Method for evaluating one or more abilities selected from the ability to retain chlorophyll, the ability to produce chlorophyll, and the ability to grow in a plant

Abstract

To provide a method for evaluating one or more abilities out of, holding chlorophyll, producing chlorophyll, and growing chlorophyll, in plants.SOLUTION: A method comprises a step 11 for measuring, expression of one or more genes and / or activity of protein derived from the gene, the genes being selected from, an ALKBH8, a homolog of ALKBH8, TGA9, a homolog of TGA9, GRF15, a homolog of GRF15, At3g61540, a homolog of At3g61540, At2g01818, and a homolog of At2g01818. The plant having low expression or activity, is concluded as a plant in which the one or more abilities are high.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a method for evaluating one or more abilities selected from the ability to retain chlorophyll, the ability to produce chlorophyll, and the ability to grow in a plant.

[0002] The present invention also relates to markers for assessing one or more of the above capabilities in plants. The present invention also relates to a method for selecting a plant having high one or more of the above abilities. The present invention also relates to a method for breeding a plant having high one or more of the above abilities.

[0003] The present invention also relates to a method for improving one or more of the above-mentioned capabilities in a plant. The present invention also relates to plants in which one or more of the above abilities are improved. [Background technology]

[0004] It is predicted that a food crisis will occur due to the world's growing population in the future, and increasing crop production is a challenge that humanity must address. In response, methods for improving crop growth and yield through genetic modification have been developed. For example, disrupting the ABP gene can increase cytokinin levels in plants, thereby increasing the yield per plant (Patent Document 1). Activating the γ-glutamylcysteine ​​synthetase gene and increasing the amount of glutathione in plants can improve plant growth and yield (Patent Document 2). Furthermore, overexpressing a group of genes containing a ZIM motif but not a CCT motif or a GATA-type zinc finger domain in plants can increase plant growth and yield (Patent Document 3).

[0005] The ALKBH8 gene is widely conserved in various organisms, and in Arabidopsis thaliana it has been reported as a gene involved in the modification of tRNA (Non-Patent Document 1). Specifically, the ALKBH8 protein encodes methylated uridine (mcm 5U: 5-methoxycarbonyl-methyluridine) is hydroxylated to give (S)-mchm 5 U: It is involved in converting 5-(S)-[methoxycarbonyl-(hydroxy)methyl]uridine. On the other hand, no function other than that involved in modifying this tRNA has been reported for the ALKBH8 gene. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Special Edition 2014-501521 [Patent Document 2] WO2008 / 087932 [Patent Document 3] Patent Publication No. 2008-271805 [Non-patent literature]

[0007] [Non-Patent Document 1] Leihne et al. (2011) Nucleic Acids Res 39:7688-7701 Summary of the Invention [Problem to be solved by the invention]

[0008] Previously reported methods for promoting plant growth each have their own issues. For example, plant hormones must be carefully controlled depending on the growth stage, and increasing the amount of cytokinin, as in Patent Document 1, can only increase yield in certain crops.

[0009] As in Patent Document 2 or Patent Document 3, the creation of recombinant crops using methods in which a group of genes is introduced into a plant and overexpressed requires skill, and in order to make the crops practical, various regulations must be cleared, which is time-consuming and costly.

[0010] On the other hand, if the ability to retain chlorophyll for a long period of time or the ability to produce chlorophyll could be improved, it would be possible to effectively promote plant growth; however, no satisfactory technology has been provided to date to improve these abilities. [Means for solving the problem]

[0011] The present inventors have made the surprising discovery that suppressing the expression of one or more genes selected from plant ALKBH8, ALKBH8 homologs, TGA9, TGA9 homologs, GRF15, GRF15 homologs, At3g61540, At3g61540 homologs, At2g01818, and At2g01818 homologs and / or the activity of proteins derived from said genes can prolong the retention of chlorophyll in leaves, increase the chlorophyll content, and promote plant growth, thereby completing the present invention. The present invention encompasses the following inventions.

[0012] (1) A method for evaluating one or more abilities selected from the ability to retain chlorophyll, the ability to produce chlorophyll, and the ability to grow in a plant, comprising: Step 11: Measuring the expression of one or more genes selected from ALKBH8, ALKBH8 homologs, TGA9, TGA9 homologs, GRF15, GRF15 homologs, At3g61540, At3g61540 homologs, At2g01818, and At2g01818 homologs in a plant and / or the activity of a protein derived from said gene. A method comprising: (2) The ALKBH8 or ALKBH8 homolog gene is selected from the following (1A) to (1F): (1A) a base sequence set forth in SEQ ID NO: 1, 2, 3, or 48; (1B) a base sequence encoding an amino acid sequence set forth in SEQ ID NO: 4, 5, 6, or 49, or an amino acid sequence having 70% or more sequence identity to the amino acid sequence; (1C) The following IDs in KEGG (Kyoto Encyclopedia of Genes and Genomes):dosa:Os04t0602700-00、dosa:Os02t0750500-01、osa:4330741、osa:4336893、ath:AT1G36310、ath:AT1G31600、aly:ARALYDRAFT_473619、aly:ARAL YDRAFT_473716、crb:CARUB_v10009616mg、crb:CARUB_v10009306mg、csat :104757812、csat:104787286、csat:104777497、csat:104742087、csat:1 04741763、csat:104777194、eus:EUTSA_v10004311mg、eus:EUTSA_v10006 757mg、brp:103867438、brp:103848760、brp:103833805、bna:111205055、 bna:106411643、bna:106447489、bna:106399512、bna:106411644、bna:10 6412207、bna:106411637、bna:106418960、boe:106342941、boe:10630818 4、boe:106307572、thj:104798532、thj:104813129、cpap:110812544、cpa p:110818506、cit:102616373、cit:102625137、cic:CICLE_v10001407mg、 cic:CICLE_v10004016mg, tcc:18606930, tcc:18592225, gra:105773784, gra:105764921, ghi:107962467, ghi:107918198, ghi:107946865, ghi:10 7946842、ghi:107933507、dzi:111275453、dzi:111295045、dzi:11131235 5、egr:104444193、egr:104456636、gmx:100784028、gmx:100777529、gmx: 100807751、pvu:PHAVU_002G123600g、pvu:PHAVU_009G027100g、vra:1067 59935、vra:106777615、var:108335758、var:108329993、ccaj:109815811、ccaj:109796724、mtr:MTR_2g460880、mtr:MTR_5g026030、cam:101494433、cam:101511642、adu:107460794、adu:107487073、aip:107606112、aip:107642443、lja:Lj0g3v0064359.1、lja:Lj0g3v0046209.1、lja:Lj2g3v1573010.1、lja:Lj2g3v1573010.2、lang:109348998、lang:109357026、fve:101305382、fve:101301875、pper:18776672、pper:18788824、pmum:103336690、pmum:103332964、pavi:110755353、pavi:110753860、mdm:103441609、mdm:103416044、mdm:103401476、pxb:103934470、pxb:103955543、zju:107422678、zju:107430520、csv:101210859、csv:101216715、cmo:103485433、cmo:103487636、mcha:111008031、mcha:111020230、cmax:111477075、cmax:111487507、cmax:111489322、rcu:8263009、rcu:8280124、jcu:105643385、jcu:105638853、hbr:110640684、hbr:110644165、hbr:110667825、pop:POPTR_0003s10550g、pop:POPTR_0002s09190g、 pop:POPTR_0975s00200g、jre:109007265、jre:109021366、jre:109019322、jre:109012454、vvi:100252728、vvi:100262638、sly:101260324、sly:101263510、spen:107015589、spen:107005551、sot:102591578、sot:102587122、cann:107867583、cann:107850720、nta:107782177、nta:107784057、nta:107778504、nta:107777356、nta:107799087、nta:107802481、nsy:104228789、nsy:1 04246141、nsy:104231727、nto:104097471、nto:104088120、nto:1041044 03、ini:109170629、ini:109170202、sind:105159340、sind:105164991、 oeu:111404274、oeu:111377742、oeu:111378936、oeu:111383327、han:11 0868608、han:110918392、bvg:104892444、bvg:104905687、soe:1107748 79、soe:110797769、nnu:104610319、nnu:104600059、obr:102703096、bdi :100835069、bdi:100827681、ats:109741229、ats:109760172、sbi:8066 423、sbi:8072284、zma:103627749、zma:100282025、sita:101777379、sit a:101775180、pda:103713010、pda:103715224、egu:105053522、egu:105 033011、mus:103996326、mus:103990395、dct:110097186、dct:110106918 、aof:109832664、aof:109825376、atr:18422692、atr:18422136、smo:SE LMODRAFT_78730、smo:SELMODRAFT_78643、smo:SELMODRAFT_112315、smo: SELMODRAFT_112271、ppp:PHYPADRAFT_106932、ppp:PHYPADRAFT_15669、vcn:VOLCADRAFT_108374、mng:MNEG_3576、olu:OSTLU_88555、ota:OT_ost ta13g02300、ota:OT_ostta11g00670、bpg:Bathy04g02310、bpg:Bathy13g02890、mis:MICPUN_86885、mis:MICPUN_60739、mpp:MICPUCDRAFT_41620、mpp:MICPUCDRAFT_17097, apro:F751_1394, or gsl:Gasu_35150, The base sequence of the gene to which (1D) an amino acid sequence encoded by a gene assigned the ID described in (1C) in KEGG, or a base sequence encoding an amino acid sequence having 70% or more sequence identity with the amino acid sequence; (1E) A nucleotide sequence having 70% or more sequence identity with the nucleotide sequence of (1A), (1B), (1C), or (1D), (1F) A base sequence complementary to the base sequence of (1A), (1B), (1C), (1D) or (1E), The method according to (1), wherein the DNA or RNA comprises any one of the base sequences listed above. (3) The TGA9 or TGA9 homolog gene is selected from the following (2A) to (2F): (2A) a base sequence set forth in SEQ ID NO: 34, 35, 36, 37, or 38; (2B) a base sequence encoding an amino acid sequence set forth in SEQ ID NO: 39, 40, 41, 42, or 43, or an amino acid sequence having 70% or more sequence identity to the amino acid sequence; (2C) The following IDs in KEGG (Kyoto Encyclopedia of Genes and Genomes): aly:9325751, csat:104755012, crb:17900540, eus:EUTSA_v10007490mg, bna:106412233, boe:106344082, brp:103871579, or rsz:108860791 The base sequence of the gene to which (2D) An amino acid sequence encoded by a gene assigned the ID described in (2C) in KEGG, or a base sequence encoding an amino acid sequence having 70% or more sequence identity with the amino acid sequence; (2E) A nucleotide sequence having 70% or more sequence identity with the nucleotide sequence of (2A), (2B), (2C), or (2D); (2F) A base sequence complementary to the base sequence of (2A), (2B), (2C), (2D) or (2E); The method according to (1), wherein the DNA or RNA comprises any one of the base sequences listed above. (4) The GRF15 or GRF15 homolog gene is selected from the following (3A) to (3F): (3A) the base sequence set forth in SEQ ID NO: 26; (3B) a base sequence encoding the amino acid sequence set forth in SEQ ID NO: 27 or an amino acid sequence having 70% or more sequence identity to the amino acid sequence; (3C) The following IDs in KEGG (Kyoto Encyclopedia of Genes and Genomes): bna:106429451, boe:106342871, or brp:103863788 The base sequence of the gene to which (3D) An amino acid sequence encoded by a gene assigned the ID described in (3C) in KEGG, or a base sequence encoding an amino acid sequence having 70% or more sequence identity with the amino acid sequence; (3E) A nucleotide sequence having 70% or more sequence identity with the nucleotide sequence of (3A), (3B), (3C), or (3D); (3F) A base sequence complementary to the base sequence of (3A), (3B), (3C), (3D) or (3E); The method according to (1), wherein the DNA or RNA comprises any one of the base sequences listed above. (5) At3g61540 or a homologous gene of At3g61540 is one of the following (4A) to (4F): (4A) the base sequence set forth in SEQ ID NO: 28; (4B) a base sequence encoding the amino acid sequence set forth in SEQ ID NO: 29 or an amino acid sequence having 70% or more sequence identity to the amino acid sequence; (4C) The following IDs in KEGG (Kyoto Encyclopedia of Genes and Genomes): aly:9314457, crb:17884851, csat:104788446, eus:EUTSA, rsz:108814367, bna:106406599, boe:106296419, or brp:103830306 The base sequence of the gene to which (4D) an amino acid sequence encoded by a gene assigned the ID described in (4C) in KEGG, or a base sequence encoding an amino acid sequence having 70% or more sequence identity with the amino acid sequence; (4E) A nucleotide sequence having 70% or more sequence identity with the nucleotide sequence of (4A), (4B), (4C), or (4D); (4F) A base sequence complementary to the base sequence of (4A), (4B), (4C), (4D) or (4E); The method according to (1), wherein the DNA or RNA comprises any one of the base sequences listed above. (6) At2g01818 or a homologous gene of At2g01818 is one of the following (5A) to (5F): (5A) the base sequence set forth in SEQ ID NO: 44 or 45; (5B) a base sequence encoding the amino acid sequence set forth in SEQ ID NO: 46 or 47, or an amino acid sequence having 70% or more sequence identity to the amino acid sequence; (5C) The following IDs in KEGG (Kyoto Encyclopedia of Genes and Genomes): aly:9311163, eus:EUTSA_v10005480mg, csat:104710111, crb:17885477, brp:103854038, bna:106432932, boe:106325697, or rsz:108809176 The base sequence of the gene to which (5D) an amino acid sequence encoded by a gene assigned the ID described in (5C) in KEGG, or a base sequence encoding an amino acid sequence having 70% or more sequence identity with the amino acid sequence; (5E) A base sequence having 70% or more sequence identity with the base sequence of (5A), (5B), (5C), or (5D); (5F) A base sequence complementary to the base sequence of (5A), (5B), (5C), (5D) or (5E); The method according to (1), wherein the DNA or RNA comprises any one of the base sequences listed above. (7) The method according to any one of (1) to (6), wherein step 11 comprises carrying out a nucleic acid amplification reaction using DNA or RNA obtained from a plant as a template to measure the expression of the gene. (8) A marker for evaluating one or more abilities selected from the ability to retain chlorophyll, the ability to produce chlorophyll, and the ability to grow in a plant, A marker consisting of all or part of one or more genes selected from ALKBH8, ALKBH8 homologs, TGA9, TGA9 homologs, GRF15, GRF15 homologs, At3g61540, At3g61540 homologs, At2g01818, and At2g01818 homologs, or proteins derived from the genes. (9) A method for selecting a plant having high ability in one or more of an ability to retain chlorophyll, an ability to produce chlorophyll, and an ability to grow, comprising the steps of: Step 21: measuring the expression of one or more genes selected from ALKBH8, ALKBH8 homologs, TGA9, TGA9 homologs, GRF15, GRF15 homologs, At3g61540, At3g61540 homologs, At2g01818, and At2g01818 homologs in a plant, and / or the activity of a protein derived from said gene; Step 22: selecting a plant in which the expression of the gene and / or the activity of the protein derived from the gene measured in step 21 is lower than that of a control plant; A method comprising: (10) The method according to (9), wherein step 21 comprises measuring the expression of the gene by carrying out a nucleic acid amplification reaction using DNA or RNA obtained from a plant as a template. (11) A method for breeding a plant having high abilities selected from an ability to retain chlorophyll, an ability to produce chlorophyll, and an ability to grow, comprising: a step 31 of crossing a first parental plant individual with a second parental individual in which the expression of one or more genes selected from ALKBH8, ALKBH8 homologs, TGA9, TGA9 homologs, GRF15, GRF15 homologs, At3g61540, At3g61540 homologs, At2g01818, and At2g01818 homologs and / or the activity of proteins derived from said genes is the same as or lower than that of the first parental individual to obtain offspring; a step 32 of measuring the expression of said gene and / or the activity of a protein derived from said gene in said offspring; Step 33: selecting the offspring when the expression of the gene and / or the activity of the protein derived from the gene in the offspring measured in step 32 is the same as or lower than the expression of the gene and / or the activity of the protein derived from the gene in the second parent individual; A method comprising: (12) The method according to (11), wherein step 31 comprises measuring the expression of the gene by performing a nucleic acid amplification reaction using DNA or RNA obtained from the offspring as a template. (13) A method for improving one or more abilities selected from an ability to retain chlorophyll, an ability to produce chlorophyll, and an ability to grow in a plant, comprising: Step 41: Suppressing the expression of one or more genes selected from ALKBH8, ALKBH8 homologs, TGA9, TGA9 homologs, GRF15, GRF15 homologs, At3g61540, At3g61540 homologs, At2g01818, and At2g01818 homologs in a plant, and / or the activity of proteins derived from the genes. A method comprising: (14) Step 41 is (I) disruption of the gene in the genomic DNA of a plant; (II) introducing antisense DNA for the gene into a plant; and (III) introducing into a plant an antagonist to the protein derived from the gene; The method according to (13), comprising any one or more of the following: (15) A plant (excluding Arabidopsis thaliana) in which the expression of one or more genes selected from ALKBH8, ALKBH8 homologs, TGA9, TGA9 homologs, GRF15, GRF15 homologs, At3g61540, At3g61540 homologs, At2g01818, and At2g01818 homologs, and / or the activity of proteins derived from said genes, is suppressed. [Effects of the Invention]

[0013] According to the methods described in (1) to (7) above, one or more abilities selected from the ability to retain chlorophyll, the ability to produce chlorophyll, and the ability to grow in a plant can be easily evaluated.

[0014] The markers described in (8) above are useful for evaluating one or more of the above abilities in plants.

[0015] According to the method described in (9) or (10) above, a plant having high one or more of the above abilities can be selected.

[0016] According to the method described in (11) or (12) above, it is possible to breed a plant that is high in one or more of the above abilities.

[0017] According to the method described in (13) or (14) above, one or more of the above abilities in a plant can be improved. The plant described in (15) above is a plant in which one or more of the above abilities are enhanced. [Brief explanation of the drawings]

[0018] [Figure 1]Figure 1 shows the positions of T-DNA insertions in Arabidopsis T-DNA insertion mutants (SALK_083838C, SALK_094502C) and primers in the coding regions of four splice variants (At1g31600.1, At1g31600.2, At1g31600.3, and At1g31600.4) of the Arabidopsis ALKBH8 gene. [Figure 2] Figure 2 shows the results of PCR using DNA from Arabidopsis T-DNA insertion mutants (SALK_083838C, SALK_094502C) and the wild-type strain as templates, with primer set 1 for detecting the At1g31600 gene or primer set 2 for detecting the border region of the T-DNA insertion. [Figure 3] FIG. 3 shows photographs of the Arabidopsis wild-type strain and two ALKBH8 disruptant strains (alkbh8-1, alkbh8-2) 10 weeks after sowing. [Figure 4] FIG. 4 shows the position of the T-DNA insertion in the Arabidopsis T-DNA insertion mutant (SALK_112825) in the coding region of the Arabidopsis GRF15 (At2g10450) gene, and the positions of the primers. [Figure 5] Figure 5 shows the results of PCR using DNA from an Arabidopsis T-DNA insertion mutant (SALK_112825) and a wild-type strain as templates, with primer set 1 for detecting the GRF15 (At2g10450) gene or primer set 2 for detecting the border region of the T-DNA insertion. [Figure 6] FIG. 6 shows the positions of T-DNA insertions in the Arabidopsis T-DNA insertion mutant (SALK_135878) and primer positions in the coding regions of the Arabidopsis KPHMT2 (At3g61530) and At3g61540 genes. [Figure 7]Figure 7 shows the results of PCR using DNA from an Arabidopsis T-DNA insertion mutant (SALK_135878) and a wild-type strain as templates, and primer set 1 for detecting the KPHMT2 (At3g61530) gene and the At3g61540 gene, or primer set 2 for detecting the border region where T-DNA was inserted. [Figure 8] Figure 8 shows the positions of T-DNA insertions in the Arabidopsis T-DNA insertion mutant (SALK_091349) and primer positions in the coding regions of five splicing variants (At1g08320.1, At1g08320.2, At1g08320.3, At1g08320.4, and At1g08320.5) of the Arabidopsis TGA9 (At1g08320) gene. [Figure 9] Figure 9 shows the results of PCR using DNA from an Arabidopsis T-DNA insertion mutant (SALK_091349) and a wild-type strain as templates, and primer set 1 for detecting the TGA9 (At1g08320) gene or primer set 2 for detecting the border region where the T-DNA was inserted. [Figure 10] FIG. 10 shows the positions of T-DNA insertions in the Arabidopsis T-DNA insertion mutant (SAIL_568_E01) and primer positions in the coding regions of two splicing variants (At2g01818.1 and At2g01818.2) of the Arabidopsis At2g01818 gene. [Figure 11] Figure 11 shows the results of PCR using DNA from an Arabidopsis T-DNA insertion mutant (SAIL_568_E01) and a wild-type strain as templates, and primer set 1 for detecting the At2g01818 gene or primer set 2 for detecting the border region where the T-DNA was inserted. [Figure 12] FIG. 12 shows the appearance of Arabidopsis rosette leaves when the evaluation score indicating the degree of chlorophyll maintenance is 0, 1, 2, or 3. [Figure 13]Figure 13 shows the average green leaf index per plant (n = 9) for the Arabidopsis wild-type strain (Col) and the GRF15, At3g61540, TGA9, and At2g01818 gene disruption strains at 48, 56, 63, and 70 days after sowing (DAS). [Figure 14] Figure 14 shows the average total number of rosette leaves (green leaves) per plant (n = 9 per plant) with chlorophyll content scores ranging from 1 to 3 at the end of growth, 10 weeks (70 days) after sowing, for the Arabidopsis wild-type plant (Col) and the GRF15, At3g61540, TGA9, and At2g01818 gene disruption plants, and the relative ratio of the number of green leaves for each gene disruption plant to the number of green leaves for the wild-type plant, which is set at 100. [Figure 15] Figure 15 shows the locations of the OsALKBH8_seq_f primer, OsALKBH8_seq_r primer, and the guide RNA (gOsALKBH8-1) used for genome editing in the coding region of the Os04g0602700 gene, which is an ortholog of the ALKBH8 gene in rice. [Figure 16] FIG. 16 shows the SPAD values ​​(an index correlated with the amount of chlorophyll in leaves) of the rice OsALKBH8 gene disruptant and wild-type rice at 8 weeks of age. DETAILED DESCRIPTION OF THE INVENTION

[0019] The type of "plant" targeted by the present invention is not particularly limited, and may be various plants such as dicotyledonous plants and monocotyledonous plants.

[0020] Examples of dicotyledonous plants include plants of the genus Ipomoea, Ipomoea, Cuscuta, Dianthus, Chickweed, Alpine clover, Eucalyptus, Trifolium, Fleabane, Brassica napus, Celeriac, Corylus, Corylus pratense, Silene, Celeriac, Psammophila, Caryophyllaceae, Casuarina, Houttuynia cordata, Piperaceae, Cranberry, Salicaceae, Myrica rubra, Juglandaceae, Birch, and beech. Family: Ulmaceae, Moraceae, Urticaceae, Podocarpus, Proteaceae, Ragwort, Sandalwood, Mistletoe, Aristolochiae, Mistletoe, Aristolochiae, Miacaceae, Aconitaceae, Polygonaceae, Chenopodiaceae, Amaranthaceae, Nyctaginaceae, Saccharinaceae, Phytolacca, Bucconaceae, Portulacaceae, Magnoliaceae, Trochuraceae, Juglans, Nymphaeaceae, Brunswick, Ranunculaceae, Akebia, Berberidaceae, Menispermaceae, Wintersweet, Lauraceae, Ke Plants of the family Lamiaceae, Capparaceae, Brassicaceae, Sundew, Pitcher, Crassulaceae, Saxifragaceae, Pittosporaceae, Witch hazel, Platanaceae, Rosaceae, Fabaceae, Oxalidaceae, Geraniaceae, Linaceae, Tribulus, Rutaceae, Cerataceae, Meliaceae, Polygalaceae, Euphorbiaceae, Botryllaceae, Buxaceae, Crowberry, Deutzia, Anacardiaceae, Ilex, Celastrus, Tribulus, Scutellaria, and Maple Plants of the following families are included: Aesculaceae, Sapindaceae, Balsaminaceae, Balsaminaceae, Rhamnaceae, Vitaceae, Bombacaceae, Tilia, Mallow, Sterculiaceae, Aroniaceae, Theaceae, Hypericaceae, Lythrum, Tuna, Violets, Atractylodes, Stalactaceae, Passifloraceae, Begoniaceae, Cactaceae, Thymelaeaceae, Elaeaceae, Lythrum, Pomegranate, Rhizophoraceae, Cucurbitaceae, Melastomataceae, Trachycarpus, Onagraceae,Plants of the following families are included: Araceae, Horsetail, Araliaceae, Umbelliferae, Cornaceae, Atractylodes, Chloraceae, Araceae, Ericaceae, Ardisiaceae, Primulaceae, Plumageae, Ebenaceae, Cymbidium, Styrax, Oleaceae, Buddleaceae, Gentianaceae, Apocynaceae, Asclepiadaceae, Polemoniaceae, Boraginaceae, Crocus Examples of plants include plants of the family Pinaceae, Lamiaceae, Solanaceae, Scrophulariaceae, Bignoniaceae, Pedaliaceae, Orobanchaceae, Gesneriaceae, Utriculariaceae, Acanthaceae, Pithunaceae, Plantaginaceae, Rubiaceae, Caprifoliaceae, Membranaceae, Valerianaceae, Scabiosa, Cucurbitaceae, Campanulaceae, and Asteraceae.

[0021] Examples of monocotyledonous plants include plants of the genus Lemna, Lemna, Cattleya, Cymbidium, Dendrobium, Phalaenopsis, Vanda, Paphiopedilum, Orchids, Typha, Eclipta, Potamogeton, Lilium, Lilium, Cyperaceae, Palmaceae, Araceae, Eriophyaceae, Daylily, Pontedria, Juncaceae, Santalum, Liliaceae, Amaryllidaceae, Dioscorea, Iridaceae, Musaceae, Zingiberaceae, Cannaceae, and Lepidoptera. As the monocotyledonous plants, grasses are preferred, and as the grasses, rice is particularly preferred.

[0022] <1. Ability to retain chlorophyll> In the present invention, the "ability to retain chlorophyll" refers to the ability to retain chlorophyll produced in the plant body, such as leaves, for a certain period of time. A plant that can retain chlorophyll in the plant body for a longer period of time can be said to have a high ability to retain chlorophyll.

[0023] Higher plants have chlorophyll a and chlorophyll b. In the present invention, chlorophyll refers to at least one of chlorophyll a and chlorophyll b, and preferably both.

[0024] 2. Ability to produce chlorophyll In the present invention, the "ability to produce chlorophyll" refers to the ability to produce chlorophyll in the plant body, such as leaves. A plant that can produce a large amount of chlorophyll in the plant body, particularly in the leaves, can be said to have a high ability to produce chlorophyll.

[0025] <3. Ability to grow> In the present invention, the "growth" of a plant typically refers to an increase in one or more of the following items at any stage of the plant's growth (seedling, vegetative growth, reproductive growth): biomass weight, plant height, leaf area, and stem number. A plant that shows a greater increase in these items can be said to have high growth potential.

[0026] 4. Genes 4.1. ALKBH8 or ALKBH8 homologs ALKBH8 (AlkB homolog 8) is a member of the ALKBH family, consisting of an RNA recognition motif (RRM), an AlkB domain with hydroxylase activity, and a Trm9-like domain with methyltransferase activity. Arabidopsis ALKBH8 (At1g31600) has four splicing variants: At1g31600.1, At1g31600.2, At1g31600.3, and At1g31600.4. The nucleotide sequences of the coding regions of At1g31600.1 and At1g31600.3 are shown in SEQ ID NO: 1, At1g31600.2, and At1g31600.4, respectively. The amino acid sequences of At1g31600.1 and At1g31600.3 are shown in SEQ ID NO: 4, the amino acid sequence of At1g31600.2 is shown in SEQ ID NO: 5, and the amino acid sequence of At1g31600.4 is shown in SEQ ID NO: 6.

[0027] The rice homolog (ortholog) of the ALKBH8 gene, Os04g0602700 (hereafter referred to as OsALKBH8), has been assigned the ID dosa:Os04t0602700-00 in KEGG (http: / / www.genome.jp / kegg / ), and its nucleotide sequence and the amino acid sequence encoded by this nucleotide sequence are registered. The nucleotide sequence of OsALKBH8 is shown in SEQ ID NO: 48, and the amino acid sequence is shown in SEQ ID NO: 49.

[0028] In the present invention, "ALKBH8 or an ALKBH8 homolog gene" is not limited to the ALKBH8 gene derived from Arabidopsis thaliana, but also includes genes having equivalent functions and mutants thereof.

[0029] The genes assigned the following IDs are registered in KEGG (Kyoto Encyclopedia of Genes and Genomes) (http: / / www.genome.jp / kegg / ) as ALKBH8 or ALKBH8 homolog genes. KEGG registers the nucleotide sequence and the amino acid sequence encoded by the nucleotide sequence for each gene.

[0030] [Table 1] TIFF0007783695000002.tif234144 TIFF0007783695000003.tif224143 TIFF0007783695000004.tif237144 TIFF0007783695000005.tif243143 TIFF0007783695000006.tif71143

[0031] More preferably, the gene of ALKBH8 or an ALKBH8 homolog is DNA or RNA that includes or consists of any of the base sequences (1A) to (1F) above.

[0032] Here, when the gene for ALKBH8 or an ALKBH8 homolog is RNA, thymine (T) in any of the above base sequences (1A) to (1F) is replaced with uracil (U).

[0033] A more preferred embodiment of (1B) above is a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO: 4, 5, 6, or 49, or a nucleotide sequence encoding an amino acid sequence having preferably 80% or more, more preferably 90% or more, more preferably 95% or more, more preferably 97% or more, more preferably 98% or more, and more preferably 99% or more sequence identity to the amino acid sequence set forth in SEQ ID NO: 4, 5, 6, or 49. The protein consisting of the amino acid sequence encoded by the nucleotide sequence of (1B) above may have ALKBH8 activity. In the present invention, ALKBH8 activity refers to the activity of a protein containing methylated uridine (mcm) at a position corresponding to the amino acid sequence of the methylated uridine (mcm). 5 U: 5-methoxycarbonyl-methyluridine) is hydroxylated to give (S)-mchm 5 U: refers to the activity of converting to 5-(S)-[methoxycarbonyl-(hydroxy)methyl]uridine. Preferably, the ALKBH8 activity of a protein consisting of the amino acid sequence encoded by the base sequence of (1B) above is 80% or more, more preferably 90% or more, more preferably 95% or more, and more preferably 100% or more of the ALKBH8 activity of a protein consisting of the amino acid sequence set forth in SEQ ID NO: 4, 5, 6, or 49.

[0034] A more preferred embodiment of (1D) above is a nucleotide sequence encoding an amino acid sequence encoded by a gene assigned the ID described in (1C) above in KEGG, or a nucleotide sequence encoding an amino acid sequence having preferably 80% or more, more preferably 90% or more, more preferably 95% or more, more preferably 97% or more, more preferably 98% or more, and more preferably 99% or more sequence identity to the amino acid sequence encoded by a gene assigned the ID described in (1C) above in KEGG. The protein consisting of the amino acid sequence encoded by the nucleotide sequence described in (1D) above may have ALKBH8 activity. Preferably, the ALKBH8 activity of the protein consisting of the amino acid sequence encoded by the nucleotide sequence described in (1D) above is 80% or more, more preferably 90% or more, more preferably 95% or more, and more preferably 100% or more of the ALKBH8 activity of a protein consisting of the amino acid sequence encoded by a gene assigned the ID described in (1C) above in KEGG.

[0035] A more preferred embodiment of (1E) above is a nucleotide sequence having preferably 80% or more, more preferably 90% or more, more preferably 95% or more, more preferably 97% or more, more preferably 98% or more, and more preferably 99% or more sequence identity with the nucleotide sequence of (1A), (1B), (1C), or (1D) above. The nucleotide sequence of (1E) above may be any nucleotide sequence as long as the protein comprising the amino acid sequence encoded by it has ALKBH8 activity. Preferably, the ALKBH8 activity of the protein comprising the amino acid sequence encoded by the nucleotide sequence of (1E) above is 80% or more, more preferably 90% or more, more preferably 95% or more, and more preferably 100% or more of the ALKBH8 activity of the protein comprising the amino acid sequence encoded by the nucleotide sequence of (1A), (1B), (1C), or (1D) above.

[0036] In the above (1B), (1D), and (1E), the identity values ​​are calculated using software (e.g., FASTA, DNASIS, and BLAST) that calculates the identity between multiple amino acid sequences or nucleotide sequences with default settings. The identity value of nucleotide sequences is calculated by aligning a pair of nucleotide sequences to maximize the degree of identity, calculating the number of matching bases, and calculating the ratio of the number of matching bases to the total number of bases in the compared nucleotide sequences. Details of the method for determining identity are described, for example, in Altschul et al., Nuc. Acids. Res. 25, 3389-3402, 1977 and Altschul et al., J. Mol. Biol. 215, 403-410, 1990. The identities in (2B), (2D), (2E), (3B), (3D), (3E), (4B), (4D), (4E), (5B), (5D) and (5E) described below are calculated in the same manner.

[0037] <4.2. TGA9 or TGA9 homologs> TGA9 is a basic region leucine zipper transcription factor. Arabidopsis TGA9 (At1g08320) has five splicing variants: At1g08320.1, At1g08320.2, At1g08320.3, At1g08320.4, and At1g08320.5. The nucleotide sequences of the coding regions of At1g08320.1, At1g08320.2, At1g08320.3, At1g08320.4, and At1g08320.5 are shown in SEQ ID NOs: 34, 35, 36, 37, and 38, respectively, and the amino acid sequences are shown in SEQ ID NOs: 39, 40, 41, 42, and 43, respectively.

[0038] In the present invention, the term "TGA9 or a TGA9 homolog gene" is not limited to the Arabidopsis-derived TGA9 gene, but also includes genes having equivalent functions and mutants thereof.

[0039] The genes assigned the following IDs are registered in KEGG (Kyoto Encyclopedia of Genes and Genomes) (http: / / www.genome.jp / kegg / ) as TGA9 or TGA9 homolog genes. KEGG registers the nucleotide sequence and the amino acid sequence encoded by the nucleotide sequence for each gene.

[0040] [Table 2]

[0041] More preferably, the gene of TGA9 or a TGA9 homolog is DNA or RNA that includes or consists of any of the base sequences (2A) to (2F) above.

[0042] Here, when the gene of TGA9 or a TGA9 homolog is RNA, thymine (T) in any of the base sequences (2A) to (2F) above is read as uracil (U).

[0043] A more preferred embodiment of (2B) above is a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO: 39, 40, 41, 42, or 43, or a nucleotide sequence encoding an amino acid sequence having preferably 80% or more, more preferably 90% or more, more preferably 95% or more, more preferably 97% or more, more preferably 98% or more, and more preferably 99% or more sequence identity to the amino acid sequence set forth in SEQ ID NO: 39, 40, 41, 42, or 43. A protein consisting of the amino acid sequence encoded by the nucleotide sequence of (2B) above may have TGA9 activity. Preferably, the TGA9 activity of a protein consisting of the amino acid sequence encoded by the nucleotide sequence of (2B) above is 80% or more, more preferably 90% or more, more preferably 95% or more, and more preferably 100% or more of the TGA9 activity of a protein consisting of the amino acid sequence set forth in SEQ ID NO: 39, 40, 41, 42, or 43.

[0044] A more preferred embodiment of (2D) above is a nucleotide sequence encoding an amino acid sequence encoded by a gene assigned the ID described in (2C) above in KEGG, or a nucleotide sequence encoding an amino acid sequence having preferably 80% or more, more preferably 90% or more, more preferably 95% or more, more preferably 97% or more, more preferably 98% or more, and more preferably 99% or more sequence identity to the amino acid sequence encoded by a gene assigned the ID described in (2C) above in KEGG. The protein consisting of the amino acid sequence encoded by the nucleotide sequence described in (2D) above may have TGA9 activity. Preferably, the TGA9 activity of the protein consisting of the amino acid sequence encoded by the nucleotide sequence described in (2D) above is 80% or more, more preferably 90% or more, more preferably 95% or more, and more preferably 100% or more of the TGA9 activity of the protein consisting of the amino acid sequence encoded by a gene assigned the ID described in (2C) above in KEGG.

[0045] A more preferred embodiment of (2E) above is a base sequence having preferably 80% or more, more preferably 90% or more, more preferably 95% or more, more preferably 97% or more, more preferably 98% or more, and more preferably 99% or more sequence identity with the base sequence of (2A), (2B), (2C), or (2D) above. The base sequence of (2E) above may be any as long as the protein consisting of the amino acid sequence encoded by it has TGA9 activity. Preferably, the TGA9 activity of the protein consisting of the amino acid sequence encoded by the base sequence of (2E) above is 80% or more, more preferably 90% or more, more preferably 95% or more, and more preferably 100% or more of the TGA9 activity of the protein consisting of the amino acid sequence encoded by the base sequence of (2A), (2B), (2C), or (2D).

[0046] <4.3. GRF15 or GRF15 homologs> GRF15 is a growth regulator. The nucleotide sequence of the coding region of the Arabidopsis thaliana GRF15 (At2g10450) gene is shown in SEQ ID NO: 26, and the amino acid sequence of Arabidopsis thaliana GRF15 encoded by the gene is shown in SEQ ID NO: 27.

[0047] In the present invention, "GRF15 or a gene homologous to GRF15" is not limited to the GRF15 gene derived from Arabidopsis thaliana, but also includes genes having equivalent functions and mutants thereof.

[0048] The genes assigned the following IDs are registered in KEGG (Kyoto Encyclopedia of Genes and Genomes) (http: / / www.genome.jp / kegg / ) as GRF15 or GRF15 homolog genes. KEGG registers the nucleotide sequence and the amino acid sequence encoded by the nucleotide sequence for each gene.

[0049] [Table 3]

[0050] More preferably, the gene of GRF15 or a GRF15 homolog is DNA or RNA that includes or consists of any of the base sequences (3A) to (3F) above.

[0051] Here, when the gene of GRF15 or a homologue of GRF15 is RNA, thymine (T) in any of the base sequences (3A) to (3F) above is read as uracil (U).

[0052] A more preferred embodiment of (3B) above is a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO: 27, or a nucleotide sequence encoding an amino acid sequence having preferably 80% or more, more preferably 90% or more, more preferably 95% or more, more preferably 97% or more, more preferably 98% or more, and more preferably 99% or more sequence identity to the amino acid sequence set forth in SEQ ID NO: 27. The protein consisting of the amino acid sequence encoded by the nucleotide sequence of (3B) above may have GRF15 activity. Preferably, the GRF15 activity of the protein consisting of the amino acid sequence encoded by the nucleotide sequence of (3B) above is 80% or more, more preferably 90% or more, more preferably 95% or more, and more preferably 100% or more of the GRF15 activity of the protein consisting of the amino acid sequence set forth in SEQ ID NO: 27.

[0053] A more preferred embodiment of (3D) above is a nucleotide sequence encoding an amino acid sequence encoded by a gene assigned the ID described in (3C) above in KEGG, or a nucleotide sequence encoding an amino acid sequence having preferably 80% or more, more preferably 90% or more, more preferably 95% or more, more preferably 97% or more, more preferably 98% or more, and more preferably 99% or more sequence identity to the amino acid sequence encoded by a gene assigned the ID described in (3C) above in KEGG. The protein consisting of the amino acid sequence encoded by the nucleotide sequence described in (3D) above may have GRF15 activity. Preferably, the GRF15 activity of the protein consisting of the amino acid sequence encoded by the nucleotide sequence described in (3D) above is 80% or more, more preferably 90% or more, more preferably 95% or more, and more preferably 100% or more of the GRF15 activity of a protein consisting of the amino acid sequence encoded by a gene assigned the ID described in (3C) above in KEGG.

[0054] A more preferred embodiment of (3E) above is a base sequence having preferably 80% or more, more preferably 90% or more, more preferably 95% or more, more preferably 97% or more, more preferably 98% or more, and more preferably 99% or more sequence identity with the base sequence of (3A), (3B), (3C), or (3D) above. The base sequence of (3E) above may be any as long as a protein consisting of the amino acid sequence encoded by it has GRF15 activity. Preferably, the GRF15 activity of a protein consisting of the amino acid sequence encoded by the base sequence of (3E) above is 80% or more, more preferably 90% or more, more preferably 95% or more, and more preferably 100% or more of the GRF15 activity of a protein consisting of the amino acid sequence encoded by the base sequence of (3A), (3B), (3C), or (3D).

[0055] <4.4. At3g61540 or At3g61540 homologs> The protein encoded by At3g61540 is a member of the α / β-hydrolase superfamily of proteins. The nucleotide sequence of the coding region of the Arabidopsis thaliana At3g61540 gene is shown in SEQ ID NO: 28, and the amino acid sequence of Arabidopsis thaliana At3g61540 encoded by the gene is shown in SEQ ID NO: 29.

[0056] In the present invention, "At3g61540 or a gene homologous to At3g61540" is not limited to the At3g61540 gene derived from Arabidopsis thaliana, but also includes genes having equivalent functions and mutants thereof.

[0057] The genes assigned the following IDs are registered in KEGG (Kyoto Encyclopedia of Genes and Genomes) (http: / / www.genome.jp / kegg / ) as At3g61540 or At3g61540 homolog genes. KEGG registers the nucleotide sequence and the amino acid sequence encoded by the nucleotide sequence for each gene.

[0058] [Table 4]

[0059] More preferably, the gene of At3g61540 or a homologue of At3g61540 is a DNA or RNA containing or consisting of any of the base sequences (4A) to (4F) above.

[0060] Here, when the gene of At3g61540 or a homologue of At3g61540 is RNA, thymine (T) in any of the base sequences (4A) to (4F) above is read as uracil (U).

[0061] A more preferred embodiment of (4B) above is a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO: 29, or a nucleotide sequence encoding an amino acid sequence having preferably 80% or more, more preferably 90% or more, more preferably 95% or more, more preferably 97% or more, more preferably 98% or more, and more preferably 99% or more sequence identity to the amino acid sequence set forth in SEQ ID NO: 29. The protein consisting of the amino acid sequence encoded by the nucleotide sequence of (4B) above may have At3g61540 activity. Preferably, the At3g61540 activity of the protein consisting of the amino acid sequence encoded by the nucleotide sequence of (4B) above is 80% or more, more preferably 90% or more, more preferably 95% or more, and more preferably 100% or more of the At3g61540 activity of the protein consisting of the amino acid sequence set forth in SEQ ID NO: 29.

[0062] A more preferred embodiment of (4D) above is a nucleotide sequence encoding an amino acid sequence encoded by a gene assigned the ID described in (4C) above in KEGG, or a nucleotide sequence encoding an amino acid sequence having preferably 80% or more, more preferably 90% or more, more preferably 95% or more, more preferably 97% or more, more preferably 98% or more, and more preferably 99% or more sequence identity to the amino acid sequence encoded by a gene assigned the ID described in (4C) above in KEGG. The protein consisting of the amino acid sequence encoded by the nucleotide sequence described in (4D) above may have At3g61540 activity. Preferably, the At3g61540 activity of the protein consisting of the amino acid sequence encoded by the nucleotide sequence described in (4D) above is 80% or more, more preferably 90% or more, more preferably 95% or more, and more preferably 100% or more of the At3g61540 activity of the protein consisting of the amino acid sequence encoded by a gene assigned the ID described in (4C) above in KEGG.

[0063] A more preferred embodiment of (4E) above is a nucleotide sequence having preferably 80% or more, more preferably 90% or more, more preferably 95% or more, more preferably 97% or more, more preferably 98% or more, and more preferably 99% or more sequence identity with the nucleotide sequence of (4A), (4B), (4C), or (4D) above. The nucleotide sequence of (4E) above may be any nucleotide sequence as long as a protein comprising the amino acid sequence encoded by it has At3g61540 activity. Preferably, the At3g61540 activity of a protein comprising the amino acid sequence encoded by the nucleotide sequence of (4E) above is 80% or more, more preferably 90% or more, more preferably 95% or more, and more preferably 100% or more of the At3g61540 activity of a protein comprising the amino acid sequence encoded by the nucleotide sequence of (4A), (4B), (4C), or (4D) above.

[0064] <4.5. At2g01818 or At2g01818 homologs> The protein encoded by At2g01818 is a member of the PLATZ transcription factor family. The Arabidopsis At2g01818 gene has two splicing variants, At2g01818.1 and At2g01818.2. The nucleotide sequences of the coding regions of At2g01818.1 and At2g01818.2, respectively, are shown in SEQ ID NOs: 44 and 45, and their amino acid sequences are shown in SEQ ID NOs: 46 and 47.

[0065] In the present invention, "At2g01818 or a gene homologous to At2g01818" is not limited to the At2g01818 gene derived from Arabidopsis thaliana, but also encompasses genes having equivalent functions and mutants thereof.

[0066] The genes assigned the following IDs are registered in KEGG (Kyoto Encyclopedia of Genes and Genomes) (http: / / www.genome.jp / kegg / ) as At2g01818 or At2g01818 homolog genes. KEGG registers the nucleotide sequence and the amino acid sequence encoded by the nucleotide sequence for each gene.

[0067] [Table 5]

[0068] More preferably, the gene of At2g01818 or a homologue of At2g01818 is DNA or RNA that includes or consists of any of the base sequences (5A) to (5F) above.

[0069] Here, when the gene of At2g01818 or a homologue of At2g01818 is RNA, thymine (T) in any of the base sequences (5A) to (5F) above is read as uracil (U).

[0070] A more preferred embodiment of (5B) above is a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO: 46 or 47, or a nucleotide sequence encoding an amino acid sequence having preferably 80% or more, more preferably 90% or more, more preferably 95% or more, more preferably 97% or more, more preferably 98% or more, and more preferably 99% or more sequence identity to the amino acid sequence set forth in SEQ ID NO: 46 or 47. A protein consisting of the amino acid sequence encoded by the nucleotide sequence of (5B) above may have At2g01818 activity. Preferably, the At2g01818 activity of a protein consisting of the amino acid sequence encoded by the nucleotide sequence of (5B) above is 80% or more, more preferably 90% or more, more preferably 95% or more, and more preferably 100% or more of the At2g01818 activity of a protein consisting of the amino acid sequence set forth in SEQ ID NO: 46 or 47.

[0071] A more preferred embodiment of (5D) above is a nucleotide sequence encoding an amino acid sequence encoded by a gene assigned the ID described in (5C) above in KEGG, or a nucleotide sequence encoding an amino acid sequence having preferably 80% or more, more preferably 90% or more, more preferably 95% or more, more preferably 97% or more, more preferably 98% or more, and more preferably 99% or more sequence identity to the amino acid sequence encoded by a gene assigned the ID described in (5C) above in KEGG. A protein consisting of an amino acid sequence encoded by the nucleotide sequence described in (5D) above may have At2g01818 activity. Preferably, the At2g01818 activity of a protein consisting of an amino acid sequence encoded by the nucleotide sequence described in (5D) above is 80% or more, more preferably 90% or more, more preferably 95% or more, and more preferably 100% or more of the At2g01818 activity of a protein consisting of an amino acid sequence encoded by a gene assigned the ID described in (5C) above in KEGG.

[0072] A more preferred embodiment of (5E) above is a nucleotide sequence having preferably 80% or more, more preferably 90% or more, more preferably 95% or more, more preferably 97% or more, more preferably 98% or more, and more preferably 99% or more sequence identity with the nucleotide sequence of (5A), (5B), (5C), or (5D) above. The nucleotide sequence of (5E) above may be any nucleotide sequence as long as a protein comprising the amino acid sequence encoded by it has At2g01818 activity. Preferably, the At2g01818 activity of a protein comprising the amino acid sequence encoded by the nucleotide sequence of (5E) above is 80% or more, more preferably 90% or more, more preferably 95% or more, and more preferably 100% or more of the At2g01818 activity of a protein comprising the amino acid sequence encoded by the nucleotide sequence of (5A), (5B), (5C), or (5D) above.

[0073] 5. Methods for assessing plant performance One aspect of the present invention is A method for evaluating one or more abilities selected from the ability to retain chlorophyll, the ability to produce chlorophyll, and the ability to grow in a plant, comprising: Step 11: Measuring the expression of one or more genes selected from ALKBH8, ALKBH8 homologs, TGA9, TGA9 homologs, GRF15, GRF15 homologs, At3g61540, At3g61540 homologs, At2g01818, and At2g01818 homologs in a plant and / or the activity of a protein derived from said gene. The present invention relates to a method comprising:

[0074] The present inventors were surprised to find that plants in which the expression of the gene and / or the activity of the protein derived from the gene is suppressed have improved abilities to retain chlorophyll, to produce chlorophyll, and to grow, compared to plants in which the expression and activity are not suppressed (e.g., wild-type plants), and have thus completed the present invention.

[0075] If the expression of the gene and / or the activity of the protein derived from the gene of the test plant measured in step 11 is suppressed compared to a control plant (e.g., a wild-type plant corresponding to the test plant or a plant used as material for breeding), the test plant can be determined to be a plant with higher one or more of the abilities, and if the expression or activity is the same or higher than that of the control plant, the test plant can be determined to be a plant with the same or lower one or more abilities than the target plant.

[0076] In the present invention, the term "protein derived from the gene" refers to a protein produced through transcription and translation from one or more genes selected from ALKBH8, ALKBH8 homologs, TGA9, TGA9 homologs, GRF15, GRF15 homologs, At3g61540, At3g61540 homologs, At2g01818, and At2g01818 homologs.

[0077] In the present invention, the expression of the gene may be measured by quantifying the mRNA corresponding to the gene in the plant cells, by quantifying the protein derived from the gene in the plant cells, or by quantifying or detecting the presence or absence of the nucleotide sequence of the genomic DNA corresponding to the gene in the plant cells (if the nucleotide sequence of the genomic DNA corresponding to the gene is not present, it can be concluded that the gene is not expressed, and if the nucleotide sequence of the genomic DNA corresponding to the gene is less than that of the wild type, it can be concluded that the expression of the gene is low).

[0078] In the present invention, the activity of a protein derived from one or more genes selected from ALKBH8 and ALKBH8 homologs can be measured using the above-mentioned ALKBH8 activity as an indicator.

[0079] In the present invention, the activity of a protein derived from one or more genes selected from TGA9 and TGA9 homologs can be measured using the above-mentioned TGA9 activity as an index.

[0080] In the present invention, the activity of a protein derived from one or more genes selected from GRF15 and GRF15 homologs can be measured using the above-mentioned GRF15 activity as an indicator.

[0081] In the present invention, the activity of a protein derived from one or more genes selected from At3g61540 and At3g61540 homologs can be measured using the above-mentioned At3g61540 activity as an index.

[0082] In the present invention, the activity of a protein derived from one or more genes selected from At2g01818 and At2g01818 homologs can be measured using the above-mentioned At2g01818 activity as an indicator.

[0083] Step 11 preferably includes measuring the expression of the gene by performing a nucleic acid amplification reaction using DNA or RNA obtained from a plant as a template. Examples of DNA obtained from a plant include genomic DNA and cDNA prepared from mRNA. Examples of RNA obtained from a plant include mRNA. The primer set for the nucleic acid amplification reaction may be designed to specifically amplify a region characteristic of the base sequence of the gene. The nucleic acid amplification reaction may be performed by PCR (polymerase chain reaction) or an isothermal amplification method such as LAMP.

[0084] <6. Marker> Another aspect of the present invention is A marker for evaluating one or more abilities selected from the ability to retain chlorophyll, the ability to produce chlorophyll, and the ability to grow in a plant, The present invention relates to a marker consisting of all or part of one or more genes selected from ALKBH8, ALKBH8 homologs, TGA9, TGA9 homologs, GRF15, GRF15 homologs, At3g61540, At3g61540 homologs, At2g01818, and At2g01818 homologs, or a protein derived from the gene.

[0085] As described above, plants in which the expression of the gene and / or the activity of the protein derived from the gene are suppressed have improved abilities to retain chlorophyll, produce chlorophyll, and grow compared to plants in which the expression and activity are not suppressed (e.g., wild-type plants). Therefore, all or part of the gene, or the protein derived from the gene, is useful as a marker for evaluating one or more of the abilities.

[0086] Particularly preferred markers are mRNA corresponding to the gene, a fragment of the mRNA, cDNA prepared from the mRNA, or a fragment of the cDNA.

[0087] <7. How to select plants> Another aspect of the present invention is A method for selecting a plant having high ability in one or more of an ability to retain chlorophyll, an ability to produce chlorophyll, and an ability to grow, comprising the steps of: Step 21: measuring the expression of one or more genes selected from ALKBH8, ALKBH8 homologs, TGA9, TGA9 homologs, GRF15, GRF15 homologs, At3g61540, At3g61540 homologs, At2g01818, and At2g01818 homologs in a plant, and / or the activity of a protein derived from said gene; Step 22: selecting a plant in which the expression of the gene and / or the activity of the protein derived from the gene measured in step 21 is lower than that of a control plant; The present invention relates to a method comprising:

[0088] Step 21 can be carried out in the same manner as step 11.

[0089] Plants in which the expression of the gene and / or the activity of the protein derived from the gene measured in step 21 is lower than that of a control plant are plants that have higher one or more of the above abilities than the control plant, and are therefore selected in step 22.

[0090] Typical examples of the "control plant" in step 22 are the corresponding wild-type plant and the plant (parent individual, etc.) used as the material for breeding.

[0091] 8. Plant breeding methods Another aspect of the present invention is A method for breeding a plant having high abilities selected from an ability to retain chlorophyll, an ability to produce chlorophyll, and an ability to grow, comprising: a step 31 of crossing a first parental plant individual with a second parental individual in which the expression of one or more genes selected from ALKBH8, ALKBH8 homologs, TGA9, TGA9 homologs, GRF15, GRF15 homologs, At3g61540, At3g61540 homologs, At2g01818, and At2g01818 homologs and / or the activity of proteins derived from said genes is the same as or lower than that of the first parental individual to obtain offspring; a step 32 of measuring the expression of said gene and / or the activity of a protein derived from said gene in said offspring; Step 33: selecting the offspring when the expression of the gene and / or the activity of the protein derived from the gene in the offspring measured in step 32 is the same as or lower than the expression of the gene and / or the activity of the protein derived from the gene in the second parent individual; The present invention relates to a method comprising:

[0092] The second parent individual in step 31 is typically a plant individual in which the expression of the gene and / or the activity of the protein derived from the gene is suppressed, more preferably a plant individual in which the expression and / or activity is suppressed compared to the wild type.

[0093] In this method, a plant individual in which the expression of the gene and / or the activity of the protein derived from the gene is suppressed is used as a second parent individual, and by crossing it with the first parent individual and selecting from the offspring obtained, plant individuals in which the expression and / or activity is similar to or lower than that of the second parent individual, it is possible to efficiently breed plants with high one or more of the above abilities.

[0094] Step 32 can be carried out in the same manner as step 11.

[0095] Progeny whose expression of the gene and / or activity of the protein derived from the gene measured in step 32 is the same or lower than that of the second parent individual are plants whose one or more abilities are equal to or higher than that of the second parent individual, and are therefore selected in step 33.

[0096] The progeny selected in step 33 may be used to cross with additional plant individuals to obtain the next generation of progeny.

[0097] <9. How to improve plant performance> Another aspect of the present invention is A method for improving one or more abilities selected from an ability to retain chlorophyll, an ability to produce chlorophyll, and an ability to grow in a plant, comprising: Step 41: Suppressing the expression of one or more genes selected from ALKBH8, ALKBH8 homologs, TGA9, TGA9 homologs, GRF15, GRF15 homologs, At3g61540, At3g61540 homologs, At2g01818, and At2g01818 homologs in a plant, and / or the activity of proteins derived from the genes. The present invention relates to a method comprising:

[0098] In step 41, suppressing the expression of the gene and / or the activity of the protein derived from the gene includes partially or completely deleting the base sequence of the gene on the chromosome of the plant or the base sequence (control sequence) that has the function of controlling the expression of the gene, reducing the expression level of the gene, degrading the transcription product of the gene, suppressing the activity of the translation product (protein) of the gene, degrading the protein, etc.

[0099] For example, methods for partially or completely deleting the base sequence or regulatory sequence of the gene, or methods for reducing the expression of the gene, include gene mutation using drugs, radiation, or ultraviolet light, T-DNA methods, genome editing methods, transposon methods, transgene methods, post-transcriptional gene silencing methods, RNAi methods, nonsense mediated decay (NMD), ribozyme methods, antisense methods, miRNA methods (micro-RNA), siRNA methods (small interfering RNA), antibody methods, and the like.

[0100] Methods for suppressing the activity of the protein include introducing an antagonist against the protein into a plant to cause competitive inhibition or antagonistic inhibition.

[0101] Step 41 particularly preferably comprises: (I) disruption of the gene in the genomic DNA of a plant; (II) introducing antisense DNA for the gene into a plant; and (III) introducing into a plant an antagonist to the protein derived from the gene; Contains one or more of the following: The disruption of the gene in (I) refers to the partial or complete deletion of the base sequence or regulatory sequence of the gene.

[0102] The timing of performing step 41 is not important. For example, it may be at the seed stage, any growth stage (seedling, vegetative growth, reproductive growth), or callus stage. Furthermore, the part of the plant where step 41 is performed is also not important. For example, it may be performed only on specific cells or cell tissues such as the reproductive meristem or the shoot apex.

[0103] <10.Plants> Another aspect of the present invention is The present invention relates to a plant (excluding Arabidopsis thaliana) in which the expression of one or more genes selected from ALKBH8, ALKBH8 homologs, TGA9, TGA9 homologs, GRF15, GRF15 homologs, At3g61540, At3g61540 homologs, At2g01818, and At2g01818 homologs, and / or the activity of proteins derived from said genes, are suppressed.

[0104] The plant of this embodiment is useful because it has high ability to retain chlorophyll, high ability to produce chlorophyll, and high ability to grow.

[0105] The plant of this embodiment can be produced by a method including step 41 above. The present invention will be described below with reference to specific examples, which, however, are not intended to limit the scope of the present invention. [Example]

[0106] (1) The Arabidopsis ALKBH8 gene is identified by the AGI code At1g31600. This At1g31600 gene was reported by Leihne et al. (2011) Nucleic Acids Res 39: 7688-7701 as a gene involved in tRNA modification.

[0107] The nucleotide sequence of the coding region of the Arabidopsis thaliana ALKBH8 (At1g31600) gene is shown in SEQ ID NOs: 1, 2, and 3, and the amino acid sequence of ALKBH8 encoded by the gene is shown in SEQ ID NOs: 4, 5, and 6.

[0108] (2) Arabidopsis T-DNA tagged lines The Arabidopsis T-DNA insertion mutants SALK_083838C and SALK_094502C, obtained from the Nottingham Arabidopsis Stock Center (http: / / arabidopsis.info / ), were used, in which T-DNA was inserted into the At1g31600 gene.

[0109] (3) Identification of T-DNA insertion site and homozygous lines As shown in Figure 1, SALK_083838C (referred to as "alkbh8-1") and SALK_094502C (referred to as "alkbh8-2") have a T-DNA inserted into the At1g31600 gene. First, the obtained seeds were cultivated, and the following procedure was used to confirm that the strains had a homozygous T-DNA insertion.

[0110] Specifically, alkbh8-1 strain, alkbh8-2 strain, and wild-type Arabidopsis thaliana (Col) were sown in 6cm x 6cm x 5cm pots with a bottom layer of vermiculite (Nitta), a middle layer of Kureha horticultural soil (Kureha), and a top layer of vermiculite in a ratio of 80ml:40ml:40ml.Four weeks later, rosette leaves were collected from each strain and DNA was extracted. The primer set 2 (primer set 2) for detecting the T-DNA insertion borders in alkbh8-1 and alkbh8-2 strains was 5'-TGGTTCACGTAGTGGGCCATCG-3' (LBa1; SEQ ID NO: 7) and 5'-TGAGCTTCCGTCGTTTGTT-3' (AtALKBH8_seq_f; SEQ ID NO: 8). The primer set 1 (primer set 1) for detecting the At1g31600 gene was AtALKBH8_seq_f and 5'-TCGTGCTTCACCGGATAATAAG-3' (AtALKBH8_seq_r; SEQ ID NO: 9) (Figure 1). As a control, PCR was also performed with wild-type Arabidopsis (Col) using the same primer set. For both primer sets, PCR was performed with one cycle at 94°C (1 min), followed by 35 cycles at 94°C (30 s), 60°C (30 s), and 72°C (2 min).

[0111] In both mutant lines (alkbh8-1 and alkbh8-2), PCR using primer set 2 confirmed amplification of the T-DNA insertion boundary region, but PCR using primer set 1 did not confirm amplification of the At1g31600 gene (Figure 2). In contrast, PCR using primer set 2 did not confirm amplification of the T-DNA insertion boundary region in wild-type Arabidopsis (Col) DNA, but PCR using primer set 1 confirmed amplification of the At1g31600 gene (Figure 2). Based on these band patterns, we determined that the T-DNA had been homozygously inserted into the At1g31600 gene in these mutants. [Example]

[0112] Growth analysis <Experiment 1> The growth of wild-type Arabidopsis thaliana (Col) was compared with that of the ALKBH8 disruptant strains (alkbh8-1 and alkbh8-2 strains) described in Example 1. The plants were cultivated under a nutrient solution of approximately 100 μmol / m 2 The planting conditions were a 16-hour light / 8-hour dark cycle with a light intensity of / s and a temperature of 22°C. Vermiculite and Kureha horticultural soil were used as the growing medium. Specifically, 6cm x 6cm x 5cm pots were layered with vermiculite in the bottom layer, Kureha horticultural soil in the middle layer, and vermiculite on top in a ratio of 80ml:40ml:40ml. Kureha horticultural soil contained 0.47mg / ml nitrogen, 1.18mg / ml phosphorus, and 0.71mg / ml potassium. Arabidopsis plants grown under these conditions typically show no signs of nitrogen deficiency, even without additional fertilization. Plants were grown at a density of three plants per pot. Submerged water was used as needed to prevent the soil from drying out. Nine pots per plot were used for measuring aboveground weight, and six plants per plot were used for measuring chlorophyll content.

[0113] Growth was assessed by measuring shoot weight and leaf chlorophyll content at 4 weeks. The results showed that the shoot weight of the wild-type plant was 489 mg / pot at 4 weeks, while that of the ALKBH8-disrupted plants was 601 mg / pot and 610 mg / pot, respectively, confirming that the ALKBH8-disrupted plants had a 23-25% greater shoot weight than the wild-type plant. Furthermore, the chlorophyll content per leaf area, calculated as chlorophyll a, was 21.6 nmol / cm in the wild-type plant. 2 In contrast, the ALKBH8 disruptant had a level of 23.5 nmol / cm 2 and 22.6 nmol / cm 2 The chlorophyll b concentration was 5.4 nmol / cm compared to the wild type. 2 In contrast, the ALKBH8 disruptant had a 5.9 nmol / cm 2 and 5.7 nmol / cm 2 This was a 6-9% increase compared to the wild-type strain. Thus, it was confirmed that disrupting the ALKBH8 gene increases the chlorophyll content in the leaves, resulting in increased photosynthesis and promoting plant growth.

[0114] [Table 6]

[0115] <Experiment 2> Next, the amount of fertilizer was increased, and the growth of the Arabidopsis thaliana (Col) wild-type strain and the ALKBH8 disruptant strain described in Example 1 were compared. Light intensity, light / dark period, and temperature were the same as in Experiment 1. The soil for cultivation was layered in 6 cm × 6 cm × 5 cm pots, with vermiculite in the bottom layer, Kureha horticulture soil in the middle layer, and vermiculite in the top layer, in a ratio of 40 ml:80 ml:40 ml. The plants were grown at a density of three plants per pot. Water was applied by subirrigation as needed to prevent the soil from drying out. From the first to fifth week, 1 mM oxidized glutathione was applied by subirrigation once a week. In addition, in the fourth week, a liquid fertilizer "Hyponex Concentrate" (Hyponex Japan) diluted 1:1000 with water was applied by subirrigation. The number of plants per plot was eight pots (24 plants).

[0116] Growth was assessed by measuring the number of branches and final above-ground weight at 10 weeks. Results showed that the wild-type plant had an average of 3.7 branches per plant at 10 weeks, while the ALKBH8-disrupted plants had 4.7 and 4.3 branches, representing an 8-13% increase compared to the wild-type. Final above-ground weight was 1268 mg / pot for the wild-type plant, while the ALKBH8-disrupted plants had 1298 mg / pot and 1343 mg / pot, representing a 2-6% increase compared to the wild-type plant.

[0117] These results indicate that disrupting the ALKBH8 gene can further increase plant growth rates, even under conditions of sufficient fertilization.

[0118] [Table 7]

[0119] <Experiment 3> The chlorophyll content of Arabidopsis thaliana (Col) wild-type strains and the ALKBH8 disruptant strain described in Example 1 was compared over time. The experimental conditions (light intensity, light / dark cycle, and temperature) were the same as in Experiments 1 and 2. As in Experiment 2, plants were grown in 6 cm × 6 cm × 5 cm pots, layered with vermiculite in the bottom layer, Kureha horticulture soil in the middle layer, and vermiculite in the top layer in a ratio of 40 ml:80 ml:40 ml. Plants were grown at a density of three plants per pot. Water was applied by subirrigation as needed to prevent the soil from drying out. In addition, from the fourth week, a liquid fertilizer "Hyponex Concentrate" (Hyponex Japan) diluted 1:1000 with water was applied by subirrigation. Furthermore, plants for which chlorophyll content was measured were subirrigated with 1 mM oxidized glutathione once a week from the first to fifth weeks.

[0120] Rosette leaves were sampled from eight plants at 4, 5, and 6 weeks after sowing, and the chlorophyll a and chlorophyll b contents per leaf weight were measured. In addition, the number of rosette leaves retaining chlorophyll was counted at 10 weeks in two pots in each plot.

[0121] At 6 weeks, the wild-type plant had chlorophyll a and chlorophyll b contents of 2.07 nmol / mg and 0.56 nmol / mg, respectively, while the ALKBH8-disrupted plants, alkbh8-1, had chlorophyll a and chlorophyll b contents of 2.16 nmol / mg and 0.59 nmol / mg, respectively, and alkbh8-2, had chlorophyll contents of 2.23 nmol / mg and 0.61 nmol / mg, respectively. Furthermore, at 10 weeks, almost all rosette leaves of the wild-type plant had yellowed and chlorophyll was not being retained, whereas the rosette leaves of the ALKBH8-disrupted plants had green leaves of 2.3 per plant (alkbh8-1) and 1.0 per plant (alkbh8-2), clearly indicating that chlorophyll was being retained.

[0122] Therefore, it was revealed that disrupting ALKBH8 can maintain chlorophyll content for a long period of time.

[0123] [Table 8]

[0124] [Table 9] [Example]

[0125] (1) Arabidopsis GRF15 gene disruption mutant (1-1) The Arabidopsis GRF15 gene is identified by the AGI code: At2g10450. The nucleotide sequence of the coding region of the Arabidopsis thaliana GRF15 (At2g10450) gene is shown in SEQ ID NO: 26, and the amino acid sequence of Arabidopsis thaliana GRF15 encoded by the gene is shown in SEQ ID NO: 27.

[0126] (1-2) Arabidopsis T-DNA tagged lines The Arabidopsis T-DNA insertion mutant, SALK_112825, obtained from the Nottingham Arabidopsis Stock Center (http: / / arabidopsis.info / ), was used, in which T-DNA was inserted into the At2g10450 gene.

[0127] (1-3) Confirmation of T-DNA insertion site and homozygous line As shown in Figure 4, SALK_112825 has a T-DNA inserted into the GRF15 (At2g10450) gene. First, the obtained seeds were cultivated and the strain was confirmed to be a homozygous T-DNA insert using the following procedure.

[0128] Specifically, SALK_112825 and wild-type Arabidopsis (Col) were seeded in 6 cm × 6 cm × 5 cm pots layered with vermiculite (Nittai) in the bottom layer, Kureha horticultural soil (Kureha) in the middle layer, and vermiculite in the top layer (80 ml:40 ml:40 ml). After 4 weeks, rosette leaves were harvested from each plant and DNA was extracted. GRF15_seq_f (SEQ ID NO: 10) and LBa1 (SEQ ID NO: 7) were used as primer set 2 to detect the T-DNA insertion boundary in SALK_112825, and GRF15_seq_f (SEQ ID NO: 10) and GRF15_seq_r (SEQ ID NO: 11) were used as primer set 1 to detect the GRF15 (At2g10450) gene (Figure 4). As a control, PCR was also performed on wild-type Arabidopsis (Col) using the primer sets described above. For both primer sets, PCR was performed using one cycle at 94°C (1 min), followed by 35 cycles at 94°C (30 s), 60°C (30 s), and 72°C (3 min).

[0129] In the DNA of the SALK_112825 strain, PCR using primer set 2 confirmed amplification of the T-DNA insertion boundary region, but PCR using primer set 1 did not confirm amplification of the GRF15 (At2g10450) gene (Figure 5). On the other hand, in the DNA of wild-type Arabidopsis thaliana (Col), PCR using primer set 2 did not confirm amplification of the T-DNA insertion boundary region, but PCR using primer set 1 confirmed amplification of the GRF15 (At2g10450) gene (Figure 5). From these band patterns, we determined that the T-DNA in the SALK_112825 strain is homozygous for the GRF15 (At2g10450) gene.

[0130] (2) Arabidopsis At3g61540 gene disruption mutant (2-1) The nucleotide sequence of the coding region of the gene (At3g61540 gene) identified by the Arabidopsis thaliana AGI code: At3g61540 is shown in SEQ ID NO: 28, and the amino acid sequence of the Arabidopsis thaliana At3g61540 gene encoded by this gene is shown in SEQ ID NO: 29.

[0131] The KPHMT2 gene, which is located adjacent to the upstream side of the At3g61540 gene on the Arabidopsis thaliana genomic DNA and whose transcription direction is opposite to that of the At3g61540 gene, is identified by the AGI code At3g61530. Two splicing variants, At3g61530.1 and At3g61530.2, are known for the Arabidopsis KPHMT2 (At3g61530) gene. The nucleotide sequences of their coding regions are shown in SEQ ID NOs: 30 and 31, and their amino acid sequences are shown in SEQ ID NOs: 32 and 33.

[0132] (2-2) Arabidopsis T-DNA tagged lines The Arabidopsis T-DNA insertion mutant, SALK_135878, obtained from the Nottingham Arabidopsis Stock Center (http: / / arabidopsis.info / ), was used, in which T-DNA was inserted into the At3g61540 gene.

[0133] (2-3) Confirmation of T-DNA insertion site and homozygous line As shown in Figure 6, SALK_135878 has a T-DNA insertion within the coding region of the At3g61540 gene near the start codon of the At3g61540 gene and in the untranslated region of the KPHMT2 (At3g61530) gene upstream of the start codon. First, the obtained seeds were cultivated, and the strain was confirmed to have a homozygous T-DNA insertion by the following procedure.

[0134] Specifically, SALK_135878 and wild-type Arabidopsis thaliana (Col) were seeded in the pots described in (1-3) above. After 4 weeks, rosette leaves were collected from each plant and DNA was extracted. LBa1 (SEQ ID NO: 7) and At3g61540_seq_r (SEQ ID NO: 13) were used as primer sets (primer set 2) to detect the T-DNA insertion boundary region in SALK_135878, and KPHMT2_seq_r (SEQ ID NO: 12) and At3g61540_seq_r (SEQ ID NO: 13) were used as primer sets (primer set 1) to detect the KPHMT2 (At3g61530) and At3g61540 genes (Figure 6). As a control, PCR was also performed on wild-type Arabidopsis thaliana (Col) using the same primer sets. The PCR reaction conditions were as described in (1-3) above.

[0135] In the DNA of the SALK_135878 strain, PCR using primer set 2 confirmed amplification of the T-DNA insertion boundary region, but PCR using primer set 1 did not confirm amplification of the KPHMT2(At3g61530)-At3g61540 gene (Figure 7). In contrast, in the DNA of wild-type Arabidopsis thaliana (Col), PCR using primer set 2 did not confirm amplification of the T-DNA insertion boundary region, but PCR using primer set 1 confirmed amplification of the KPHMT2(At3g61530)-At3g61540 gene (Figure 7). Based on these band patterns, we determined that the T-DNA in the SALK_135878 strain was homozygously inserted into the At3g61540 gene.

[0136] (3) Arabidopsis TGA9 gene disruption mutant (3-1) The Arabidopsis TGA9 gene is identified by the AGI code: At1g08320. The Arabidopsis thaliana TGA9 (At1g08320) gene has five known splicing variants: At1g08320.1, At1g08320.2, At1g08320.3, At1g08320.4, and At1g08320.5. The nucleotide sequences of the respective coding regions are shown in SEQ ID NOs: 34, 35, 36, 37, and 38, and the respective amino acid sequences are shown in SEQ ID NOs: 39, 40, 41, 42, and 43.

[0137] (3-2) Arabidopsis T-DNA tagged lines The Arabidopsis T-DNA insertion mutant SALK_091349, obtained from the Nottingham Arabidopsis Stock Center (http: / / arabidopsis.info / ), was used, in which T-DNA was inserted into the At1g08320 gene.

[0138] (3-3) Confirmation of T-DNA insertion site and homozygous line As shown in Figure 8, SALK_091349 has a T-DNA inserted into the TGA9 (At1g08320) gene. First, the obtained seeds were cultivated and the strain was confirmed to have a homozygous T-DNA insertion by the following procedure.

[0139] Specifically, the SALK_091349 strain and wild-type Arabidopsis thaliana (Col) were sown in the pots described in (1-3) above. After 4 weeks, rosette leaves were collected from each strain and DNA was extracted. TGA9_seq_f (SEQ ID NO: 14) and LBa1 (SEQ ID NO: 7) were used as primer set 2 to detect the T-DNA insertion boundary region in the SALK_091349 strain, and TGA9_seq_f (SEQ ID NO: 14) and TGA9_seq_r (SEQ ID NO: 15) were used as primer set 1 to detect the TGA9 (At1g08320) gene (Figure 8). As a control, PCR was also performed on wild-type Arabidopsis thaliana (Col) using the same primer set. The PCR reaction conditions were as described in (1-3) above.

[0140] In the DNA of the SALK_091349 strain, amplification of the T-DNA insertion boundary region was confirmed by PCR using primer set 2, but amplification of the TGA9 (At1g08320) gene was not confirmed by PCR using primer set 1 (Figure 9). On the other hand, in the DNA of wild-type Arabidopsis thaliana (Col), amplification of the T-DNA insertion boundary region was not confirmed by PCR using primer set 2, but amplification of the TGA9 (At1g08320) gene was confirmed by PCR using primer set 1 (Figure 9). From these band patterns, we determined that the T-DNA had been homozygously inserted into the TGA9 (At1g08320) gene in the SALK_091349 strain.

[0141] (4) Arabidopsis At2g01818 gene disruption mutant (4-1) The gene identified by the Arabidopsis AGI code: At2g01818 (At2g01818 gene) has two known splicing variants, At2g01818.1 and At2g01818.2. The nucleotide sequences of the respective coding regions are shown in SEQ ID NOs: 44 and 45, and the respective amino acid sequences are shown in SEQ ID NOs: 46 and 47.

[0142] (4-2) Arabidopsis T-DNA tagged lines The Arabidopsis T-DNA insertion mutant SAIL_568_E01, obtained from the Nottingham Arabidopsis Stock Center (http: / / arabidopsis.info / ), was used as the At2g01818 gene-introduced T-DNA insertion mutant.

[0143] (4-3) Confirmation of T-DNA insertion site and homozygous line As shown in Figure 10, SAIL_568_E01 has a T-DNA inserted into the At2g01818 gene. First, the obtained seeds were cultivated, and the strain was confirmed to be a homozygous T-DNA inserted strain by the following procedure.

[0144] Specifically, SAIL_568_E01 and wild-type Arabidopsis thaliana (Col) were sown in the pots described in (1-3) above. After 4 weeks, rosette leaves were collected from each plant and DNA was extracted. AT2G01818_seq_f (SEQ ID NO: 16) and LB3 (SEQ ID NO: 18) were used as primer sets (primer set 2) to detect the T-DNA insertion boundary region in SAIL_568_E01, and AT2G01818_seq_f (SEQ ID NO: 16) and AT2G01818_seq_r (SEQ ID NO: 17) were used as primer sets (primer set 1) to detect the At2g01818 gene (Figure 10). As a control, PCR was also performed on wild-type Arabidopsis thaliana (Col) using the same primer sets. The PCR reaction conditions were as described in (1-3) above.

[0145] In the DNA of the SAIL_568_E01 line, amplification of the border region where the T-DNA was inserted was confirmed by PCR using primer set 2, but amplification of the At2g01818 gene was not confirmed by PCR using primer set 1 (Figure 11). On the other hand, in the DNA of wild-type Arabidopsis thaliana (Col), amplification of the border region where the T-DNA was inserted was not confirmed by PCR using primer set 2, but amplification of the At2g01818 gene was confirmed by PCR using primer set 1 (Figure 11). From these band patterns, it was determined that the T-DNA in the SAIL_568_E01 line was homozygously inserted into the At2g01818 gene.

[0146] [Table 10] [Example]

[0147] The number of rosette leaves retaining chlorophyll was compared over time between the Arabidopsis thaliana (Col) wild-type strain and the GRF15, At3g61540, TGA9, and At2g01818 gene disruption strains described in Example 3.

[0148] The test conditions (light intensity, light / dark period, temperature) were the same as in Experiments 1, 2, and 3 of Example 2. As in Experiment 1 of Example 2, the soil was layered in a ratio of 80 ml:40 ml:40 ml, with vermiculite in the bottom layer of a 6 cm x 6 cm x 5 cm pot, Kureha horticultural soil in the middle layer, and vermiculite in the top layer. The plants were cultivated at a planting density of three plants per pot. Water was applied to the bottom as needed to prevent the soil from drying out.

[0149] The number of rosette leaves that retained chlorophyll was counted 7 weeks (48 days), 8 weeks (56 days), 9 weeks (63 days), and 10 weeks (70 days) after sowing. The degree to which chlorophyll was maintained for each fully expanded rosette leaf was scored as a continuous value ranging from 0 to 3, and the sum of the scores for each individual leaf on one plant was calculated as the "green leaf index." A score of 0, 1, 2, and 3 indicate the following conditions, respectively. 0: No green color remaining 1: Only part of the leaf remains green 2: The green color is fading 3: Dark green remains Only fully expanded rosette leaves were counted. Evaluation was performed on n = 9 plants for each of the wild-type and gene-disruption lines, and the average green leaf index per plant was calculated. FIG. 12 shows the appearance of Arabidopsis rosette leaves when the evaluation score indicating the degree of chlorophyll maintenance is 0, 1, 2, or 3.

[0150] FIG. 13 shows the average green leaf index per plant (n=9) at 48, 56, 63, and 70 days after sowing (DAS) for the Arabidopsis thaliana wild-type strain (Col) and the GRF15, At3g61540, TGA9, and At2g01818 gene disruption strains described in Example 3.

[0151] FIG. 14 shows the average total number of rosette leaves (green leaves) per plant with a chlorophyll content evaluation score of 1 to 3 (n=9 per plant) at the end of growth, 10 weeks (70 days) after sowing, for the Arabidopsis wild-type plant (Col) and the GRF15, At3g61540, TGA9, and At2g01818 gene disruption plants described in Example 3, and the relative ratio of the number of green leaves for each gene disruption plant to the number of green leaves for the wild-type plant, which is set at 100.

[0152] The results shown in Figures 13 and 14 indicate that in Arabidopsis thaliana GRF15 gene, At3g61540 gene, TGA9 gene, or At2g01818 gene disruption strains, the decrease in chlorophyll over time is suppressed compared to the wild-type strain, and chlorophyll is maintained even at the end of growth. [Example]

[0153] (1) Generation of rice ALKBH8 gene disruptants The rice ortholog of the ALKBH8 gene, Os04g0602700 (hereafter referred to as OsALKBH8), has been assigned the ID dosa:Os04t0602700-00 in KEGG (http: / / www.genome.jp / kegg / ), and its nucleotide sequence and the amino acid sequence encoded by this nucleotide sequence are registered. The nucleotide sequence of OsALKBH8 is shown in SEQ ID NO: 48, and the amino acid sequence is shown in SEQ ID NO: 49.

[0154] Two guide RNA transcription sequences for genome editing (SEQ ID NO: 19 and SEQ ID NO: 20) were designed for OsALKBH8. Based on the designed sequences, two complementary oligo DNAs were synthesized and annealed. The pU6-ccdB-gRNA vector was digested with the restriction enzyme BbsI and purified by gel extraction using the QIAquick Gel Extraction Kit (QIAGEN). The restriction enzyme-digested vector and guide RNA transcription sequence were ligated using Takara Ligation Mix (Takara Bio). The vector was transformed into Ecos Competent E. coli DH5α (Nippon Gene) and cultured overnight at 37°C on an LB medium plate containing 50 μg / mL ampicillin. A colony from the plate was scraped and added to 2 mL of liquid LB medium containing 50 μg / mL ampicillin, followed by overnight incubation at 37°C with shaking at 180 rpm. The plasmid was purified using the QIAprep Spin Miniprep Kit (QIAGEN). Sequence analysis was performed using the OsU6-2_F2 primer (SEQ ID NO: 21) and M13_F primer (SEQ ID NO: 22) to confirm the sequence. Plasmids with confirmed sequences and the pZH-gYSA-PubiMMCas9 vector were digested with the restriction enzyme I-SceI. The fragments excised from the plasmid and the vector backbone were purified by gel extraction using a QIAquick Gel Extraction Kit. They were ligated using Takara Ligation Mix and transformed into Ecos Competent E. coli DH5α. The cells were grown overnight at 37°C on LB medium plates containing 50 μg / mL spectinomycin. A scraped portion of an E. coli colony was used as a template for PCR using the M13_R primer (SEQ ID NO: 50), PZmUbi_R primer (SEQ ID NO: 23), and OsU6-2_F2 primer (SEQ ID NO: 21). E. coli carrying the vector containing the target fragment was cultured overnight in liquid LB medium containing spectinomycin at 37°C with shaking at 180 rpm, and the plasmid was purified using the QIAprep Spin Miniprep Kit. Sequence analysis was performed using the three primers mentioned above to confirm that the target sequence had been inserted.

[0155] Rice (Nipponbare) seeds were treated with 70% ethanol for 1 minute, 20% sodium hypochlorite for 1 hour, and then washed five times with sterile water. The seeds were placed on callus induction medium plates and incubated at 30°C with a 16-hour light / 8-hour dark period. After 3 weeks of incubation, the resulting callus was transferred to another callus induction medium.

[0156] Competent cells of Agrobacterium strain EHA101 were thawed on ice and the plasmid pZH-gOsALKBH8-PubiMMCas9, prepared as described above, was added. Using a Gene Pulser II (Bio-Rad) set to 18 kV and 25 μF, the bacterial solution was placed in a cuvette with a 0.1 cm electrode gap and electroporation was performed. SOC medium was added to the cuvette, mixed with the bacterial solution, and then transferred to a test tube and cultured at 28°C and 180 rpm for 3 hours. After culturing, the bacterial solution was spread onto an LB medium plate containing 50 μg / mL kanamycin and 50 μg / mL spectinomycin and incubated at 28°C. After two days of culture, colonies were scraped and added to liquid LB medium containing 50 μg / mL kanamycin and 50 μg / mL spectinomycin and cultured at 28°C and 180 rpm with shaking. After one day of culture, the culture was centrifuged at 1,700 × g for 10 minutes. Liquid transformation medium was added and the bacteria were suspended to an OD600 of approximately 0.1. 5 mL of liquid transformation medium was placed in a sterile dish, and the callus cultured on callus induction medium for 3 days was transferred thereto. The bacterial suspension was then poured over the callus and allowed to stand for 5 minutes. The callus was then transferred to co-cultivation medium and cultured at 28°C in the dark.

[0157] After culturing for three days, the calli were washed five times with 20 mL of sterile water. They were then washed three more times with 20 mL of sterile water containing 500 μg / mL carbenicillin. The calli were transferred to selection medium and cultured at 30°C under a 16-hour light / 8-hour dark cycle. After culturing for two weeks, the surviving calli were transferred to regeneration medium and cultured under the same conditions. Thereafter, the calli were transferred to new regeneration medium every two weeks. Calli from which shoots had differentiated were transferred to hormone-free medium and cultured under the same conditions.

[0158] Vermiculite was filled into No. 3 pots (9 cm diameter, 8 cm height) and allowed to absorb water. Plants on hormone-free medium whose leaves had grown to the top of the test tube were then transplanted into the pots. A support was set up, and a nylon bag was placed over the top and secured with a rubber band. The plants were cultured under a 14-hour light period (32°C) and 10-hour dark period (25°C). Slits were gradually made in the bag over the course of about a week to allow the plants to acclimate.

[0159] A mixture of 10 L of vermiculite and 2 L of King Soil Special No. 1 (Kasanen Kogyo) was placed in a 1 / 2000a Wagner pot. After cultivating the plants in the No. 3 pots for approximately two weeks, they were transplanted into Wagner pots. They were cultivated under a 12-hour light period (30°C) and 12-hour dark period (25°C). Approximately two months after transplanting, the plants were treated with a 2000-fold diluted solution of Hyponex concentrate once every two weeks. Three and a half months after transplanting, the cultivation conditions were changed to 11 hours of light (28°C) and 13 hours of dark (23°C), and after four months, the conditions were changed to 10 hours of light (27°C) and 14 hours of dark (22°C).

[0160] Leaf pieces approximately 1 mm square were collected from T0 plants under cultivation, soaked in extraction buffer, and crushed. A 20-fold diluted sample of the extract was used as a template for PCR using the OsALKBH8_seq_f primer (SEQ ID NO: 24) and OsALKBH8_seq_r primer (SEQ ID NO: 25). The resulting product was sequenced using the same primers. T0 plants with confirmed insertions or deletions were grown to maturity, self-crossed, and T1 seeds were harvested. The T1 seeds were then sown and cultivated, and the T1 plants were self-crossed to obtain T2 seeds for testing. The locations of the OsALKBH8_seq_f primer, OsALKBH8_seq_r primer, and guide RNA (gOsALKBH8-1) in the OsALKBH8 gene are shown in Figure 15.

[0161] [Table 11]

[0162] The compositions and preparation methods of the medium and extraction buffer used to create the above-mentioned rice ALKBH8 gene disruptant are shown below.

[0163] <Callus induction medium> [Table 12]

[0164] The pH was adjusted to 5.8 with a few drops of 1 M KOH, and the mixture was diluted to 80 mL with ion-exchanged water. 640 mg of agar was added, and the mixture was sterilized in an autoclave at 121°C for 20 minutes. 1.5 mL of the mixture was dispensed into two 24-well plates to serve as callus induction medium.

[0165] <Liquid medium for transformation> [Table 13]

[0166] The pH was adjusted to 5.6 with a few drops of 1M KOH, and the volume was adjusted to 200mL with ion-exchanged water. The medium was sterilized in an autoclave at 121°C for 20 minutes. After cooling, 20µL of a 100mg / mL acetosyringone in DMSO solution was added and mixed.

[0167] <Co-culture medium> [Table 14]

[0168] The pH was adjusted to 5.6 with a few drops of 1M KOH, and the volume was adjusted to 200mL with ion-exchanged water. 1.6g of agar was added, and the mixture was sterilized in an autoclave at 121°C for 20 minutes. After the medium was cooled to below 60°C, 20µL of a 100mg / mL acetosyringone-DMSO solution was added. 40mL aliquots were dispensed into rectangular Petri dishes (96 x 96 x 15mm) to prepare the transformation medium.

[0169] <Selective medium> [Table 15]

[0170] The pH was adjusted to 5.8 with a few drops of 1M KOH, and the volume was adjusted to 200mL with ion-exchanged water. 1.6g of agar was added, and the medium was sterilized in an autoclave at 121°C for 20 minutes. After cooling the medium to below 60°C, 100µL of 100mg / mL hygromycin and 1000µL of 100mg / mL carbenicillin were added and mixed. 40mL aliquots were dispensed into rectangular Petri dishes (96 x 96 x 15mm) to serve as selection medium.

[0171] <Regeneration medium> [Table 16]

[0172] The pH was adjusted to 5.8 with a few drops of 5N HCl, and the volume was then adjusted to 200 mL with ion-exchanged water. 1.6 g of agar was added, and the medium was sterilized in an autoclave at 121°C for 20 minutes. After cooling the medium to below 60°C, 100 μL of 100 mg / mL hygromycin and 400 μL of 100 mg / mL carbenicillin were added and mixed. 40 mL of the medium was dispensed into rectangular Petri dishes (96 x 96 x 15 mm) to serve as the regeneration medium.

[0173] <Hormone-free medium> [Table 17]

[0174] The pH was adjusted to 5.8 with a few drops of 1M KOH, and the volume was adjusted to 50mL with ion-exchanged water. 400mg of agar was added, and the mixture was sterilized in an autoclave at 121°C for 20 minutes. 25mL of the mixture was dispensed into two culture test tubes (4cm diameter, 13cm height).

[0175] <Extraction buffer> [Table 18]

[0176] (2) Measurement of chlorophyll content in rice OsALKBH8 gene disruptants A mixture of 10 L of vermiculite and 2 L of King Soil Special No. 1 (Kasanen Kogyo) was filled into 1 / 2000a Wagner pots. In early June, after allowing the soil to absorb sufficient water, T2 seeds of the rice OsALKBH8 gene disruptant created in (1) above and wild-type rice seeds were sown, six per pot, and cultivated in a special screened greenhouse. After two weeks of cultivation, the plants were thinned to three per pot. At eight weeks of age, the SPAD value was measured near the center of the second fully expanded leaf from the top of the main stem using a SPAD-502Plus chlorophyll meter (Konica Minolta). The SPAD value is an index that correlates with the amount of chlorophyll in plant leaves.

[0177] The results are shown in Figure 16. It was confirmed that the OsALKBH8 gene disruptant rice line had a 17% higher SPAD value at 8 weeks of age and a higher amount of chlorophyll than wild-type rice. [Industrial Applicability]

[0178] The present invention can be used in the fields of agriculture and horticulture. [Sequence List Free Text]

[0179] SEQ ID NOs: 7 to 25, 50: primers

Claims

1. A method for evaluating one or more abilities selected from the ability to retain chlorophyll, the ability to produce chlorophyll, and the ability to grow in a plant, comprising: a measuring step of measuring the expression of one or more genes selected from the following (1A) to (1F), (2A) to (2F), (3A) to (3F), (4A) to (4F), and (5A) to (5F) in a plant and / or the activity of a protein derived from said gene; Methods including: (1A) a base sequence set forth in SEQ ID NO: 1, 2, 3, or 48; (1B) an amino acid sequence set forth in SEQ ID NO: 4, 5, 6, or 49, or a base sequence encoding an amino acid sequence having 90% or more sequence identity to the amino acid sequence and having ALKBH8 activity; (1C) The following ID in KEGG (Kyoto Encyclopedia of Genes and Genomes): dosa:Os04t0602700-00、dosa:Os02t0750500-01、osa:4330741、osa:4336893、ath:AT1G36310、ath:AT1G31600、aly:ARALYDRAFT_473619、aly:ARAL YDRAFT_473716、crb:CARUB_v10009616mg、crb:CARUB_v10009306mg、csat :104757812、csat:104787286、csat:104777497、csat:104742087、csat:1 04741763、csat:104777194、eus:EUTSA_v10004311mg、eus:EUTSA_v10006 757mg、brp:103867438、brp:103848760、brp:103833805、bna:111205055、 bna:106411643、bna:106447489、bna:106399512、bna:106411644、bna:10 6412207、bna:106411637、bna:106418960、boe:106342941、boe:10630818 4、boe:106307572、thj:104798532、thj:104813129、cpap:110812544、cpa p:110818506、cit:102616373、cit:102625137、cic:CICLE_v10001407mg、 cic:CICLE_v10004016mg, tcc:18606930, tcc:18592225, gra:105773784, gra:105764921, ghi:107962467, ghi:107918198, ghi:107946865, ghi:10 7946842、ghi:107933507、dzi:111275453、dzi:111295045、dzi:11131235 5、egr:104444193、egr:104456636、gmx:100784028、gmx:100777529、gmx: 100807751、pvu:PHAVU_002G123600g、pvu:PHAVU_009G027100g、vra:1067 59935、vra:106777615、var:108335758、var:108329993、ccaj:109815811、ccaj:109796724、mtr:MTR_2g460880、mtr:MTR_5g026030、cam:101494433、cam:101511642、adu:107460794、adu:107487073、aip:107606112、aip:107642443、lja:Lj0g3v0064359.1、lja:Lj0g3v0046209.1、lja:Lj2g3v1573010.1、lja:Lj2g3v1573010.2、lang:109348998、lang:109357026、fve:101305382、fve:101301875、pper:18776672、pper:18788824、pmum:103336690、pmum:103332964、pavi:110755353、pavi:110753860、mdm:103441609、mdm:103416044、mdm:103401476、pxb:103934470、pxb:103955543、zju:107422678、zju:107430520、csv:101210859、csv:101216715、cmo:103485433、cmo:103487636、mcha:111008031、mcha:111020230、cmax:111477075、cmax:111487507、cmax:111489322、rcu:8263009、rcu:8280124、jcu:105643385、jcu:105638853、hbr:110640684、hbr:110644165、hbr:110667825、pop:POPTR_0003s10550g、pop:POPTR_0002s09190g、 pop:POPTR_0975s00200g、jre:109007265、jre:109021366、jre:109019322、jre:109012454、vvi:100252728、vvi:100262638、sly:101260324、sly:101263510、spen:107015589、spen:107005551、sot:102591578、sot:102587122、cann:107867583、cann:107850720、nta:107782177、nta:107784057、nta:107778504、nta:107777356、nta:107799087、nta:107802481、nsy:104228789、nsy:1 04246141、nsy:104231727、nto:104097471、nto:104088120、nto:1041044 03、ini:109170629、ini:109170202、sind:105159340、sind:105164991、 oeu:111404274、oeu:111377742、oeu:111378936、oeu:111383327、han:11 0868608、han:110918392、bvg:104892444、bvg:104905687、soe:1107748 79、soe:110797769、nnu:104610319、nnu:104600059、obr:102703096、bdi :100835069、bdi:100827681、ats:109741229、ats:109760172、sbi:8066 423、sbi:8072284、zma:103627749、zma:100282025、sita:101777379、sit a:101775180、pda:103713010、pda:103715224、egu:105053522、egu:105 033011、mus:103996326、mus:103990395、dct:110097186、dct:110106918 、aof:109832664、aof:109825376、atr:18422692、atr:18422136、smo:SE LMODRAFT_78730、smo:SELMODRAFT_78643、smo:SELMODRAFT_112315、smo: SELMODRAFT_112271、ppp:PHYPADRAFT_106932、ppp:PHYPADRAFT_15669、vcn:VOLCADRAFT_108374、mng:MNEG_3576、olu:OSTLU_88555、ota:OT_ost ta13g02300、ota:OT_ostta11g00670、bpg:Bathy04g02310、bpg:Bathy13g02890、mis:MICPUN_86885、mis:MICPUN_60739、mpp:MICPUCDRAFT_41620、mpp:MICPUCDRAFT_17097, apro:F751_1394, or gsl:Gasu_35150, The base sequence of the gene to which (1D) an amino acid sequence encoded by a gene assigned the ID described in (1C) in KEGG, or a base sequence encoding an amino acid sequence having 90% or more sequence identity with the amino acid sequence and having ALKBH8 activity; (1E) A base sequence having 90% or more sequence identity with the base sequence of (1A) and encoding an amino acid sequence having ALKBH8 activity, or a base sequence having 90% or more sequence identity with the base sequence of (1C) and encoding an amino acid sequence having ALKBH8 activity; (1F) A base sequence complementary to the base sequence of (1A), (1B), (1C), (1D) or (1E); (2A) a base sequence set forth in SEQ ID NO: 34, 35, 36, 37, or 38; (2B) a base sequence encoding an amino acid sequence set forth in SEQ ID NO: 39, 40, 41, 42, or 43, or an amino acid sequence having 90% or more sequence identity to the amino acid sequence and having TGA9 activity; (2C) The following ID in KEGG (Kyoto Encyclopedia of Genes and Genomes): aly:9325751, csat:104755012, crb:17900540, eus:EUTSA_v10007490mg, bna:106412233, boe:106344082, brp:103871579, or rsz:108860791 The base sequence of the gene to which (2D) an amino acid sequence encoded by a gene assigned the ID described in (2C) in KEGG, or a base sequence encoding an amino acid sequence having 90% or more sequence identity with the amino acid sequence and having TGA9 activity; (2E) A nucleotide sequence having 90% or more sequence identity with the nucleotide sequence of (2A) or (2C) and encoding an amino acid sequence having TGA9 activity; or (2F) A base sequence complementary to the base sequence of (2A), (2B), (2C), (2D) or (2E); (3A) the base sequence set forth in SEQ ID NO: 26; (3B) a base sequence encoding the amino acid sequence set forth in SEQ ID NO: 27, or an amino acid sequence having 90% or more sequence identity to the amino acid sequence and having GRF15 activity; (3C) The following ID in KEGG (Kyoto Encyclopedia of Genes and Genomes): bna:106429451, boe:106342871, or brp:103863788 The base sequence of the gene to which (3D) an amino acid sequence encoded by a gene assigned the ID described in (3C) in KEGG, or a base sequence encoding an amino acid sequence having 90% or more sequence identity with the amino acid sequence and having GRF15 activity; (3E) A nucleotide sequence having 90% or more sequence identity with the nucleotide sequence of (3A) or (3C) and encoding an amino acid sequence having GRF15 activity; or (3F) A base sequence complementary to the base sequence of (3A), (3B), (3C), (3D) or (3E); (4A) the base sequence set forth in SEQ ID NO: 28; (4B) an amino acid sequence set forth in SEQ ID NO: 29, or a base sequence encoding an amino acid sequence having 90% or more sequence identity to the amino acid sequence and having At3g61540 activity; (4C) the following ID in KEGG (Kyoto Encyclopedia of Genes and Genomes): aly:9314457, crb:17884851, csat:104788446, eus:EUTSA, rsz:108814367, bna:106406599, boe:106296419, or brp:103830306 The base sequence of the gene to which (4D) an amino acid sequence encoded by a gene assigned the ID described in (4C) in KEGG, or a base sequence encoding an amino acid sequence having 90% or more sequence identity with the amino acid sequence and having At3g61540 activity; (4E) A base sequence having 90% or more sequence identity with the base sequence of (4A) or (4C) and encoding an amino acid sequence having At3g61540 activity; (4F) A base sequence complementary to the base sequence of (4A), (4B), (4C), (4D) or (4E); (5A) the base sequence set forth in SEQ ID NO: 44 or 45; (5B) a base sequence encoding an amino acid sequence set forth in SEQ ID NO: 46 or 47, or an amino acid sequence having 90% or more sequence identity to the amino acid sequence and having At2g01818 activity; (5C) The following ID in KEGG (Kyoto Encyclopedia of Genes and Genomes): aly:9311163, eus:EUTSA_v10005480mg, csat:104710111, crb:17885477, brp:103854038, bna:106432932, boe:106325697, or rsz:108809176 The base sequence of the gene to which (5D) an amino acid sequence encoded by a gene assigned the ID described in (5C) in KEGG, or a base sequence encoding an amino acid sequence having 90% or more sequence identity with the amino acid sequence and having At2g01818 activity; (5E) A base sequence having 90% or more sequence identity with the base sequence of (5A) or (5C) and encoding an amino acid sequence having At2g01818 activity; (5F) A base sequence complementary to the base sequence of (5A), (5B), (5C), (5D) or (5E).

2. The method according to claim 1, wherein the one or more genes are DNA and / or RNA comprising any one of the base sequences (1A) to (1F), (2A) to (2F), (3A) to (3F), (4A) to (4F), and (5A) to (5F).

3. The method according to claim 1 or 2, wherein the measuring step comprises measuring the expression of the gene by carrying out a nucleic acid amplification reaction using DNA or RNA obtained from the plant as a template.

4. Use of the compound as a marker for evaluating one or more abilities selected from the ability to retain chlorophyll, the ability to produce chlorophyll, and the ability to grow in a plant, comprising: Use of all or part of one or more genes selected from the following (1A) to (1F), (2A) to (2F), (3A) to (3F), (4A) to (4F), and (5A) to (5F), or proteins derived from said genes: (1A) a base sequence set forth in SEQ ID NO: 1, 2, 3, or 48; (1B) an amino acid sequence set forth in SEQ ID NO: 4, 5, 6, or 49, or a base sequence encoding an amino acid sequence having 90% or more sequence identity to the amino acid sequence and having ALKBH8 activity; (1C) The following ID in KEGG (Kyoto Encyclopedia of Genes and Genomes): dosa:Os04t0602700-00、dosa:Os02t0750500-01、osa:4330741、osa:4336893、ath:AT1G36310、ath:AT1G31600、aly:ARALYDRAFT_473619、aly:ARAL YDRAFT_473716、crb:CARUB_v10009616mg、crb:CARUB_v10009306mg、csat :104757812、csat:104787286、csat:104777497、csat:104742087、csat:1 04741763、csat:104777194、eus:EUTSA_v10004311mg、eus:EUTSA_v10006 757mg、brp:103867438、brp:103848760、brp:103833805、bna:111205055、 bna:106411643、bna:106447489、bna:106399512、bna:106411644、bna:10 6412207、bna:106411637、bna:106418960、boe:106342941、boe:10630818 4、boe:106307572、thj:104798532、thj:104813129、cpap:110812544、cpa p:110818506、cit:102616373、cit:102625137、cic:CICLE_v10001407mg、 cic:CICLE_v10004016mg, tcc:18606930, tcc:18592225, gra:105773784, gra:105764921, ghi:107962467, ghi:107918198, ghi:107946865, ghi:10 7946842、ghi:107933507、dzi:111275453、dzi:111295045、dzi:11131235 5、egr:104444193、egr:104456636、gmx:100784028、gmx:100777529、gmx: 100807751、pvu:PHAVU_002G123600g、pvu:PHAVU_009G027100g、vra:1067 59935、vra:106777615、var:108335758、var:108329993、ccaj:109815811、ccaj:109796724、mtr:MTR_2g460880、mtr:MTR_5g026030、cam:101494433、cam:101511642、adu:107460794、adu:107487073、aip:107606112、aip:107642443、lja:Lj0g3v0064359.1、lja:Lj0g3v0046209.1、lja:Lj2g3v1573010.1、lja:Lj2g3v1573010.2、lang:109348998、lang:109357026、fve:101305382、fve:101301875、pper:18776672、pper:18788824、pmum:103336690、pmum:103332964、pavi:110755353、pavi:110753860、mdm:103441609、mdm:103416044、mdm:103401476、pxb:103934470、pxb:103955543、zju:107422678、zju:107430520、csv:101210859、csv:101216715、cmo:103485433、cmo:103487636、mcha:111008031、mcha:111020230、cmax:111477075、cmax:111487507、cmax:111489322、rcu:8263009、rcu:8280124、jcu:105643385、jcu:105638853、hbr:110640684、hbr:110644165、hbr:110667825、pop:POPTR_0003s10550g、pop:POPTR_0002s09190g、 pop:POPTR_0975s00200g、jre:109007265、jre:109021366、jre:109019322、jre:109012454、vvi:100252728、vvi:100262638、sly:101260324、sly:101263510、spen:107015589、spen:107005551、sot:102591578、sot:102587122、cann:107867583、cann:107850720、nta:107782177、nta:107784057、nta:107778504、nta:107777356、nta:107799087、nta:107802481、nsy:104228789、nsy:1 04246141、nsy:104231727、nto:104097471、nto:104088120、nto:1041044 03、ini:109170629、ini:109170202、sind:105159340、sind:105164991、 oeu:111404274、oeu:111377742、oeu:111378936、oeu:111383327、han:11 0868608、han:110918392、bvg:104892444、bvg:104905687、soe:1107748 79、soe:110797769、nnu:104610319、nnu:104600059、obr:102703096、bdi :100835069、bdi:100827681、ats:109741229、ats:109760172、sbi:8066 423、sbi:8072284、zma:103627749、zma:100282025、sita:101777379、sit a:101775180、pda:103713010、pda:103715224、egu:105053522、egu:105 033011、mus:103996326、mus:103990395、dct:110097186、dct:110106918 、aof:109832664、aof:109825376、atr:18422692、atr:18422136、smo:SE LMODRAFT_78730、smo:SELMODRAFT_78643、smo:SELMODRAFT_112315、smo: SELMODRAFT_112271、ppp:PHYPADRAFT_106932、ppp:PHYPADRAFT_15669、vcn:VOLCADRAFT_108374、mng:MNEG_3576、olu:OSTLU_88555、ota:OT_ost ta13g02300、ota:OT_ostta11g00670、bpg:Bathy04g02310、bpg:Bathy13g02890、mis:MICPUN_86885、mis:MICPUN_60739、mpp:MICPUCDRAFT_41620、mpp:MICPUCDRAFT_17097, apro:F751_1394, or gsl:Gasu_35150, The base sequence of the gene to which (1D) an amino acid sequence encoded by a gene assigned the ID described in (1C) in KEGG, or a base sequence encoding an amino acid sequence having 90% or more sequence identity with the amino acid sequence and having ALKBH8 activity; (1E) A base sequence having 90% or more sequence identity with the base sequence of (1A) and encoding an amino acid sequence having ALKBH8 activity, or a base sequence having 90% or more sequence identity with the base sequence of (1C) and encoding an amino acid sequence having ALKBH8 activity; (1F) A base sequence complementary to the base sequence of (1A), (1B), (1C), (1D) or (1E); (2A) a base sequence set forth in SEQ ID NO: 34, 35, 36, 37, or 38; (2B) a base sequence encoding an amino acid sequence set forth in SEQ ID NO: 39, 40, 41, 42, or 43, or an amino acid sequence having 90% or more sequence identity to the amino acid sequence and having TGA9 activity; (2C) The following ID in KEGG (Kyoto Encyclopedia of Genes and Genomes): aly:9325751, csat:104755012, crb:17900540, eus:EUTSA_v10007490mg, bna:106412233, boe:106344082, brp:103871579, or rsz:108860791 The base sequence of the gene to which (2D) an amino acid sequence encoded by a gene assigned the ID described in (2C) in KEGG, or a base sequence encoding an amino acid sequence having 90% or more sequence identity with the amino acid sequence and having TGA9 activity; (2E) A nucleotide sequence having 90% or more sequence identity with the nucleotide sequence of (2A) or (2C) and encoding an amino acid sequence having TGA9 activity; or (2F) A base sequence complementary to the base sequence of (2A), (2B), (2C), (2D) or (2E); (3A) the base sequence set forth in SEQ ID NO: 26; (3B) a base sequence encoding the amino acid sequence set forth in SEQ ID NO: 27, or an amino acid sequence having 90% or more sequence identity to the amino acid sequence and having GRF15 activity; (3C) The following ID in KEGG (Kyoto Encyclopedia of Genes and Genomes): bna:106429451, boe:106342871, or brp:103863788 The base sequence of the gene to which (3D) an amino acid sequence encoded by a gene assigned the ID described in (3C) in KEGG, or a base sequence encoding an amino acid sequence having 90% or more sequence identity with the amino acid sequence and having GRF15 activity; (3E) A nucleotide sequence having 90% or more sequence identity with the nucleotide sequence of (3A) or (3C) and encoding an amino acid sequence having GRF15 activity; or (3F) A base sequence complementary to the base sequence of (3A), (3B), (3C), (3D) or (3E); (4A) the base sequence set forth in SEQ ID NO: 28; (4B) an amino acid sequence set forth in SEQ ID NO: 29, or a base sequence encoding an amino acid sequence having 90% or more sequence identity to the amino acid sequence and having At3g61540 activity; (4C) the following ID in KEGG (Kyoto Encyclopedia of Genes and Genomes): aly:9314457, crb:17884851, csat:104788446, eus:EUTSA, rsz:108814367, bna:106406599, boe:106296419, or brp:103830306 The base sequence of the gene to which (4D) an amino acid sequence encoded by a gene assigned the ID described in (4C) in KEGG, or a base sequence encoding an amino acid sequence having 90% or more sequence identity with the amino acid sequence and having At3g61540 activity; (4E) A base sequence having 90% or more sequence identity with the base sequence of (4A) or (4C) and encoding an amino acid sequence having At3g61540 activity; (4F) A base sequence complementary to the base sequence of (4A), (4B), (4C), (4D) or (4E); (5A) the base sequence set forth in SEQ ID NO: 44 or 45; (5B) a base sequence encoding an amino acid sequence set forth in SEQ ID NO: 46 or 47, or an amino acid sequence having 90% or more sequence identity to the amino acid sequence and having At2g01818 activity; (5C) The following ID in KEGG (Kyoto Encyclopedia of Genes and Genomes): aly:9311163, eus:EUTSA_v10005480mg, csat:104710111, crb:17885477, brp:103854038, bna:106432932, boe:106325697, or rsz:108809176 The base sequence of the gene to which (5D) an amino acid sequence encoded by a gene assigned the ID described in (5C) in KEGG, or a base sequence encoding an amino acid sequence having 90% or more sequence identity with the amino acid sequence and having At2g01818 activity; (5E) A base sequence having 90% or more sequence identity with the base sequence of (5A) or (5C) and encoding an amino acid sequence having At2g01818 activity; (5F) A base sequence complementary to the base sequence of (5A), (5B), (5C), (5D) or (5E).

5. A method for selecting a plant having high ability in one or more of an ability to retain chlorophyll, an ability to produce chlorophyll, and an ability to grow, comprising: a measuring step of measuring the expression of one or more genes selected from the following (1A) to (1F), (2A) to (2F), (3A) to (3F), (4A) to (4F), and (5A) to (5F) and / or the activity of a protein derived from said gene in a plant; a selection step of selecting a plant in which the expression of the gene and / or the activity of the protein derived from the gene measured in the measurement step is lower than that of a control plant; Methods including: (1A) a base sequence set forth in SEQ ID NO: 1, 2, 3, or 48; (1B) an amino acid sequence set forth in SEQ ID NO: 4, 5, 6, or 49, or a base sequence encoding an amino acid sequence having 90% or more sequence identity to the amino acid sequence and having ALKBH8 activity; (1C) The following ID in KEGG (Kyoto Encyclopedia of Genes and Genomes): dosa:Os04t0602700-00、dosa:Os02t0750500-01、osa:4330741、osa:4336893、ath:AT1G36310、ath:AT1G31600、aly:ARALYDRAFT_473619、aly:ARAL YDRAFT_473716、crb:CARUB_v10009616mg、crb:CARUB_v10009306mg、csat :104757812、csat:104787286、csat:104777497、csat:104742087、csat:1 04741763、csat:104777194、eus:EUTSA_v10004311mg、eus:EUTSA_v10006 757mg、brp:103867438、brp:103848760、brp:103833805、bna:111205055、 bna:106411643、bna:106447489、bna:106399512、bna:106411644、bna:10 6412207、bna:106411637、bna:106418960、boe:106342941、boe:10630818 4、boe:106307572、thj:104798532、thj:104813129、cpap:110812544、cpa p:110818506、cit:102616373、cit:102625137、cic:CICLE_v10001407mg、 cic:CICLE_v10004016mg, tcc:18606930, tcc:18592225, gra:105773784, gra:105764921, ghi:107962467, ghi:107918198, ghi:107946865, ghi:10 7946842、ghi:107933507、dzi:111275453、dzi:111295045、dzi:11131235 5、egr:104444193、egr:104456636、gmx:100784028、gmx:100777529、gmx: 100807751、pvu:PHAVU_002G123600g、pvu:PHAVU_009G027100g、vra:1067 59935、vra:106777615、var:108335758、var:108329993、ccaj:109815811、ccaj:109796724、mtr:MTR_2g460880、mtr:MTR_5g026030、cam:101494433、cam:101511642、adu:107460794、adu:107487073、aip:107606112、aip:107642443、lja:Lj0g3v0064359.1、lja:Lj0g3v0046209.1、lja:Lj2g3v1573010.1、lja:Lj2g3v1573010.2、lang:109348998、lang:109357026、fve:101305382、fve:101301875、pper:18776672、pper:18788824、pmum:103336690、pmum:103332964、pavi:110755353、pavi:110753860、mdm:103441609、mdm:103416044、mdm:103401476、pxb:103934470、pxb:103955543、zju:107422678、zju:107430520、csv:101210859、csv:101216715、cmo:103485433、cmo:103487636、mcha:111008031、mcha:111020230、cmax:111477075、cmax:111487507、cmax:111489322、rcu:8263009、rcu:8280124、jcu:105643385、jcu:105638853、hbr:110640684、hbr:110644165、hbr:110667825、pop:POPTR_0003s10550g、pop:POPTR_0002s09190g、 pop:POPTR_0975s00200g、jre:109007265、jre:109021366、jre:109019322、jre:109012454、vvi:100252728、vvi:100262638、sly:101260324、sly:101263510、spen:107015589、spen:107005551、sot:102591578、sot:102587122、cann:107867583、cann:107850720、nta:107782177、nta:107784057、nta:107778504、nta:107777356、nta:107799087、nta:107802481、nsy:104228789、nsy:1 04246141、nsy:104231727、nto:104097471、nto:104088120、nto:1041044 03、ini:109170629、ini:109170202、sind:105159340、sind:105164991、 oeu:111404274、oeu:111377742、oeu:111378936、oeu:111383327、han:11 0868608、han:110918392、bvg:104892444、bvg:104905687、soe:1107748 79、soe:110797769、nnu:104610319、nnu:104600059、obr:102703096、bdi :100835069、bdi:100827681、ats:109741229、ats:109760172、sbi:8066 423、sbi:8072284、zma:103627749、zma:100282025、sita:101777379、sit a:101775180、pda:103713010、pda:103715224、egu:105053522、egu:105 033011、mus:103996326、mus:103990395、dct:110097186、dct:110106918 、aof:109832664、aof:109825376、atr:18422692、atr:18422136、smo:SE LMODRAFT_78730、smo:SELMODRAFT_78643、smo:SELMODRAFT_112315、smo: SELMODRAFT_112271、ppp:PHYPADRAFT_106932、ppp:PHYPADRAFT_15669、vcn:VOLCADRAFT_108374、mng:MNEG_3576、olu:OSTLU_88555、ota:OT_ost ta13g02300、ota:OT_ostta11g00670、bpg:Bathy04g02310、bpg:Bathy13g02890、mis:MICPUN_86885、mis:MICPUN_60739、mpp:MICPUCDRAFT_41620、mpp:MICPUCDRAFT_17097, apro:F751_1394, or gsl:Gasu_35150, The base sequence of the gene to which (1D) an amino acid sequence encoded by a gene assigned the ID described in (1C) in KEGG, or a base sequence encoding an amino acid sequence having 90% or more sequence identity with the amino acid sequence and having ALKBH8 activity; (1E) A base sequence having 90% or more sequence identity with the base sequence of (1A) and encoding an amino acid sequence having ALKBH8 activity, or a base sequence having 90% or more sequence identity with the base sequence of (1C) and encoding an amino acid sequence having ALKBH8 activity; (1F) A base sequence complementary to the base sequence of (1A), (1B), (1C), (1D) or (1E); (2A) a base sequence set forth in SEQ ID NO: 34, 35, 36, 37, or 38; (2B) a base sequence encoding an amino acid sequence set forth in SEQ ID NO: 39, 40, 41, 42, or 43, or an amino acid sequence having 90% or more sequence identity to the amino acid sequence and having TGA9 activity; (2C) The following ID in KEGG (Kyoto Encyclopedia of Genes and Genomes): aly:9325751, csat:104755012, crb:17900540, eus:EUTSA_v10007490mg, bna:106412233, boe:106344082, brp:103871579, or rsz:108860791 The base sequence of the gene to which (2D) an amino acid sequence encoded by a gene assigned the ID described in (2C) in KEGG, or a base sequence encoding an amino acid sequence having 90% or more sequence identity with the amino acid sequence and having TGA9 activity; (2E) A nucleotide sequence having 90% or more sequence identity with the nucleotide sequence of (2A) or (2C) and encoding an amino acid sequence having TGA9 activity; or (2F) A base sequence complementary to the base sequence of (2A), (2B), (2C), (2D) or (2E); (3A) the base sequence set forth in SEQ ID NO: 26; (3B) a base sequence encoding the amino acid sequence set forth in SEQ ID NO: 27, or an amino acid sequence having 90% or more sequence identity to the amino acid sequence and having GRF15 activity; (3C) The following ID in KEGG (Kyoto Encyclopedia of Genes and Genomes): bna:106429451, boe:106342871, or brp:103863788 The base sequence of the gene to which (3D) an amino acid sequence encoded by a gene assigned the ID described in (3C) in KEGG, or a base sequence encoding an amino acid sequence having 90% or more sequence identity with the amino acid sequence and having GRF15 activity; (3E) A nucleotide sequence having 90% or more sequence identity with the nucleotide sequence of (3A) or (3C) and encoding an amino acid sequence having GRF15 activity; or (3F) A base sequence complementary to the base sequence of (3A), (3B), (3C), (3D) or (3E); (4A) the base sequence set forth in SEQ ID NO: 28; (4B) an amino acid sequence set forth in SEQ ID NO: 29, or a base sequence encoding an amino acid sequence having 90% or more sequence identity to the amino acid sequence and having At3g61540 activity; (4C) the following ID in KEGG (Kyoto Encyclopedia of Genes and Genomes): aly:9314457, crb:17884851, csat:104788446, eus:EUTSA, rsz:108814367, bna:106406599, boe:106296419, or brp:103830306 The base sequence of the gene to which (4D) an amino acid sequence encoded by a gene assigned the ID described in (4C) in KEGG, or a base sequence encoding an amino acid sequence having 90% or more sequence identity with the amino acid sequence and having At3g61540 activity; (4E) A base sequence having 90% or more sequence identity with the base sequence of (4A) or (4C) and encoding an amino acid sequence having At3g61540 activity; (4F) A base sequence complementary to the base sequence of (4A), (4B), (4C), (4D) or (4E); (5A) the base sequence set forth in SEQ ID NO: 44 or 45; (5B) a base sequence encoding an amino acid sequence set forth in SEQ ID NO: 46 or 47, or an amino acid sequence having 90% or more sequence identity to the amino acid sequence and having At2g01818 activity; (5C) The following ID in KEGG (Kyoto Encyclopedia of Genes and Genomes): aly:9311163, eus:EUTSA_v10005480mg, csat:104710111, crb:17885477, brp:103854038, bna:106432932, boe:106325697, or rsz:108809176 The base sequence of the gene to which (5D) an amino acid sequence encoded by a gene assigned the ID described in (5C) in KEGG, or a base sequence encoding an amino acid sequence having 90% or more sequence identity with the amino acid sequence and having At2g01818 activity; (5E) A base sequence having 90% or more sequence identity with the base sequence of (5A) or (5C) and encoding an amino acid sequence having At2g01818 activity; (5F) A base sequence complementary to the base sequence of (5A), (5B), (5C), (5D) or (5E).

6. The method according to claim 5 , wherein the measuring step comprises measuring the expression of the gene by carrying out a nucleic acid amplification reaction using DNA or RNA obtained from the plant as a template.

7. A method for breeding a plant having high ability in one or more selected from an ability to retain chlorophyll, an ability to produce chlorophyll, and an ability to grow, comprising: a crossbreeding step of crossing a first parental plant individual with a second parental plant individual in which the expression of one or more genes selected from the following (1A) to (1F), (2A) to (2F), (3A) to (3F), (4A) to (4F), and (5A) to (5F) and / or the activity of a protein derived from said gene is the same as or lower than that of the first parental plant individual to obtain offspring; a measuring step of measuring the expression of the gene and / or the activity of a protein derived from the gene in the offspring; a selection step of selecting the offspring when the expression of the gene and / or the activity of the protein derived from the gene in the offspring measured in the measurement step is the same as or lower than the expression of the gene and / or the activity of the protein derived from the gene in the second parent individual; Methods including: (1A) a base sequence set forth in SEQ ID NO: 1, 2, 3, or 48; (1B) an amino acid sequence set forth in SEQ ID NO: 4, 5, 6, or 49, or a base sequence encoding an amino acid sequence having 90% or more sequence identity to the amino acid sequence and having ALKBH8 activity; (1C) The following ID in KEGG (Kyoto Encyclopedia of Genes and Genomes): dosa:Os04t0602700-00、dosa:Os02t0750500-01、osa:4330741、osa:4336893、ath:AT1G36310、ath:AT1G31600、aly:ARALYDRAFT_473619、aly:ARAL YDRAFT_473716、crb:CARUB_v10009616mg、crb:CARUB_v10009306mg、csat :104757812、csat:104787286、csat:104777497、csat:104742087、csat:1 04741763、csat:104777194、eus:EUTSA_v10004311mg、eus:EUTSA_v10006 757mg、brp:103867438、brp:103848760、brp:103833805、bna:111205055、 bna:106411643、bna:106447489、bna:106399512、bna:106411644、bna:10 6412207、bna:106411637、bna:106418960、boe:106342941、boe:10630818 4、boe:106307572、thj:104798532、thj:104813129、cpap:110812544、cpa p:110818506、cit:102616373、cit:102625137、cic:CICLE_v10001407mg、 cic:CICLE_v10004016mg, tcc:18606930, tcc:18592225, gra:105773784, gra:105764921, ghi:107962467, ghi:107918198, ghi:107946865, ghi:10 7946842、ghi:107933507、dzi:111275453、dzi:111295045、dzi:11131235 5、egr:104444193、egr:104456636、gmx:100784028、gmx:100777529、gmx: 100807751、pvu:PHAVU_002G123600g、pvu:PHAVU_009G027100g、vra:1067 59935、vra:106777615、var:108335758、var:108329993、ccaj:109815811、ccaj:109796724、mtr:MTR_2g460880、mtr:MTR_5g026030、cam:101494433、cam:101511642、adu:107460794、adu:107487073、aip:107606112、aip:107642443、lja:Lj0g3v0064359.1、lja:Lj0g3v0046209.1、lja:Lj2g3v1573010.1、lja:Lj2g3v1573010.2、lang:109348998、lang:109357026、fve:101305382、fve:101301875、pper:18776672、pper:18788824、pmum:103336690、pmum:103332964、pavi:110755353、pavi:110753860、mdm:103441609、mdm:103416044、mdm:103401476、pxb:103934470、pxb:103955543、zju:107422678、zju:107430520、csv:101210859、csv:101216715、cmo:103485433、cmo:103487636、mcha:111008031、mcha:111020230、cmax:111477075、cmax:111487507、cmax:111489322、rcu:8263009、rcu:8280124、jcu:105643385、jcu:105638853、hbr:110640684、hbr:110644165、hbr:110667825、pop:POPTR_0003s10550g、pop:POPTR_0002s09190g、 pop:POPTR_0975s00200g、jre:109007265、jre:109021366、jre:109019322、jre:109012454、vvi:100252728、vvi:100262638、sly:101260324、sly:101263510、spen:107015589、spen:107005551、sot:102591578、sot:102587122、cann:107867583、cann:107850720、nta:107782177、nta:107784057、nta:107778504、nta:107777356、nta:107799087、nta:107802481、nsy:104228789、nsy:1 04246141、nsy:104231727、nto:104097471、nto:104088120、nto:1041044 03、ini:109170629、ini:109170202、sind:105159340、sind:105164991、 oeu:111404274、oeu:111377742、oeu:111378936、oeu:111383327、han:11 0868608、han:110918392、bvg:104892444、bvg:104905687、soe:1107748 79、soe:110797769、nnu:104610319、nnu:104600059、obr:102703096、bdi :100835069、bdi:100827681、ats:109741229、ats:109760172、sbi:8066 423、sbi:8072284、zma:103627749、zma:100282025、sita:101777379、sit a:101775180、pda:103713010、pda:103715224、egu:105053522、egu:105 033011、mus:103996326、mus:103990395、dct:110097186、dct:110106918 、aof:109832664、aof:109825376、atr:18422692、atr:18422136、smo:SE LMODRAFT_78730、smo:SELMODRAFT_78643、smo:SELMODRAFT_112315、smo: SELMODRAFT_112271、ppp:PHYPADRAFT_106932、ppp:PHYPADRAFT_15669、vcn:VOLCADRAFT_108374、mng:MNEG_3576、olu:OSTLU_88555、ota:OT_ost ta13g02300、ota:OT_ostta11g00670、bpg:Bathy04g02310、bpg:Bathy13g02890、mis:MICPUN_86885、mis:MICPUN_60739、mpp:MICPUCDRAFT_41620、mpp:MICPUCDRAFT_17097, apro:F751_1394, or gsl:Gasu_35150, The base sequence of the gene to which (1D) an amino acid sequence encoded by a gene assigned the ID described in (1C) in KEGG, or a base sequence encoding an amino acid sequence having 90% or more sequence identity with the amino acid sequence and having ALKBH8 activity; (1E) A base sequence having 90% or more sequence identity with the base sequence of (1A) and encoding an amino acid sequence having ALKBH8 activity, or a base sequence having 90% or more sequence identity with the base sequence of (1C) and encoding an amino acid sequence having ALKBH8 activity; (1F) A base sequence complementary to the base sequence of (1A), (1B), (1C), (1D) or (1E); (2A) a base sequence set forth in SEQ ID NO: 34, 35, 36, 37, or 38; (2B) a base sequence encoding an amino acid sequence set forth in SEQ ID NO: 39, 40, 41, 42, or 43, or an amino acid sequence having 90% or more sequence identity to the amino acid sequence and having TGA9 activity; (2C) The following ID in KEGG (Kyoto Encyclopedia of Genes and Genomes): aly:9325751, csat:104755012, crb:17900540, eus:EUTSA_v10007490mg, bna:106412233, boe:106344082, brp:103871579, or rsz:108860791 The base sequence of the gene to which (2D) an amino acid sequence encoded by a gene assigned the ID described in (2C) in KEGG, or having 90% or more sequence identity with the amino acid sequence; a base sequence encoding an amino acid sequence having TGA9 activity; (2E) A nucleotide sequence having 90% or more sequence identity with the nucleotide sequence of (2A) or (2C) and encoding an amino acid sequence having TGA9 activity; or (2F) A base sequence complementary to the base sequence of (2A), (2B), (2C), (2D) or (2E); (3A) the base sequence set forth in SEQ ID NO: 26; (3B) a base sequence encoding the amino acid sequence set forth in SEQ ID NO: 27, or an amino acid sequence having 90% or more sequence identity to the amino acid sequence and having GRF15 activity; (3C) The following ID in KEGG (Kyoto Encyclopedia of Genes and Genomes): bna:106429451, boe:106342871, or brp:103863788 The base sequence of the gene to which (3D) an amino acid sequence encoded by a gene assigned the ID described in (3C) in KEGG, or a base sequence encoding an amino acid sequence having 90% or more sequence identity with the amino acid sequence and having GRF15 activity; (3E) A nucleotide sequence having 90% or more sequence identity with the nucleotide sequence of (3A) or (3C) and encoding an amino acid sequence having GRF15 activity; or (3F) A base sequence complementary to the base sequence of (3A), (3B), (3C), (3D) or (3E); (4A) the base sequence set forth in SEQ ID NO: 28; (4B) an amino acid sequence set forth in SEQ ID NO: 29, or a base sequence encoding an amino acid sequence having 90% or more sequence identity to the amino acid sequence and having At3g61540 activity; (4C) the following ID in KEGG (Kyoto Encyclopedia of Genes and Genomes): aly:9314457, crb:17884851, csat:104788446, eus:EUTSA, rsz:108814367, bna:106406599, boe:106296419, or brp:103830306 The base sequence of the gene to which (4D) an amino acid sequence encoded by a gene assigned the ID described in (4C) in KEGG, or a base sequence encoding an amino acid sequence having 90% or more sequence identity with the amino acid sequence and having At3g61540 activity; (4E) A base sequence having 90% or more sequence identity with the base sequence of (4A) or (4C) and encoding an amino acid sequence having At3g61540 activity; (4F) A base sequence complementary to the base sequence of (4A), (4B), (4C), (4D) or (4E); (5A) the base sequence set forth in SEQ ID NO: 44 or 45; (5B) a base sequence encoding an amino acid sequence set forth in SEQ ID NO: 46 or 47, or an amino acid sequence having 90% or more sequence identity to the amino acid sequence and having At2g01818 activity; (5C) The following ID in KEGG (Kyoto Encyclopedia of Genes and Genomes): aly:9311163, eus:EUTSA_v10005480mg, csat:104710111, crb:17885477, brp:103854038, bna:106432932, boe:106325697, or rsz:108809176 The base sequence of the gene to which (5D) an amino acid sequence encoded by a gene assigned the ID described in (5C) in KEGG, or a base sequence encoding an amino acid sequence having 90% or more sequence identity with the amino acid sequence and having At2g01818 activity; (5E) A base sequence having 90% or more sequence identity with the base sequence of (5A) or (5C) and encoding an amino acid sequence having At2g01818 activity; (5F) A base sequence complementary to the base sequence of (5A), (5B), (5C), (5D) or (5E).

8. The method according to claim 7 , wherein the mating step comprises measuring the expression of the gene by performing a nucleic acid amplification reaction using DNA or RNA obtained from the offspring as a template.

9. A method for improving one or more abilities selected from an ability to retain chlorophyll, an ability to produce chlorophyll, and an ability to grow in a plant, comprising: A method for suppressing the expression of one or more genes selected from the following (1A) to (1F), (2A) to (2F), (3A) to (3F), (4A) to (4F), and (5A) to (5F) in a plant and / or the activity of a protein derived from said gene(s): (1A) a base sequence set forth in SEQ ID NO: 1, 2, 3, or 48; (1B) an amino acid sequence set forth in SEQ ID NO: 4, 5, 6, or 49, or a base sequence encoding an amino acid sequence having 90% or more sequence identity to the amino acid sequence and having ALKBH8 activity; (1C) The following ID in KEGG (Kyoto Encyclopedia of Genes and Genomes): dosa:Os04t0602700-00、dosa:Os02t0750500-01、osa:4330741、osa:4336893、ath:AT1G36310、ath:AT1G31600、aly:ARALYDRAFT_473619、aly:ARAL YDRAFT_473716、crb:CARUB_v10009616mg、crb:CARUB_v10009306mg、csat :104757812、csat:104787286、csat:104777497、csat:104742087、csat:1 04741763、csat:104777194、eus:EUTSA_v10004311mg、eus:EUTSA_v10006 757mg、brp:103867438、brp:103848760、brp:103833805、bna:111205055、 bna:106411643、bna:106447489、bna:106399512、bna:106411644、bna:10 6412207、bna:106411637、bna:106418960、boe:106342941、boe:10630818 4、boe:106307572、thj:104798532、thj:104813129、cpap:110812544、cpa p:110818506、cit:102616373、cit:102625137、cic:CICLE_v10001407mg、 cic:CICLE_v10004016mg, tcc:18606930, tcc:18592225, gra:105773784, gra:105764921, ghi:107962467, ghi:107918198, ghi:107946865, ghi:10 7946842、ghi:107933507、dzi:111275453、dzi:111295045、dzi:11131235 5、egr:104444193、egr:104456636、gmx:100784028、gmx:100777529、gmx: 100807751、pvu:PHAVU_002G123600g、pvu:PHAVU_009G027100g、vra:1067 59935、vra:106777615、var:108335758、var:108329993、ccaj:109815811、ccaj:109796724、mtr:MTR_2g460880、mtr:MTR_5g026030、cam:101494433、cam:101511642、adu:107460794、adu:107487073、aip:107606112、aip:107642443、lja:Lj0g3v0064359.1、lja:Lj0g3v0046209.1、lja:Lj2g3v1573010.1、lja:Lj2g3v1573010.2、lang:109348998、lang:109357026、fve:101305382、fve:101301875、pper:18776672、pper:18788824、pmum:103336690、pmum:103332964、pavi:110755353、pavi:110753860、mdm:103441609、mdm:103416044、mdm:103401476、pxb:103934470、pxb:103955543、zju:107422678、zju:107430520、csv:101210859、csv:101216715、cmo:103485433、cmo:103487636、mcha:111008031、mcha:111020230、cmax:111477075、cmax:111487507、cmax:111489322、rcu:8263009、rcu:8280124、jcu:105643385、jcu:105638853、hbr:110640684、hbr:110644165、hbr:110667825、pop:POPTR_0003s10550g、pop:POPTR_0002s09190g、 pop:POPTR_0975s00200g、jre:109007265、jre:109021366、jre:109019322、jre:109012454、vvi:100252728、vvi:100262638、sly:101260324、sly:101263510、spen:107015589、spen:107005551、sot:102591578、sot:102587122、cann:107867583、cann:107850720、nta:107782177、nta:107784057、nta:107778504、nta:107777356、nta:107799087、nta:107802481、nsy:104228789、nsy:1 04246141、nsy:104231727、nto:104097471、nto:104088120、nto:1041044 03、ini:109170629、ini:109170202、sind:105159340、sind:105164991、 oeu:111404274、oeu:111377742、oeu:111378936、oeu:111383327、han:11 0868608、han:110918392、bvg:104892444、bvg:104905687、soe:1107748 79、soe:110797769、nnu:104610319、nnu:104600059、obr:102703096、bdi :100835069、bdi:100827681、ats:109741229、ats:109760172、sbi:8066 423、sbi:8072284、zma:103627749、zma:100282025、sita:101777379、sit a:101775180、pda:103713010、pda:103715224、egu:105053522、egu:105 033011、mus:103996326、mus:103990395、dct:110097186、dct:110106918 、aof:109832664、aof:109825376、atr:18422692、atr:18422136、smo:SE LMODRAFT_78730、smo:SELMODRAFT_78643、smo:SELMODRAFT_112315、smo: SELMODRAFT_112271、ppp:PHYPADRAFT_106932、ppp:PHYPADRAFT_15669、vcn:VOLCADRAFT_108374、mng:MNEG_3576、olu:OSTLU_88555、ota:OT_ost ta13g02300、ota:OT_ostta11g00670、bpg:Bathy04g02310、bpg:Bathy13g02890、mis:MICPUN_86885、mis:MICPUN_60739、mpp:MICPUCDRAFT_41620、mpp:MICPUCDRAFT_17097, apro:F751_1394, or gsl:Gasu_35150, The base sequence of the gene to which (1D) an amino acid sequence encoded by a gene assigned the ID described in (1C) in KEGG, or a base sequence encoding an amino acid sequence having 90% or more sequence identity with the amino acid sequence and having ALKBH8 activity; (1E) A base sequence having 90% or more sequence identity with the base sequence of (1A) and encoding an amino acid sequence having ALKBH8 activity, or a base sequence having 90% or more sequence identity with the base sequence of (1C) and encoding an amino acid sequence having ALKBH8 activity; (1F) A base sequence complementary to the base sequence of (1A), (1B), (1C), (1D) or (1E); (2A) a base sequence set forth in SEQ ID NO: 34, 35, 36, 37, or 38; (2B) a base sequence encoding an amino acid sequence set forth in SEQ ID NO: 39, 40, 41, 42, or 43, or an amino acid sequence having 90% or more sequence identity to the amino acid sequence and having TGA9 activity; (2C) The following ID in KEGG (Kyoto Encyclopedia of Genes and Genomes): aly:9325751, csat:104755012, crb:17900540, eus:EUTSA_v10007490mg, bna:106412233, boe:106344082, brp:103871579, or rsz:108860791 The base sequence of the gene to which (2D) an amino acid sequence encoded by a gene assigned the ID described in (2C) in KEGG, or a base sequence encoding an amino acid sequence having 90% or more sequence identity with the amino acid sequence and having TGA9 activity; (2E) A nucleotide sequence having 90% or more sequence identity with the nucleotide sequence of (2A) or (2C) and encoding an amino acid sequence having TGA9 activity; or (2F) A base sequence complementary to the base sequence of (2A), (2B), (2C), (2D) or (2E); (3A) the base sequence set forth in SEQ ID NO: 26; (3B) a base sequence encoding the amino acid sequence set forth in SEQ ID NO: 27, or an amino acid sequence having 90% or more sequence identity to the amino acid sequence and having GRF15 activity; (3C) The following ID in KEGG (Kyoto Encyclopedia of Genes and Genomes): bna:106429451, boe:106342871, or brp:103863788 The base sequence of the gene to which (3D) an amino acid sequence encoded by a gene assigned the ID described in (3C) in KEGG, or a base sequence encoding an amino acid sequence having 90% or more sequence identity with the amino acid sequence and having GRF15 activity; (3E) A nucleotide sequence having 90% or more sequence identity with the nucleotide sequence of (3A) or (3C) and encoding an amino acid sequence having GRF15 activity; or (3F) A base sequence complementary to the base sequence of (3A), (3B), (3C), (3D) or (3E); (4A) the base sequence set forth in SEQ ID NO: 28; (4B) an amino acid sequence set forth in SEQ ID NO: 29, or a base sequence encoding an amino acid sequence having 90% or more sequence identity to the amino acid sequence and having At3g61540 activity; (4C) the following ID in KEGG (Kyoto Encyclopedia of Genes and Genomes): aly:9314457, crb:17884851, csat:104788446, eus:EUTSA, rsz:108814367, bna:106406599, boe:106296419, or brp:103830306 The base sequence of the gene to which (4D) an amino acid sequence encoded by a gene assigned the ID described in (4C) in KEGG, or a base sequence encoding an amino acid sequence having 90% or more sequence identity with the amino acid sequence and having At3g61540 activity; (4E) A base sequence having 90% or more sequence identity with the base sequence of (4A) or (4C) and encoding an amino acid sequence having At3g61540 activity; (4F) A base sequence complementary to the base sequence of (4A), (4B), (4C), (4D) or (4E); (5A) the base sequence set forth in SEQ ID NO: 44 or 45; (5B) a base sequence encoding an amino acid sequence set forth in SEQ ID NO: 46 or 47, or an amino acid sequence having 90% or more sequence identity to the amino acid sequence and having At2g01818 activity; (5C) The following ID in KEGG (Kyoto Encyclopedia of Genes and Genomes): aly:9311163, eus:EUTSA_v10005480mg, csat:104710111, crb:17885477, brp:103854038, bna:106432932, boe:106325697, or rsz:108809176 The base sequence of the gene to which (5D) an amino acid sequence encoded by a gene assigned the ID described in (5C) in KEGG, or a base sequence encoding an amino acid sequence having 90% or more sequence identity with the amino acid sequence and having At2g01818 activity; (5E) A base sequence having 90% or more sequence identity with the base sequence of (5A) or (5C) and encoding an amino acid sequence having At2g01818 activity; (5F) A base sequence complementary to the base sequence of (5A), (5B), (5C), (5D) or (5E).

10. The suppression step (I) disruption of the gene in the genomic DNA of a plant; (II) introducing antisense DNA for the gene into a plant; and (III) introducing into a plant an antagonist to the protein derived from the gene.

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