention, reference is made to a Capsicum annuum plant comprisingthe trait of multiple flowers per node. i.e. an increased number of flowers per node relative to aCapsicum annuum plant lacking said genetic determinant and / or the trait of an improved pepperfruit yield.

[0030] "Average" refers herein to the arithmetic mean.

[0031] As used herein, the term “plant” includes the whole plant or any parts or derivatives thereof,such as plant organs (e.g., harvested or non-harvested fruits, leaves, seed, flowers, etc.), plant cells, plant protoplasts, plant cell or tissue cultures from which whole plants can be regenerated, plant calli, plant cell clumps, and plant cells that are intact in plants, or parts of plants, such as embryos, pollen, ovules, ovaries, fruits (e.g., harvested tissues or organs, such as harvested pep- per fruits or parts thereof), flowers, leaves, seeds, clonally propagated plants, roots, root-stocks, stems, root tips and the like. Also any developmental stage is included, such as seedlings, imma- ture and mature, etc.

[0032] A "plant line" or "breeding line" refers to a plant and its progeny. As used herein, the term"inbred line" refers to a plant line which has been repeatedly selfed, preferably more than three time, more preferably more than 6 times.

[0033] In one preferred aspect, the Capsicum annuum plant is a cultivated Capsicum annuumplant. The term "cultivar" (or “cultivated” plant”) is used herein to denote a plant having a biological status other than a "wild" status, which "wild" status indicates the original non-cultivated, non- domesticated, or natural state of a plant or accession, and the term cultivated does not include such wild, or weedy plants. The term cultivar does include material with good agronomic charac- teristics, such as breeding material, research material, breeding lines, elite breeding lines, syn- thetic population, hybrid, founder stock / base population, inbred lines, cultivars (open pollinated of hybrid cultivar), segregating population, mutant / genetic stock, and advanced / improved cultivar.Nunhems Netherlands B.V. 230055WO017 The so-called heirloom varieties or cultivars, i.e. open pollinated varieties or cultivars commonly grown during earlier periods in human history and often adapted to specific geographic regions,are in one aspect of the invention encompassed herein as cultivated plants. In one aspect theterm cultivar also includes landraces, i.e. plants (or populations) selected and cultivated locally by humans over many years and adapted to a specific geographic environment and sharing a common gene pool.

[0034] The term "hybrid" plant (or hybrid seed) refers to a plant or seed obtained from crossing twoinbred parent lines. The term "F1 hybrid" plant (or "F1 hybrid" seed or "F1 seed") refers to a first- generation plant or seed obtained from crossing two inbred parent lines.

[0035] "Plant variety" is a group of plants within the same botanical taxon of the lowest gradeknown, which (irrespective of whether the conditions for the recognition of plant breeder’s rights are fulfilled or not) can be defined on the basis of the expression of characteristics that result froma certain genotype or a combination of genotypes, can be distinguished from any other group ofplants by the expression of at least one of those characteristics, and can be regarded as an entity, because it can be multiplied without any change. Therefore, the term “plant variety” cannot be used to denote a group of plants, even if they are of the same kind, if they are all characterized by the presence of one locus or gene (or a series of phenotypical characteristics due to this single locus or gene), but which can otherwise differ from one another enormously as regards the other loci or genes.

[0036] "Backcrossing" refers to a breeding method by which a (single) trait, such as the multipleflowering trait of the present invention, can be transferred from one genetic background (also referred to as "donor" generally, but not necessarily, this is an inferior genetic background) into another genetic background (also referred to as "recurrent parent"; generally, but not necessarily, this is a superior genetic background). An offspring of a cross (e.g. an F1 plant obtained by cross- ing a first plant of a certain plant species comprising the genetic determinant of the present in- vention with a second plant of the same plant species or of a different plant species that can be crossed with said first plant species wherein said second plant species does not comprise the genetic determinant of the present invention; or an F2 plant or F3 plant, etc., obtained by selfing the F1) is "backcrossed" to a parent plant of said second plant species. After repeated backcross- ing, the trait of the donor genetic background, e.g. the genetic determinant conferring the multiple flowers phenotype, will have been incorporated into the recurrent genetic background. The terms "gene converted" or "conversion plant" or "single locus conversion" in this context refer to plants which are developed by backcrossing wherein essentially all of the desired morphological and / or physiological characteristics of the recurrent parent are recovered in addition to the one or more genes transferred from the donor parent. The plants grown from the seeds produced by back-crossing of the F1 plants with the second parent plant line is referred to as the "BC1F1 genera-tion". Plants from the BC1F1 population may be selfed resulting in the BC1F2 generation or back-crossed again with the cultivated parent plant line to provide the BC2F1 generation. The terms"F1, F2, etc." refer to the consecutive related generations following a cross between two parent plants or parent lines. The plants grown from the seeds produced by crossing two plants or lines is called the F1 generation. Selfing the F1 plants results in the F2 generation, etc. An "M1 popu- lation" is a plurality of mutagenized seeds / plants of a certain plant line. "M2, M3, M4, etc." refers to the consecutive generations obtained following selfing of a first mutagenized seed / plant (M1).Nunhems Netherlands B.V. 230055WO018

[0037] The term Solanaceae refers to a family of plants, which include genera (especially the ge-nus Solanum and the genus Capsicum) that comprise fruit and vegetable species which are andbred by humans, such as e.g. Solanum lycopersicum (tomato), Capsicum annuum (pepper), So-lanum melongena (eggplant) and Solanum muricatum (pepino). "Solanaceous plants" or "plantsof the family Solanaceae" are plants of the botanical family Solanaceae, i.e. any plant of the family Solanaceae, including wild solanaceous plants and cultivated solanaceous plants. The botanicalfamily Solanaceae consists about 98 genera of which the genera Solanum and Capsicum are thecommercially most relevant as they comprise many domesticated species that are widely culti- vated and used as food crops with high economic importance.

[0038] The genus Capsicum consists of 20 to 27 species, five of which are domesticated: C. an-nuum, C. baccatum, C. chinense, C. frutescens, and C. pubescens. Phylogenetic relationships between species have been investigated using bio-geographical, morphological, chemosystem- atic, hybridization, and genetic data. Fruits of Capsicum, often named as "peppers" or "pepper fruits", can vary tremendously in colour, shape, and size both between and within species. Chem- osystematic studies helped distinguish the difference between varieties and species.

[0039] Capsicum annuum L. plants are herbaceous plants of the family Solanaceae that are ofparticular relevance in the context of the present invention. Capsicum annuum plants reach about0.5-1.5 meters (about 20-60 inches). Single white flowers bear the pepper fruit which is green when unripe, changing principally to red, although some varieties may ripen to brown or purple.While the species can tolerate most climates, they are especially productive in warm and dryclimates. Cultivated plants of the species Capsicum annuum include different types of peppers,such as bell peppers, cayenne peppers, paprika, and jalapeños. "Capsicum annuum chromo-some 2" refers to the Capsicum annuum chromosome 2, as known in the art (see Capsicumannuum cv CM334 genome chromosomes (release 1.55) and Capsicum annuum UCD10X ge-nome chromosomes (v1.0) and Capsicum annuum Dempsey genome chromosomes (v1.0)."Orthologous chromosome 2" refers to the corresponding chromosome of relatives of Capsicumannuum. Analogous, "Capsicum annuum chromosome 12" refers to the Capsicum annuum chro-mosome 12, as known in the art (see Capsicum annuum cv CM334 genome chromosomes (re- lease 1.55) and Capsicum annuum UCD10X genome chromosomes (v1.0) and Capsicum an-nuum Dempsey genome chromosomes (v1.0). As used herein, the term “Capsicum annuumplant” refers to any plant that belongs to the botanical species Capsicum annuum, and accordinglymay comprise introgression fragments from other plant species of the genus Capsicum, including,but not limited to C. baccatum, C. chinense, C. frutescens, and C. pubescens, allowing for theincorporation of desirable traits associated with said other plant species from the genus Capsi-cum, such as enhanced flavor profiles or resistance to specific pests and diseases. It is readilyapparent to a person skilled in the art that the term “Capsicum annuum plant” does not compriseinterspecific hybrids between Capsicum annuum and other Capsicum species, such as Capsicumchinense. This is because such an interspecific hybrid between Capsicum annuum and Capsicumchinense often is sterile, e.g. due to chromosomal incompatibility. Such chromosomal incompati-bilities are also known as crossing barriers playing a major role in differentiating species from oneto another; see e.g. Lanteri & Pickersgill (1993) Euphitica 67:155-160 and Wu et al (2009) TheorAppl Genet 118:1279-1293. In one embodiment, the Capsicum annuum plant of the present in-vention comprises no more than 40 % of the genome of an other plant species of the genusCapsicum, such as Capsicum chinense, preferably no more than 24 % of the genome of an otherplant species of the genus Capsicum, such as Capsicum chinense, more preferably no more thanNunhems Netherlands B.V. 230055WO019 12 % of the genome of an other plant species of the genus Capsicum, such as Capsicum chinense, even more preferably no more than 6 % of the genome of an other plant species of thegenus Capsicum, such as Capsicum chinense, and most preferably no more than 3 % of thegenome of an other plant species of the genus Capsicum, such as Capsicum chinense.

[0040] QTL MF2.1 as further described herein and QTL MF2.2 as further described herein accord-ingly in combination are capable of conferring a multiple flowers per node phenotype in a Capsi-cum annuum plant.

[0041] QTL MF2.1 as provided by the present invention, and which is capable of conferring multipleflowers per node when comprised in the genome of a Capsicum annuum plant together with QTLMF2.2, is located between 158,316,753 bp and 163,630,465 bp on chromosome 2 of C. annuumDempsey V1.1 reference genome. This means that in the context of the present invention it waspossible to map the locus of QTL MF2.1 as comprised on chromosome 2 to a specific region ofsaid chromosome 2, wherein said region of chromosome 2 is defined by the flanking markersSNP_1 and SNP_2 as further described herein. In one aspect, QTL MF2.1 as comprised in theCapsicum annuum plant according to the present invention comprises an adenine genotype forSNP marker SNP_1 at a position corresponding to position 51 in SEQ ID NO: 9 or at position 51of a sequence comprising at least 95% (more preferably at least 96%, at least 97%, at least 98%,at least 99%, or even 100%) identity to SEQ ID NO: 9 and / or cytosine genotype for SNP markerSNP_2 at a position corresponding to position 51 in SEQ ID NO: 11 or at position 51 of a sequencecomprising at least 95% (more preferably at least 96%, at least 97%, at least 98%, at least 99%,or even 100%) identity to SEQ ID NO: 11.

[0042] Preferably, QTL MF2.1 as provided by the present invention, and which is capable of con-ferring multiple flowers per node when comprised in the genome of a Capsicum annuum planttogether with QTL MF2.2, is located between 158,316,753 bp and 162,234,480 bp on chromo-some 2 of C. annuum Dempsey V1.1 reference genome. This means that in the context of thepresent invention it was possible to fine-map the locus of QTL MF2.1 as comprised on chromo-some 2 to an even more specific region of said chromosome 2, wherein said region of chromo-some 2 is defined by the flanking markers SNP_1 and SNP_7 as further described herein. In oneaspect, QTL MF2.1 as comprised in the Capsicum annuum plant according to the present inven-tion comprises an adenine genotype for SNP marker SNP_1 at a position corresponding to posi- tion 51 in SEQ ID NO: 9 or at position 51 of a sequence comprising at least 95% (more preferablyat least 96%, at least 97%, at least 98%, at least 99%, or even 100%) identity to SEQ ID NO: 9and / or guanine genotype for SNP marker SNP_7 at a position corresponding to position 51 inSEQ ID NO: 25 or at position 51 of a sequence comprising at least 95% (more preferably at least96%, at least 97%, at least 98%, at least 99%, or even 100%) identity to SEQ ID NO: 25.

[0043] QTL MF2.2 as provided by the present invention, and which is capable of conferring multipleflowers per node when comprised in the genome of a Capsicum annuum plant together with QTLMF2.1, is located between 137,347,441 bp and 151,147,982 on chromosome 2 of C. annuumDempsey V1.1 reference genome. This means that in the context of the present invention it waspossible to map the locus of QTL MF2.2 as comprised on chromosome 2 to a specific region of said chromosome 2, wherein said region of chromosome 2 is defined by the flanking markersSNP_3 and SNP_4 as further described herein. In one aspect, QTL MF2.2 as comprised in theCapsicum annuum plant according to the present invention comprises an adenine genotype forNunhems Netherlands B.V. 230055WO0110 SNP marker SNP_3 at a position corresponding to position 101 in SEQ ID NO: 13 or at position 101 of a sequence comprising at least 95% (more preferably at least 96%, at least 97%, at least98%, at least 99%, or even 100%) identity to SEQ ID NO: 13 and / or cytosine genotype for SNPmarker SNP_4 at a position corresponding to position 101 in SEQ ID NO: 15 or at position 101 ofa sequence comprising at least 95% (more preferably at least 96%, at least 97%, at least 98%,at least 99%, or even 100%) identity to SEQ ID NO: 15.

[0044] Preferably, QTL MF2.2 as provided by the present invention, and which is capable of con-ferring multiple flowers per node when comprised in the genome of a Capsicum annuum planttogether with QTL MF2.1, is located between 143,111,884 bp and 147,107,830 on chromosome2 of C. annuum Dempsey V1.1 reference genome. This means that in the context of the presentinvention it was possible to fine-map the locus of QTL MF2.2 as comprised on chromosome 2 toan even more specific region of said chromosome 2, wherein said region of chromosome 2 isdefined by the flanking markers SNP_8 and SNP_9 as further described herein. In one aspect,QTL MF2.2 as comprised in the Capsicum annuum plant according to the present invention com-prises an cytosine genotype for SNP marker SNP_8 at a position corresponding to position 51 inSEQ ID NO: 27 or at position 51 of a sequence comprising at least 95% (more preferably at least96%, at least 97%, at least 98%, at least 99%, or even 100%) identity to SEQ ID NO: 27 and / orguanine genotype for SNP marker SNP_9 at a position corresponding to position 51 in SEQ IDNO: 29 or at position 51 of a sequence comprising at least 95% (more preferably at least 96%, atleast 97%, at least 98%, at least 99%, or even 100%) identity to SEQ ID NO: 29.

[0045] “Sequence identity" or “identity” or "sequence similarity" can be determined by alignment oftwo peptide or two nucleotide sequences using global or local alignment algorithms. Sequences may then be referred to as "substantially identical" or "essentially similar" when they share at least a certain minimal percentage of sequence identity (as defined further below) after optimally align- ment by, for example, the program GAP or BESTFIT or the Emboss program "Needle" (using default parameters, see below). These programs use the Needleman and Wunsch global align- ment algorithm to align two sequences over their entire length, maximizing the number of matches and minimizing the number of gaps. Generally, the default parameters are used, with a gap cre- ation penalty = 10 and gap extension penalty = 0.5 (both for nucleotide and protein alignments). For nucleotides the default scoring matrix used is DNAFULL and for proteins the default scoringmatrix is Blosum62 (Henikoff & Henikoff, 1992, PNAS 89, 10915- 10919). Sequence alignmentsand scores for percentage sequence identity may for example be determined using computer programs, such as EMBOSS (http: / / www.ebi.ac.uk / Tools / psa / emboss_needle / ). Alternatively, se- quence similarity or identity may be determined by searching against databases such as FASTA, BLAST, etc. Hits are preferably aligned pairwise to compare sequence identity, preferably over the full length of the sequences.

[0046] As used herein, two nucleotide sequences have "substantial sequence identity" if the per-centage sequence identity is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 98.3%, 98.7%, 99.0%, 99.2%, 99.3%, 99.5%,99.7%, 99.9% or even 100%, preferably as determined over their entire lengths (as determined by Emboss "needle" using default parameters, i.e. gap creation penalty = 10, gap extension pen- alty = 0.5, using scoring matrix DNAFULL for nucleic acids).Nunhems Netherlands B.V. 230055WO0111

[0047] The term "hybridisation" as used herein is used to indicate hybridisation of nucleic acids atappropriate conditions of stringency as would be readily evident to those skilled in the art depend-ing upon the nature of the probe sequence and target sequences. Conditions of hybridisation andwashing are well known in the art, and the adjustment of conditions depending upon the desired stringency by varying incubation time, temperature and / or ionic strength of the solution are readily accomplished. See, for example, Sambrook, J. et al., Molecular Cloning: A Laboratory Manual, 2nd edition, Cold Spring Harbor Press, Cold Spring Harbor, New York, 1989. The choice of con-ditions is dictated by the length of the sequences being hybridised, in particular, the length of theprobe sequence, the relative G-C content of the nucleic acids and the amount of mismatches to be permitted. Low stringency conditions are preferred when partial hybridisation between strands that have lesser degrees of complementarity is desired. When perfect or near perfect comple- mentarity is desired, high stringency conditions are preferred. When reference is made to a nu- cleic acid sequence (e.g. DNA or genomic DNA) having "substantial sequence identity to" a ref- erence sequence or having a sequence identity of at least 80%, e.g. at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,98.3%, 98.7%, 99.0%, 99.2%, 99.3%, 99.5%, 99.7%, 99.9% or even 100% nucleic acid sequenceidentity to a reference sequence, in one aspect said nucleotide sequence is considered substan-tially identical to the given nucleotide sequence and can be identified using stringent hybridisation conditions. In another aspect, the nucleic acid sequence comprises one or more mutations com- pared to the given nucleotide sequence but still can be identified using stringent hybridization conditions.

[0048] "Stringent hybridisation conditions" can be used to identify nucleotide sequences, which aresubstantially identical to a given nucleotide sequence. Stringent conditions are sequence depend- ent and will be different in different circumstances. Generally, stringent conditions are selected to be about 5°C lower than the thermal melting point (Tm) for the specific sequences at a defined ionic strength and pH. The Tm is the temperature (under defined ionic strength and pH) at which 50% of the target sequence hybridises to a perfectly matched probe. Typically stringent conditions will be chosen in which the salt concentration is about 0.02 molar at pH 7 and the temperature is at least 60°C. Lowering the salt concentration and / or increasing the temperature increases strin- gency. Stringent conditions for RNA-DNA hybridisations (Northern blots using a probe of e.g.100 nucleotides) are for example those which include at least one wash in 0.2X SSC at 63°C for 20min, or equivalent conditions. Stringent conditions for DNA-DNA hybridisation (Southern blots using a probe of e.g.100 nucleotides) are for example those which include at least one wash (usually 2) in 0.2X SSC at a temperature of at least 50°C, usually about 55°C, for 20 min, or equivalent conditions. See also Sambrook et al. (1989) and Sambrook and Russell (2001).

[0049] As used herein, the phrase "hybridizes” to a DNA or RNA molecule is used to indicate thata molecule recognizes and hybridizes to another nucleic acid molecule by base pairing, meaning that there is enough sequence similarity between the two nucleic acid molecules to effect hybrid- ization under appropriate conditions.

[0050] The present invention accordingly provides a Capsicum annuum plant having multiple flow-ers per node, which allows a further increase of the pepper fruit yield that may be obtained whencultivating the Capsicum annuum plant of the present invention. In one aspect, QTL MF2.1 andQTL MF2.2 comprised in the Capsicum annuum plant according to according to the presentNunhems Netherlands B.V. 230055WO0112invention confers an increased number of flowers per node relative to a plant comprising QTLMF2.1 and lacking QTL MF2.2; or a plant comprising QTL MF2.2 and lacking QTL MF2.1.

[0051] The Capsicum annuum plant according to the present invention accordingly comprises inits genome QTL MF2.1 as further described herein and QTL MF2.2 as further described herein.In one aspect, the Capsicum annuum plant according to the present invention comprises in itsgenome an introgression on chromosome 2 conferring multiple flowers per node, relative to a plant lacking said introgression, wherein said introgression comprises QTL MF2.1 as describedherein and QTL MF2.2 as described herein. In one aspect, both QTL MF2.1 and QTL MF2.2 arecomprised in one single introgression fragment, i.e. in one and the same genomic fragment that is transduced from the donor parent to the recipient parent. In one aspect, QTL MF2.1 and QTLMF2.2 are comprised in two introgression fragments, i.e. QTL MF2.1 is comprised in a first ge-nomic fragment that is transduced from the donor parent to the recipient parent and QTL MF2.2is comprised in a second genomic fragment that is transduced from the donor parent to the recip-ient parent, wherein said first genomic fragment and said second genomic fragment are different.

[0052] As used herein, the terms "genomic introgression", "introgression", "introgressed" and "in-trogressing" refer to both a natural and artificial process whereby a genomic fragment of one species, variety or cultivar, termed donor parent, is transduced into the genome of another spe- cies, variety or cultivar, termed recipient parent, for example by crossing the donor and recipient parent. The process may optionally be completed by backcrossing the resulting plants to the re- cipient parent, which is than termed recurrent parent. An introgression fragment (also named herein as an “introgression”) is present outside of its natural genomic context, meaning that aplant harbouring an introgression fragment from e.g. Capsicum chinense is not a C. chinenseplant. As used herein, the term “recombinant introgression” refers to a genomic introgression which has undergone a recombination to remove an allele that is genetically linked to the desiredtrait, which in the context of the present invention is the multiple flowers per node phenotype. Inone aspect, the recombinant introgression fragment as described herein is obtained by a sponta- neous meiotic recombination event. In a further aspect, the recombinant introgression fragment as described herein is obtained by a technical method, such as targeted mutagenesis method, e.g. the method as described in WO2021 / 228700 A1.

[0053] In one aspect, QTL MF2.1 and QTL MF2.2 are as present in, or as obtainable from, or asobtained from, or as comprised in the genome of a Capsicum annuum plant designated: PP193-22, a representative sample of which has been deposited under accession number NCIMB 44384.

[0054] In one aspect, QTL MF2.1 and / or QTL MF2.2 as comprised in the Capsicum annuum plantaccording to the present invention is comprised in an introgression fragment, preferably in an introgression fragment from Capsicum chinense. In one aspect, QTL MF2.1 and QTL MF2.2 ascomprised in the Capsicum annuum plant according to the present invention is comprised in twoseparate introgressions. In an alternative aspect, QTL MF2.1 and QTL MF2.2 as comprised inthe Capsicum annuum plant according to the present invention is comprised in one introgression(i.e. the introgression comprises both MF2.1 and MF2.2).

[0055] In a further aspect, the introgression comprising QTL MF2.1 and / or QTL MF2.2 may be afunctional fragment of the introgression fragment as present in, or as obtainable from, or as ob-tained from, or as comprised in the genome of a Capsicum annuum plant designated: PP193-22,Nunhems Netherlands B.V. 230055WO0113 a representative sample of which has been deposited under accession number NCIMB 44384. A “functional fragment” of the introgression fragment comprising QTL MF2.1 and / or QTL MF2.2, denotes an introgression fragment that is smaller than the introgression fragment as present inCapsicum annuum plant designated: PP193-22, a representative sample of which has been de-posited under accession number NCIMB 44384, which still exerts the desired function, i.e. whichconfers multiple flowers per node relative to a plant lacking said QTL MF2.1 and QTL MF2.2. Anassay for determining of whether a functional fragment confers multiple flowers per node relativeto a plant lacking said QTL MF2.1 and QTL MF2.2 is provided herein in the Examples section, see Example 1.

[0056] In one aspect, the Capsicum annuum plant according to present invention further comprisesa mutant allele of the Pun1 / AT3 gene, wherein said mutant allele results in a reduced expressionor no expression of the wild type Pun1 / AT3 gene and / or wherein the mutant allele encodes aprotein having a decreased function or loss-of-function when compared to the wild type Pun1 / AT3protein and wherein the wild type Pun1 / AT3 gene encodes a protein comprising at least 80%, e.g.81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, 98.3%, 98.7%, 99.0%, 99.2%, 99.3%, 99.5%, 99.7%, 99.9% or even 100% sequence aminoacid sequence identity to SEQ ID NO: 1.

[0057] "Wild type allele" (WT) refers herein to an allele of a gene encoding a fully functional protein,also referred to herein as “wild type protein”, which in the context of the present invention is de- fined as a protein that does not show reduced protein function. The term "reduced protein func- tion" as used herein refers to a reduced level, activity and expression of said protein. Accordingly, a “reduced protein function” includes reduced expression, preferably no expression, suppression or temporal or spatial misexpression of the encoded protein and / or expression of a protein with “reduced function”, which is defined herein as a protein having a reduction in biological effects or activity when compared to the wild type protein, preferably having loss-of-function. The term “wild type allele” accordingly also comprises “functional variants”, which refers to a version of a gene that is not identical to the reference version of said gene, but that does not result in a reduced protein function when compared to the wild type allele.

[0058] Whether a certain variant of the herein specifically described wild type Pun1 / AT3 gene rep-resents a functional variant can be determined by using routine methods, including, but not limitedto, phenotypic testing for acyltransferase activity and in silico prediction of amino acid changesthat affect protein function. For instance, a web-based computer program SIFT (Sorting Intolerant From Tolerant) is a program that predicts whether an amino acid substitution affects protein func-tion; see world wide web at sift.bii.a-star.edu.sg / . Functionally important amino acids will be con-served in the protein family, and so changes at well-conserved positions tend to be predicted as not tolerated or deleterious; see also Ng and Henikoff (2003) Nucleic Acids Res 31(13): 3812– 3814. For example, if a position in an alignment of a protein family only contains the amino acid isoleucine, it is presumed that substitution to any other amino acid is selected against and that isoleucine is necessary for protein function. Therefore, a change to any other amino acid will be predicted to be deleterious to protein function. If a position in an alignment contains the hydro- phobic amino acids isoleucine, valine and leucine, then SIFT assumes, in effect, that this position can only contain amino acids with hydrophobic character. At this position, changes to other hy- drophobic amino acids are usually predicted to be tolerated but changes to other residues (suchNunhems Netherlands B.V. 230055WO0114 as charged or polar) will be predicted to affect protein function. An alternative tool useful for the prediction of protein function is Provean; see world wide web at provean.jcvi.org / index.php.

[0059] "Mutant allele" refers herein to an allele of a gene that results in a reduced protein functionwhen compared to the wild type protein. In the context of the present invention, a mutant allele may be naturally occurring mutation or a technically induced mutation. The term "naturally occur-ring mutation", as used herein, refers to a genetic variation that has arisen spontaneously in na-ture without any human intervention. The term "technically induced mutation", as used herein, refers to a genetic variation that is the direct result from a human intervention.

[0060] The terms "technically induced mutant", as used herein, is a non-naturally occurring mutantcreated by man. A technically induced mutant can be produced through mutagenesis. “Mutagen- esis” or “induced variation”, as used herein, refers to the process in which plant cells (e.g., aplurality of diploid dicotyledonous crop plant seeds or other parts, such as pollen, etc.) are sub-jected to a technique which induces mutations in the DNA of the cells, such as contact with a mutagenic agent, such as a chemical substance (such as ethylmethylsulfonate (EMS), ethylnitro- sourea (ENU), etc.) or ionizing radiation (neutrons (such as in fast neutron mutagenesis, etc.), alpha rays, gamma rays (such as that supplied by a Cobalt 60 source), X-rays, UV-radiation, etc.), or a combination of two or more of these. While mutations created by irradiation are often large deletions or other gross lesions such as translocations or complex rearrangements, mutations created by chemical mutagens are often more discrete lesions such as point mutations. For ex- ample, EMS alkylates guanine bases, which results in base mispairing: an alkylated guanine will pair with a thymine base, resulting primarily in G / C to A / T transitions. Mutagenesis can comprise random mutagenesis, or can comprise targeted mutagenesis, such as genome editing. Mutagen- esis can also result in epimutations that cause epigenetic silencing.

[0061] Genome editing, also called gene editing, genome engineering, as used herein, refers tothe targeted modification of genomic DNA in which the DNA may be inserted, deleted, modified or replaced in the genome. Genome editing may use sequence-specific enzymes (such as endo- nuclease, nickases, base conversion enzymes) and / or donor nucleic acids (e.g. dsDNA, oligo’s)to introduce desired changes in the DNA. Sequence-specific nucleases that can be programmedto recognize specific DNA sequences include meganucleases (MGNs), zinc-finger nucleases (ZFNs), TAL-effector nucleases (TALENs) and RNA-guided or DNA-guided nucleases such as Cas9, Cpf1, CasX, CasY, C2c1, C2c3, certain Argonaut-based systems (see e.g. Osakabe and Osakabe, Plant Cell Physiol.2015 Mar;56(3):389-400; Ma et al., Mol Plant.2016 Jul 6;9(7):961- 74; Bortesie et al., Plant Biotech J, 2016, 14; Murovec et al., Plant Biotechnol J. 15:917-926, 2017; Nakade et al., Bioengineered Vol 8, No.3:265-273, 2017; Burstein et al., Nature 542, 37– 241; Komor et al., Nature 533, 420–424, 2016; all incorporated herein by reference). Donor nu- cleic acids can be used as a template for repair of the DNA break induced by a sequence specific nuclease. Donor nucleic acids can also be used as such for genome editing without DNA break induction to introduce a desired change into the genomic DNA.

[0062] As used herein the term "non-pungent Capsicum annuum plant "refers to a Capsicum an-nuum plant producing fruits having an average total capsaicinoid content (preferably capsaicin and dihydrocapsaicin content) of less than 150 µg / g FW (fresh weight), more preferably of less than 100 µg / g FW, most preferably of less than 50 µg / g FW. Preferably, the capsaicin of the fruit is determined using HPLC analysis using standard methods. As used herein the term "pungentNunhems Netherlands B.V. 230055WO0115Capsicum annuum plant" refers to a Capsicum annuum plant producing fruits having a higheraverage total capsaicinoid content than that of a non-pungent Capsicum annuum plant as defined herein. Accordingly, the term "pungent Capsicum annuum plant” as used herein preferably refersto a Capsicum annuum plant producing fruits having an average total capsaicinoid content (pref-erably capsaicin and dihydrocapsaicin content) of at least 150 µg / g FW (fresh weight), more pref- erably of at least 100 µg / g FW, most preferably of at least 50 µg / g FW.

[0063] Accordingly, the term "wild type Pun1 / AT3 allele" or "wild type allele of the Pun1 / AT3 gene"refers to an allele of the Pun1 / AT3 gene encoding a fully functional Pun1 / AT3 protein, also re-ferred to herein as “wild type Pun1 / AT3 protein” or “Pun1 / AT3 protein”, which allows the normalcapsaicin biosynthesis. The Pun1 / AT3 gene that encodes an acyltransferase that has been pro-posed to be the putative capsaicinoid synthase (CS) responsible for the last catalytic step of cap-saicinoid biosynthesis. Preferably, a plant comprising the wild type Pun1 / AT3 allele does not havea reduced level, activity and expression as a plant comprising the (non-mutated) reference ver-sion of the Pun1 / AT3 gene. The Pun1 / AT3 gene for pungency in pepper encodes a putative acyl-transferase; see Stewart et al. (2005) Plant J 42(5):675-688. doi: 10.1111 / j.1365-313X.2005.02410.x. Such a wild type Pun1 / AT3 allele in the species Capsicum annuum for in-stance is the wild type genomic DNA which encodes the wild type Capsicum annuum Pun1 / AT3cDNA (mRNA) sequence depicted in SEQ ID NO:2. The protein sequence encoded by this wildtype Capsicum annuum Pun1 / AT3 cDNA has 440 amino acids and is depicted in SEQ ID NO:1,which corresponds to NCBI reference sequence AAV66310.1 (protein GenBank) and SequenceID: AY819028.1. The wild type Pun1 / AT3 allele further comprises functional variants of the wildtype genomic DNA which encodes the wild type Pun1 / AT3 cDNA and amino acid sequences asdescribed herein. One example of such a functional variant of the wild type genomic DNA of theCapsicum annuum Pun1 / AT3 gene is the wild type genomic DNA which encodes the wild typeCapsicum chinense Pun1 / AT3 cDNA (mRNA) sequence depicted in SEQ ID NO:4. The proteinsequence encoded by this wild type Capsicum chinense Pun1 / AT3 cDNA also has 440 aminoacids and is depicted in SEQ ID NO:3, which corresponds to NCBI reference sequence AAV66309.1 (protein GenBank) and Sequence ID: AY819027.1. Another example of such a func-tional variant of the wild type genomic DNA of the Capsicum annuum Pun1 / AT3 gene is the wildtype genomic DNA which encodes the wild type Capsicum frutescens Pun1 / AT3 cDNA (mRNA)sequence depicted in SEQ ID NO:24. The protein sequence encoded by this wild type Capsicumfrutescens Pun1 / AT3 cDNA also has 440 amino acids and is depicted in SEQ ID NO:23, whichcorresponds to NCBI reference sequence AAV66308 (protein GenBank) and Sequence ID:AY819026. Whether a certain variant of the herein specifically described wild type Pun1 / AT3 al-lele represents a functional variant can be determined by using routine methods, including, but not limited to, testing of acyltransferase and / or capsaicinoid synthase enzymatic activity and phe-notypic testing for total capsaicinoid content and in silico prediction of amino acid changes thataffect protein function. For instance, a web-based computer program SIFT (Sorting Intolerant From Tolerant) is a program that predicts whether an amino acid substitution affects protein func-tion; see world wide web at sift.bii.a-star.edu.sg / . Functionally important amino acids will be con-served in the protein family, and so changes at well-conserved positions tend to be predicted as not tolerated or deleterious; see also Ng and Henikoff (2003) Nucleic Acids Res 31(13): 3812– 3814. For example, if a position in an alignment of a protein family only contains the amino acid isoleucine, it is presumed that substitution to any other amino acid is selected against and that isoleucine is necessary for protein function. Therefore, a change to any other amino acid will beNunhems Netherlands B.V. 230055WO0116 predicted to be deleterious to protein function. If a position in an alignment contains the hydro- phobic amino acids isoleucine, valine and leucine, then SIFT assumes, in effect, that this positioncan only contain amino acids with hydrophobic character. At this position, changes to other hy-drophobic amino acids are usually predicted to be tolerated but changes to other residues (such as charged or polar) will be predicted to affect protein function. An alternative tool useful for the prediction of protein function is Provean; see world wide web at provean.jcvi.org / index.php.

[0064] Accordingly, the present invention provides a Capsicum annuum plant comprising in its ge-nome a mutant allele of the Pun1 / AT3 gene, wherein said mutant allele confers a non-pungentfruit phenotype when present in homozygous form. The wild type Pun1 / AT3 gene encodes a pro-tein comprising at least 80% amino acid sequence identity to SEQ ID NO:1, e.g.81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 98.3%,98.7%, 99.0%, 99.2%, 99.3%, 99.5%, 99.7%, 99.9% or even 100% sequence identity to SEQ IDNO: 1, as determined using methods discloses elsewhere herein.

[0065] The mutant allele of the Pun1 / AT3 gene according to the present invention confers a non-pungent fruit phenotype when present in homozygous form. In one aspect, the Capsicum annuumplant according to the present invention is homozygous for a mutant allele of the Pun1 / AT3 geneas described herein.

[0066] In one aspect, the mutant allele of the of the wild type Pun1 / AT3 gene leads to a disruptionof the normal (wild type) protein function of the protein encoded by Pun1 / AT3 gene. The mutantallele as described herein thus may result in reduced expression or no expression of the wild typePun1 / AT3 gene. The mutant allele as described herein may also encode a protein having a de-creased function or loss-of-function when compared to the wild type protein. Thus, the mutantallele that confers a non-pungent fruit phenotype when present in homozygous form may be as-sociated with a reduced expression or even a loss of expression of an otherwise functionalPun1 / AT3 gene product. In a non-limiting example, such a reduced expression or loss of expres-sion may be the result of one or more mutations in a regulatory region of the Pun1 / AT3 gene, e.g.in a promoter sequence of the Pun1 / AT3 gene. In a further non-limiting example, such a reducedexpression or loss of expression may be the result of one or more mutations in a transcriptionfactor that is required for normal (wild type) expression of the Pun1 / AT3 gene product (e.g. afunctional variant of the wild type Pun1 / AT3 protein). In a further non-limiting example, such areduced expression or loss of expression may be the result of posttranscriptional gene silencing or RNAi. Means and methods to determine the expression level of a given gene are well known in the art including, but not limited to, quantitative reverse transcription polymerase chain reaction (quantitative RT-PCR) for the detection and quantification of a specific mRNA and enzyme-linked immunosorbent assay (ELISA) for the detection and quantification of a specific protein. The mu-tant allele of the Pun1 / AT3 gene according to the present invention that confers a non-pungentfruit phenotype when present in homozygous form may be associated with the expression of aprotein having a decreased function or loss-of-function when compared to the wild type protein(e.g. a non-functional variant of the wild type Pun1 / AT3 protein). In a non-limiting example, sucha decreased function or loss-of-function may be the result of a mutation in the coding region ofthe Pun1 / AT3 gene, resulting e.g. in one or more amino acids being replaced (e.g. through aframe-shift mutation or due to a missense mutation), inserted or deleted compared to the wildtype protein. Means and methods to determine protein function are well known in the art including,but not limited to phenotypic testing assays for normal protein function (e.g. by determining theNunhems Netherlands B.V. 230055WO0117 total capsaicinoid content of the fruits), bioassays capable of quantification of enzymatic activity and in silico prediction of amino acid changes that affect protein function, as further describedherein above. Preferably, the mutant allele of the of the wild type Pun1 / AT3 gene contains adeletion spanning the promoter and first exon of the predicted coding region as described in Stewart et al. (2005) loc. cit.

[0067] In one aspect, the mutant allele of the wild type Pun1 / AT3 gene comprises the pun1 geno-type, which comprises a 2529 bp-deletion in the 5’ upstream region of the Pun1 / AT3 gene asfurther described by Stewart et al. (2005) Plant J doi: 10.1111 / j.1365-313X.2005.02410.x, whichhas the effect that the wild type Pun1 / AT3 protein is not transcribed or translated. The pun1 mu-tant allele of the Pun1 / AT3 gene accordingly represents a mutant allele having reduced expres-sion or no expression of the wild type Pun1 / AT3 gene. In one embodiment, the mutant allele ofthe wild type Pun1 / AT3 gene comprises the pun1 genotype is the mutant Pun1 / AT3 allele that ispresent in Capsicum annuum PP183-72 deposited as NCIMB 44383.

[0068] In one aspect, the mutant allele of the of the wild type Pun1 / AT3 gene comprises the pun12genotype, which comprises a 4-bp deletion that causes a frame-shift mutation, resulting in a trun-cated protein as further described by Stewart et al. (2007) J Exp Bot doi: 10.1093 / jxb / erl243. Thepun12 mutant allele of the Pun1 / AT3 gene accordingly represents a mutant allele which encodesa protein having a decreased function or loss-of-function when compared to the wild type protein.The pun12 mutant allele of the Pun1 / AT3 gene accordingly may encode a protein comprising atleast 80% amino acid sequence identity to SEQ ID NO: 5, e.g.81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 98.3%, 98.7%, 99.0%,99.2%, 99.3%, 99.5%, 99.7%, 99.9% or even 100% sequence identity to SEQ ID NO: 5, as de-termined using methods discloses elsewhere herein.

[0069] In one aspect, the mutant allele of the of the wild type Pun1 / AT3 gene comprises the pun13genotype, which comprises a large deletion in the second exon region leading to the loss of 70amino acids in the Pun1 / AT3 protein as described by Stellari et al. (2010) doi:10.1038 / hdy.2009.131. The pun13 mutant allele of the Pun1 / AT3 gene accordingly represents amutant allele which encodes a protein having a decreased function or loss-of-function when com-pared to the wild type protein. The pun13 mutant allele of the Pun1 / AT3 gene accordingly mayencode a protein comprising at least 80% amino acid sequence identity to SEQ ID NO: 7, e.g.81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, 98.3%, 98.7%, 99.0%, 99.2%, 99.3%, 99.5%, 99.7%, 99.9% or even 100% sequence iden-tity to SEQ ID NO: 7, as determined using methods discloses elsewhere herein.

[0070] In one aspect, the mutant allele of the of the wild type Pun1 / AT3 gene comprises the pun14genotype, which comprises a single adenine nucleotide insertion in the second exon region asdescribed by Kirii et al. (2017) doi: 10.2503 / hortj.MI-148 and which causes a frameshift mutationleading to a truncation of the protein with a loss of 50 amino acid residues and a loss of proteinfunction. The pun14 mutant allele of the Pun1 / AT3 gene accordingly represents a mutant allelewhich encodes a protein having a decreased function or loss-of-function when compared to thewild type protein. The pun14 mutant allele of the Pun1 / AT3 gene accordingly may encode a pro-tein comprising at least 80% amino acid sequence identity to SEQ ID NO: 21, e.g. 81%, 82%,83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,Nunhems Netherlands B.V. 230055WO011898.3%, 98.7%, 99.0%, 99.2%, 99.3%, 99.5%, 99.7%, 99.9% or even 100% sequence identity toSEQ ID NO: 21, as determined using methods discloses elsewhere herein.

[0071] As described herein, QTL MF2.1 and / or QTL MF2.2 as comprised in the Capsicum annuumplant according to the present invention may be comprised in an introgression fragment, prefera-bly in an introgression fragment from Capsicum chinense. In the context of the present invention, it was found that an introgression fragment comprising both QTL MF2.1 and QTL MF2.2 furthercomprises the wild type Pun1 / AT3 pungency allele, which is genetically linked to said QTL MF2.1and QTL MF2.2.

[0072] In one aspect, the introgression (introgression fragment) comprising both MF2.1 and MF2.2may be a recombinant introgression (recombinant introgression fragment). In one aspect, QTLMF2.1, the mutant allele of the Pun1 / AT3 gene and QTL MF2.2 are comprised in a recombinantintrogression on chromosome 2. This means that the introgression comprising QTL MF2.1 andQTL MF2.2, which confer the desired multiple flowers per node phenotype, has undergone arecombination to remove the genetically linked wild type Pun1 / AT3 allele, which confers a pun-gent Capsicum annuum plant phenotype.

[0073] QTL MF2.1 and QTL MF2.2 as comprised in the Capsicum annuum plant according to thepresent invention are preferably comprised in a recombinant introgression on chromosome 2,wherein said recombinant introgression comprises a mutant allele of the Pun1 / AT3 gene, whereinsaid mutant allele results in a reduced expression or no expression of the wild type Pun1 / AT3 gene and / or wherein the mutant allele encodes a protein having a decreased function or loss-of-function when compared to the wild type Pun1 / AT3 protein and wherein the wild type Pun1 / AT3gene encodes a protein comprising at least 80% sequence amino acid sequence identity to SEQ ID NO: 1.

[0074] In one aspect, the recombinant introgression fragment as described herein is obtained bya spontaneous meiotic recombination event. In a further aspect, the recombinant introgressionfragment as described herein is obtained by technical method, such as a targeted mutagenesismethod, e.g. the method as described in WO2021 / 228700 A1.

[0075] In one aspect, the Capsicum annuum plant according to present invention is a non-pungentCapsicum annuum plant, wherein said non-pungent Capsicum annuum plant preferably producesfruits having an average total capsaicinoid content (preferably capsaicin and dihydro-capsaicincontent) of less than 150 µg / g FW (fresh weight), more preferably of less than 100 µg / g FW (freshweight), even more preferably of less than 50 µg / g FW (fresh weight). In one aspect, the non-pungent Capsicum annuum plant comprises the recombinant introgression of the present inven-tion, wherein said recombinant introgression preferably comprises a recombinant introgression on chromosome 2, wherein said recombinant introgression on chromosome 2 comprises a mutantallele of the Pun1 / AT3 gene as described herein. In the context of the present invention, themutant allele of the Pun1 / AT3 gene preferably is: the pun1 mutant allele as described hereinwhich comprises a 2529 bp-deletion in the 5’ upstream region of the Pun1 / AT3 gene; the pun12mutant allele as described herein which comprises a 4-bp deletion that causes a frame-shift mu-tation, resulting in a truncated protein; the pun13 mutant allele as described herein which com-prises a large deletion in the second exon region leading to the loss of 70 amino acids in theNunhems Netherlands B.V. 230055WO0119Pun1 / AT3 protein; or the pun14 mutant allele as described herein which comprises a single ade-nine nucleotide insertion in the second exon region.

[0076] The multiple flowering QTLs QTL MF2.1 and QTL MF2.2 according to the present inventionthus confer a multiple flowers per node phenotype when said QTL MF2.1 and QTL MF2.2 arepresent in in the genome of a Capsicum annuum plant. A representative sample of Capsicumannuum seeds comprising QTL MF2.1 and QTL MF2.2 as described herein has been depositedand from the deposit, or from descendants of this deposit, QTL MF2.1 and QTL MF2.2 of the present invention can be easily transferred into any other plant that can be crossed with the Cap-sicum annuum plant, or descendants thereof, grown from the deposited seeds. Alternatively,other donors can be identified which comprise the same QTL MF2.1 and / or QTL MF2.2, e.g.comprising the same SNP haplotypes for QTL MF2.1 and / or QTL MF2.2.

[0077] In one aspect, the introgression on chromosome 2 as comprised in the Capsicum annuumplant according to present invention is as present in, or as obtainable from, or as obtained from,or as comprised in the genome of a Capsicum annuum plant designated: Capsicum annuum plantdesignated: PP183-72, a representative sample of which has been deposited under accession number NCIMB 44383.

[0078] In a further aspect, the recombinant introgression comprising QTL MF2.1 and QTL MF2.2may be a functional fragment of the recombinant introgression as present in, or as obtainablefrom, or as obtained from, or as comprised in the genome of a Capsicum annuum plant desig-nated: PP183-72, a representative sample of which has been deposited under accession number NCIMB 44383. A “functional fragment” of the recombinant introgression fragment comprising QTL MF2.1 and QTL MF2.2, denotes an introgression fragment that is smaller than the introgressionfragment as present in Capsicum annuum plant designated: PP183-72, a representative sampleof which has been deposited under accession number NCIMB 44383, which still exerts the de-sired function, i.e. which confers multiple flowers per node relative to a plant lacking said QTLMF2.1 and QTL MF2.2.

[0079] The plant according to the present invention comprising in its genome an introgression onchromosome 2 conferring multiple flowers per node, relative to a plant lacking said introgression, comprises at least one copy of QTL MF2.1 as described herein and at least one copy of QTLMF2.2 as described herein. QTL MF2.1 and QTL MF2.2 accordingly are dominant. This meansthat at least one copy of QTL MF2.1 and at least one copy of QTL MF2.2 needs to be comprisedin the genome of a Capsicum annuum plant in order to achieve the multiple flowers per nodephenotype conferred by said QTL MF2.1 and QTL MF2.2. In one aspect, the Capsicum annuum plant according to the present invention is heterozygous for the introgression on chromosome 2.In one aspect, the Capsicum annuum plant according to the present invention is heterozygousfor QTL MF2.1 as described herein and is heterozygous for QTL MF2.2 as described herein. Inone aspect, the Capsicum annuum plant according to the present invention is homozygous forQTL MF2.1 as described herein and is heterozygous for QTL MF2.2 as described herein. In oneaspect, the Capsicum annuum plant according to the present invention is heterozygous for QTLMF2.1 as described herein and is homozygous for QTL MF2.2 as described herein. In one aspect,the Capsicum annuum plant according to the present invention is homozygous for QTL MF2.1 asdescribed herein and is homozygous for QTL MF2.2 as described herein. In one aspect, the Cap-sicum annuum plant according to the present invention is homozygous for the introgression asNunhems Netherlands B.V. 230055WO0120described herein. In one aspect, the Capsicum annuum plant according to the present inventionis homozygous for the recombinant introgression as described herein. In one aspect, the Capsi-cum annuum plant according to the present invention is homozygous for the mutant allele of thePun1 / AT3 gene as described herein.

[0080] QTL MF2.1 as further described herein and QTL MF2.2 as further described herein accord-ingly are capable of conferring the multiple flowers per node phenotype in a Capsicum annuumplant. In one aspect, the Capsicum annuum plant according to the present invention comprisesQTL ft12.1 on chromosome 12, wherein said QTL ft12.1 is located between about 7,392,029 bpand about 13,736,988 bp on chromosome 12 of the C. annuum Dempsey V1.1 reference genomeand wherein said QTL ft12.1 prevents suppression of the multiple flowers allele in the offspring ofsaid Capsicum annuum plant. In the context of the present invention, it was found that epistaticinteractions between QTL MF2.1 and QTL MF2.2 as described herein and the suppressor alleleof QTL ft12.1 results into severe linkage drag in later inbreeding stages. Particularly, it was foundthat segregation during inbreeding that is required to obtain inbred parental lines for producingF1 hybrid seed is distorted when the suppressor allele of QTL ft12.1 as described herein is pre-sent in the genome, resulting in a dramatically decreased proportion of the desired multiple flow-ers allele in the offspring.

[0081] QTL ft12.1 according to the present invention prevents suppression of the multiple flowersallele in the offspring of a Capsicum annuum plant according to the present invention when pre-sent in heterozygous form or when present in homozygous form. In one aspect, accordingly, theCapsicum annuum plant according to the present invention is homozygous for QTL ft12.1 asdescribed herein.

[0082] The Capsicum annuum plant according to the present invention accordingly preferably doesnot comprise in its genome the suppressor allele of QTL ft12.1 as further described herein. QTLft12.1 as provided by the present invention, and which is capable of preventing the suppressionof the multiple flowers allele as provided by the present invention is located between 7,392,029bp and 13,736,988 bp on chromosome 12 of C. annuum Dempsey V1.1 reference genome. Thismeans that in the context of the present invention it was possible to map the locus of the sup-pressor QTL ft12.1 as comprised on chromosome 12 to a specific region of said chromosome 12,wherein said region of chromosome 12 is defined by the flanking markers SNP_5 and SNP_6 asfurther described herein.

[0083] In one aspect, the suppressor allele of QTL ft12.1 which is preferably not comprised in theCapsicum annuum plant according to the present invention comprises a cytosine genotype forSNP marker SNP_5 at a position corresponding to position 51 in SEQ ID NO: 17 and / or thyminegenotype for SNP marker SNP_6 at a position corresponding to position 51 in SEQ ID NO: 19.Hence, the presence of the null allele of QTL ft12.1 according to the present invention (i.e. theabsence of the suppressor allele of QTL ft12.1) may be associated with a thymine genotype forSNP marker SNP_5 at a position corresponding to position 51 in SEQ ID NO: 18 and / or a cytosinegenotype for SNP marker SNP_6 at a position corresponding to position 51 in SEQ ID NO” 20.

[0084] In one aspect, QTL ft12.1 is as present in, or as obtainable from, or as obtained from, or ascomprised in the genome of a Capsicum annuum plant designated: PP193-22, a representativeNunhems Netherlands B.V. 230055WO0121 sample of which has been deposited under accession number NCIMB 44384; or PP183-72, a representative sample of which has been deposited under accession number NCIMB 44383.

[0085] As further described in the Examples, QTL MF2.1 and QTL MF2.2 as described herein wereidentified by crossing a Capsicum chinense accession and a Capsicum annuum plant. The Cap-sicum annuum plant according to the present invention accordingly can be obtained by crossingand subsequent selection, preferably by means of a technical process such as marker-based selection, without that the accordingly obtained plant comprises a technically induced mutation.

[0086] In one aspect, the Capsicum annuum plant according to the present invention comprises atechnically induced mutation. Said technically induced mutation preferably comprises a modifica-tion in the genome created with untargeted mutagenesis techniques (such as EMS mutagenesis)preferably in combination with reverse screening (such as TILLING), or with genome editing tech-nologies. Preferably, the technically induced mutation as comprised in the Capsicum annuumplant according to the present invention is the mutant allele of the Pun1 / AT3 gene as describedherein. Other examples of technically induced mutations in the Capsicum annuum plant accordingto the present invention include, but are not limited to mutations in the SSPER-1 gene as de-scribed in WO 2020 / 120242 A1, mutations in the cl gene encoding the Capsicum stay-green(CaSGR) protein and mutations in the wt gene encoding the phytoene synthase (PSY) protein asdescribed in WO 2020 / 254655 A1.

[0087] In one aspect, accordingly, the technically induced mutation comprised in the Capsicumannuum plant according to the present invention may be a mutation resulting in a mutant allele ofthe Pun1 / AT3 gene as described herein and / or a mutation resulting in a mutant allele of thePun1 / AT3 gene as described herein. In one aspect of the invention therefore provides a Capsi-cum annuum plant comprising a mutant allele of the Pun1 / AT3 protein-encoding gene character-ized in that the mutant Pun1 / AT3 allele comprises or effects one or more of the mutations selectedfrom the group consisting of:(a) a deletion, truncation, insertion, point mutation, nonsense mutation, missense or non-syn-onymous mutation, splice-site mutation, frame shift mutation in the genomic sequence;(b) a mutation in one or more regulatory sequences;(c) a deletion, truncation, insertion, point mutation, nonsense mutation, missense or non-syn-onymous mutation, splice-site mutation, frame shift mutation in the coding sequence;(d) a deletion, truncation, insertion, point mutation, nonsense mutation, missense or non-syn-onymous mutation, splice-site mutation, frame shift mutation in the pre-mRNA or mRNA; and / or(e) a deletion, truncation, insertion or replacement of one or more amino acids in thePun1 / AT3 protein.

[0088] The above mutant allele results in decreased activity of the mutant Pun1 / AT3 protein com-pared to the wild type Pun1 / AT3 protein in the Capsicum annuum plant. The decreased activityis due to a knock-out of expression of the Pun1 / AT3 gene, a knock-down of expression of thegene, a loss of function of the encoded mutant Pun1 / AT3 protein or a decrease of function of themutant Pun1 / AT3 protein.

[0089] In one aspect, the present invention provides a plant comprising a mutant allele of the wildtype Pun1 / AT3 gene, wherein the mutant allele as described herein encodes a protein that istruncated when compared to the wild type protein, wherein said truncation causes a loss-of-Nunhems Netherlands B.V. 230055WO0122 function of the encoded protein. In one aspect, the mutant allele of the wild type Pun1 / AT3 gene encodes a protein that is truncated with a loss of at least the final 30 amino acid residues, at least the final 40 amino acid residues, at least the final 50 amino acid residues, or at least the final 60 amino acid residues.

[0090] The plants of the present invention may be any Capsicum annuum plant as de-scribedherein, comprising in its genome QTL MF2.1 as described herein and QTL MF2.2 as describedherein (optionally further comprising the mutant allele of the Pun1 / AT3 gene as described hereinand / or QTL ft12.1 as described herein). In one aspect, the present invention provides a plant asdescribed herein that further is an inbred plant, a dihaploid plant or a hybrid plant. In one aspect, accordingly, the present invention provides that the plant of the present invention is an inbred plant. Such an inbred plant is highly homozygous, for instance by repeated selfing crossing steps. Such an inbred plant may be very useful as a parental plant for the production of F1 hybrid seed. In one aspect, the disclosure provides for haploid plants and / or dihaploid (double haploid) plants of plant of the invention are encompassed herein, which comprise in its genome QTL MF2.1 as described herein and QTL MF2.2 as described herein. Haploid and dihaploid plants can for ex- ample be produced by anther or microspore culture and regeneration into a whole plant. For dihaploid production chromosome doubling may be induced using known methods, such as col-chicine treatment or the like. So, in one aspect a Capsicum annuum plant is provided, comprisingthe multiple flowers per node phenotype as described, wherein the plant is a dihaploid plant. Thepresent invention further provides hybrid plants, which may have advantages such as improveduniformity, vitality and / or disease tolerance. In one aspect, the Capsicum annuum plant accordingto present invention is a F1 hybrid. More preferably, the Capsicum annuum plant according topresent invention is a single cross F1 hybrid plant.

[0091] The present invention further provides seed produced by the Capsicum annuum plant ac-cording to the present invention, wherein the seed comprises QTL MF2.1 as described hereinand QTL MF2.2 as described herein, wherein said seed optionally further comprises the mutantallele of the Pun1 / AT3 gene as described herein and / or QTL ft12.1 as described herein.

[0092] The present invention further provides seed from which the Capsicum annuum plant ac-cording to the present invention can be grown.

[0093] Furthermore, the invention provides a plurality of seed according to the present invention.

[0094] A seed of the invention can be distinguished from other seeds due to the presence of QTLMF2.1 and QTL MF2.2 as described herein (and optionally the presence of the mutant allele ofthe Pun1 / AT3 gene as described herein and / or QTL ft12.1 as described herein), either phenotyp-ically (based on the multiple flowers phenotype of the present invention) and / or using molecularmethods to detect the presence of QTL MF2.1 and QTL MF2.2 as described herein (and optionallythe presence of the mutant allele of the Pun1 / AT3 gene as described herein and / or QTL ft12.1 asdescribed herein) in the cells or tissues, such as molecular genotyping methods to detect QTLMF2.1 and QTL MF2.2 as described herein (and optionally the mutant allele of the Pun1 / AT3gene as described herein and / or QTL ft12.1 as described herein), or sequencing. Accordingly,seed of the invention can be distinguished from other seeds due to the presence of QTL MF2.1 as described herein and QTL MF2.2 as described herein, either phenotypically (based on themultiple flowers phenotype of the present invention) and / or using molecular methods to detectNunhems Netherlands B.V. 230055WO0123 the presence of QTL MF2.1 and QTL MF2.2, in the cells or tissues, such as molecular genotypingmethods to detect QTL MF2.1 and QTL MF2.2 of the present invention, or sequencing. Seedsinclude for example seeds produced by a plant of the invention which is heterozygous for QTL MF2.1 and / or QTL MF2.2 as described herein (and optionally the mutant allele of the Pun1 / AT3gene as described herein and / or QTL ft12.1 as described herein), and optionally selection of thoseseeds which comprise one or two copies of QTL MF2.1 and / or QTL MF2.2 as described herein(and optionally the mutant allele of the Pun1 / AT3 gene as described herein and / or QTL ft12.1 asdescribed herein), e.g. by non-destructive seed sampling methods and analysis of the presence of QTL MF2.1 and / or QTL MF2.2 as described herein (and optionally the mutant allele of thePun1 / AT3 gene as described herein and / or QTL ft12.1 as described herein), or seed producedafter cross-pollination, e.g. pollination of a plant of the invention with pollen from another Capsi-cum plant, preferably from another Capsicum annuum plant, or pollination of another Capsicumannuum plant with pollen of a plant of the invention). Accordingly, seeds include for exampleseeds produced by a plant of the invention which is heterozygous for QTL MF2.1 and QTL MF2.2, and optionally selection of those seeds which comprise one or two copies of QTL MF2.1 and QTL MF2.2, e.g. by non-destructive seed sampling methods and analysis of the presence of QTL MF2.1 and QTL MF2, or seed produced after cross-pollination, e.g. pollination of a plant of theinvention with pollen from another Capsicum plant, preferably from another Capsicum annuumplant, or pollination of another Capsicum annuum plant with pollen of a plant of the invention).

[0095] The present invention further provides seeds obtained from the methods of producing plantsas described herein.

[0096] In one aspect, a plurality of seed is packaged into a container (e.g. a bag, a carton, a canetc.). Containers may be any size. The seeds may be pelleted prior to packing (to form pills or pellets) and / or treated with various compounds, including seed coatings.

[0097] Furthermore, the invention provides progeny comprising or retaining the multiple flowersphenotype of the present invention (conferred by QTL MF2.1 and QTL MF2.2 as described herein,optionally in combination with the mutant allele of the Pun1 / AT3 gene as described herein), suchas progeny obtained by, e.g., selfing one or more times and / or cross-pollinating a plant of the invention with another capsicum plant of a different variety or breeding line of the same plantspecies (or of a plant species that can be crossed with the Capsicum annuum plant of the presentinvention), or with a Capsicum annuum plant of the invention one or more times. In one aspectthe invention relates to for a progeny plant comprising QTL MF2.1 and QTL MF2.2 as describedherein (optionally in combination with the mutant allele of the Pun1 / AT3 gene as describedherein), such as a progeny plant that is produced from a Capsicum annuum plant of the presentinvention comprising QTL MF2.1 and QTL MF2.2 as described herein (optionally further compris-ing the mutant allele of the Pun1 / AT3 gene as described herein and / or QTL ft12.1 as describedherein) by one or more methods selected from the group consisting of: selfing, crossing, mutation, double haploid production or transformation. Mutation preferable are human induced (i.e. tech- nical) mutations or somaclonal mutations. In one aspect, plants or plant parts such as seeds or plant cells of the invention may also be mutated (by e.g. irradiation, chemical mutagenesis, heat treatment, TILLING, etc.) and / or mutated seeds or plants may be selected (e.g. somaclonal vari- ants, etc.) in order to change one or more characteristics of the plants. Similarly, plants of the invention may be transformed and regenerated, whereby one or more chimeric genes are intro- duced into the plants. Transformation can be carried out using standard methods, such asNunhems Netherlands B.V. 230055WO0124Agrobacterium tumefaciens mediated transformation or biolistics, followed by selection of thetransformed cells and regeneration into plants. A desired trait (e.g. genes conferring pest or dis- ease resistance, herbicide, fungicide or insecticide tolerance, etc.) can be introduced into the plants, or progeny thereof, by transforming a plant of the invention or progeny thereof with a transgene that confers the desired trait, wherein the transformed plant retains QTL MF2.1 and QTL MF2.2 as described herein (optionally in combination with the mutant allele of the Pun1 / AT3gene as described herein), the multiple flowers phenotype conferred by it and contains the desiredtrait.

[0098] The terms "progeny", "progenies" and "descendants", as used herein, refer to any and alloffspring that are derivable from or obtainable from a plant of the invention that comprises the multiple flowers per node phenotype described herein. Progeny may be derived by cell culture or by tissue culture, or by producing seeds of a plant. The term progeny may also encompass plants derived from crossing of at least one parent plant according to the invention with another plant of the same or another variety or (breeding) line. A progeny is directly derived from, obtained from, obtainable from or derivable from the parent plant by, e.g., traditional breeding methods (selfing and / or crossing) or regeneration or transformation. However, the term "progeny" generally en- compasses further generations such as second, third, fourth, fifth, sixth, seventh or more gener-ations, i.e., generations of plants which are derived from, obtained from, obtainable from or deriv-able from the former generation by, e.g., traditional breeding methods, regeneration or genetic transformation techniques. For example, a second-generation progeny can be produced from a first generation progeny by any of the methods mentioned above.

[0099] In a further aspect a plant part, obtained from (obtainable from) a plant of the invention isprovided herein, and a container or a package comprising said plant part. The present inventionaccordingly further provides a plant cell, tissue or plant part of the Capsicum annuum plant ac-cording to the present invention or of the seed according to present invention comprising QTLMF2.1 and QTL MF2.2 as described herein (wherein said plant cell, tissue or plant part optionallyfurther comprises the mutant allele of the Pun1 / AT3 gene as described herein and / or QTL ft12.1as described herein). Said plant part preferably is a leaf, anther, pistil, stem, petiole, root, ovule,pollen, microspore, protoplast, callus, tissue, seed, flower, cotyledon, hypocotyl, embryo or cell.

[0100] In a further aspect, the plant part is a plant cell. In still a further aspect, the plant part is anon-regenerable cell or a regenerable cell. In another aspect the plant cell is a somatic cell. A non-regenerable cell is a cell which cannot be regenerated into a whole plant through in vitro culture. The non-regenerable cell may be in a plant or plant part (e.g. leaves) of the invention.The non-regenerable cell may be a cell in a seed, or in the seed coat of said seed. Mature plantorgans, including a mature leaf, a mature stem or a mature root, contain at least one non-regen- erable cell.

[0101] In a further aspect the plant cell is a reproductive cell, such as an ovule or a cell which ispart of a pollen. In an aspect, the pollen cell is the vegetative (non-reproductive) cell, or the sperm cell (Tiezzi, Electron Microsc. Review, 1991). Such a reproductive cell is haploid. When it is re- generated into whole a plant, it comprises the haploid genome of the starting plant. If chromosome doubling occurs (e.g. through chemical treatment), a double haploid plant can be regenerated. In one aspect the plant of the invention comprising QTL MF2.1 and QTL MF2.2 as described herein(optionally further comprising the mutant allele of the Pun1 / AT3 gene as described herein and / orNunhems Netherlands B.V. 230055WO0125QTL ft12.1 as described herein) is a haploid or a double haploid Capsicum annuum plant accord-ing to the present invention.

[0102] The present invention further provides a vegetatively propagated plant, wherein said plantis propagated from a plant part according to the present invention. "Vegetative propagation" or"clonal propagation" refers to propagation of plants from vegetative tissue, e.g. by propagatingplants from cuttings or by in vitro propagation. In vitro propagation involves in vitro cell or tissueculture and regeneration of a whole plant from the in vitro culture. Clones (i.e. genetically identicalvegetative propagations) of the original plant can thus be generated by in vitro culture. "Cell cul-ture" or "tissue culture" refers to the in vitro culture of cells or tissues of a plant. "Regeneration"refers to the development of a plant from cell culture or tissue culture or vegetative propagation. “Non-propagating cell” refers to a cell which cannot be regenerated into a whole plant; see Feher (2019) Frontier in Plant Science doi: 10.3389 / fpls.2019.00536.

[0103] The present invention further provides a haploid plant or dihaploid plant derived from theCapsicum annuum plant according to the present invention. Preferably, said haploid plant ordihaploid plant derived from the Capsicum annuum plant according to the present invention com-prises QTL MF2.1 and QTL MF2.2 as described herein (wherein said haploid plant or dihaploidplant optionally further comprises the mutant allele of the Pun1 / AT3 gene as described hereinand / or QTL ft12.1 as described herein). In one aspect, accordingly, the present invention providesthat the plant of the present invention is an inbred plant. Such an inbred plant is highly homozy- gous, for instance by repeated selfing crossing steps. Such an inbred plant may be very useful as a parental plant for the production of F1 hybrid seed. In one aspect, the disclosure provides for haploid plants and / or dihaploid (double haploid) plants of plant of the invention are encom-passed herein comprising QTL MF2.1 and QTL MF2.2 as described herein (optionally furthercomprising the mutant allele of the Pun1 / AT3 gene as described herein and / or QTL ft12.1 asdescribed herein). Haploid and dihaploid plants can for example be produced by anther or micro-spore culture and regeneration into a whole plant. For dihaploid production chromosome doubling may be induced using known methods, such as colchicine treatment or the like. So, in one aspecta Capsicum annuum plant is provided, comprising QTL MF2.1 and QTL MF2.2 as described,wherein the plant is a dihaploid plant.

[0104] Moreover, there is provided an in vitro cell culture or tissue culture of the Capsicum annuumplant of the invention in which the cell- or tissue culture is derived from a plant part describedherein, such as, for example and without limitation, a leaf, a pollen, an embryo, cotyledon, hypo- cotyls, callus, a root, a root tip, an anther, a flower, a seed or a stem, or a part of any of them, or a meristematic cell, a somatic cell, or a reproductive cell.

[0105] The present invention further provides a vegetatively propagated plant, wherein said plantis propagated from a plant part according to the present invention. Thus, when said cells or tissuesare regenerated or grown into a whole Capsicum annuum plant, the plant comprises QTL MF2.1and QTL MF2.2 as described herein (optionally further comprising the mutant allele of thePun1 / AT3 gene as described herein and / or QTL ft12.1 as described herein) capable of conferringa multiple flowers phenotype.Nunhems Netherlands B.V. 230055WO0126

[0106] Thus, also an in vitro cell culture and / or tissue culture of cells or tissues of plants of theinvention is provided. The cell or tissue culture can be treated with shooting and / or rooting mediato regenerate a Capsicum annuum plant.

[0107] Also vegetative or clonal propagation of plants according to the invention is encompassedherein. Many different vegetative propagation techniques exist. Cuttings (nodes, shoot tips, stems, etc.) can for example be used for in vitro culture as described above. Also other vegetative propagation techniques exist and can be sued, such as grafting, or air layering. In air layering a piece of stem is allowed to develop roots while it is still attached to the parent plant and once enough roots have developed the clonal plant is separated from the parent.

[0108] Thus, in one aspect a method is provided comprising:(a) obtaining a part of a plant of the invention (e.g. cells or tissues, e.g. cuttings),(b) vegetatively propagating said plant part to generate an identical plant from the plant part.

[0109] Thus, also the use of vegetative plant parts of plants of the invention for clonal / vegetativepropagation is an aspect of the invention. In one aspect a method is provided for vegetativelyreproducing a Capsicum annuum plant of the invention comprising QTL MF2.1 and QTL MF2.2as described herein (optionally further comprising the mutant allele of the Pun1 / AT3 gene asdescribed herein and / or QTL ft12.1 as described herein) is provided. Also a vegetatively producedCapsicum annuum plant comprising QTL MF2.1 and QTL MF2.2 as described herein (optionallyfurther comprising the mutant allele of the Pun1 / AT3 gene as described herein and / or QTL ft12.1as described herein) is provided.

[0110] In another aspect a Capsicum annuum plant according to the invention, comprising QTLMF2.1 and QTL MF2.2 as described herein (optionally further comprising the mutant allele of thePun1 / AT3 gene as described herein and / or QTL ft12.1 as described herein) is propagated bysomatic embryogenesis techniques.

[0111] Also provided is a Capsicum annuum plant regenerated from any of the above-describedplant parts, or regenerated from the above-described cell or tissue cultures, said regenerated plant comprising in its genome QTL MF2.1 and QTL MF2.2 as described herein (optionally furthercomprising the mutant allele of the Pun1 / AT3 gene as described herein and / or QTL ft12.1 asdescribed herein).This plant can also be referred to as a vegetative propagation of plants of the invention.

[0112] The invention also relates to a food product comprising or consisting of a plant part de-scribed herein. The term "food" is any substance consumed to provide nutritional support for the body. It is usually of plant or animal origin, and contains essential nutrients, such as carbohy- drates, fats, proteins, vitamins, or minerals. The substance is ingested by an organism and as- similated by the organism's cells in an effort to produce energy, maintain life, or stimulate growth. The term food includes both substance consumed to provide nutritional support for the human and animal body.

[0113] In one aspect plants, plant parts and cells according to the present invention are obtainedby a technical method such as a marker assisted selection method as described herein. In oneaspect plants, plant parts and cells according to the present invention are not exclusively obtainedby means of an essentially biological process, e.g. as defined by Rule 28(2) EPC. In one aspect,Nunhems Netherlands B.V. 230055WO0127accordingly, the present invention provides the Capsicum annuum plant as further describedherein, wherein said plant comprises a technically induced mutation, such as a randomly induced mutation, a targeted gene modification or a transgene. The technically induced mutation prefera-bly may be the mutant allele of the Pun1 / AT3 gene according to the present invention or may beany other technically induced mutation. In one aspect, a process for the production of plants or animals is essentially biological if it consists entirely of natural phenomena such as crossing or selection e.g. as defined by Rule 26(5) EPC and Article 2(2) of the Biotech Directive 98 / 44 / EC.

[0114] The term “exclusively” in the context of the proviso that a plant or plant part is not exclusivelyobtained by means of an essentially biological process is used herein to mean that a plant or plant part originating from a technical process or characterised by a technical intervention in the ge- nome is not covered by the exclusion from patentability even if in addition an essentially biological process (such as crossing and selection) is applied in its production or propagation. Accordingly,the progeny of a plant or a plant part according to the present invention comprising at least onecopy of QTL MF2.1 and QTL MF2.2 as described herein (and optionally at least one copy of themutant allele of the Pun1 / AT3 gene as described herein and / or at least one copy of QTL ft12.1as described herein) does not fall outside the scope of the claims merely because exclusively anessentially biological process was used to obtain said progeny.

[0115] The present invention further provides methods wherein a Capsicum annuum plant as de-scribed herein comprising QTL MF2.1 and QTL MF2.2 as described herein (optionally in combi-nation with the mutant allele of the Pun1 / AT3 gene as described herein and / or QTL ft12.1 asdescribed herein) is used and / or obtained.

[0116] The present invention further provides a method for identifying and / or selecting a Capsicumannuum plant or plant part comprising determining whether said plant or plant part comprises inits genome QTL MF2.1 and QTL MF2.2 according to the present invention or the recombinant introgression on chromosome 2 according to the present invention and optionally determiningwhether said plant or plant part comprises in its genome the QTL ft12.1 on chromosome 12 ac-cording to the present invention. Such method preferably comprises identifying at least onemarker within 50 cM or less, preferably 40 cM or less, 30 cM or less, 20cM or less, 10 cM or less, 9 cM or less, 8 cM or less, 7 cM or less, 6 cM or less, 5 cM or less, 4 cM or less, 3 cM or less, 2cM or less, 1 cM or less, 0.75 cM or less, 0.5 cM or less, or even 0.25 cM or less, of the locus ofQTL MF2.1 as further described herein and / or at least one marker within 50 cM or less, preferably40 cM or less, 30 cM or less, 20cM or less, 10 cM or less, 9 cM or less, 8 cM or less, 7 cM or less, 6 cM or less, 5 cM or less, 4 cM or less, 3 cM or less, 2 cM or less, 1 cM or less, 0.75 cM or less,0.5 cM or less, or even 0.25 cM or less, of the locus of the QTL MF2.2 as further described herein.Such method may alternatively comprise identifying at least one marker within 50 cM or less,preferably 40 cM or less, 30 cM or less, 20cM or less, 10 cM or less, 9 cM or less, 8 cM or less,7 cM or less, 6 cM or less, 5 cM or less, 4 cM or less, 3 cM or less, 2 cM or less, 1 cM or less,0.75 cM or less, 0.5 cM or less, or even 0.25 cM or less of the locus of the recombinant introgres-sion on chromosome 2 according to the present invention. Such method may optionally comprise identifying at least one marker within 50 cM or less, preferably 40 cM or less, 30 cM or less, 20cM or less, 10 cM or less, 9 cM or less, 8 cM or less, 7 cM or less, 6 cM or less, 5 cM or less, 4 cM or less, 3 cM or less, 2 cM or less, 1 cM or less, 0.75 cM or less, 0.5 cM or less, or even 0.25 cMor less of the locus of QTL ft12.1 on chromosome 12 according to the present invention.In one aspect, the marker for QTL MF2.1 in the method for identifying and / or selecting a CapsicumNunhems Netherlands B.V. 230055WO0128annuum plant or plant part is one or more of SNP_1, SNP_2 and SNP_7 as further describedherein. In one aspect, the marker for QTL MF2.2 in the method for identifying and / or selecting aCapsicum annuum plant or plant part is one or more of SNP_3, SNP_4, SNP_8 and SNP_9 asfurther described herein. In one aspect, the marker for QTL ft12.1 in the method for identifyingand / or selecting a Capsicum annuum plant or plant part is one or more of SNP_5 and SNP_6 asfurther described herein.

[0117] In addition, the present invention provides a marker for determining the presence or ab-sence of QTL MF2.1 as described herein, which is capable of conferring multiple flowers per nodewhen comprised in the genome of a Capsicum annuum plant together with QTL MF2.2 as de-scribed herein, whereby said marker is a SNP marker selected from the group consisting of:SNP_1 comprising an adenine at nucleotide 51 of SEQ ID NO: 9 or at nucleotide 51 of a se-quence comprising at least 95% (preferably at least 96%, at least 97%, at least 98% and most preferably at least 99%) sequence identity to SEQ ID NO: 9;SNP_2 comprising a cytosine at nucleotide 51 of SEQ ID NO: 11 or at nucleotide 51 of a se-quence comprising at least 95% (preferably at least 96%, at least 97%, at least 98% and most preferably at least 99%) sequence identity to SEQ ID NO: 11: andSNP_7 comprising an guanine at nucleotide 51 of SEQ ID NO: 25 or at nucleotide 51 of a se-quence comprising at least 95% (preferably at least 96%, at least 97%, at least 98% and most preferably at least 99%) sequence identity to SEQ ID NO: 25.

[0118] In addition, the present invention provides a marker for determining the presence or ab-sence of QTL MF2.2 as described herein, which is capable of conferring multiple flowers per nodewhen comprised in the genome of a Capsicum annuum plant together with QTL MF2.1 as de-scribed herein, whereby said marker is a SNP marker selected from the group consisting of:SNP_3 comprising an adenine at nucleotide 101 of SEQ ID NO: 13 or at nucleotide 51 of a se-quence comprising at least 95% (preferably at least 96%, at least 97%, at least 98% and most preferably at least 99%) sequence identity to SEQ ID NO: 13;SNP_4 comprising a cytosine at nucleotide 101 of SEQ ID NO: 15 or at nucleotide 51 of a se-quence comprising at least 95% (preferably at least 96%, at least 97%, at least 98% and most preferably at least 99%) sequence identity to SEQ ID NO: 15:SNP_8 comprising a cytosine at nucleotide 51 of SEQ ID NO: 27 or at nucleotide 51 of a se-quence comprising at least 95% (preferably at least 96%, at least 97%, at least 98% and mostpreferably at least 99%) sequence identity to SEQ ID NO: 27; andSNP_9 comprising a guanine at nucleotide 51 of SEQ ID NO: 29 or at nucleotide 51 of a se-quence comprising at least 95% (preferably at least 96%, at least 97%, at least 98% and most preferably at least 99%) sequence identity to SEQ ID NO: 29.

[0119] In addition, the present invention provides a marker for determining the presence or ab-sence of QTL ft12.1 as described herein, which prevents suppression of the multiple flowers allelein the offspring of the Capsicum annuum plant, whereby said marker is a SNP marker selectedfrom the group consisting of:SNP_5 comprising a thymine at nucleotide 51 of SEQ ID NO: 17 or at nucleotide 51 of a se-quence comprising at least 95% (preferably at least 96%, at least 97%, at least 98% and mostpreferably at least 99%) sequence identity to SEQ ID NO: 17; andSNP_6 comprising a cytosine at nucleotide 51 of SEQ ID NO: 19 or at nucleotide 51 of aNunhems Netherlands B.V. 230055WO0129 sequence comprising at least 95% (preferably at least 96%, at least 97%, at least 98% and most preferably at least 99%) sequence identity to SEQ ID NO: 19.

[0120] The method for identifying and / or selecting a Capsicum annuum plant or plant part accord-ing to the present invention comprises screening at the DNA, RNA (or cDNA) or protein level using known methods, in order to detect the presence of QTL MF2.1 and / or QTL MF2.2 as de- scribed herein, the recombinant introgression on chromosome 2 as described herein, and / or QTLft12.1 as described herein. There are many methods to detect the presence of a specific geneticdeterminant, which may be a specific QTL, introgression, mutant allele or mutant gene (e.g. QTLMF2.1, QTL MF2.2, the introgression on chromosome 2, the recombinant introgression on chro-mosome 2, the mutant allele of the Pun1 / AT3 gene and QTL ft12.1 as described herein).

[0121] In one aspect, the method for identifying and / or selecting a Capsicum annuum plant or plantpart according to the present invention comprises the step of identifying at least one marker within 50 cM of one or more of the following marker intervals:the marker interval from marker SNP_1 to marker SNP_2; and / orthe marker interval from marker SNP_3 to marker SNP_4; and optionallythe marker interval from marker SNP_5 to marker SNP_6 as described herein, e.g. as definedin Table 2. Preferably, the method for identifying and / or selecting a Capsicum annuum plant orplant part according to the present invention comprises the step of identifying at least one marker within 10 cM of one or more of the following marker intervals: the marker interval from marker SNP_1 to marker SNP_2; and / or the marker interval from marker SNP_3 to marker SNP_4; and optionally the marker interval from marker SNP_5 to marker SNP_6 as described herein, e.g. as definedin Table 2.

[0122] Distances between loci are usually measured by frequency of crossing-over between locion the same chromosome. The further apart two loci are, the more likely that a crossover will occur between them. Conversely, if two loci are close together, a crossover is less likely to occur between them. As a rule, one centimorgan (cM) is equal to 1% recombination between loci (mark- ers). When a locus, preferably a QTL, can be indicated by multiple markers the genetic distance between the end-point markers is indicative of the size of the locus, respectively the QTL. Markers that define the QTL may be markers that are linked to the QTL or markers that are in linkage disequilibrium with the QTL. As used herein, the term "linked to” or "genetically linked" when used in the context of markers and / or genomic regions means that the two linked loci (e.g. a markerand a QTL) are inherited as a single genetic unit in at least 50% of the cases. Thus, in this respect,the term linked can be a separation of 50 cM or less, preferably 40 cM or less, 30 cM or less, 20cM or less, 10 cM or less, 9 cM or less, 8 cM or less, 7 cM or less, 6 cM or less, 5 cM or less, 4 cM or less, 3 cM or less, 2 cM or less, 1 cM or less, 0.75 cM or less, 0.5 cM or less, or even 0.25 cM or less. As used herein, the term "linkage disequilibrium” describes a non-random seg- regation of genetic loci or traits (or both).

[0123] In one aspect, the method for identifying and / or selecting a Capsicum annuum plant or plantpart according to the present invention accordingly comprises identifying and / or selecting a Cap-sicum annuum plant or plant part comprising in its genome QTL MF2.1 located between158,316,753 bp and 163,630,465 bp on chromosome 2 of C. annuum Dempsey V1.1 referencegenome (preferably located between 158,316,753 bp and 162,234,480 bp on chromosome 2 ofNunhems Netherlands B.V. 230055WO0130C. annuum Dempsey V1.1 reference genome); and QTL MF2.2 located between 137,347,441 bpand 151,147,982 bp on chromosome 2 of C. annuum Dempsey V1.1 reference genome (prefer-ably located between 143,111,884 bp and 147,107,830 on chromosome 2 of C. annuum Demp-sey V1.1 reference genome), wherein said QTL MF2.1 and QTL MF2.2 preferably are as presentin, or as obtainable from, or as obtained from, or as comprised in the genome of a Capsicumannuum plant designated: PP193-22, a representative sample of which has been deposited underaccession number NCIMB 44384.

[0124] More preferably, the method for identifying and / or selecting a Capsicum annuum plant orplant part according to the present invention comprises identifying and / or selecting a Capsicumannuum plant or plant part comprising in its genome QTL MF2.1 as described herein and QTLMF2.2 as described herein, wherein QTL MF2.1 and QTL MF2.2 as comprised in the Capsicumannuum plant according to the present invention are comprised in a recombinant introgressionon chromosome 2, wherein said recombinant introgression comprises a mutant allele of thePun1 / AT3 gene, wherein said mutant allele results in a reduced expression or no expression ofthe wild type Pun1 / AT3 gene and / or wherein the mutant allele encodes a protein having a de-creased function or loss-of-function when compared to the wild type Pun1 / AT3 protein andwherein the wild type Pun1 / AT3 gene encodes a protein comprising at least 80% sequence aminoacid sequence identity to SEQ ID NO: 1.

[0125] Even more preferably, the method for identifying and / or selecting a Capsicum annuumplant or plant part according to the present invention comprises identifying and / or selecting aCapsicum annuum plant or plant part wherein the recombinant introgression on chromosome 2is as present in, or as obtainable from, or as obtained from, or as comprised in the genome of aCapsicum annuum plant designated: Capsicum annuum plant designated: PP183-72, a repre-sentative sample of which has been deposited under accession number NCIMB 44383.

[0126] As used herein, the term “molecular marker” or short “marker” refers to a measurable, ge-netic characteristic with a fixed position in the genome, which is normally inherited in a Mendelian fashion, and which can be used for mapping of a trait of interest. A molecular marker may be a short DNA sequence, such as a sequence surrounding a single base-pair change, i.e. a single nucleotide polymorphism or SNP, or a long DNA sequence, such as microsatellites or Simple Sequence Repeats (SSRs). The nature of the marker is dependent on the molecular analysis used and can be detected at the DNA, RNA or protein level. Genetic mapping can be performed using molecular markers such as, but not limited to, RFLP (restriction fragment length polymor- phisms) Botstein et al. (1980) Am J Hum Genet 32:314-331; Tanksley et al. (1989), Bio / Technol- ogy 7:257-263, RAPD (random amplified polymorphic DNA) Williams et al. (1990), NAR 18:6531- 6535; and AFLP (Amplified Fragment Length Polymorphism) Vos et al. (1995) NAR 23:4407- 4414. Appropriate primers or probes are dictated by the mapping method used.

[0127] The term “marker allele" refers to the version of the marker that is present in a particularplant at one of the chromosomes. Typically, a marker can exist as or can be said to have or to comprise two marker alleles. The term “haplotype”, as used herein, refers to a specific combina- tion of marker alleles as present within a certain plant or group of (related) plants. As described herein, a marker allele can be the version of the marker that is present in the plant according to the present invention having the multiple flowers phenotype (multiple flowers marker allele). The version of the same marker that is present in a wild type Capsicum annuum plant which does notNunhems Netherlands B.V. 230055WO0131 have the multiple flowers phenotype of the present invention can be referred to as single flower marker allele.

[0128] Suitable molecular markers are, for example SNP markers (Single Nucleotide Polymor-phisms), AFLP markers, microsatellites, minisatellites, Random Amplified Polymorphic DNA’s (RAPD) markers, RFLP markers, Sequence Characterized Amplified Regions (SCAR) markers, and others, such as TRAP markers described by Hu et al. 2007, Genet Resour Crop Evol 54: 1667-1674).

[0129] Methods and assays for marker detection, or for analyzing the genomic DNA for the pres-ence of a marker, are widely known in the art. The presence of a marker can, for example be detected in hybridization-based methods (e.g. allele-specific hybridization), using Taqman, In- vader, PCR-based methods, oligonucleotide ligation based methods, or sequencing-based meth- ods.

[0130] For example, if there is a single nucleotide difference (single nucleotide polymorphism,SNP) between the single flower marker allele and the multiple flowers marker allele, a SNP gen-otyping assay can be used to detect whether a plant or plant part or cell comprises the singleflower marker nucleotide or the multiple flower marker nucleotide in its genome. For example theSNP can easily be detected using a KASP-assay (see world wide web at kpbioscience.co.uk) or other SNP genotyping assays. For developing a KASP-assay, for example 70 base pairs up-stream and 70 base pairs downstream of the SNP can be selected and two allele-specific forwardprimers and one allele specific reverse primer can be designed. See e.g. Allen et al.2011, Plant Biotechnology J.9, 1086-1099, especially p097-1098 for KASP-assay method.

[0131] AFLP analysis is a DNA fingerprinting technique which detects multiple DNA restrictionfragments by means of PCR amplification. The AFLP technology usually comprises the following steps: (i) the restriction of the DNA with two restriction enzymes, preferably a hexa-cutter and a tetra-cutter, such as EcoRI, PstI and MseI; (ii) the ligation of double-stranded adapters to the ends of the restriction fragments, such as EcoRI, PstI and MseI adaptors; (iii) the amplification of a subset of the restriction fragments using two primers complementary to the adapter and restriction site sequences, and extended at their 3' ends by one to three “selective” nucleotides, i.e., the selective amplification is achieved by the use of primers that extend into the restriction fragments, amplifying only those fragments in which the primer extensions match the nucleotides flanking the restriction sites. AFLP primers thus have a specific sequence and each AFLP primer has a specific code; (iv) gel electrophoresis of the amplified restriction fragments on denaturing slab gels or capillaries; (v) the visualization of the DNA fingerprints by means of autoradiography, phosphor-imaging, or other methods. Using this method, sets of restriction fragments may be visualized by PCR without knowledge of nucleotide sequence. An AFLP marker, as used herein, is a DNA fragment of a specific size, which is generated and visualized as a band on a gel by carrying out an AFLP analysis. Each AFLP marker is designated by the primer combination used to amplify it, followed by the approximate size (in base pairs) of the amplified DNA fragment. It is understood that the size of these fragments may vary slightly depending on laboratory conditions and equipment used. Every time reference is made herein to an AFLP marker by referring to a primer combination and the specific size of a fragment, it is to be understood that such size is approximate and comprises or is intended to include the slight variations observed in different labs. Each AFLP marker represents a certain locus in the genome.Nunhems Netherlands B.V. 230055WO0132

[0132] The term “primer” or “oligonucleotide” as used herein encompasses any nucleic acid that iscapable of priming the synthesis of a nascent nucleic acid in a template-dependent process, such as PCR. Typically, primers are oligonucleotides from 10 to 30 nucleotides (such as 15 to 30 nu- cleotides), but longer sequences can be employed. Primers may be provided in double-stranded form, though the single-stranded form is preferred. Probes can be used as primers, but are de- signed to bind to the target DNA or RNA and need not be used in an amplification process. Typi- cally, the probe or primer shall be capable of detecting a genetic determinant of the present in- vention (e.g. one or more of QTL MF2.1, QTL MF2.2, the introgression on chromosome 2, therecombinant introgression on chromosome 2, the mutant allele of the Pun1 / AT3 gene and QTLft12.1 as described herein), e.g. by binding to it.

[0133] The term “recognizing” as used herein when referring to specific primers, refers to the factthat the specific primers specifically hybridize to a specific nucleic acid sequence under suitable conditions, such as the conditions set forth in the method (such as the conditions of the PCR identification protocol), whereby the specificity is determined by the presence of positive and neg- ative controls.

[0134] The term “SSR” refers to Simple Sequence Repeats or microsatellite (Tautz et al. (1989),NAR 17:6463-6471). Short Simple Sequence stretches occur as highly repetitive elements in all eukaryotic genomes. Simple sequence loci usually show extensive length polymorphisms. These simple sequence length polymorphisms (SSLP) can be detected by polymerase chain reaction (PCR) analysis and be used for identity testing, population studies, linkage analysis and genome mapping.

[0135] Equally other genotyping assays can be used. For example, a TaqMan SNP genotypingassay, a High Resolution Melting (HRM) assay, SNP- genotyping arrays (e.g. Fluidigm, Illumina,etc.) or DNA sequencing may equally be used.

[0136] It is understood that molecular markers can be converted into other types of molecularmarkers. When referring to a specific molecular marker in the present invention, it is understood that the definition encompasses other types of molecular markers used to detect the genetic var-iation originally identified by the specific molecular markers. For example, if an AFLP marker isconverted into another molecular marker using known methods, this other marker is included in the definition. For example, AFLP markers can be converted into sequence-specific markers such as, but not limited to STS (sequenced-tagged-site) or SCAR (sequence-characterized-amplified- region) markers using standard technology as described in Meksem et al. (2001), Mol GenGenomics 265(2):207-214; Negi et al. (2000), TAG 101:146-152, Barret et al. (1989), TAG97:828-833, Xu et al. (2001), Genome 44(1):63-70, Dussel et al. (2002), TAG 105:1190-1195 or Guo et al. (2003), TAG 103:1011-1017. For example, Dussel et al. (2002), TAG 105:1190-1195converted AFLP markers linked to the trait of the present invention into PCR-based sequencetagged site markers such as indel (insertion / deletion) markers and CAPS (cleaved amplified pol- ymorphic sequence) markers.

[0137] A molecular marker linked to genetic determinant according to the present invention (e.g.QTL MF2.1, QTL MF2.2, the introgression on chromosome 2, the recombinant introgression onchromosome 2, the mutant allele of the Pun1 / AT3 gene and QTL ft12.1 as described herein)refers to a molecular marker in a region in the genome that inherits with said genetic determinantNunhems Netherlands B.V. 230055WO0133 as a single genetic unit in at least 50% of the cases. Thus, in this respect, the term linked can be a separation of 50 cM, 40 cM, 30 cM, 20 cM, 10 cM, 9 cM, 8 cM, 7 cM, 6 cM, 5 cM, 4 cM, 3 cM, 2 cM, 1 cM, 0.75 cM, 0.5 cM, or even 0.25 cM.

[0138] A “molecular marker linked to QTL MF2.1 and / or QTL MF2.2”, or a “molecular marker linkedto the presence of QTL MF2.1 and / or QTL MF2.2“ can also be a marker located in the introgres- sion comprising said QTL MF2.1 and / or QTL MF2.2, preferably a marker located in the recombi-nant introgression as further described herein.

[0139] Molecular markers accordingly may be used to aid in the identification of the plants (or plantparts or nucleic acids obtained therefrom) comprising QTL MF2.1 and QTL MF2.2 as describedherein (optionally in combination with the recombinant introgression as described herein). Forexample, one can develop one or more suitable molecular markers which are closely genetically (and preferably also physically) linked to QTL MF2.1 and / or QTL MF2.2 as described hereinand / or QTL ft12.1 as described herein). This can be done by crossing a Capsicum annuum plantaccording to the present invention (preferably having the multiple flowers phenotype) with a wildtype plant and developing a segregating population (e.g. F2 or backcross population) from thatcross. The segregating population can then be phenotyped for the multiple flowers phenotypeand genotyped using e.g. molecular markers such as SNPs (Single Nucleotide Polymorphisms), AFLPs (Amplified Fragment Length Polymorphisms; see, e.g., EP 534858), or others, and bysoftware analysis molecular markers which co-segregate with the multiple flowers phenotype inthe segregating population can be identified and their order and genetic distance (centimorgan distance, cM) to the gene or locus of interest (i.e. one or more of QTL MF2.1, QTL MF2.2 andQTL ft12.1 as described herein) can be identified. Molecular markers which are closely linked tothe gene or locus of interest, e.g. markers at a 50 cM distance, preferably at a 10 cM distance,can then be used in detecting and / or selecting plants (e.g. plants of the invention or progeny of a plant of the invention) or plant parts comprising or retaining said gene or locus of interest. Such closely linked molecular markers can replace phenotypic selection (or be used in addition to phe- notypic selection) in breeding programs, i.e. in Marker Assisted Selection (MAS). Preferably, linked markers are used in MAS. More preferably, flanking markers are used in MAS, i.e. one marker on either side of the gene or locus of interest, which preferably is an introgression on chromosome 2 comprising QTL MF2.1 and QTL MF2.2 as described herein, more preferably a recombinant introgression on chromosome 2 comprising QTL MF2.1 and QTL MF2.2, whereinsaid recombinant introgression comprises a mutant allele of the Pun1 / AT3 gene as describedherein.

[0140] In yet a further aspect, a kit is provided for the detection of one or more genetic determinantas provided by the present invention in a Capsicum DNA sample, wherein said genetic determi-nant preferably is QTL MF2.1, QTL MF2.2, the introgression on chromosome 2, the recombinantintrogression on chromosome 2, the mutant allele of the Pun1 / AT3 gene, or QTL ft12.1 as de-scribed herein. In one aspect, accordingly, a kit for the detection of QTL MF2.1 and QTL MF2.2 according to the present invention is provided, wherein said kit comprises at least one primer or probe which specifically recognizes a molecular marker linked to QTL MF2.1 and QTL MF2.2 as described herein. In one aspect, a kit for the detection of QTL MF2.1 and QTL MF2.2 according to the present invention is provided, wherein said kit comprises at least one primer or probe which specifically recognizes a molecular marker linked to the introgression on chromosome 2 compris- ing said QTL MF2.1 and QTL MF2.2 as described herein. In one aspect, a kit for the detection ofNunhems Netherlands B.V. 230055WO0134 QTL MF2.1 and QTL MF2.2 according to the present invention is provided, wherein said kit com- prises at least one primer or probe which specifically recognizes a molecular marker linked to the recombinant introgression on chromosome 2 comprising said QTL MF2.1 and QTL MF2.2 asdescribed herein. In one aspect, a kit for the detection of the mutant allele of the Pun1 / AT3 geneaccording to the present invention is provided, wherein said kit comprises at least one primer or probe which specifically recognizes a molecular marker linked to the mutant allele of thePun1 / AT3 gene as described herein. In one aspect, a kit for the detection of QTL ft12.1 accordingto the present invention is provided, wherein said kit comprises at least one primer or probe whichspecifically recognizes a molecular marker linked to QTL ft12.1 as described herein. The kit forthe detection of a genetic determinant according to the present invention as provided preferablycomprises at least one, two, three, or more of: at least one primer or probe which specificallyrecognizes a molecular marker linked to QTL MF2.1 and QTL MF2.2 as described herein; at least one primer or probe which specifically recognizes a molecular marker linked to the introgression on chromosome 2 comprising said QTL MF2.1 and QTL MF2.2 as described herein; at least one primer or probe which specifically recognizes a molecular marker linked to the recombinant intro- gression on chromosome 2 comprising said QTL MF2.1 and QTL MF2.2 as described herein; at least one primer or probe which specifically recognizes a molecular marker linked to the mutantallele of the Pun1 / AT3 gene as described herein; and at least one primer or probe which specifi-cally recognizes a molecular marker linked to QTL ft12.1 as described herein.

[0141] A “kit”, as used herein, refers to a set of reagents for the purpose of performing the methodof the invention, more particularly, the identification of a genetic determinant as provided by the present invention (e.g. one or more of QTL MF2.1, QTL MF2.2, the introgression on chromosome2, the recombinant introgression on chromosome 2, the mutant allele of the Pun1 / AT3 gene andQTL ft12.1 as described herein) in biological samples.

[0142] In particular, the methods and kits according to the invention are suitable to determine thepresence of detection genetic determinant according to the present invention. The presence of detection genetic determinant according to the present invention can be determined using at least one molecular marker, wherein said one molecular marker is linked to the presence of a genetic determinant as provided by the present invention (i.e. one or more of QTL MF2.1, QTL MF2.2,the introgression on chromosome 2, the recombinant introgression on chromosome 2, the mutantallele of the Pun1 / AT3 gene and QTL ft12.1 gene as described herein). A “biological sample” canbe a plant or part of a plant such as a plant tissue or a plant cell. “Providing genomic DNA” as used herein refers to providing a sample comprising genomic DNA from the plant.

[0143] The sample can refer to a tissue sample which has been obtained from said plant, such as,for example, a leaf sample, comprising genomic DNA from said plant. The sample can further refer to genomic DNA which is obtained from a tissue sample, such as genomic DNA which has been obtained from a tissue, such as a leaf sample. Providing genomic DNA can include, but does not need to include, purification of genomic DNA from the tissue sample. Providing genomic DNA thus also includes obtaining tissue material from a plant or larger piece of tissue and pre- paring a crude extract or lysate therefrom.

[0144] In one aspect, the kit comprises at least one oligonucleotide for identifying a genetic deter-minant as provided by the present invention. Preferably, said genetic determinant as provided bythe present invention is one or more of QTL MF2.1, QTL MF2.2, the introgression on chromosomeNunhems Netherlands B.V. 230055WO01352, the recombinant introgression on chromosome 2, the mutant allele of the Pun1 / AT3 gene andQTL ft12.1 as described herein.

[0145] The present invention further provides the use of a genetic marker specific for QTL MF2.1and QTL MF2.2 according to the present invention or the recombinant introgression on chromo-some 2 according to the present invention for selecting a Capsicum annuum plant having anincreased number of flowers per node phenotype.

[0146] The present invention accordingly provides the use of a genetic marker specific for a geneticdeterminant of the present invention (i.e. one or more of QTL MF2.1, QTL MF2.2, the introgres-sion on chromosome 2, the recombinant introgression on chromosome 2, the mutant allele of thePun1 / AT3 gene and QTL ft12.1 as described herein) for selecting a Capsicum annuum planthaving a multiple flowers phenotype of the present invention.

[0147] The present invention further provides a method for generating or producing a Capsicumannuum plant having a multiple flowers per node phenotype according to the present invention orplant part thereof, comprising providing a seed mixture harvested from a cross between a firstCapsicum annuum plant and a second Capsicum plant, wherein the first Capsicum annuum plantis a Capsicum annuum plant according to the present invention as further defined herein; andselecting seeds comprising in its genome QTL MF2.1 and QTL MF2.2 according to the present invention or the recombinant introgression on chromosome 2 according to the present inventionand optionally determining whether said plant or plant part comprises in its genome the QTL ft12.1on chromosome 12 according to the present invention. In one aspect, said method for generatingor producing a Capsicum annuum plant having a multiple flowers per node phenotype accordingto the present invention or plant part thereof comprises the step of identifying at least one markerwithin 50 cM (or within 40 cM, 30 cM, 20 cM, 10 cM, 9 cM, 8 cM, 7 cM, 6 cM, 5 cM, 4 cM, 3 cM, 2.5 cM, 2 cM, 1 cM, or even less) of one or more of the following marker intervals: the marker interval from marker SNP_1 to marker SNP_2; and / or the marker interval from marker SNP_3 to marker SNP_4; and optionally the marker interval from marker SNP_5 to marker SNP_6 as described herein, e.g. as definedin Table 2. In one aspect, said method for generating or producing a Capsicum annuum planthaving a multiple flowers per node phenotype according to the present invention or plant partthereof comprises that said first Capsicum annuum plant is derived from a seed as depositedunder accession number NCIMB 44384 or under accession number NCIMB 44383. In one aspect,the marker for QTL MF2.1 in the method for generating or producing a Capsicum annuum planthaving a multiple flowers per node phenotype is one or more of SNP_1, SNP_2 and SNP_7 asfurther described herein. In one aspect, the marker for QTL MF2.2 in the method for generatingor producing a Capsicum annuum plant having a multiple flowers per node phenotype is one ormore of SNP_3, SNP_4, SNP_8 and SNP_9 as further described herein. In one aspect, themarker for QTL ft12.1 in the method for identifying and / or selecting a Capsicum annuum plant orplant part is one or more of SNP_5 and SNP_6 as further described herein.

[0148] The present invention further provides a method for producing a Capsicum annuum planthaving a multiple flowers per node phenotype according to the present invention, said methodcomprising the step(s) of: (i) crossing a first Capsicum annuum plant and a second Capsicumplant, wherein the first Capsicum annuum plant is a Capsicum annuum plant according to theNunhems Netherlands B.V. 230055WO0136 present invention; optionally harvesting seed from the crossing of (i) and selecting seed compris- ing QTL MF2.1 and QTL MF2.2 or the recombinant introgression on chromosome 2 in its genome.

[0149] In one aspect, the method for producing a Capsicum annuum plant having a multiple flowersper node phenotype according to the present invention comprising the step of selecting seedcomprising QTL MF2.1 and QTL MF2.2 or the recombinant introgression on chromosome 2 in itsgenome by identifying at least one marker within 50 cM, preferably within 10 cM, of one or moreof the following marker intervals: the marker interval from marker SNP_1 to marker SNP_2; and / orthe marker interval from marker SNP_3 to marker SNP_4 as described herein, e.g. as defined inTable 2.

[0150] The plants according to the present invention accordingly can be used in a conventionalbreeding scheme to produce more plants with the same characteristics or to introduce the multiple flowers phenotype according to the invention in other varieties of the same or related plant spe- cies, or in hybrid plants. The obtained plants can further be used for creating propagating material. Plants according to the invention can further be used to produce gametes, seeds (including crushed seeds and seed cakes), seed oil, embryos, either zygotic or somatic, progeny or hybrids of plants obtained by methods of the invention. Seeds obtained from the plants according to the invention are also encompassed by the invention.

[0151] “Creating propagating material”, as used herein, relates to any means know in the art toproduce further plants, plant parts or seeds and includes inter alia vegetative reproduction meth- ods (e.g. air or ground layering, division, (bud) grafting, micropropagation, stolons or runners, storage organs such as bulbs, corms, tubers and rhizomes, striking or cutting, twin-scaling), sex- ual reproduction (crossing with another plant) and asexual reproduction (e.g. apomixis, somatic hybridization).

[0152] The present invention further provides a method for increasing the number of flowers pernode in a Capsicum annuum plant, said method comprising introgressing QTL MF2.1 and QTLMF2.2 according to the present invention or the recombinant introgression on chromosome 2according to the present invention into said Capsicum annuum plant.

[0153] The present invention further provides the use of QTL MF2.1 and QTL MF2.2 according tothe present invention or the recombinant introgression on chromosome 2 according to the presentinvention for increasing the number of flowers per node in a Capsicum annuum plant.

[0154] Suitable to the invention is a method to produce pepper fruits from Capsicum annuum plantshaving a multiple flowers phenotype, comprising the steps of sowing seeds from the Capsicumannuum plants according to the invention comprising QTL MF2.1 and QTL MF2.2 as describedherein (optionally in combination with the recombination as described herein and / or QTL ft12.1 as described herein), growing said plants in the field and harvesting.

[0155] It is to be understood that this invention is not limited to the particular methodology or pro-tocols. It is also to be understood that the terminology used herein is for the purpose of describing particular aspects only, and is not intended to limit the scope of the present invention which will be limited only by the appended claims. It must be noted that as used herein and in the appended claims, the singular forms "a," "and," and "the" include plural reference unless the context clearly dictates otherwise. Thus, for example, reference to "a vector" is a reference to one or more vectorsNunhems Netherlands B.V. 230055WO0137 and includes equivalents thereof known to those skilled in the art, and so forth. The term "about" is used herein to mean approximately, roughly, around, or in the region of. When the term "about" is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the numerical values set forth. In general, the term "about" is used herein to modify a numerical value above and below the stated value by a variance of 20 percent, preferably 10 percent up or down (higher or lower). As used herein, the word "or" means any one member of a particular list and also includes any combination of members of that list. The words "com-prise," "comprising," "include," "including," and "includes" when used in this specification and inthe following claims are intended to specify the presence of one or more stated features, integers, components, or steps, but they do not preclude the presence or addition of one or more other features, integers, components, steps, or groups thereof.

[0156] Representative samples of seeds of a Capsicum annuum plant comprising the multiple flow-ers phenotype according to the present invention were deposited by Nunhems Netherlands B.V.on 16 April 2024 at the NCIMB Ltd. (Ferguson Building, Craibstone Estate, Bucksburn Aberdeen,Scotland AB219YA, UK) according to the Budapest Treaty, under the Expert Solution (EPC 2000, Rule 32(1)). Seeds were given the following deposit numbers: NCIMB 44384 for Capsicum an-nuum PP193-22 and NCIMB 44383 for Capsicum annuum PP183-72.

[0157] The Applicant requests that samples of the biological material and any material derivedtherefrom be only released to a designated Expert in accordance with Rule 32(1) EPC or related legislation of countries or treaties having similar rules and regulation, until the mention of the grant of the patent, or for 20 years from the date of filing if the application is refused, withdrawn or deemed to be withdrawn.

[0158] Access to the deposit will be available during the pendency of this application to personsdetermined by the Director of the U.S. Patent Office to be entitled thereto upon request. Subject to 37 C.F.R. § 1.808(b), all restrictions imposed by the depositor on the availability to the public of the deposited material will be irrevocably removed upon the granting of the patent. The deposit will be maintained for a period of 30 years, or 5 years after the most recent request, or for theenforceable life of the patent whichever is longer and will be replaced if it ever becomes nonviableduring that period. Applicant does not waive any rights granted under this patent on this applica- tion or under the Plant Variety Protection Act (7 USC 2321 et seq.). SEQUENCESSEQ ID NO: 1 C. annuum PUN1 / AT3 protein acyltransferase; AAV66310.1MAFALPSSLV SVCNKSFIKP SSLTPSTLRF HKLSFIDQSL SNMYIPCAFF YPKVQQRLED SKNSDELSHI AHLLQTSLSQ TLVSYYPYAG KLKDNATVDC NDMGAEFLSV RIKCSMSEIL DHPHASLAES IVLPKDLPWA NNCEGGNLLV VQVSKFDCGG IAISVCFSHK IGDGCSLLNF LNDWSSVTRD HTTTTLVPSP RFVGDSVFST QKYGSLITPQ ILSDLNQCVQ KRLIFPTDKL DALRAKVAEE SGVKNPTRAE VVSALLFKCA TKASSSMLPS KLVHFLNIRT MIKPRLPRNA IGNLSSIFSI EATNMQDMEL PTLVRNLRKE VEVAYKKDQV EQNELILEVV ESMREGKLPF ENMDGYKNVY TCSNLCKYPY YTVDFGWGRP ERVCLGNGPS KNAFFLKDYK AGQGVEARVM LHKQQMSEFE RNEELFEFIANunhems Netherlands B.V. 230055WO0138SEQ ID NO: 2 C. annuum PUN1 / AT3 acyltransferase CDNA; AY819028.1ATGGCTTTTG CATTACCATC ATCACTTGTT TCAGTTTGTA ACAAATCTTT TATCAAACCT TCCTCTCTCA CCCCCTCTAC ACTTAGATTT CACAAGCTAT CTTTCATCGA TCAATCTTTA AGTAATATGT ATATCCCTTG TGCATTTTTT TACCCTAAAG TACAACAAAG ACTAGAAGAC TCCAAAAATT CTGATGAGCT TTCCCATATA GCCCACTTGC TACAAACATC TCTATCACAA ACTCTAGTCT CTTACTATCC TTATGCTGGA AAGTTGAAGG ACAATGCTAC TGTTGACTGT AACGATATGG GAGCTGAGTT CTTGAGTGTT CGAATAAAAT GTTCCATGTC TGAAATTCTT GATCATCCTC ATGCATCTCT TGCAGAGAGC ATAGTTTTGC CCAAGGATTT GCCTTGGGCG AATAATTGTG AAGGTGGTAA TTTGCTTGTA GTTCAAGTAA GTAAGTTTGA TTGTGGGGGA ATAGCCATCA GTGTATGCTT TTCGCACAAG ATTGGTGATG GTTGCTCTCT GCTTAATTTC CTTAATGATT GGTCTAGCGT TACTCGTGAT CATACGACAA CAACTTTAGT TCCATCTCCT AGATTTGTAG GAGATTCAGT CTTCTCTACA CAAAAATATG GTTCTCTCAT TACGCCACAA ATTTTGTCCG ATCTCAACCA GTGCGTACAG AAAAGACTCA TTTTTCCTAC AGATAAGTTA GATGCACTTC GAGCTAAGGT GGCAGAAGAA TCAGGAGTAA AAAATCCAAC AAGGGCTGAA GTTGTTAGCG CTCTTCTTTT CAAATGTGCA ACAAAGGCAT CATCATCAAT GCTACCATCA AAGTTGGTTC ACTTCTTAAA CATACGTACT ATGATCAAAC CTCGTCTACC ACGAAATGCC ATTGGAAATC TCTCGTCTAT TTTCTCCATA GAAGCAACTA ACATGCAGGA CATGGAGTTG CCAACGTTGG TTCGTAATTT AAGGAAGGAA GTTGAGGTGG CATACAAGAA AGACCAAGTC GAACAAAATG AACTGATCCT AGAAGTAGTA GAATCAATGA GAGAAGGGAA ACTGCCATTT GAAAATATGG ATGGCTATAA GAATGTGTAT ACTTGCAGCA ATCTTTGCAA ATATCCATAC TACACTGTAG ATTTTGGATG GGGAAGACCT GAAAGGGTGT GTCTAGGAAA TGGTCCCTCC AAGAATGCCT TCTTCTTGAA AGATTACAAA GCTGGGCAAG GCGTGGAGGC GCGGGTGATG TTGCACAAGC AACAAATGTC TGAATTTGAA CGCAATGAGG AACTCTTTGA ATTTATTGCC TAASEQ ID NO: 3 C. chinense PUN1 protein acyltransferase; AAV66309.1MAFALPSSLV SVCDKSFIKP SSLTPSKLRF HKLSFIDQSL SNMYIPCAFF YPKVQQRLED SKNSDELSHI AHLLQTSLSQ TLVSYYPYAG KLKDNATVDC NDMGAEFLSV RIKCSMSEIL DHPHASLAES IVLPKDLPWA NNCEGGNLLV VQVSKFDCGG IAISVCFSHK IGDGCSLLNF LNDWSSVTRD HTTTALVPSP RFVGDSVFST KKYGSLITPQ ILSDLNECVQ KRLIFPTDKL DALRAKVAEE SGVKNPTRAE VVSALLFKCA TKASSSMLPS KLVHFLNIRT MIKPRLPRNA IGNLSSIFSI EATNMQDMEL PTLVRNLRKE VEVAYKKDQV EQNELILEVV ESMREGKLPF ENMDGYENVY TCSNLCKYPY YTVDFGWGRP ERVCLGNGPS KNAFFLKDYK AGQGVEARVM LHKQQMSEFE RNEELLEFIASEQ ID NO: 4 C. chinense PUN1 acyltransferase CDNA; AY819027.1ATGGCTTTTG CATTACCATC ATCACTTGTT TCAGTTTGTG ACAAATCTTT TATCAAACCT TCCTCTCTCA CCCCCTCTAA ACTTAGATTT CACAAGCTAT CTTTCATCGA TCAATCTTTA AGTAATATGT ATATCCCTTG TGCATTTTTT TACCCTAAAG TACAACAAAG ACTAGAAGAC TCCAAAAATT CTGATGAGCT TTCCCATATA GCCCACTTGC TACAAACATC TCTATCACAA ACTCTAGTCT CTTACTATCC TTATGCAGGA AAGTTGAAGG ACAATGCTAC TGTTGACTGT AACGATATGG GAGCTGAGTT CTTGAGTGTT CGAATAAAAT GTTCCATGTC TGAAATTCTT GATCATCCTC ATGCATCTCT TGCAGAGAGC ATAGTTTTGC CCAAGGATTT GCCTTGGGCG AATAATTGTG AAGGTGGTAA TTTGCTTGTA GTTCAAGTAA GTAAGTTTGA TTGTGGGGGA ATAGCCATCA GTGTATGCTT TTCGCACAAG ATTGGTGATG GTTGCTCTCT GCTTAATTTC CTTAATGATT GGTCTAGCGT TACTCGTGAT CATACGACAA CAGCTTTAGT TCCATCTCCT AGATTTGTAG GAGATTCTGT CTTCTCTACA AAAAAATATG GTTCTCTTAT TACGCCACAA ATTTTGTCCG ATCTCAACGA GTGCGTACAG AAAAGACTCA TTTTTCCTAC AGATAAGTTA GATGCACTTC GAGCTAAGGT GGCAGAAGAA TCAGGAGTAA AAAATCCAAC AAGGGCAGAA GTTGTTAGCG CTCTTCTTTT CAAATGTGCA ACAAAGGCAT CATCATCAAT GCTACCATCA AAGTTGGTTC ACTTCTTAAA CATACGTACT ATGATCAAAC CTCGTCTACC ACGAAATGCC ATTGGAAATC TCTCGTCTAT TTTCTCCATA GAAGCAACTA ACATGCAGGA CATGGAGTTGNunhems Netherlands B.V. 230055WO0139 CCAACGTTGG TTCGTAATTT AAGGAAGGAA GTTGAGGTGG CATACAAGAA AGACCAAGTC GAACAAAATG AACTGATCCT AGAAGTAGTA GAATCAATGA GAGAAGGGAA ACTGCCATTT GAAAATATGG ATGGCTATGA GAATGTGTAT ACTTGCAGCA ATCTTTGCAA ATATCCGTAC TACACTGTAG ATTTTGGATG GGGAAGACCT GAAAGAGTGT GTCTAGGAAA TGGTCCCTCC AAGAATGCCT TCTTCTTGAA AGATTACAAA GCTGGGCAAG GCGTGGAGGC GCGGGTGATG TTGCACAAGC AACAAATGTC TGAATTTGAA CGCAATGAGG AACTCCTTGA GTTCATTGCC TAASEQ ID NO: 5 non-pungent C. chinense pun1 protein acyltransferase;EF104910 MAFALPSSLV SVCDKSFIKP SSLTPSTLRF HKLSFIDQSL SNMYIPCAFF YPKVQQRLED SKNSDELSHI AHLLQTSLSQ TLVSYYPYAG KLKDNATVDC NDMGAEFLSV RIKCSMSEIL DHPHASLAES IVLPKDLPWA NNCEGGNLLV VQVSLIVGE*SEQ ID NO: 6 non-pungent C. chinense pun1 acyltransferase CDNA;EF104910 ATGGCTTTTG CATTACCATC ATCACTTGTT TCAGTTTGTG ACAAATCTTT TATCAAACCT TCCTCTCTCA CCCCCTCTAC ACTTAGATTT CACAAGCTAT CTTTCATCGA TCAATCTTTA AGTAATATGT ACATCCCTTG TGCATTTTTT TACCCTAAAG TACAACAAAG ACTAGAAGAC TCCAAAAATT CTGATGAGCT TTCCCATATA GCCCACTTGC TACAAACATC TCTATCACAA ACTCTAGTCT CTTACTATCC TTATGCAGGA AAGTTGAAGG ACAATGCTAC TGTTGACTGT AACGATATGG GAGCTGAGTT CTTGAGTGTT CGAATAAAAT GTTCCATGTC TGAAATTCTT GATCATCCTC ATGCATCTCT TGCAGAGAGC ATAGTTTTGC CCAAGGATTT GCCTTGGGCG AATAATTGTG AAGGTGGTAA TTTGCTTGTA GTTCAAGTAA GTTTGATTGT GGGGGAATAGSEQ ID NO: 7 non-pungent C.^frutescens pun1 protein acyltransferase;FJ871985 MAFALPSSLV SXCDKSFIKP SSLTPSTXRF HKLSFIDQSL SNMYIPCAFF YPKVQQRLED SKNSDELSHI AHLLQTSLSQ TLVSYYPYAG KLKDNATVDC NDMGAEFLSV RIKCSMSEIL DHPHASLAES IVLPKDLPWA NNCEGGNLLV VQVSKFDCGG IAISVCFSHK IGDGCSLLNF LNDWSSVTRD HTTTTLVPSP RFVGDSVFST KKYGSLITPQ ILSDLNECVQ KRLIFPTDKL DALRAKVAEE SGVKNPTRAE VVSALLFKCA TKASSSMLPS KLVHFLNIRT MIKPRLPRNA IGNLSSIFSI EATNMQDMEL PTLVRNLRKE VEVAYKKDQV EQNELILEVV ESMREGKLPF ENMDGYENVY TVGFIVCSEQ ID NO: 8 non-pungent C.^frutescens pun1 acyltransferase CDNA;FJ871985 ATGGCTTTTG CATTACCATC ATCACTTGTT TCARTTTGTG ACAAATCTTT TATYAAACCT TCCTCTCTCA CCCCCTCTAC ACWYAGATTT CACAAGCTAT CTTTCATCGA TCAATCTTTA AGTAATATGT AYATCCCTTG TGCATTTTTT TACCCTAAAG TACAACAAAG ACTAGAAGAC TCCAAAAATT CTGATGAGCT TTCCCATATA GCCCACTTGC TRCAAACATC TCTATCACAA ACTCTAGTCT CTTACTATCC TTATGCAGGA AAGTTGAARG ACAATGCTAC TGTTGACTGT AACGATATGG GAGCTGAGTT CTTGAGTGTT CGAATAAAAT GTTCCATGTC TGAAATTCTT GATCATCCTC ATGCATCTCT TGCAGAGAGC ATAGTTTTGC CCAAGGATTT GCCTTGGGCG AATAATTGTG AAGGTGGTAA TTTGCTTGTA GTTCAAGTAA GTAAGTTTGA TTGTGGGGGA ATAGCCATCA GTGTATGCTT TTCGCACAAG ATTGGTGATG GTTGCTCTCT GCTTAATTTC CTTAATGATT GGTCTAGCGT TACTCGTGAT CATACRACAA CAACTTTAGT TCCATCTCCT AGATTTGTAG GAGATTCTGT CTTCTCTACA AAAAAATATG GTTCTCTTAT TACGCCACAA ATTTTGTCCG ATCTCAACGA GTGCGTACAG AAAAGACTCA TTTTTCCTAC AGATAAGTTA GATGCACTTC GAGCTAAGGT GGCAGAAGAA TCAGGAGTAA AAAATCCAAC AAGGGCAGAA GTTGTTAGCG CTCTTCTTTT CAAATGTGCA ACAAAGGCAT CATCATCAAT GCTACCATCA AAGTTGGTTC ACTTCTTAAA CATACGTACT ATGATCAAAC CTCGTCTACC ACGAAATGCC ATTGGAAATC TCTCGTCTAT TTTCTCCATA GAAGCAACTA ACATGCAGGA CATGGAGTTG CCAACGTTGG TTCGTAATTT AAGGAAGGAA GTTGAGGTGG CATACAAGAA AGACCAAGTC GAACAAAATG AACTGATCCT AGAAGTAGTA GAATCAATGA GAGAAGGGAA ACTGCCATTTNunhems Netherlands B.V. 230055WO0140 GAAAATATGG ATGGCTATGA GAATGTGTAT ACTGTTGGGT TCATAGTGTG TSEQ ID NO: 9 Start MF2.1 SNP_1 MFPosition in Dempsey_1.1: chr02 - 158,316,753 bpTCAAGGGATTGAAGATTTGGTGCTGAACCAAATGTTGGAGGTGGATGAATA CTGGA-GACRAAACAACATGGAAGCACACAAAATTAGGTTGGGATATCAAMSEQ ID NO: 10 Start MF2.1 SNP_1 wtPosition in Dempsey_1.1: chr02 - 158,316,753 bpTCAAGGGATTGAAGATTTGGTGCTGAACCAAATGTTGGAGGTGGATGAATG CTGGA-GACRAAACAACATGGAAGCACACAAAATTAGGTTGGGATATCAAMSEQ ID NO: 11 Stop MF2.1 SNP_2 MFPosition in Dempsey_1.1: chr02 - 163,630,465 bpACTTCCTTTTTATTTGTAAATATGGGCCCATAACTCACAGGTCCCGGGATC GAAACCTGCCTCTTTCCAGGGGNCCATAGGAAAGAGCCTCTCTACCTCACSEQ ID NO: 12 Stop MF2.1 SNP_2 wtPosition in Dempsey_1.1: chr02 - 163,630,465 bpACTTCCTTTTTATTTGTAAATATGGGCCCATAACTCACAGGTCCCGGGATA GAAACCTGCCTCTTTCCAGGGGNCCATAGGAAAGAGCCTCTCTACCTCACSEQ ID NO: 13 Start MF2.2 SNP_3 MFPosition in Dempsey_1.1: chr02 - 137,347,441 bpAGAACGGCTAGCTATCAGACCCATCCATGGAGAAGATTCTCAAGGTGGAAGCCGTKCACAG- TATGAGGTCAAAATTCGTAGAGCAGAGAAGTTTAAAGACA AGMTGATTTTTACACTCCCAGCAACTGGTCAGATYAYGGAAGTTAATGTCSEQ ID NO: 14 Start MF2.2 SNP_3 wtPosition in Dempsey_1.1: chr02 - 137,347,441 bpAGAACGGCTAGCTATCAGACCCATCCATGGAGAAGATTCTCAAGGTGGAAGCCGTKCACAG- TATGAGGTCAAAATTCGTAGAGCAGAGAAGTTTAAAGACC AGMTGATTTTTACACTCCCAGCAACTGGTCAGATYAYGGAAGTTAATGTCSEQ ID NO: 15 Stop MF2.2 SNP_4 MFPosition in Dempsey_1.1: chr02 - 151,147,982 bpCACATAGACCTTCTGAAGATATTGCATTTGCAATTGCTCGTTTCTTTCAGC GTGGGGTCAGCTTACAGAACTATTATATGTATTTTGATGGGACAAATTTTSEQ ID NO: 16 Stop MF2.2 SNP_4 wtPosition in Dempsey_1.1: chr02 - 151,147,982 bpCACATAGACCTTCTGAAGATATTGCATTTGCAATTGCTCGTTTCTTTCAGT GTGGGGTCAGCTTACAGAACTATTATATGTATTTTGATGGGACAAATTTTSEQ ID NO: 17 Start QTL ft12.1 SNP_5 MF – suppressor allele.Position in Dempsey_1.1: chr12 – 7,392,029ATGAAAGATGGTCTAGGGACCCACGTGACGGCGAGCTTTGCAGCTGGATTC GTGGCTGCAG-TAGCCTCAAATCCAGTCGACRTGATTAAGACACGTGTYATSEQ ID NO: 18 Start QTL ft12.1 SNP_5 wt non-suppressor allelePosition in Dempsey_1.1: chr12 – 7,392,029ATGAAAGATGGTCTAGGGACCCACGTGACGGCGAGCTTTGCAGCTGGATTT GTGGCTGCAG-TAGCCTCAAATCCAGTCGACRTGATTAAGACACGTGTYATNunhems Netherlands B.V. 230055WO0141SEQ ID NO: 19 Stop QTL ft12.1 SNP_6 MF – suppressor allelePosition in Dempsey_1.1: chr12 - 13,736,988 bpTTTCAATAATTGTGCTTCCTCAACATGATCCCTTAGAGAGAATTGCATGAT GAACTTCACAACGTGAGGCAAACATTACTAATCGAGGAAAAGCAAAGGAASEQ ID NO: 20 Stop QTL ft12.1 SNP_6 wt – non-suppressor allelePosition in Dempsey_1.1: chr12 - 13,736,988 bpTTTCAATAATTGTGCTTCCTCAACATGATCCCTTAGAGAGAATTGCATGAC- GAACTTCACAACGTGAGGCAAACATTACTAATCGAGGAAAAGCAAAGGAA SEQ ID NO: 21 non-pungent C. annuum pun1 protein acyltransferase; MAFALPSSLV SVCNKSFIKP SSLTPSTLRF HKLSFIDQSL SNMYIPCAFF YPKVQQRLED SKNSDELSHI AHLLQTSLSQ TLVSYYPYAG KLKDNATVDC NDMGAEFLSV RIKCSMSEIL DHPHASLAES IVLPKDLPWA NNCEGGNLLV VQVSKFDCGG IAISVCFSHK IGDGCSLLNF LNDWSSVTRD PTTTTLVPSP RFVGDSVFST QKYGSLITPQ ILSDLNQCVQ KRLIFPTDKL DALRAKVAEE SGVKNPTRAE VVSALLFKCA TKASSSMLPS KLVHFLNIRT MIKPRLPRNA IGNLSSIFSI EATNMQDMEL PTLVRNLRKE VEVAYKKDQV EQNELILEVV ESMREGKLPF ENMDGYKNVY NLQQSLQISI LHCRFWMGKT *SEQ ID NO: 22 non-pungent C. annuum pun1 acyltransferase CDNAATGGCTTTTG CATTACCATC ATCACTTGTT TCAGTTTGTA ACAAATCTTT TATCAAACCT TCCTCTCTCA CCCCCTCTAC ACTTAGATTT CACAAGCTAT CTTTCATCGA TCAATCTTTA AGTAATATGT ATATCCCTTG TGCATTTTTT TACCCTAAAG TACAACAAAG ACTAGAAGAC TCCAAAAATT CTGATGAGCT TTCCCATATA GCCCACTTGC TACAAACATC TCTATCACAA ACTCTAGTCT CTTACTATCC TTATGCTGGA AAGTTGAAGG ACAATGCTAC TGTTGACTGT AACGATATGG GAGCTGAGTT CTTGAGTGTT CGAATAAAAT GTTCCATGTC TGAAATTCTT GATCATCCTC ATGCATCTCT TGCAGAGAGC ATAGTTTTGC CCAAGGATTT GCCTTGGGCG AATAATTGTG AAGGTGGTAA TTTGCTTGTA GTTCAAGTAA GTAAGTTTGA TTGTGGGGGA ATAGCCATCA GTGTATGCTT TTCGCACAAG ATTGGTGATG GTTGCTCTCT GCTTAATTTC CTTAATGATT GGTCTAGCGT TACTCGTGAT CCTACGACAA CAACTTTAGT TCCATCTCCT AGATTTGTAG GAGATTCAGT CTTCTCTACA CAAAAATATG GTTCTCTCAT TACGCCACAA ATTTTGTCCG ATCTCAACCA GTGCGTACAG AAAAGACTCA TTTTTCCTAC AGATAAGTTA GATGCACTTC GAGCTAAGGT GGCAGAAGAA TCAGGAGTAA AAAATCCAAC AAGGGCTGAA GTTGTTAGCG CTCTTCTTTT CAAATGTGCA ACAAAGGCAT CATCATCAAT GCTACCATCA AAGTTGGTTC ACTTCTTAAA CATACGTACT ATGATCAAAC CTCGTCTACC ACGAAATGCC ATTGGAAATC TCTCGTCTAT TTTCTCCATA GAAGCAACTA ACATGCAGGA CATGGAGTTG CCAACGTTGG TTCGTAATTT AAGGAAGGAA GTTGAGGTGG CATACAAGAA AGACCAAGTC GAACAAAATG AACTGATCCT AGAAGTAGTA GAATCAATGA GAGAAGGGAA ACTGCCATTT GAAAATATGG ATGGCTATAA GAATGTGTAT AACTTGCAGC AATCTTTGCA AATATCCATA CTACACTGTA GATTTTGGAT GGGGAAGACC TGA SEQ ID NO 23: C. frutescens PUN1-1 / AT3 protein acyltransferase; AAV66308 MAFALPSSLV SICDKSFIKP SSLTPSTLRF HKLSFIDQSL SNMYIPCAFF YPKVQQRLED SKNSDELSHI AHLLQTSLSQ TLVSYYPYAG KLKDNATVDC NDMGAEFLSV RIKCSMSEIL DHPHASLAES IVLPKDLPWA NNCEGGNLLV VQVSKFDCGG IAISVCFSHK IGDGCSLLNF LNDWSSVTRD HTTTTLVPSP RFVGDSVFST KKYGSLITPQ ILSDLNECVQ KRLIFPTDKL DALRAKVAEE SGVKNPTRAE VVSALLFKCA TKASSSMLPS KLVHFLNIRT MIKPRLPRNA IGNLSSIFSI EATNMQDMEL PTLVRNLRKE VEVAYKKDQV EQNELILEVV ESMREGKLPF ENMDGYENVY TCSNLCKYPY YTVDFGWGRP ERVCLGNGPS KNAFFLKDYK AGQGVEARVM LHKQQMSEFE RNEELLEFIASEQ ID NO: 24 C. frutescens PUN1-1 / AT3 acyltransferase CDNA; AY819026ATGGCTTTTG CATTACCATC ATCACTTGTT TCAATTTGTG ACAAATCTTT TATTAAACCT TCCTCTCTCA CCCCCTCTAC ACTTAGATTT CACAAGCTAT CTTTCATCGA TCAATCTTTA AGTAATATGT ATATCCCTTG TGCATTTTTT TACCCTAAAG TACAACAAAG ACTAGAAGACNunhems Netherlands B.V. 230055WO0142 TCCAAAAATT CTGATGAGCT TTCCCATATA GCCCACTTGC TACAAACATC TCTATCACAA ACTCTAGTCT CTTACTATCC TTATGCAGGA AAGTTGAAAG ACAATGCTAC TGTTGACTGT AACGATATGG GAGCTGAGTT CTTGAGTGTT CGAATAAAAT GTTCCATGTC TGAAATTCTT GATCATCCTC ATGCATCTCT TGCAGAGAGC ATAGTTTTGC CCAAGGATTT GCCTTGGGCG AATAATTGTG AAGGTGGTAA TTTGCTTGTA GTTCAAGTAA GTAAGTTTGA TTGTGGGGGA ATAGCCATCA GTGTATGCTT TTCGCACAAG ATTGGTGATG GTTGCTCTCT GCTTAATTTC CTTAATGATT GGTCTAGCGT TACTCGTGAT CATACGACAA CAACTTTAGT TCCATCTCCT AGATTTGTAG GAGATTCTGT CTTCTCTACA AAAAAATATG GTTCTCTTAT TACGCCACAA ATTTTGTCCG ATCTCAACGA GTGCGTACAG AAAAGACTCA TTTTTCCTAC AGATAAGTTA GATGCACTTC GAGCTAAGGT GGCAGAAGAA TCAGGAGTAA AAAATCCAAC AAGGGCAGAA GTTGTTAGCG CTCTTCTTTT CAAATGTGCA ACAAAGGCAT CATCATCAAT GCTACCATCA AAGTTGGTTC ACTTCTTAAA CATACGTACT ATGATCAAAC CTCGTCTACC ACGAAATGCC ATTGGAAATC TCTCGTCTAT TTTCTCCATA GAAGCAACTA ACATGCAGGA CATGGAGTTG CCAACGTTGG TTCGTAATTT AAGGAAGGAA GTTGAGGTGG CATACAAGAA AGACCAAGTC GAACAAAATG AACTGATCCT AGAAGTAGTA GAATCAATGA GAGAAGGGAA ACTGCCATTT GAAAATATGG ATGGCTATGA GAATGTGTAT ACTTGCAGCA ATCTTTGCAA ATATCCATAC TACACTGTAG ATTTTGGATG GGGAAGACCT GAAAGGGTGT GTCTAGGAAA TGGTCCCTCC AAGAATGCCT TCTTCTTGAA AGATTACAAA GCTGGGCAAG GCGTGGAGGC GCGGGTGATG TTGCACAAGC AACAAATGTC TGAATTTGAA CGCAATGAGG AACTCCTTGA GTTCATTGCC TAASEQ ID NO: 25 Stop MF2.1 SNP_7 MFPosition in Dempsey_1.1: chr02 - 162,234,480 bpCTGAGTTCCATTTTTACATGCTGTCCCTGCTCATTTTCTCACTTTGNTATC TTTCTGTGAATGAATCTTTTTCTCATATATTAGTGCGCATCTTTTCTCCASEQ ID NO: 26 Stop MF2.1 SNP_7 wtPosition in Dempsey_1.1: chr02 - 162,234,480 bpCTGAGTTCCATTTTTACATGCTGTCCCTGCTCATTTTCTCACTTTGNTATT TTTCTGTGAATGAATCTTTTTCTCATATATTAGTGCGCATCTTTTCTCCASEQ ID NO: 27 Start MF2.2 SNP_8 MFPosition in Dempsey_1.1: chr02 - 143,111,884 bpGAACTTAAAGATGCAACCTGACACACAAGAGTCTAAAGGATCCTTTGATTC GACTTCACAGCTACAATTTAACAATAAGCTTGATCATGCTTTTCCTGAGGSEQ ID NO: 28 Start MF2.2 SNP_8 wtPosition in Dempsey_1.1: chr02 - 143,111,884 bpGAACTTAAAGATGCAACCTGACACACAAGAGTCTAAAGGATCCTTTGATTT GACTTCACAGCTACAATTTAACAATAAGCTTGATCATGCTTTTCCTGAGGSEQ ID NO: 29 Stop MF2.2 SNP_9 MFPosition in Dempsey_1.1: chr02 - 147,107,830 bpTCGATTAATGCATGAAAATGCAGTAATCATACATTGTGATGCTGACAACCG AGGNTGTAAAGATTACAGAAAGGTGTATTCCAATATATAGTAGAAAAAATSEQ ID NO: 30 Stop MF2.2 SNP_9 wtPosition in Dempsey_1.1: chr02 - 147,107,830 bpTCGATTAATGCATGAAAATGCAGTAATCATACATTGTGATGCTGACAACCA AGGNTGTAAAGATTACAGAAAGGTGTATTCCAATATATAGTAGAAAAAATNunhems Netherlands B.V. 230055WO0143 EXAMPLES

[0159] Unless stated otherwise in the Examples, all recombinant techniques are carried out ac-cording to standard protocols as described in “Sambrook J and Russell DW (eds.) (2001) Molec- ular Cloning: A Laboratory Manual, 3rd Edition, Cold Spring Harbor Laboratory Press, New York” and in “Ausubel FA, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA and Struhl K (eds.) (2006) Current Protocols in Molecular Biology. John Wiley & Sons, New York”.

[0160] Standard materials and references are described in “Croy RDD (ed.) (1993) Plant MolecularBiology LabFax, BIOS Scientific Publishers Ltd., Oxford and Blackwell Scientific Publications,Oxford” and in “Brown TA, (1998) Molecular Biology LabFax, 2nd Edition, Academic Press, SanDiego”. Standard materials and methods for polymerase chain reactions (PCR) can be found in “McPherson MJ and Møller SG (2000) PCR (The Basics), BIOS Scientific Publishers Ltd., Oxford”and in “PCR Applications Manual, 3rd Edition (2006), Roche Diagnostics GmbH, Mannheim orwww.roche-applied-science.com“.

[0161] It should be understood that a number of parameters in any lab protocol such as the PCRprotocols in the below Examples may need to be adjusted to specific laboratory conditions, and may be modified slightly to obtain similar results. For instance, use of a different method for preparation of DNA or the selection of other primers in a PCR method may dictate other optimal conditions for the PCR protocol. These adjustments will however be apparent to a person skilled in the art and are furthermore detailed in current PCR application manuals. EXAMPLE 1Phenotyping of flowering behaviour in Capsicum annuum plants

[0162] Phenotyping of the plants of interest is carried out over the entire life cycle of the plants.The life cycle of the plant is from seedling to full maturity towards the end of the growing cycle. The growing cycle includes sowing around March / April, followed by transplanting around April / May, with the plants continuing to grow until the end of the season around November. Grow- ing conditions during phenotype data collection are best described as medium-tech greenhouse conditions, Almeria, Spain.

[0163] Phenotype observations are recorded 3 times per growing season, aiming at early, middle,and late plant stage.

[0164] The early plant stage, or 1st evaluation comprises data records approximately for themonths May and June. The middle plant stage, or 2nd evaluation comprises data records approx- imately for the months July and August. The late plant stage or, 3rd evaluation comprises data records approximately for the months September October and November.

[0165] Values used for data analyses per evaluation 1 2 or 3, are percentages of nodes havingmore than 1 flower. On every single plant, per branched node a visual tag is applied whenever the node of the branch shows more than 1 flower. For every evaluation, (1,2 and 3), a differentcolour visual tag was used. Applying the coloured tags secures the possibility of data collectionNunhems Netherlands B.V. 230055WO0144at a later timepoint even when flowers have dropped / aborted for any reason. A Capsicum annuumplant is categorized to have the “multiple flowers” flowering phenotype when several nodes showmore than 1 flower per node compared to a Capsicum annuum line without QTLs MF2.1 andMF2.2. Co-segregation between the described MF2.1 and or MF2.2 QTLs and the described phenotype can be established by means of QTL mapping. EXAMPLE 2Identification of QTLs capable of inducing a multiple flowers phenotype in Capsicum an-nuum plants

[0166] A cross was made between a Capsicum chinense line having a multiple flowers phenotypeand which is indicated herein as DONOR01 and an inbred Charliston Capsicum annuum linewhich has a solitary flowers phenotype. The BC1F1 generation which is obtained by backcrossingbetween the F1 and the Charliston Capsicum annuum line was phenotyped as described in Ex-ample 1. From the BC1F1 generation a subset of plants were selected based on the percentageof multiple flowers per internode, fertility and good horticultural value. The resulting subset ofBC1F1 plants was selfed to provide a BC1F2 population which again was phenotyped as de- scribed in Example 1 and subsequently used for QTL mapping.

[0167] The Molecular markers linked to the multiple flowers phenotype accordingly were developedby linkage mapping (Doerge, 2002; Koornneef et al., 2004). Marker-trait associations wereidentified using interval mapping implemented in Rqtl (Doerge, 2002). First a Genome WideMapping (GW) was performed using a marker density of approximately 1 marker every 15 cM.Once QTLs were identified a several additional rounds of mapping or saturation mapping wasperformed by adding further SNP markers within the QTL intervals identified by GW mapping.Three rounds of fine mapping were performed by evaluating BC1F3, BC1F4 and BC1F5 recom-binants within the chromosomal regions of interest. The identified QTLs MF2.1 and MF2.2 asso-ciated with the multiple flowers phenotype according to the present invention are indicated herein below in Table 1.

[0168] It was further surprisingly found that epistatic interactions between QTL MF2.1 and QTLMF2.2 and a suppressor allele may result into severe linkage drag in later inbreeding stages.Particularly, it was found that segregation during inbreeding is distorted when the suppressorallele of QTL ft12.1 as described herein is present in the genome, results in a dramatically de-creased proportion of the desired multiple flowers allele in the offspring. The identified QTL ft12.1associated with the suppression of the multiple flowers phenotype is indicated herein below inTable 1.Table 1. QTLs associated to multiple flowers phenotypeexplained LOD Position in reference genome Demspey_1.1 variance value (95% confidence interval) QTL MF2.1 21% 11.9 between chr02:158316753 and chr02:163630465,preferably between chr02:158316753 and chr02:162234840Nunhems Netherlands B.V. 230055WO0145 QTL MF2.2 53.6% 45.3 between chr02:137347441 and chr02:151147982,preferably between chr02:143111884 andchr02:147107830 QTL ft12.1 30.7% 23.9 between chr12:7392029 and chr12:13736988

[0169] The herein-described QTL MF2.1, QTL MF2.2 and QTL ft12.1 can also be identified usingthe flanking markers as indicated herein below in Table 2.Table 2. Nucleotide sequences of the identified flanking markers(“MF” means multiple flowers allele; “wt” means wild type allele; “sup” means suppressor allele;“null” means null allele, i.e. not the suppressor allele)pheno- SEQ ID Sequence type NO: SNP_1 MF 9 TCAAGGGATTGAAGATTTGGTGCTGAACCAAATGTTGGAGGTGGATGAATACTGGAGACRAAACAACATGGAAGCACACAAAATTAGGTTGGGATATCAAM SNP_1 wt 10 TCAAGGGATTGAAGATTTGGTGCTGAACCAAATGTTGGAGGTGGATGAATGCTGGAGACRAAACAACATGGAAGCACACAAAATTAGGTTGGGATATCAAM SNP_2 MF 11 ACTTCCTTTTTATTTGTAAATATGGGCCCATAACTCACAGGTCCCGGGATCGAAACCTGCCTCTTTCCAGGGGNCCATAGGAAAGAGCCTCTCTACCTCAC SNP_2 wt 12 ACTTCCTTTTTATTTGTAAATATGGGCCCATAACTCACAGGTCCCGGGATAGAAACCTGCCTCTTTCCAGGGGNCCATAGGAAAGAGCCTCTCTACCTCAC SNP_3 MF 13 AGAACGGCTAGCTATCAGACCCATCCATGGAGAAGATTCTCAAGGTGGAA-GCCGTKCACAGTATGAGGTCAAAATTCGTAGAGCAGAGAAGTTTAAAGACA AGMTGATTTTTACACTCCCAGCAACTGGTCAGATYAYGGAAGTTAATGTC SNP_3 wt 14 AGAACGGCTAGCTATCAGACCCATCCATGGAGAAGATTCTCAAGGTGGAA-GCCGTKCACAGTATGAGGTCAAAATTCGTAGAGCAGAGAAGTTTAAAGACC AGMTGATTTTTACACTCCCAGCAACTGGTCAGATYAYGGAAGTTAATGTC SNP_4 MF 15 CACATAGACCTTCTGAAGATATTGCATTTGCAATTGCTCGTTTCTTTCAGCGTGGGGTCAGCTTACAGAACTATTATATGTATTTTGATGGGACAAATTTT SNP_4 wt 16 CACATAGACCTTCTGAAGATATTGCATTTGCAATTGCTCGTTTCTTTCAGTGTGGGGTCAGCTTACAGAACTATTATATGTATTTTGATGGGACAAATTTT SNP_5 null 18 ATGAAAGATGGTCTAGGGACCCACGTGACGGCGAGCTTTGCAGCTGGATTTGTGGCTGCAGTAGCCTCAAATCCAGTCGACRTGATTAAGACACGTGTYAT SNP_5 sup 17 ATGAAAGATGGTCTAGGGACCCACGTGACGGCGAGCTTTGCAGCTGGATTCGTGGCTGCAGTAGCCTCAAATCCAGTCGACRTGATTAAGACACGTGTYAT SNP_6 null 20 TTTCAATAATTGTGCTTCCTCAACATGATCCCTTAGAGAGAATTGCATGACGAACTTCACAACGTGAGGCAAACATTACTAATCGAGGAAAAGCAAAGGAA SNP_6 sup 19 TTTCAATAATTGTGCTTCCTCAACATGATCCCTTAGAGAGAATTGCATGATGAACTTCACAACGTGAGGCAAACATTACTAATCGAGGAAAAGCAAAGGAA SNP_7 MF 25 CTGAGTTCCATTTTTACATGCTGTCCCTGCTCATTTTCTCACTTTGNTATCTTTCTGTGAATGAATCTTTTTCTCATATATTAGTGCGCATCTTTTCTCCA SNP_7 wt 26 CTGAGTTCCATTTTTACATGCTGTCCCTGCTCATTTTCTCACTTTGNTATTTTTCTGTGAATGAATCTTTTTCTCATATATTAGTGCGCATCTTTTCTCCA SNP_8 MF 27 GAACTTAAAGATGCAACCTGACACACAAGAGTCTAAAGGATCCTTTGATTCGACTTCACAGCTACAATTTAACAATAAGCTTGATCATGCTTTTCCTGAGG SNP_8 wt 28 GAACTTAAAGATGCAACCTGACACACAAGAGTCTAAAGGATCCTTTGATTTGACTTCACAGCTACAATTTAACAATAAGCTTGATCATGCTTTTCCTGAGGNunhems Netherlands B.V. 230055WO0146 SNP_9 MF 29 TCGATTAATGCATGAAAATGCAGTAATCATACATTGTGATGCTGACAACCGAGGNTGTAAAGATTACAGAAAGGTGTATTCCAATATATAGTAGAAAAAAT SNP_9 wt 30 TCGATTAATGCATGAAAATGCAGTAATCATACATTGTGATGCTGACAACCAAGGNTGTAAAGATTACAGAAAGGTGTATTCCAATATATAGTAGAAAAAAT EXAMPLE 3Breeding of non-pungent Capsicum annuum plant comprising QTL MF2.1 and QTL MF2.2

[0170] In the context of the present invention, it was accordingly found that QTL MF2.1 and QTLMF2.2 are located at chromosome 2, wherein both QTL MF2.1 and QTL MF2.2 are closely linkedto the Pun1 locus. Pungency in pepper is qualitatively controlled by the Pun1 locus, which en-codes a putative acyltransferase enzyme. Conventional non-pungent chili pepper have a non- functional Capsaicin synthase gene, e.g. comprising a 2529 or 4 bp-deletion in this gene leadingto a truncated protein. Therefore, both QTL MF2.1 and QTL MF2.2 cannot be readily introgressedfrom the Capsicum chinense donor line (DONOR01) without also introgressing the chinense pun-gency capsaicin synthase gene, resulting in Capsicum annuum progeny that produces hot pepperfruits. When setting out to provide a non-pungent Capsicum annuum plant having the multipleflowers phenotype of the present invention, it appeared not possible to introgress the QTL capableof inducing the multiple flowers phenotype from a donor C. chinense plant without also introgress-ing the C. chinense PUN1 gene leading to a pungent pepper fruit phenotype. Only in the contextof the present invention, it was surprisingly found that the Pun1 locus is located in between QTLMF2.1 and MF2.2 as described herein and that a double recombinant non-pungent C. annuumplant comprising in its genome QTL MF2.1 and QTL MF2.2 could be obtained, which requiredmultiple generations having a population sizes larger than 260 plants per generation.

[0171] Capsicum annuum plants having the multiple flowers phenotype according to the presentinvention and which produce non-pungent pepper fruits were obtained as follows. In the BC1F3generation as described in Example 1, a recombination event was identified which resulted inQTL MF2.1 to be homozygous present, while QTL MF2.2 and the chinense pungency Capsaicinsynthase gene were heterozygous present. In subsequent BC1F4 generation a recombinationevent was identified which resulted in QTL MF2.1 to be homozygous present, QTL MF2.2 to beheterozygous present and the Capsaicin synthase gene homozygous non-pungent. This recom- binant BC1F4 plant, was self-pollinated to obtain BC1F5 seeds which are homozygous for QTLMF2.1, homozygous for the non-pungent Capsaicin synthase gene and homozygous for QTLMF2.2.

[0172] BC1F5 seeds were self-pollinated to produce the BC1F6 generation from which the seeddeposits NCIMB 44383 and NCIMB 44384 are selected.EXAMPLE 4 Mutagenesis of AT3 gene in Pun1 locus

[0173] The Capsaicin synthase gene (CS) also known as Acyltransferase (AT3), is responsible forthe last catalytic step of capsaicin biosynthesis. Loss of function of the Pun1 / AT3 gene results innon-pungency; see e.g. Kirii et al. (2017) doi: 10.2503 / hortj.MI-148.Nunhems Netherlands B.V. 230055WO0147

[0174] Mutant Capsicum annuum plants having the multiple flowers phenotype according to thepresent invention and having a loss of function mutation in the AT3 gene in Pun1 locus may be obtained by random mutagenesis followed by reverse screening from TILLING mutant population.

[0175] A highly homozygous inbred line homozygous for QTL MF2.1 and QTL MF2.2 as describedherein can be used for mutagenesis treatment with the following protocol. After seed imbibitionon damp Whatman® paper for 24h, -20,000 seeds, divided in 8 batches of 2500 respectively, is soaked in 100 ml of ultrapure water and ethyl methanesulfonate (EMS) at a concentration of 1% in conical flasks. The flasks are gently shaken for 16h at room temperature. Finally, EMS is rinsed out under flowing water. Following EMS treatment, seeds are directly sown in the greenhouse. Out of the seeds that germinate, a sufficient number of plantlets are trans-planted in the field. From these plantlets, at least one fruit is harvested from the surviving and plant bearing plants. For instance, from each remaining M1 mutant plant one fruits is harvested and its seeds isolated.From the obtained population, named M2 population, specific families may be excluded from thepopulation due to low seed set.

[0176] DNA is extracted from a pool of 10 seeds originating from each M2 seed lot. Per mutantline, 10 seeds are pooled in a Micronic® deepwell tube; http: / / www.micronic.com from a 96 deep-well plate, 2 stainless balls are added to each tube. The tubes and seeds are frozen in liquid nitrogen for 1 minute and seeds are immediately ground to a fine powder in a Deepwell shaker (Vaskon 96 grinder, Belgium; http: / / www.vaskon.com) for 2 minutes at 16,8 Hz (80% of the max-imum speed). 300 μ l Agowa® Lysis buffer P from the AGOWA® Plant DNA Isolation Kithttp: / / www.agowa.de is added to the sample plate and the powder is suspended in solution byshaking 1 minute at 16,8 Hz in the Deepwell shaker. Plates are centrifuged for 10 minutes at 4000rpm.75 μl of the supernatant is pipetted out to a 96 Kingfisher plate using a Janus MDT® (PerkinElmer, USA; http: / / www.perkinelmer.com) platform (96 head). The following steps are performedusing a Perkin Elmer Janus® liquid handler robot and a 96 Kingfisher® (Thermo labsystems,Finland; http: / / www.thermo.com). The supernatant containing the DNA is diluted with binding buffer (150 μl) and magnetic beads (20 μl). Once DNA is bound to the beads, two successive washing steps are carried out (Wash buffer 1: Agowa wash buffer 11 / 3, ethanol 1 / 3, isopropanol 1 / 3; Wash buffer 2: 70% ethanol, 30% Agowa wash buffer 2) and finally eluted in elution buffer (100 μl MQ, 0,025 μl Tween).

[0177] Grinding 4 C. annuum seeds generally produces enough DNA to saturate the magneticbeads, thus highly homogenous and comparable DNA concentrations of all samples are obtained. Comparing with lambda DNA references, a concentration of 30 ng / μl for each sample is esti- mated. Two times diluted DNA was 4 fold flat pooled.2 μl pooled DNA was used in multiplex PCRs for mutation detection analysis.

[0178] High Resolution Melt curve analysis (HRM) is proven to be sensitive and high-throughputmethods in human and plant genetics. HRM is a non-enzymatic screening technique. During the PCR amplification dye (LCGreen+ dye, Idaho Technology Inc., UT, USA) molecules intercalate between each annealed base pair of the double stranded DNA molecule. When captured in the molecule, the dye emits fluorescence at 510 nm after excitation at 470 nm. A camera in a fluo- rescence detector (LightScanner, Idaho Technology Inc., UT, USA) records the fluorescence in- tensity while the DNA sample is progressively heated. At a temperature dependent on the se- quence specific stability of the DNA helices, the double stranded PCR product starts to melt,Nunhems Netherlands B.V. 230055WO0148 releasing the dye. The release of dye results in decreased fluorescence that is recorded as a melting curve by the fluorescence detector. Pools containing a mutation form hetero duplexes in the post-PCR fragment mix. These are identified as differential melting temperature curves in comparison to homo duplexes.

[0179] Primers useful to amplify gene fragments for HRM are designed using a computer program(Primer3, http: / / primer3.sourceforge.net / ). The length of the amplification product is limited be- tween 200 and 400 base pairs. Quality of the primers is determined by a test PCR reaction that should yield a single product.

[0180] Polymerase Chain Reaction (PCR) to amplify gene fragments can be performed as follows.10ng of genomic DNA is mixed with 4µl reaction buffer (5x Reaction Buffer), 2µl 10xLC dye ((LCGreen+ dye, Idaho Technology Inc., UT, USA), 5pmole of forward and reverse primers each, 4nmole dNTPs (Life Technologies, NY, USA) and 1 unit DNA polymerase (Hot Start II DNA Pol- ymerase) in a total volume of 10µl. Reaction conditions were: 30s 98°C, then 40 cycles of 10s. 98°C, 15s 60°C, 25s of 72°C and finally 60s at 72°C.

[0181] The presence of a particular mutation in individual plants is confirmed by repeating the HRManalysis on DNA from the individual M2 seed lots of the identified corresponding DNA pool. When the presence of the mutation, based on the HRM profile, is confirmed in one of the four individual M2 family DNA samples, the PCR fragments are sequenced to identify the mutation in the gene.

[0182] Once the mutation is known the effect of such a mutation can be predicted, e.g. by using acomputer program CODDLe (for Choosing codons to Optimize Discovery of Deleterious Lesions, http: / / www.proweb.org / coddle / ) that identifies the region(s) of a user-selected gene and of its cod- ing sequence where the anticipated point mutations are most likely to result in deleterious effects on the gene's function.

[0183] Seeds from M2 families that contain mutations with predicted effect on protein activity aresown for phenotypic analysis of the plants. Homozygous mutants are selected or obtained after selfing and subsequent selection. The effect of the mutation on the corresponding protein and phenotype of the plant is subsequently determined, for instance by using the methods as de- scribed herein. EXAMPLE 5 Characterization of the multiple flowers trait according to the present invention

[0184] Test F1 hybrids in 3 different elite Capsicum annuum backgrounds were made with variousgenetic status for QTLs MF2.1 and MF2.2.

[0185] These results as described in Table 3 indicate that both QTL MF2.1 and QTL MF2.2 arerequired to obtain the multiple flowers trait according to the present invention. Among these 3backgrounds pungent and non-pungent backgrounds were used. Non-pungent materials were obtained through double recombination event described in Example 3.Nunhems Netherlands B.V. 230055WO0149 C C C C C C C C C C C C C C S T .a.a.a.a.a.a.a.a.a.a.a.a.a.cp a e b n n n n n n n hlenu n n n n n n n c u nu nu nu nu n n n n n n ninie 3u u u u u u uu uu uu uu uu u u e s . m m m mmmm m m mmumumn R see suJa C C S J C C S S S J C C H Pltslaa h e a mlaa h em e e a a h a epp y e e a nrnliip y e e a m mlanrlii i ip ye a b p h nrlia p en o ns cetoh n ns ca o etohc cen nsne e o a ha ha o etorttx xnrelix xnrlir rnro ypyps s x s s s x slislis(likeicmemseto e em n mmsetototoecm n n n )hichici icica r ahacrlisrhh hic ac tlisa a a h oratelirlisrlisrl rnto s tnto oto isizn n nto antion 70 C P B 7 C P B B B 7 C P 6 PIX(PC 0 5 6 PIX(PC(PC(PD D C 06 PIXO e 3IX1IXIXIXs -M8 -0x1 -0F 5 3 1 4 -M8 -01 x 1 -0F 1 3 - 1 1 0 F2 -0F 5 3 2 -M8 -0N c 1 Ori xBx1 1 xBxR 0pBCB / CD BCB / D / D / ti &D& 1oC 1 O C 1 C O O O n 1 F 1 N 1 F 1 N N N F 4 F 4 4 O F 3 F O O O R 3 3 R R B B C R 0 0 0 0 C 1 1 1 1 1 ) ) ) F 1 3 ) F3 M T M T M TrS T T TrS p M p M E E E O d PF es F es F ees c e es es ees c e op a op alitlitlitri es e22t2 2 oleoleF 2u p u pe e e gc d. . t . tmc tFtFtFmcp pinig1F 21F1 bte1 1 1 btelatini latni Ja CacharipreM M Mnid aM M M nidog ioglayalde F F F a n n peenrl tio ioF F F n 2 n stn n 2 2 t 2 2 t n noo . 2 1.1.1.1.1.1o e n r ++ ++ / + + + - / - / - / +- / - - / - - / - + / A A - ny n - / y - - / - - / - + / M A + F2lle.2les+ / + - / - - / - - / - + / + + + A A + +ta- / - / - / - ny n - / y / + / + -+ / A P + T u 3 ntu1 s / forde + + + + + + + + A A s / - / - / - / + / - / - / - / +ny n - / y - - / - - / - + / M + F c 2r.i 1bed - / - - / - - / - - / - - / - - / - - / - - / - + / + - / +- / - - / - - / - + / FQ+T1 T 2 L .1s M M M M S S S M usF F S S S M P F o o oul suo o o h l u tiplu tilu tiltilitalitalu italtipop w plop wlitalu italitaltielon w lp p peleleleryfloflofloflo fr rpreereer r rpo e lyoflyflelf ssrinriy y y legssngf f ftyrinwowowlo ede l l ldofwowowlope g w w w we e e w e w e r e e e srsrsrsrsrsrse e e r e srsrsrsrsNunhems Netherlands B.V. 230055WO0150 CLAIMS1. A Capsicum annuum plant comprising in its genome QTL MF2.1 located between158,316,753 bp and 163,630,465 bp on chromosome 2 of C. annuum Dempsey V1.1reference genome; and QTL MF2.2 located between 137,347,441 bp and 151,147,982 bp on chromosome 2 of C. annuum Dempsey V1.1 reference genome, wherein said QTLMF2.1 and QTL MF2.2 confer multiple flowers per node relative to a plant lacking saidQTL MF2.1 and QTL MF2.2.2. The Capsicum annuum plant according to claim 1, wherein QTL MF2.1 and QTL MF2.2confer an increased number of flowers per node relative to:a plant comprising QTL MF2.1 and lacking QTL MF2.2; or a plant comprising QTL MF2.2 and lacking QTL MF2.1.3. The Capsicum annuum plant according to claim 1 or 2, wherein QTL MF2.1 and QTLMF2.2 are as present in, or as obtainable from, or as obtained from, or as comprised in the genome of a Capsicum annuum plant designated: PP193-22, a representative sam-ple of which has been deposited under accession number NCIMB 44384.4. The Capsicum annuum plant according to any one of claims 1 to 3, wherein said plantcomprises a mutant allele of the Pun1 / AT3 gene, wherein said mutant allele results in areduced expression or no expression of the wild type Pun1 / AT3 gene and / or wherein themutant allele encodes a protein having a decreased function or loss-of-function when compared to the wild type Pun1 / AT3 protein and wherein the wild type Pun1 / AT3 geneencodes a protein comprising at least 80% sequence amino acid sequence identity to SEQ ID NO: 1.5. The Capsicum annuum plant according to claim 4, wherein QTL MF2.1, the mutant alleleof the Pun1 / AT3 gene and QTL MF2.2 are comprised in a recombinant introgression onchromosome 2.6. The Capsicum annuum plant according to claim 5, wherein said recombinant introgres-sion is as present in, or as obtainable from, or as obtained from, or as comprised in the genome of a Capsicum annuum plant designated: PP183-72, a representative sampleof which has been deposited under accession number NCIMB 44383.7. The Capsicum annuum plant according to any one of claims 1 to 5, wherein said plant ishomozygous for QTL MF2.1 and is homozygous for QTL MF2.2.8. The Capsicum annuum plant according to any one of claims 1 to 7, wherein said plantcomprises QTL ft12.1 on chromosome 12, wherein said QTL ft12.1 is located betweenabout 7,392,029 bp and about 13,736,988 bp on chromosome 12 of the C. annuum Dempsey V1.1 reference genome and wherein said QTL ft12.1 prevents suppression ofthe multiple flowers allele in the offspring of said Capsicum annuum plant.Nunhems Netherlands B.V. 230055WO01519. The Capsicum annuum plant according to any one of claims 1 to 8, wherein said QTLft12.1 is as present in, or as obtainable from, or as obtained from, or as comprised in thegenome of a Capsicum annuum plant designated: PP193-22, a representative sampleof which has been deposited under accession number NCIMB 44384; or PP183-72, a representative sample of which has been deposited under accession number NCIMB 44383.10. The Capsicum annuum plant according to any one of claims 1 to 9, wherein said plant isa non-pungent Capsicum annuum plant, wherein said non-pungent Capsicum annuumplant preferably produces fruits having an average total capsaicinoid content (preferably capsaicin and dihydro-capsaicin content) of less than 150 µg / g FW (fresh weight).11. The Capsicum annuum plant according to any one of claims 1 to 10, wherein said plantcomprises a technically induced mutation, preferably a modification in the genome cre- ated with genome editing technologies.12. The Capsicum annuum plant according to any one of claims 1 to 11, wherein said plantis a F1 hybrid.13. Seed produced by the Capsicum annuum plant according to any of the preceding claims,wherein the seed comprises QTL MF2.1 and QTL MF2.2.14. A seed from which the Capsicum annuum plant according to any one of claims 1 to 12can be grown.15. A plant cell, tissue or plant part of the Capsicum annuum plant according to any one ofclaims 1 to 12 or of the seed according to claim 13 or 14, comprising QTL MF2.1 andQTL MF2.2.16. A haploid plant or dihaploid plant derived from the Capsicum annuum plant according toany one of claims 1 to 12.17. A method for identifying and / or selecting a Capsicum annuum plant or plant part com-prising determining whether said plant or plant part comprises in its genome QTL MF2.1 and QTL MF2.2 as defined in any one of claims 1 to 3 or the recombinant introgression on chromosome 2 as defined in claims 5 or 6 and optionally determining whether saidplant or plant part comprises in its genome the QTL ft12.1 on chromosome 12 as definedin any one of claims 8 to 9.18. The method according to claim 17, which method comprises the step of identifying atleast one marker within 50 cM of one or more of the following marker intervals: the marker interval from marker SNP_1 to marker SNP_2; and / orthe marker interval from marker SNP_3 to marker SNP_4; and optionallythe marker interval from marker SNP_5 to marker SNP_6 as defined in Table 2.

Claims

Nunhems Netherlands B.V. 230055WO015219. A method for producing a Capsicum annuum plant having a multiple flowers per nodephenotype, said method comprising the step(s) of: (i) crossing a first Capsicum annuum plant and a second Capsicum plant, whereinthe first Capsicum annuum plant is a plant as defined in any one of claims 1 to12; (ii) optionally harvesting seed from the crossing of (i) and selecting seed comprisingQTL MF2.1 and QTL MF2.2 or the recombinant introgression on chromosome 2 in its genome.

20. The method according to claim 19, further comprising the step of selecting seed com-prising QTL MF2.1 and QTL MF2.2 or the recombinant introgression on chromosome 2 in its genome by identifying at least one marker within 50 cM of one or more of the fol- lowing marker intervals: the marker interval from marker SNP_1 to marker SNP_2; and / orthe marker interval from marker SNP_3 to marker SNP_4 as defined in Table 2.

21. Method for increasing the number of flowers per node in a Capsicum annuum plant, saidmethod comprising introgressing QTL MF2.1 and QTL MF2.2 as defined in any one of claims 1-3 or the recombinant introgression on chromosome 2 as defined in claims 5 or6 into said Capsicum annuum plant.

22. Use of QTL MF2.1 and QTL MF2.2 as defined in any one of claims 1-3 or the recombi-nant introgression on chromosome 2 as defined in claims 4 or 5 for increasing the num-ber of flowers per node in a Capsicum annuum plant.

23. Use of a genetic marker specific for QTL MF2.1 and QTL MF2.2 as defined in any oneof claims 1-3 or the recombinant introgression on chromosome 2 as defined in claims 5 or 6 for selecting a Capsicum annuum plant having an increased number of flowers pernode phenotype.Nunhems Netherlands B.V. 230055WO0153 PEPPER PLANT HAVING MULTIPLE FLOWERS PER NODE ABSTRACTThe present invention relates to a Capsicum annuum plant comprising QTL MF2.1 and QTLMF2.2 conferring multiple flowers per node. The present invention further relates to a seed pro- duced by the plant according to the present invention, a seed from which a plant according to present invention can be grown, a fruit produced by a plant according to the present invention and a part of a plant according to the present invention. The present invention further relates to a method of identifying and / or selecting a plant or plant part according to the present invention. Thepresent invention further relates to a method for producing a Capsicum annuum plant having themultiple flowers per node phenotype according to the present invention. The present inventionfurther relates to a method for increasing the number of flowers per node in a Capsicum annuumplant and the use of QTL MF2.1 and QTL MF2.2 for increasing the number of flowers per node ina Capsicum annuum plant. The present invention further relates to genetic markers specific forthe QTLs according to the present invention and the use thereof for selecting a Capsicum annuumplant having an increased number of flowers per node.Nunhems Netherlands B.V. 230055W0015219. A method for producing a Capsicum annuum plant having a multiple flowers per node phenotype, said method comprising the step(s) of:(i) crossing a first Capsicum annuum plant and a second Capsicum plant, wherein the first Capsicum annuum plant is a plant as defined in any one of claims 1 to 12;(ii) optionally harvesting seed from the crossing of (i) and selecting seed comprising QTL MF2.1 and QTL MF2.2 or the recombinant introgression on chromosome 2 in its genome.

20. The method according to claim 19, further comprising the step of selecting seed comprising QTL MF2.1 and QTL MF2.2 or the recombinant introgression on chromosome 2 in its genome by identifying at least one marker within 50 cM of one or more of the following marker intervals: the marker interval from marker SNP_1 to marker SNP_2; and / or the marker interval from marker SNP_3 to marker SNP_4 as defined in Table 2.21 . Method for increasing the number of flowers per node in a Capsicum annuum plant, said method comprising introgressing QTL MF2.1 and QTL MF2.2 as defined in any one of claims 1-3 or the recombinant introgression on chromosome 2 as defined in claims 5 or 6 into said Capsicum annuum plant.

22. Use of QTL MF2.1 and QTL MF2.2 as defined in any one of claims 1-3 or the recombinant introgression on chromosome 2 as defined in claims 4 or 5 for increasing the number of flowers per node in a Capsicum annuum plant.

23. Use of a genetic marker specific for QTL MF2.1 and QTL MF2.2 as defined in any one of claims 1-3 or the recombinant introgression on chromosome 2 as defined in claims 5 or 6 for selecting a Capsicum annuum plant having an increased number of flowers per node phenotype.