Indel molecular marker related to the aroma trait of pumpkin fruit and its application

Abstract

The application discloses an Indel molecular marker related to a pumpkin fruit taro smell character and application thereof, wherein the sequence of the Indel molecular marker is shown as SEQ ID NO:2. DNA of a pumpkin to be detected is used as a template, a forward primer with a sequence shown as SEQ ID NO:4 and a reverse primer with a sequence shown as SEQ ID NO:5 are used for PCR amplification; if the size of the amplification product is about 5100bp, the pumpkin fruit to be detected has the taro smell; if the size of the amplification product is about 500bp, the pumpkin fruit to be detected does not have the taro smell. The Indel molecular marker and the primer for specifically amplifying the Indel molecular marker can be applied to screening of pumpkin varieties with the taro smell character, or detecting whether the pumpkin varieties have the taro smell, so that the pumpkin taro smell character can be quickly and accurately judged from the genotype, and thus an assisted selection breeding technology system of the pumpkin taro smell character is established.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of crop genetic breeding, and particularly relates to an Indel molecular marker related to a pumpkin fruit taro flavor trait and application thereof. BACKGROUND

[0002] Pumpkin (Cucurbit moschata) is one of important melon crops of Cucurbitaceae, plays an important role in the dietary structure of residents, and has broad market prospects and important economic value.

[0003] The pumpkin taro flavor trait is a special flavor trait in pumpkin resources, and pumpkin varieties with rich taro flavor are favored by the market and consumers. However, the judgment of the taro flavor trait at present mainly relies on the sense of smell of breeders.

[0004] Therefore, in-depth excavation and development of the molecular marker closely linked to the taro flavor trait are beneficial to promote the breeding of new taro flavor pumpkin varieties and accelerate the process of pumpkin molecular breeding. SUMMARY

[0005] Based on this, the purpose of the present application is to provide an Indel molecular marker related to a pumpkin fruit taro flavor trait.

[0006] The specific technical scheme for realizing the above-mentioned purpose of the application includes the following.

[0007] In a first aspect of the present application, an Indel molecular marker related to a pumpkin fruit taro flavor trait is provided, and the sequence of the Indel molecular marker is shown in SEQ ID NO: 2.

[0008] In a second aspect of the present application, the application of the above-mentioned Indel molecular marker in detecting whether the pumpkin fruit has taro flavor or screening taro flavor pumpkin varieties is provided.

[0009] In a third aspect of the present application, a primer for specifically detecting the above-mentioned Indel molecular marker is provided, including a forward primer with a sequence shown in SEQ ID NO: 4 and a reverse primer with a sequence shown in SEQ ID NO: 5.

[0010] In a fourth aspect of the present application, the application of the above-mentioned primer in preparing a kit for detecting whether the pumpkin fruit has taro flavor or screening taro flavor pumpkin varieties is provided.

[0011] In a fifth aspect of the present application, a kit for detecting whether the pumpkin fruit has taro flavor or screening taro flavor pumpkin varieties is provided, including the above-mentioned primer for specifically detecting the Indel molecular marker.

[0012] In a sixth aspect of the present application, a method for screening a pumpkin variety with an earthy flavor is provided, comprising the following steps: detecting the Indel molecular marker with the sequence as shown in SEQ ID NO: 2.

[0013] In a seventh aspect of the present application, a method for detecting whether a pumpkin fruit has an earthy flavor is provided, comprising the following steps: using the DNA of a pumpkin to be tested as a template, and performing PCR amplification with a forward primer with the sequence as shown in SEQ ID NO: 4 and a reverse primer with the sequence as shown in SEQ ID NO: 5; if the size of the amplification product is about 5100 bp, then the pumpkin fruit to be tested has an earthy flavor; if the size of the amplification product is about 500 bp, then the pumpkin fruit to be tested does not have an earthy flavor.

[0014] In the present application, through the gene mapping of the earthy flavor of pumpkin, it is found that there is a significant difference in the expression of betaine aldehyde dehydrogenase (BADH) gene between the earthy-flavor pumpkin and the non-earthy-flavor pumpkin, and in the earthy-flavor pumpkin, there is a 4597 bp large fragment insertion in the betaine aldehyde dehydrogenase (BADH) gene. Therefore, the insertion mutation of the betaine aldehyde dehydrogenase (BADH) gene is related to the earthy flavor of pumpkin fruit, and can be used as an Indel molecular marker. The Indel molecular marker and the primers specific for amplifying the Indel molecular marker can be applied to screen the pumpkin variety with the earthy flavor, or to detect whether the pumpkin variety has an earthy flavor, so as to quickly and accurately determine the earthy flavor of pumpkin from the genotype, and thus to establish an assisted selection breeding technology system for the earthy flavor of pumpkin. BRIEF DESCRIPTION OF DRAWINGS

[0015] Figure 1 Fine mapping of the gene related to the earthy flavor of pumpkin in Example 1 of the present application.

[0016] Figure 2 Differential expression of the candidate gene related to the earthy flavor of pumpkin in Example 1 of the present application between two parents; wherein No. 44 is the female parent, and No. 45 is the male parent.

[0017] Figure 3 PCR detection results of different pumpkin resources in Example 3 of the present application; wherein a and b are the PCR amplification results of non-earthy-flavor pumpkin resources; and c is the PCR amplification result of an earthy-flavor pumpkin resource. DETAILED DESCRIPTION

[0018] In order to facilitate the understanding of the present application, the present application will be described more fully below. The present application can be realized in many different forms and is not limited to the embodiments described herein. On the contrary, these embodiments are provided so that the disclosure of the present application can be more thoroughly and completely understood.

[0019] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used in the description of the application herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety.

[0020] Unless otherwise indicated, the examples were performed according to routine experimental conditions, such as Sambrook et al. Molecular Cloning: A Laboratory Manual (2013), or according to the conditions suggested by the manufacturer's instructions.

[0021] In some embodiments of the present application, an Indel molecular marker associated with the sweet potato flavor trait of pumpkin fruit is disclosed, and the sequence of the Indel molecular marker is shown as SEQ ID NO: 2.

[0022] In some other embodiments of the present application, the use of the above-mentioned Indel molecular marker in screening sweet potato flavor pumpkin varieties or detecting whether the pumpkin fruit has sweet potato flavor is disclosed.

[0023] In some other embodiments of the present application, a primer for specifically detecting the above-mentioned Indel molecular marker is disclosed, including a forward primer with a sequence as shown in SEQ ID NO: 4 and a reverse primer with a sequence as shown in SEQ ID NO: 5.

[0024] In some other embodiments of the present application, the use of the above-mentioned primer for specifically detecting the above-mentioned Indel molecular marker in preparing a kit for screening sweet potato flavor pumpkin varieties or detecting whether the pumpkin fruit has sweet potato flavor is disclosed.

[0025] In some other embodiments of the present application, a kit for screening sweet potato flavor pumpkin varieties or detecting whether the pumpkin fruit has sweet potato flavor is disclosed, including the above-mentioned primer for specifically detecting the above-mentioned Indel molecular marker.

[0026] In some other embodiments of the present application, a method for screening sweet potato flavor pumpkin varieties is disclosed, including the following steps: detecting the Indel molecular marker with a sequence as shown in SEQ ID NO: 2 according to claim 1.

[0027] In some embodiments, the method for detecting is PCR.

[0028] In some embodiments, the primer used in the PCR method includes a forward primer with a sequence as shown in SEQ ID NO: 4 and a reverse primer with a sequence as shown in SEQ ID NO: 5.

[0029] In some embodiments, the method comprises the following steps:

[0030] PCR amplification is performed using a forward primer with the sequence shown in SEQ ID NO: 4 and a reverse primer with the sequence shown in SEQ ID NO: 5 as templates with DNA from the pumpkin to be tested; if the size of the amplification product is about 5100 bp, then the pumpkin to be tested is an onion-flavored pumpkin variety.

[0031] In some embodiments of the present application, a method for detecting whether a pumpkin fruit has an onion flavor is disclosed, comprising the following steps: PCR amplification is performed using a forward primer with the sequence shown in SEQ ID NO: 4 and a reverse primer with the sequence shown in SEQ ID NO: 5 as templates with DNA from the pumpkin to be tested; if the size of the amplification product is about 5100 bp, then the pumpkin fruit to be tested has an onion flavor; if the size of the amplification product is about 500 bp, then the pumpkin fruit to be tested does not have an onion flavor.

[0032] The present application will be described in detail below with reference to the accompanying drawings and specific embodiments.

[0033] Example 1: Gene mapping and candidate gene expression analysis related to the onion flavor trait of pumpkin fruit

[0034] comprising the following steps:

[0035] 1. The onion-flavored pumpkin resources (female parent: Xiangtian pumpkin) and non-onion-flavored pumpkin resources (male parent: Mihuang pumpkin) selected and preserved by the Vegetable Research Institute of the Guangdong Academy of Agricultural Sciences were used as parents, F1 was obtained by crossing the two parents, and F2 segregation population was obtained by selfing F1. F3 population was obtained by selfing F2 recombinant single plants. 2:3

[0036] 2. The F2 segregation population was used to construct an onion flavor and non-onion flavor extreme phenotype pool, each pool including 25 F2 single plants. BSA-seq technology was used to preliminarily locate the onion flavor control gene on chromosome 10, with a segment size of 2.05 Mb. Subsequently, in order to further narrow down the location segment, KASP molecular markers were developed at both ends and inside the location segment, and the genotypes of F2, F 2:3 segregation population single plants were analyzed, combined with the onion flavor judgment results, the gene location segment forming the onion flavor was narrowed down to a range of 23.3 kb, and there were 8 genes in the segment. Figure 1

[0037] 3. In order to further determine the gene affecting the formation of onion flavor, the expression of the 8 genes was detected, and it was found that only the CmoCh10G001620 gene had significantly different expression between the two parents, and the gene was significantly down-regulated in the onion-flavored pumpkin resources​​Figure 2 ) Meanwhile, according to the third generation sequencing technology, it was found that there was a significant sequence structure variation between the two parent resources in CmoChlOGOOl 620 gene, and a 4597 bp large fragment insertion (the sequence is shown as SEQ ID NO: 2) existed in the fourth intron region (physical position chr 10:734102) of the gene in the taro-flavor pumpkin resource. According to the gene annotation, it was found that CmoChlOGOOl 620 encoded betaine aldehyde dehydrogenase (BADH), and the nucleotide sequence thereof in the paternal parent honey pumpkin was shown as SEQ ID NO: 1; the sequence after the insertion of the large fragment, and the nucleotide sequence thereof in the maternal parent taro pumpkin was shown as SEQ ID NO: 3.

[0038] Therefore, the insertion mutation of the betaine aldehyde dehydrogenase gene can be used as an Indel molecular marker related to the taro-flavor trait of pumpkin.

[0039] SEQ ID NO: 1

[0040] CATCTCGATTCTCTCTTCTCTTTGCTATCAGATTCTTCCTCTTCTTTCTCCGAGATTTCTTT

[0041] GTAGCAGAGATTTTCAGAGCCAGAATCACTCAATCGATCAGTCAAATCCATGGCGGGAA

[0042] ACGGCTCGTACGATTGGGCGGACCAGTGGGATTACAGCGACTCGAAGGACGTAGTCAC

[0043] GGAGAACACGAAGAAGAGCGGCGGCGGAAACACTACGACTAAGTATAAACAGAAGGT

[0044] CGGCGAAGGCCTTGGGAAGACCAAAGCTGTGGCCTCCAATGGCGTTAAAAAGGTCAAA

[0045] CAAGGAACCTCTCTCGGCTTCCAATGGATCAAAGATAAATATAACAAAACCGCTCATAA

[0046] GAAGTAGAAATCCTTCGTATTCTACTCCTCTCTTGCTCTTCGGTTTTCTTTTTCTTATCATC

[0047] ATCGTCTCCATAGCTTATCTTTGAGCTGTATATCTTGTTTTTTCTATTTCTTGTCTGCTGGT

[0048] TATATATAGATACAGATACACCATAGATTATTCTTTTGTTAATATGATTTGATCTCTGTTCGT

[0049] TTCTTTCCTCTTGTTTTGATTCCGTTATTGTTTATCGCTGTTAAATTTCGTACTTCCTTCAG

[0050] ATTCGTTTGTTTGATTTTGATTATAATCTGAGAATTCCGCATCAGTAACATATATAAAGCAG

[0051] GGATAGAATCGCATCTTCTGATGTTTACTGGAAATCATATTAGCATACGAAGATGAAAGG

[0052] TGAAGGAGGAGATCTCGCCCTGAAAAAATTGAAAATTATTTATTTTATGAATAAAAAATCA

[0053] ATTTTGACCCCTAGATTTATTTGAATTTTTTATGTTTTGCAATTTTTGAGAAAAATTTTATAA

[0054] ATGTATTAATTTTCATTAAAATTAAATTCAAGTTCAACACCGGTTCTTCTCAACTTCTGATA

[0055] AAAAAATCTTGCATATAATAAATGTTTTTAAAAAAACAAAACATAGAGTAACAAGTTTA

[0056] AATTTAAATATATTTAAGAGTAATTGTTGCTTACCTCCACAATGTAAGTTATATATGGTTA

[0057] GATGAACACGACTCTCCACAATGATATGATATTGTCCACTTTGAGCTCATACTAATGGAG

[0058] AGATTATTTTTTTATTATAAACCTACAAATCATTTTCTAAATTAACCGATGTGGGACTTTCA

[0059] TCATCTAACATATATATCTACGTGTGTATGTATGTTTGCATGTATATGTATGTAGTGGATCCT

[0060] CCCTACCAAAAGAAAAAAAAAAATAGAATAGAATAGAATAGAATAGGTGGTGTGGCCA

[0061] GTTTTTAGCTCTTGCTTTTAATAAAGATGTATTATTTTTTTATTCTCACGTTTTTATAAAATG

[0062] CATATATTTTAATATTGATGCATCTTTAAACTTTAAAATGGGTATTTATATTATCATAAACTG

[0063] ATACTTTAAACAAACATATAAATAATTCAAACTTAAATATATCCATTAATTGCTTCAAAATT

[0064] AAATATATACATTAATTGTATTAATGGCTACTATAGGAGACAAATTTGTATCCTTCCTAAAA

[0065] AAAAAAATTCCCATTCCAAAAATACCTTTAAAATATTTAAACACTTTTTTTAGATAAAAGT

[0066] TGTTAATATTTTGTTTTAAATTTATCTTTGAACTTTCAAAACATCTATTATCTTTAAAATTGT

[0067] TAAGAAAATATCATAAGTTTTTAGAGAATTTAAGAATATTTATGAATTTTCAAAAAAAGTC

[0068] TTTTTAAAATTGATTGTTTTTTTAGGTTAAATTAAAATTATAATATTCAAGTTTCAATAAAT

[0069] TTTATAAAAAATTAAGTTTTAAGTACGAATTTCAAAATTTAGGGTTAAATATAATTTATTTT

[0070] TAATAAAAAGTACACATCTTTAATGCTTATTATGATAGCTTACTCAACGGCGTGATTTCTAT

[0071] ATAATAAAATAATGTATAATATGTTGGTGTAAGAGATTCTTTGTATTTCTATTTTCATAAATT

[0072] TTATTTTTCTTATTTAAGTTACCCAATAATTTGATTATCTTCACAGATATCATCTCGAGTTTC

[0073] GTAAAACTCAAAGAAAAAAATTGCAGAAGAAAAATCCAATACGGACGAAAAATACCGA

[0074] AAATTCCGTATTTATATTGATCAATCCAAGTCGAAGTCATTACCAATCCGACTACTTTTTA

[0075] CACTGCACTAAAGAGAAGAGCTCTGCAGAAATGGCGATTCATATTCCCAGTCGGCAGCT

[0076] ATTCATCGATGGCGAGTGGAGGGAACCAGTTCTCAAGAAACGCATCCCCATCGTCAACC

[0077] CCGCCACTGAACAAACAATCGGTCTCTTCTTCTTAACTTCTCTGTTTATGACTGTTGTTAA

[0078] TTTTGTTTGCCAACTTCTAATCGTTGTGCGGATCGGTAGACGGAAGAAGTAATTAGTTCG

[0079] AGGATCAGGTGGTTTAAAAGAGGTTGAATCTCACATTTGTGTTATTGCCAACTATGCTTC

[0080] AGGATCTATTCCCGCGGCTACTGCGGAAGATGTAGAGTTAGCAGTTGATGCTGCCCGAA

[0081] AAGCTCTTGCGAGGAATAAAGGCAAAGATTGGGCATCTGCTTCGGGGGCTCTTCGTGCC

[0082] AAGTATTTGCGTGCTATTGCTGCTAAGGTGAGTCGGTCATATGGATGTTTTACTCTTATTT

[0083] GATTGTGATCTGGTCTCTGTTTTACCATCGGACGGAGGTTATTGATCATTGATGCTTTTGT

[0084] TACTTATTTACGATTCTAGTGGCTTGAAACAATGGATTGATTTCTGATGCGATTCAATAGA

[0085] TCATGAGAAATATGATACCATCCATCTTCCCGCCATAGTTCATTGAAAAATTAGATATCTG

[0086] GATAGAGCCTTCTTTTGGTCTATGTATTGTCTTAGGAATGTTACCTTCTTGTTAATCGCCTT

[0087] GCAGGAGTTCCGTTGGAGTCGCATTTATAGTTTAAGAATCAAGTAGTTTTAGTTACTTTG

[0088] GAAGTTATATTATATATATCTTCAACTTTAATTTCTATCTTCATGGTGTTATTGTCGTCCATC

[0089] CTGAACCCTGTTTGCTGTTTTTCTCACAGATAACAGAGAGGAAATCGGAATTAGCAAAG

[0090] CTCGAAACAATAGACTGTGGAAAACCCCTGGACGAAACTGCATGGGACATTGTATGGCA

[0091] GTTTTTCTTATAGTTAAAGATATATGTTGGTTGTAACAATTTTCATTGCCGTCAATGTTTGT

[0092] TAATATTGTACAGGATGATGTTGCTGGGTGCTTGAGTACTATGCAGATCTTGCGAAGGGT

[0093] TGGATGCTAAGCAAAAAGCTCCTGTTTCCGTTCCCATGGATACATTCAAAAGCCATGTTC

[0094] TTAAAGAACCCATTGGAGTTGTTGGGTTGATTACTCCCTGGTACCTGTTGATCCATCTTCT

[0095] ACTTCTTCTTTTAGTTCTTCACGGCTATTTAGCTTCTCCGATTTTATTGGAAGTATTAGAGT

[0096] TATTGTTGTTAAACATTTTATAAATCATGGTCAACTCTTGTCACTCAACCAAGAACCTGTC

[0097] GATTTCAAACATGGTGACGGAGACTCTTCAATCCAGGTTCAAACAACTGGTCATTAACTT

[0098] AAACCTTAAGATATCAAATATTCCAAAACTTTCTTCACCAATAATGTAGAAGGGTCAAGT

[0099] CATAGTTTCAAGTGCTAATTCTTTTATATAAAATTTTCCTTATTCGTTTGAGTCATTCTTCT

[0100] GAATTTTTTCACTCTCAAAATGTGATGTTTTTCTCTTTTATATACATATATATTTTACTTGGA

[0101] TTGATTTTTTTACTACAAATAATTGTATAAAGGATTTAACGCATTATGTTATATCTATTCAAC

[0102] AAGTGTTTGATACATGTACAACAAGGATTGAATGTTGGATTGACTAACATGTGGTTCAAC

[0103] ATGTATCTAAATGCTGGGATGTTGAATACTTGTGATCATGGATATGTTACCCAACTTGAGT

[0104] AACTCTGCTTCACAGTTCCCATTTTATTGTTAATTTGAACGAGAGGCATACATTGGGACG

[0105] TTATTAGTTATTAACCTCCAGGGTCGGATCTGGTTATGTTCTCTCTTGTCCTCCTCTTTAGG

[0106] AACCAATATCATTCTAATTTTTTGATTAGCACAAACTTCGCTGATATCTGTCTTTTACATTG

[0107] AAGGAACTATCCTCTATTGATGGCTGCATGGAAAGTTGCACCTGCCTTGGCTGCTGGGTG

[0108] TGCTGCAATATTGAAGCCATCAGAACTGGCATCTGTGTAAGTGGTTATTAATTGTATGAAC

[0109] TTGATTTATTTTGTGGTATTCTTTTTAATGTTGTTACATATGAGTTTTCTTTTTCTTTCCTCA

[0110] TTTTTTTTTTTTTTGGGCAGCACCTGTTTGGAACTGGGTGAAATCTGTAAAGATGTTGGT

[0111] CTTCCATCTGGTATTTTGAATATTCTGACAGGATTTGGCCCTGAAGCTGGTGCTCCTCTAG

[0112] CATCTCATCCTCACGTTGACAAGGTATGGCTATTGCTTTTGGGCATCTACTTTGATTGTTA

[0113] AAGATAAAATGTTGTCTAATTAATAAAAAATATTGATGATAGATTGCATTTACTGGGAGTT

[0114] GTGCTACTGGAAGCAAGATTATGACAGCAGCTGCCCAACTGGTCAAGGTTGAATTTCAG

[0115] CACAGTTTACATTTACATTTGCTTAGTAGAAGTTCTATATCACCATTTGATTTATTTTATCTT

[0116] TACCTGTTTTCTTTGGCAGCCTGTAACTATGGAACTTGGTGGGAAAAGTCCAATTGTTGT

[0117] TTTTGATGATGTCGACCTTGACAAGGGTCAGTAACTTCAGAGCACAATTTTCATATCATG

[0118] AACTTTAGTTTTTCATGGTTAATTAAAAATTTACCTTGTGAAAAGGGAGCTACCTTGTAAT

[0119] CTGTGCATCAGATCTATGCCGTTCTTGATTTTGATATTATTCACTAATTTTCTTTTCCTATGA

[0120] TTTATAGCTGCCGAATGGACGATTTTTGGTTGCTTTTGGACAAATGGTCAGATTTGCAGT

[0121] GCCACATCCCGCCTGCTTGTACATGTATGTAGAATAACCTAGATGATATGCAATTAAATATT

[0122] ATTTATTTCTATCATATACGAACTTTCTCCTCCTTAATTGTGTCAAGGATGTAGATGTTTTT

[0123] TTTTTTTTTTTTTTTTTTTGCGGGATCTTCTGGGTTTAATGAAACAGAAGATGTTGTTAGC

[0124] GATCATTTTTACGATATATTTTTTTTAAAATTTAGAAATATTAAATTGTTCTCTCTGCTATTC

[0125] GATCATTTTTACGATATATTTTTTTTAAAATTTAGAAATATTAAATTGTTCTCTCTGCTATTC

[0126] GATCATTTTTACGATATATTTTTTTTAAAATTTAGAAATATTAAATTGTTCTCTCTGCTATTC

[0127] GATCATTTTTACGATATATTTTTTTTAAAATTTAGAAATATTAAATTGTTCTCTCTGCTATTC

[0128] GATCATTTTTACGATATATTTTTTTTAAAATTTAGAAATATTAAATTGTTCTCTCTGCTATTC

[0129] GATCATTTTTACGATATATTTTTTTTAAAATTTAGAAATATTAAATTGTTCTCTCTGCTATTC

[0130] GATCATTTTTACGATATATTTTTTTTAAAATTTAGAAATATTAAATTGTTCTCTCTGCTATTC

[0131] GATCATTTTTACGATATATTTTTTTTAAAATTTAGAAATATTAAATTGTTCTCTCTGCTATTC

[0132] GATCATTTTTACGATATATTTTTTTTAAAATTTAGAAATATTAAATTGTTCTCTCTGCTATTC

[0133] GATCATTTTTACGATATATTTTTTTTAAAATTTAGAAATATTAAATTGTTCTCTCTGCTATTC

[0134] GATCATTTTTACGATATATTTTTTTTAAAATTTAGAAATATTAAATTGTTCTCTCTGCTATTC

[0135] TTTACTTTTTTTTCTTTTTTTTTTTTTTTATTTGTAACTAATTAAAATTTGTCACTACGATGT

[0136] TTAGCACCTAAAGAAGGGATACTTCGTTGAACCAGCCATTGTTACTAACGTCACAACCT

[0137] CCATGCAACTATGGAGAGAAGAGGTCTTTGGACCTGTTCTATGTGTGAAGACTTTTAGTT

[0138] CTGAAGATGAAGCTATTGAATTAGCAAATGATACAATGTAAGTAAAATTCTACGCAGCTT

[0139] GGTTTAAGCTGGAAACTGGGTTCTGACGTTCAGTTGCTCTTTCTCACAGATATGGGCTAG

[0140] GTGCTGCTGTAATATCAAATGATTTAGAAAGGTGTGAGCGTGTAACCAAGGTGAGGGTT

[0141] GCAATCATCAAGCGCCTATGTAATAGTATATGCAAATGAATAATGTCTGCATTTCTGAACG

[0142] ATGTGTGCTTCCTTGTTTTCTTTCTCCTACAGGCTTTACAGGCAGGAATTGTGTGGGTTAA

[0143] TTGCTCCCAACCATGCTTCACTCAAGCCCCATGGGAGGCAACAAACGCAGTGGCTTTGG

[0144] TCGAGAACTCGGGAATGGTATATTTTGCATTTTAAACTACGTCTTAAATAAGTTTGGCTAT

[0145] ATAAAAAAACGAAAATTTAGGGTCATATTTTGAAAATTGTTCTAGAAACTACTCCATGAA

[0146] GCACTTGTTTGGGAGTTACAACATAGGTTTATGTTGAAACAGAGCATTGAAACACGATTT

[0147] TCAAATAAAATCATTAGCTGAAGAATGTAAACATTGGATGAGCAGATTTACTGAGTGTGT

[0148] GTTTGATTTGTTGGATGAACAGGGGACTTGATAATTATCTGACTGTAAAGCAGGTTACTG

[0149] AGTATGTTTCCAATGAACCATGGGGATGGTACAAGTCTCCTTCTAAATTGTAGAAGAAGA

[0150] CAACACGAGCCTCTCTTGCGTCAAAAAGTTGCAGAACCGTCCAGAAAAATAAGCACTG

[0151] CGACATCTATACTTCGTGTAGTTGGAAGAAACTCTTGCTCAAGTGTGTAGTGCTTCGCCC

[0152] TGAGTTTCCTGGTTAGTTCAGTTTAGGATGTGTCTAAGAACTCGATGAATCACTTGAAAC

[0153] AGTAGACTGTAACTCAATAGAAAATGCAGTTGTTGGTAGCCTCAAAAAGGATTGTGATC

[0154] TGAGATTTGTTTGAGATCTTTTACTTTCTTTTCAGTCTTCTCTATTACTCAAATCTTGGCTT

[0155] TGAAAAGATGAGAATAAACAAGGATTTCTTCTGTGGTAAAATATTGTAGATTCTGAGATA

[0156] TCCTTAGAATTGAGATGCTTTTTCCATGAAAATTCATCAAACTTGGCTGAAAAGGTACGA

[0157] CATCTGTCTGGAATCTCATGATAATCGCATCTTAATCCAGAAGCAACGTTGTTCATGGCTG

[0158] AGTTCTTCACCTAAACCTATATTATAATCAACCACTATATCATTATAAAATCTGATAACACT

[0159] TGATTACAATACAATATTCATCCTCATTGCCCCTTTTTCCTAAGAAGAAGAAATTTACCCTTTAG

[0160] ACATCATCAGAGATCTGTTAACCAAATCATTCCAATTATTGGTGGTAAAGTTGATAATTAC

[0161] ATTCTTCAAGATGAGCAAAGATCCAGTCAAGATAATTACAATATTCATCTTTCTCCTTCATT

[0162] CCCTCTTCTCAGACACAATGATTTCTTCCCTCATCATTCTTTATTCAAGAAACGATGCTGA

[0163] AAACTCACAAGATAATCATCTAATGCAACGAATTAAGACAGCCCAAATACTCAAAACA

[0164] AAAAGGAAAATCAACAGATGCTTAATCTTACAGGCAAAGTTTTGTTGAGCAGTATGAAG

[0165] AAGAACCCTGATGCGGTATGAAAACTTGTCGTATTATGTACAGTTCTTGGTGGGGGTGTGA

[0166] TAAAATCATGGGATCGGTAAAATTGATGCGAGGAATGATCCTGGATCCTTCCTTCAACCG

[0167] ACAAATGCACAAGGTTGAGTCAAATTGGCATTGACTTAAGTGGGCCAAACACAATCCTTG

[0168] ACGATGGGCAACCCAATATCACTAACTCTGCCATTAATTTCCACTGGTTGGCGTCCACAA

[0169] ATCTCTCTTTTTGCACAAACGCTGCCTCCAAAAGTGGAGAGAGAGAGGTAACAGAGAG

[0170] TTGGAAAGTTGATTGTTCTGGCTGGAATTTGGCCACATATCGTAAGAATCTGGCGAAGTA

[0171] CTGCATTAATTCAAATCCCGAATCCCCACGGCTTGTATTTCACAACGTTTCGTTTCGCTGT

[0172] TCTTTGTTTGTTTGTGTTTCTCCCTGAATTCAGAGATTCTGTTCACAATACCATTTCTGGG

[0173] TTGGGCATATTTACCTAAATTATCAATTTACTTGGATTTCCCATATCACTTTCTTGTTGTTG

[0174] AGATCAGTCGGCGGCGGTATTGGGTTTTAGGGATCTGGGTTTACTTCGATCAGCGATGGG

[0175] GAGCGCAGAGGAGAAGGTTGTTGCTGTGATCATGGTCGGCGGCCCCACTAAAGGTATAT

[0176] GAATTCCTAACTTCTTCCCATTCTGTT

[0177] SEQ ID NO:2

[0178] ATCAAGTTTATGCAAAACTGTGCCTCACACTATTGAGGACATATTCTATTTATATTGGCAA

[0179] AAGATTAACTAATAGATCCGATTTCATAGGAAACTAACGAAGTAAGCCTATTCTTATGGCA

[0180] TGACTAACTTCAAGCTATTCTAATGGATTGACCAATTCTAACAGTCCCCTTCAATCCCAA

[0181] CAAGTCTTGTACATGTTGAGATTGTTACAATGTGTAGTAAAAGGAGTTTTAGACAAAGTG

[0182] GTTTTGTCGGGATATCGCCTACTTGATCACTTGATGAAATGAACCGAACTTCCATTGCTTT

[0183] ACGTGCTACTTGTTCTCGAACAAAATGAAAATCAACTTCAATATGTTTCGCTTCTAGCAT

[0184] GAAATCTTGGATTTGAAGTTAAATATGTAGCTCCGAGGTTGTCACACGCAGGCGAGGAG

[0185] CCTTTGATTGATACACGCCCAGTTCCTTCAAGAATGACTTGATCCAAATCATTTCGCAAA

[0186] GTAAGATTCGCAATGGCCTTGTATTCTCGCTTCGGTGCTTGATCTTGACATCGTAGCTTTG

[0187] TTTTCTCGGAACTCCATGAGATTAGATTTGCTCCAAGAAATACAAAGAAAGCCGCTAGTT

[0188] GAACGTCGATCATCGGGACAACCAGCCCAATCAAGCATCGAGAAAACCCCGACAACAT

[0189] CATGGTTGATTTCTGAATTTTCACTCCTAATGCTAGTGTGCCTTTAACATAACGAAGAATT

[0190] CTCTTCACAAAGACCCCAATGAGCATCGGTAGGTGTATGAAGATATTGACACACTTTATT

[0191] GACAAGAAATGCCAAATCAGGCCTAGTCATTGTCAAATATTGTAGTGCTCCCCTTTGTAT

[0192] TTACGTATTTGAATTGTTCTTGGTGATAAGGGTATTCCTTGTTCTCGAATAATTTTTCAGA

[0193] GAACCCATTGGTGTAGACATAGGTTTGCATTTTTCCATGTTTACTCTTTTCAACAAATCTA

[0194] AGGCATATCGTCTCTGTGACAAGATGATACCATCTCGTGTTTTCTTGACTTCAATACCCGA

[0195] GAAAATACTCAAGACCACCCAAATCTTTTACTGCAAAATCTATTTTCAATTTCTCGAATC

[0196] AACCTTTCGGTTGCTTGATCGAGGAGCTCACAATAATTATATCATCAACATAGATGAGCAT

[0197] ATAGATAGTTATCTCTCTGTTTTTGAGAATAAAAAGAGATGAATCACCATCAAGCCTTGA

[0198] AGCCTAACTCAATAAGTTTTCCAATCAACCTTGAATGCCAAGCTTTTGGGGCTTGTTTGA

[0199] GGCCATAAAGGGCTTTCTTGAGCTTGCATATGTAATTCTTTGGTTTGGCTGAGTCTTGAA

[0200] ATCCTGGTGGTTGTCGCATGTACACTTCCTCTTTTTAATTCCATGCAAAAATGCATTTTGG

[0201] ATATCAACTTGTCTCATATTCCAGCCCTTGGTTCTCTGCTAGCGAAAGAATGACCCCGATT

[0202] GTTGACGGTTTGATCACGGGGCTAAAAGTATCAGTGTAATCAACACCAAATCTTTGCTTG

[0203] AATCCTTTGGCAACTAATCTTGCTTTTAATCTTTCAATGACCCATCTGCTTTTCTTTTCACT

[0204] TTATACACCCATTTACTATCAATGAGATTTATTCAGGTTTGGGTGGAACTAGATCCCAAGT

[0205] GGCATTTCGTTTTAGCGGCCGAGAGCTCATCATTCATTGCTCCTCTCCATCTTGGATGTTG

[0206] CATGGCTTCTGCGCAGGTTTCGAGGTTCATGAGCAGTCTCTATGATCGGAGTTGTGATAG

[0207] TTACTGGCGCTTGCGAATTTTCTTGAGGTTTCGAATATCGATAGTTCCATCGTGAATTGTT

[0208] TAGCTTGTACAATGTTATTTCTCAACCTTGTTCGCATTGGATGTTGACTATTTGCTTCAGT

[0209] TGGTTGATCAGACACATGTTCATGAGTTTGGTTTTGAGCATTGATCGAAGCGCTCTCGGC

[0210] TTCATATTCTGCAATTTCTCCGAACTTCTTGTATTATCTGCAGAAGATAAGTGACACAGAC

[0211] AAGTTGTCACTAGCAATATTATCGGTTTGACCATCATTCATATGGGAATTACTAACAACTG

[0212] GTTCAATATCTGTAAGAGCATTTTCGGTGTAAAAACTAGCAAGTTTGGCTAAAATTTGGA

[0213] AGTAGAACAGGATGATGTGGATTTGTGGTTTTGTTTGGTGGCTTAGATTCTTCAAAAGGA

[0214] AAAATATTTTCATCGAAAACCACGTCTCTAGAGATGTAAATACGTCCCGTACTTCTATTTA

[0215] AGCATTTATATCCCTTATGAGAAGAACTATAACCCAAGAATATACATCTAGTAGTTCTGAA

[0216] ACTCAGTTTCTTGTTGTTGTAAGGCCTTAAATTAGGCCAGCAAGCACAGCCCAAACACC

[0217] CTAAGCATGGAGTAGTCTGGACTTTTACCAAACAATTTATGAAGTGGTGTGTCTTGTTGT

[0218] ATGGTTCGGCTGGGCATTCTATTTATAAGAAAGCAAGCTGTGTTGAAAGCTTCATCCCAA

[0219] AGAGGATAGAGGCATGTTGGCTTGAGCGAGTAAAGCAAGGCCGGTTTCTACAATGTGTC

[0220] CGTGTTTTCTTTCGACTAACCCATTCTCGCTTGTGGTGTGTGTGAGGACAGGAGATATGA

[0221] TGTTCAATGCCTGTGGATTTGAAATAATTGTGTAACCGATGGTATTCACCCCCCAATCGAT

[0222] TGGACGGAGCGAATTTTGGTATTTAGCATAGTTTCAACATGTTTTTGAAATTGAAGGAAC

[0223] ACAGACTCAACATCAGATTTGCATCTCAGAAAGTAAATCCAAACATAACGACTAAAATC

[0224] ATCAACAAAGGAAACATAATATTTGGAATTATTTACTGACGCAATGGATGGACCCCATAC

[0225] ATCAGTGTGAATTAATTGAAGGGGTGCTGTAGATACATGCTGAGAAGAACCAAATGGCA

[0226] ATTGATGTGCTTTCCCTAATTGACAAGCATTACAAATTGAGGAAGAGGGAATATTAGTAT

[0227] CTATAGCTAAATTATTATCTTGTAGAATTCTAATGGTAATTGGAGATGCAGGGTGCCCTAG

[0228] CCTTCTGTGCCATTGTTCTTTCGAAAGTTTGGCTGTCAGCAAGGCTTGACTGAAATTATG

[0229] CGGTAGAACGTATAGGCCATTCTTACATCTACCGTGGAGCAGGAGTTTCTTCGTGACTCG

[0230] GTCCTTAACCAAAAAATAGTTTGGGTGAAATTCTACAACAAAGATTATTATCGGATGCTA

[0231] GTCTTTGTCGAAATTAGGTGCTTATTGATTTTAGGAACATATAGGATATGTTTCGTAACAA

[0232] GAGATGAACCAGAAATTAATGAATTCCCAATATGAGAGATAGACAAACCTCGCGCCATTT

[0233] GCAACCCGAATTTGGTCGGTGCCGGTGTAGCGTTCTCTCGGTGGTAAGCCTCTCTAGGT

[0234] CATTGGTGATATGATCTGTGGCTCCGGTGTCAACATACCGGTTTGTGTCACTAGTGTATCC

[0235] ACTTGTTGCCAAAGCTGCTTGTTTGAGATTATTTTCGGCTTGATATGCCCGATCAAATGTG

[0236] CCAGCATTGCAAAGCATCGTGATTGGGCTTACCACAATCGACAGACAATGCCCGAGCTG

[0237] CTTTGTGTCTTTGATGGTTGACGTCCGGGATTGTTCACAGACGCCTTGATTGTCCATGAC

[0238] CACTATTTAACCGACGACTACGACCGCGACTACCTCGACCTCCATTGGCACCTCCACGAA

[0239] TGGATATCCTATTGACATTGTTAGCAGATGAAAGTTGCTCAAATGTACCCTTACGCAAGT

[0240] GCCGCATCTCATAGCTCAAGCATGTGAGCATACACGTCGCCGACCGGTGTATACATCCGT

[0241] TCGGTTGTAATGCTTGTGACTAGAGGATCATAATCTGGACCAAGTCCTTGCAGCATGTAG

[0242] GCGATGAGTTCTTCATCCTCTATTCGCTTGCCAATGGCAGCAAGTGTATCACCAAGATGT

[0243] TTTACTTTACGAAAGTAGTCAGCAATTGTCATGTCCTTTTCTCGGATAGTAGAGAGTTCCA

[0244] TACGGATCTGCATTGCTCTTGCTCGAGATGTGGAAGCAAATTGTCGCTCCGGCGTAATCC

[0245] AGGCTTCTCGTGCAAAAGTGGTGATTCCAACCATCGTCCGAGAACCTCCTCAGTGACTG

[0246] ATGAGTTGATGAGGTGAGTACCACCGGTCCTGGGGGTACCAGACTGTGAACTCAGGGTT

[0247] GATGATAATTTTGCGATTCCCTGTTTCTTCACTTGGTTCAACGGCGATCGTCCGGCTTGGT

[0248] GCGGGCATGGATCCATCCACAAAACCAACAAGGTTTTGGCTACGCAAGTAGGGAAGGA

[0249] TTTGGGTAGACCACAGTAGATAATTTTCTTGTGTAAGCTTAACACTAATCACCTGACTGA

[0250] TCGACGGAGAAGACATTGTGTTGGTTGGGATTGACATGTCTAACAGCAGAGGCTATGGA

[0251] TCTCGTGGATCTTATTTCGGCTCTGATACCATATCAAGTTTATGCAAAACTGTGCCTCACA

[0252] CTATTGAGGACATATTCTATTTATATTGGCAAAAGATTAACTAATAGATCCGATTTCATAGG

[0253] AAACTAACGAAGTAAGCCTATTCTTATGGCATGACTAACTTCAAGCTATTCTAATGGATTG

[0254] ACCAATTCTAACAATATGT

[0255] SEQ ID NO:3

[0256] CATCTCGATTCTCTCTTCTCTTTGCTATCAGATTCTTCCTCTTCTTTCTCCGAGATTTCTTT

[0257] GTAGCAGAGATTTTCAGAGCCAGAATCACTCAATCGATCAGTCAAATCCATGGCGGGAA

[0258] ACGGCTCGTACGATTGGGCGGACCAGTGGATTACAGCGACTCGAAGGACGTAGTCAC

[0259] GGAGAACACGAAGAGGCGGCGGCGGAAACACTACGACTAAGTATAAACAGAAGGT

[0260] CGGCGAAGGCCTTGGGAAGACCAAAGCTGTGGCCTCCAATGGCGTTAAAAAGGTCAAA

[0261] CAAGGAACCTCTCTCGGCTTCCAATGGATCAAAGATAAATATAACAAAACCGCTCATAA

[0262] GAAGTAGAAATCCTTCGTATTCTACTCCTCTCTTGCTCTTCGGTTTTCTTTTTCTTATCATC

[0263] ATCGTCTCCATAGCTTATCTTTGAGCTGTATATCTTGTTTTTTCTTATTCTTTGTCTGCTGGT

[0264] TATATATAGATACAGATACACCATATAGATTATTCTTTTGTTAATATGATTTGATCTCTGTTCGT

[0265] TTCTTTCCTCTTGTTTTGATTCCGTTATTGTTTATCGCTGTTAAATTTCGTACTTCCTTCAG

[0266] ATTCGTTTGTTTGATTTTGATTATAATCTGAGAATTCCGCATCAGTAACATATATAAAGCAG

[0267] GGATAGAATCGCATCTTCTGATGTTTACTGGAAATCATATTAGCATACGAAGATGAAAGG

[0268] TGAAGGAGGAGATCTCGCCCTGAAAAAATTGAAATTATTTTATTTTATGAATAAAAAATCA

[0269] ATTTTGACCCCTAGATTTATTTGAATTTTTTATGTTTGCAATTTTTGAGAAAATATTTATAA

[0270] ATGTATTAATTTTCATTAAATTAAATTCAAGTTCAACACCGGTTCTTCTCAACTTCTGATA

[0271] AAATAAAATCTTGCATATAATAAATGTTTTTAAAAACAAAAACATAGAGTAACAAGTTTA

[0272] AATTTAAATATATTTAAGAGTAATTGTTGCTTACCTCCACAATGTAAGTATATATATGGTTA

[0273] GATGAACACGACTCTCCACAATGATATGATATTGTCCACTTTGAGCTCATACTAATGGAG

[0274] AGATTATTTTTTTATTATAAACCTACAGTCATTTTCTAAATTAACCGATGTGGGACTTTCAT

[0275] CATCTAACATATATATCTACGTGTGTATGTATGTTTGCATGTATATGTATGTAGTGGATCCTC

[0276] CCTACCAAAAGAAAAAAAAAATAGAATAGAATAGAATAGAATAGGTGGTGTGGCCAGTT

[0277] TTTAGCTCTTGCTTTTAATAAAGATGTATTATTTTTTTTATTCTCACGTTTTTATAAAATGCA

[0278] TATATTTTAATATTGATGCATCTTTAAACTTTAAAATGGGTATTTATATTATCATAAACTGAT

[0279] ACTTTAAACAAACATATAAATAATTCAAACTTAAATATATCCATTAATTGCTTCAAAATTAA

[0280] ATATATACATTAATTGTATTAATGGCTACTATAGGAGACAAATTTGTATCCTTCCTAAAAAA

[0281] AAATTCCATTCCAAAAATACCTTTAAAATATTTAAACACTTTTTTTTAGATAAAAGTTG

[0282] TTAATATTTTGTTTTAAATTTTATCTTTGAACTTTCAAAACATCTATTATCTTTAAATTGTTA

[0283] AGAAAATATCATAAGTTTTTTAGAGATTTAAGAATATTTATGAATTTTCAAAAAAGTCTTT

[0284] TTAAAATTGATTGTTTTTTTAGGTTAAAATTAAATTATAATATTCAAGTTTCAATAAATTTTA

[0285] TAAAAAATTAAGTTTTAAGTACGAATTTCAAAATTTAGGGTTAAATATAATTTATTTTTAAT

[0286] AAAAAGTACACATCTTTAATGCTTATTATGATAGCTTACTCAACGGCGTGATTTCTATATAA

[0287] TAAAATAATGTATAATATGTTGGTGTAAGAGATTCTTTGTATTTCTATTTTCATAAATTTTAT

[0288] TTTTCTTATTTAAGTTACCCAATAATTTGATTATCTTCACAGATATCATCTCGAGTTTCGTA

[0289] AAACTCAAAGAAAAAAATTGCAGAAGAAAAATCCAATACGGACGAAAAAATACCGAAAAA

[0290] TTCCGTATTTATATTGATCCAATCAAGTCGAAGTCATTACCAATCCGACTACTTTTTACACT

[0291] GCACTAAAGAGAAGAGCTCTGCAGAAATGGCGATTCATATTCCCAGTCGGCAGCTATTC

[0292] ATCGATGGCGAGTGGAGGGAACCAGTTCTCAAGAAACGCATCCCCATCGTCAACCCCGC

[0293] CACTGAACAAACAATCGGTCTCTTCTTCTTAACTTCTCTGTTTATGACTGTTGTTAATTTT

[0294] GTTTGCCAACTTCTAATCGTTGTGCGGATCGGTAGACGGAAGAAGTAATTAGTTCGAGG

[0295] ATCAGGTGGTTTAAAAGAGGTTGAATCTCACATTTGTGTTATTGCCAACTATGCTTCAGG

[0296] ATCTATTCCCGCGGCTACTGCGGAAGATGTAGAGTTAGCAGTTGATGCTGCCCGAAAAG

[0297] CTCTTGCGAGGAATAAAGGCAAAGATTGGGCATCTGCTTCGGGGGCTCTTCGTGCCAAG

[0298] TATTTGCGTGCTATTGCTGCTAAGGTGAGTCGGTCATATGGATGTTTTACTCTTATTTGATT

[0299] GTGATCTGGTCTCTGTTTTACCATCGGACGGAGGTTATTGATCATTGATGCTTTTGTTACT

[0300] TATTTACGATTCTAGTGGCTTGAAACAATGGATTGATTTCTGATGCGATTCAATAGATCAT

[0301] GAGAAATATGATACCATCCATCTTCCCGCCATAGTTCATTGAAAAATTAGATATCTGGATA

[0302] GAGCCTTCTTTTGGTCTATGTATTGTCTTAGGAATGTTACCTTCTTGTTAATCGCCTTGCA

[0303] GGAGTTCCGTTGGAGTCGCATTTATAGTTTAAGAATCAAGTAGTTTTAGTTACTTTGGAA

[0304] GTTATATTATATATATCTTCAACTTTAATTTCTATCTTCATGGTGTTATTGTCGTCCATCCTG

[0305] AACCCTGTTTGCTGTTTTTCTCACAGATAACAGAGAGGAAATCGGAATTAGCAAAGCTC

[0306] GAAACAATAGACTGTGGAAAACCCCTGGACGAAACTGCATGGGACATTGTATGGCAGTT

[0307] TTTCTTATAGTTAAAGATATATGTATCAAGTTTATGCAAAACTGTGCCTCACACTATTGAG

[0308] GACATATTCTATTTATATTGGCAAAAGATTAACTAATAGATCCGATTTCATAGGAAACTAA

[0309] CGAAGTAAGCCTATTCTTATGGCATGACTAACTTCAAGCTATTCTAATGGATTGACCAATT

[0310] CTAACAGTCCCCTTCAATCCCAACAAGTCTTGTACATGTTGAGATTGTTACAATGTGTAG

[0311] TAAAAGGAGTTTTAGACAAAGTGGTTTTGTCGGGATATCGCCTACTTGATCACTTGATGA

[0312] AATGAACCGAACTTCCATTGCTTTACGTGCTACTTGTTCTCGAACAAAATGAAAATCAAC

[0313] TTCAATATGTTTCGCTTCTAGCATGAAATCTTGGATTTGAAGTTAAATATGTAGCTCCGAG

[0314] GTTGTCACACGCAGGCGAGGAGCCTTTGATTGATACACGCCCAGTTCCTTCAAGAATGA

[0315] CTTGATCCAAATCATTTCGCAAAGTAAGATTCGCAATGGCCTTGTATTCTCGCTTCGGTG

[0316] CTTGATCTTGACATCGTAGCTTTGTTTTCTCGGAACTCCATGAGATTAGATTTGCTCCAAG

[0317] AAATACAAAGAAAGCCGCTAGTTGAACGTCGATCATCGGGACAACCAGCCCAATCAAG

[0318] CATCGAGAAAACCCCGACAACATCATGGTTGATTTCTGAATTTTCACTCCTAATGCTAGT

[0319] GTGCCTTTAACATAACGAAGAATTCTCTTCACAAAGACCCCAATGAGCATCGGTAGGTGT

[0320] ATGAAGATATTGACACACTTTATTGACAAGAAATGCCAAATCAGGCCTAGTCATTGTCAA

[0321] ATATTGTAGTGCTCCCCTTTGTATTTACGTATTTGAATTGTTCTTGGTGATAAGGGTATTCC

[0322] TTGTTCTCGAATAATTTTTCAGAGAACCCATTGGTGTAGACATAGGTTTGCATTTTTCCAT

[0323] GTTTACTCTTTTCAACAAATCTAAGGCATATCGTCTCTGTGACAAGATGATACCATCTCGT

[0324] GTTTTCTTGACTTCAATACCCGAGAAAATACTCAAGACCACCCAAATCTTTTACTGCAAA

[0325] ATCTATTTTCAATTTCTCGAATCAACCTTTCGGTTGCTTGATCGAGGAGCTCACAATAATT

[0326] ATATCATCAACATAGATGAGCATATAGATAGTTATCTCTCTGTTTTTGAGAATAAAAAGAG

[0327] ATGAATCACCATCAAGCCTTGAAGCCTAACTCAATAAGTTTTCCAATCAACCTTGAATGC

[0328] CAAGCTTTTGGGGCTTGTTTGAGGCCATAAAGGGCTTTCTTGAGCTTGCATATGTAATTC

[0329] TTTGGTTTGGCTGAGTCTTGAAATCCTGGTGGTTGTCGCATGTACACTTCCTCTTTTTAAT

[0330] TCCATGCAAAAATGCATTTTGGATATCAACTTGTCTCATATTCCAGCCCTTGGTTCTCTGC

[0331] TAGCGAAAGAATGACCCCGATTGTTGACGGTTTGATCACGGGGCTAAAAGTATCAGTGT

[0332] AATCAACACCAAATCTTTGCTTGAATCCTTTGGCAACTAATCTTGCTTTTAATCTTTCAAT

[0333] GACCCATCTGCTTTTCTTTTCACTTTATACACCCATTTACTATCAATGAGATTTATTCAGGT

[0334] TTGGGTGGAACTAGATCCCAAGTGGCATTTCGTTTTAGCGGCCGAGAGCTCATCATTCAT

[0335] TGCTCCCTCTCCATCTTGGATTGTTGCATGGCTTCTGCGCAGGTTTCGAGGTTCATGAGCAG

[0336] TTCTCATGATCGGAGTTTGTGATAGTTACTGGCGCTTGCGAATTTTCTTGAGGTTTCGAATA

[0337] TCGATAGTTCCATCGTGAATTGTTTAGCTTGTACAATGTTATTTCTCAACCTTGTTCGCATT

[0338] GGATGTTGACTATTTGCTTCAGTTGGTTGATCAGACACATGTTCATGAGTTTGGTTTTGA

[0339] GCATTGATCGAAGCGCTCTCGGCTTCATATTCTGCAATTTCTCCGAACTTCTTGTATTATC

[0340] TGCAGAAGATAAGTGACACAGACAAGTTGTCACTAGCAATATTATCGGTTTGACCATCAT

[0341] TCATATGGGAATTACTAACAACTGGTTCAATATCTGTAAGAGCATTTTCGGTGTAAAAACT

[0342] AGCAAGTTTGGCTAAAATTTGGAAGTAGAACAGGATGATGTGGATTTGTGGTTTTGTTTG

[0343] GTGGCTTAGATTCTTCAAAAGGAAAAATATTTTCATCGAAAACCACGTCTCTAGAGATGT

[0344] AAATACGTCCCGTACTTCTATTTAAGCATTTATATCCCTTATGAGAAGAACTATAACCCAA

[0345] GAATATACATCTAGTAGTTCTGAAACTCAGTTTCTTGTTGTTGTAAGGCCTTAATTAGGC

[0346] CAGCAAGCACAGCCCAAACACCCTAAGCATGGAGTAGTCTGGACTTTTACCAAACAATT

[0347] TATGAAGTGGTGTGTCTTGTTGTATGGTTCGGCTGGGCATTCTATTTATAAGAAAGCAAG

[0348] CTGTGTTGAAAGCTTCATCCCAAAGAGGATAGAGGCATGTTGGCTTGAGCGAGTAAAGC

[0349] AAGGCCGGTTTCTACAATGTGTCCGTGTTTTCTTTCGACTAACCCATTCTCGCTTGTGGT

[0350] GTGTGTGAGGACAGGAGATATGATGTTCAATGCCTGTGGATTTGAAATAATTGTGTAACC

[0351] GATGGTATTCACCCCCCAATCGATTGGACGGAGCGAATTTTGGTATTTAGCATAGTTTCA

[0352] ACATGTTTTTGAAATTGAAGGAACACAGACTCAACATCAGATTTGCATCTCAGAAAGTA

[0353] AATCCAAACATAACGACTAAAATCATCAACAAAGGAAACATAATATTTGGAATTATTTAC

[0354] TGACGCAATGGATGGACCCCATACATCAGTGTGAATTAATTGAAGGGGTGCTGTAGATAC

[0355] ATGCTGAGAAGAACCAAATGGCAATTGATGTGCTTTCCCTAATTGACAAGCATTACAAAT

[0356] TGAGGAAGAGGGAATATTAGTATCTATAGCTAAATTATTATCTTGTAGAATTCTAATGGTAA

[0357] TTGGAGATGCAGGGTGCCCTAGCCTTCTGTGCCATTGTTCTTTCGAAAGTTTGGCTGTCA

[0358] GCAAGGCTTGACTGAAATTATGCGGTAGAACGTATAGGCCATTCTTACATCTACCGTGGA

[0359] GCAGGAGTTTCTTCGTGACTCGGTCCTTAACCAAAAAATAGTTTGGGTGAAATTCTACA

[0360] ACAAAGATTATTATCGGATGCTAGTCTTTGTCGAAATTAGGTGCTTATTGATTTTAGGAAC

[0361] ATATAGGATATGTTTCGTAACAAGAGATGAACCAGAAATTAATGAATTCCCAATATGAGA

[0362] GATAGACAAACCTCGCGCCATTTGCAACCCGAATTTGGTCGGTGCCGGTGTAGCGTTCT

[0363] CTCGGTGGTAAGCCTCTCTAGGTCATTGGTGATATGATCTGTGGCTCCGGTGTCAACATA

[0364] CCGGTTTGTGTCACTAGTGTATCCACTTGTTGCCAAAGCTGCTTGTTTGAGATTATTTTCG

[0365] GCTTGATATGCCCGATCAAATGTGCCAGCATTGCAAAGCATCGTGATTGGGCTTACCACA

[0366] ATCGACAGACAATGCCCGAGCTGCTTTGTGTCTTTGATGGTTGACGTCCGGGATTGTTCA

[0367] CAGACGCCTTGATTGTCCATGACCACTATTTAACCGACGACTACGACCGCGACTACCTCG

[0368] ACCTCCATTGGCACCTCCACGAATGGATATCCTATTGACATTGTTAGCAGATGAAAGTTG

[0369] CTCAAATGTACCCTTACGCAAGTGCCGCATCTCATAGCTCAAGCATGTGAGCATACACGT

[0370] CGCCGACCGGTGTATACATCCGTTCGGTTGTAATGCTTGTGACTAGAGGATCATAATCTG

[0371] GACCAAGTCCTTGCAGCATGTAGGCGATGAGTTCTTCATCCTCTATTCGCTTGCCAATGG

[0372] CAGCAAGTGTATCACCAAGATGTTTTACTTTACGAAAGTAGTCAGCAATTGTCATGTCCT

[0373] TTTCTCGGATAGTAGAGAGTTCCATACGGATCTGCATTGCTCTTGCTCGAGATGTGGAAG

[0374] CAAATTGTCGCTCCGGCGTAATCCAGGCTTCTCGTGCAAAAGTGGTGATTCCAACCATC

[0375] GTCCGAGAACCTCCTCAGTGACTGATGAGTTGATGAGGTGAGTACCACCGGTCCTGGGG

[0376] GTACCAGACTGTGAACTCAGGGTTGATGATAATTTTGCGATTCCCTGTTTCTTCACTTGG

[0377] TTCAACGGCGATCGTCCGGCTTGGTGCGGGCATGGATCCATCCACAAAACCAACAAGGT

[0378] TTTGGCTACGCAAGTAGGGAAGGATTTGGGTAGACCACAGTAGATAATTTTCTTGTGTAA

[0379] GCTTAACACTAATCACCTGACTGATCGACGGAGAAGACATTGTGTTGGTTGGGATTGAC

[0380] ATGTCTAACAGCAGAGGCTATGGATCTCGTGGATCTTATTTCGGCTCTGATACCATATCAA

[0381] GTTTATGCAAAACTGTGCCTCACACTATTGAGGACATATTCTATTTATATTGGCAAAAGAT

[0382] TAACTAATAGATCCGATTTCATAGGAAACTAACGAAGTAAGCCTATTCTTATGGCATGACT

[0383] AACTTCAAGCTATTCTAATGGATTGACCAATTCTAACAATATGTTGGTTGTAACAATTTTC

[0384] ATTGCCGTCAATGTTTGTTAATATTGTACAGGATGATGTTGCTGGGTGCTTCGAGTACTAT

[0385] GCAGATCTTGCCGAAGGGTTGGATGCTAAGCAAAAAGCTCCTGTTTCCGTTCCCATGGA

[0386] TACATTCAAAAGCCATGTTCTTAAAGAACCCATTGGAGTTGTTGGGTTGATTACTCCCTG

[0387] GTACCTGTTGATCCATCTTCTACTTCTTCTTTTAGTTCTTCACGGCTATTTAGCTTCTCCGA

[0388] TTTTATTGGAAGTATTAGAGTTATTGTTGTTAAACATTTTATAAATCATGGTCAACTCTTGT

[0389] CACTCAACCAAGAACCTGTCGATTTCAAACATGGTGACGGAGACTCTTCAATCCAGGTT

[0390] CAAACAACTGGTCATTAACTTAAACCTTAAGATATCAAATATTCCAAAACTTTCTTCACC

[0391] AATAATGTAGAAGGGTCAAGTCATAGTTTCAAGTGCTAATTCTTTTATATAAAATTTTCCT

[0392] TATTCGTTTGAGTCATTCTTCTGAATTTTTTCACTCTCAAAATGTGATGTTTTTCTCTTTTA

[0393] TATACATATATATTTTACTTGGATTGATTTTTTTACTACAAATAATTGTATAAAGGATTTAAC

[0394] GCATTATGTTATATCTATTCAACAAGTGTTTGATACATGTACAACAAGGATTGAATGTTGG

[0395] ATTGACTAACATGTGGTTCAACATGTATCTAAATGCTGGGATGTTGAATACTTGTGATCAT

[0396] GGATATGTTACCCAACTTGAGTAACTCTGCTTCACAGTTCCCATTTTATTGTTAATTTGAA

[0397] CGAGAGGCATACATTGGGACGTTATTAGTTATTAACCTCCAGGGTCGGATCTGGTTATGTT

[0398] CTCTCTTGTCCTCCTCTTTAGGAACCAATATCATTCTAATTTTTTGATTAGCACAAACTTC

[0399] GCTGATATCTGTCTTTTACATTGAAGGAACTATCCTCTATTGATGGCTGCATGGAAAGTTG

[0400] CACCTGCCTTGGCTGCTGGGTGTGCTGCAATATTGAAGCCATCAGAACTGGCATCTGTGT

[0401] AAGTGGTTATTAATTGTATGAACTTGATTTATTTTGTGGTATTCTTTTTAATGTTGTTACATA

[0402] TGAGTTTTCTTTTTCTTTCCTCATTTTTTTTTTTGGGCAGCACCTGTTTGGAACTGGGTGA

[0403] AATCTGTAAAGATGTTGGTCTTCCATCTGGTATTTTGAATATTCTGACAGGATTTGGCCCT

[0404] GAAGCTGGTGCTCCTCTAGCATCTCATCCTCACGTTGACAAGGTATGGCTATTGCTTTTG

[0405] GGCATCTACTTTGATTGTTAAAGATAAAATGTTGTCTAATTAATAAAAAAATATTGATGAT

[0406] AGATTGCATTTACTGGGAGTTGTGCTACTGGAAGCAAGATTATGACAGCAGCTGCCCAA

[0407] CTGGTCAAGGTTGAATTTCAGCACAGTTTACATTTACATTTGCTTAGTAGAAGTTCTATAT

[0408] CACCATTTGATTTATTTTATCTTTACCTGTTTTCTTTGGCAGCCTGTAACTATGGAACTTGG

[0409] TGGGAAAAGTCCAATTGTTGTTTTTGATGATGTCGACCTTGACAAGGGTCAGTAACTTC

[0410] AGAGCACAATTTTCATATCATGAACTTTAGTTTTTCATGGTTAATTAAAAATTTACCTTGT

[0411] GAAAAGGGAGCTACCTTGTAATCTGTGCATCAGATCTATGCCGTTCTTGATTTTGATATTA

[0412] TTCACTAATTTTCTTTTCCTATGATTTATAGCTGCCGAATGGACGATTTTTGGTTGCTTTTG

[0413] GACAAATGGTCAGATTTGCAGTGCCACATCCCGCCTGCTTGTACATGTATGTAGAATAAC

[0414] CTAGATGATATGCAATTAAATATTATTTATTTCTATCATATACGAACTTTCTCCTCCTTAATT

[0415] GTGTCAAGGATGTAGATGTTTTTTTTTTTTTTTTTTTTGGCGGGATCTTCTGGGTTTAATG

[0416] AAACAGAAGATGTTGTTAGCCTAGGATAATTGTGCTTTAAGGCCAACTGATAGAGTAATT

[0417] TTCTGAAATATTTAATAAGCTGATCATTTTTACGATATATTTTTTTAAAATTTAGAAATATTA

[0418] AATTGTTCTCTCTGCTATTCCAGGAAAACATTGCTGATGAATTCTTGGATAGGCTCGTGA

[0419] AGTGGTGCCAGAACATTAAGATTTCAGATCCTTTAGAAGAAGGTTGCAGGCTTGGCCCT

[0420] GTTGTTAGTGCGGGACAGGTAAGAGAAGGATATGTTTCTATTTTTTGAGTTCTTTACGAC

[0421] AGAAAAGAAAAGCAATCCGCTGCCACAATATAAAATATAGGACACGGATATGGTAATAAT

[0422] AGGACAGAACGTGGTAGGGATACCTTGTTTTTAAAGAATATTTGTACATGTATTTATGTGT

[0423] TTCATGGATTTAACCTACAAAATGTGCTTTGCTAAGAATATTTATACAAGGTTAGTGATAA

[0424] GTTAGAGAGTGCAACTTATGCAATTCATTCTGGTTTAGAATATTATATTTGCGCTACTGTTT

[0425] GATTCTTTAACTAATTTTTGGTAATCTAAAATTCAGTATGAGAAAGTTCTGAACTTCGTCT

[0426] CAACTGCTGAGCGCGAAGGTGCAAAGCTTCTATTTGGTGGAGTTCGCCCTAAGGTAACA

[0427] CTTGATTTTTACACATCACTTTTACTTTTTTTTTTTTTTTTTTTTTTTTATTTGTAACTAATTA

[0428] AAATTTGTCACTACGATGTTTAGCACCTAAAGAAGGGATACTTCGTTGAACCAGCCATTG

[0429] TTACTAACGTCACAACCTCCATGCAACTATGGAGAGAAGAGGTCTTTGGACCTGTTCTAT

[0430] GTGTGAAGACTTTTAGTTCTGAAGATGAAGCTATTGAATTAGCAAATGATACAATGTAAG

[0431] TAAAATTCTACGCAGCTTGGTTTAAGCTGGAAACTGGGTTCTGACGTTCAGTTGCTCTTT

[0432] CTCACAGATATGGGCTAGGTGCTGCTGTAATATCAAATGATTTAGAAAGGTGTGAGCGTG

[0433] TAACCAAGGTGAGGGTTGCAATCATCAAGCGCCTATGTAATAGTATATGCAAATGAATAA

[0434] TGTCTGCATTTCTGAACGATGTGTGCTTCCTTGTTTTCTTTCTCCTACAGGCTTTACAGGC

[0435] AGGAATTGTGTGGGTTAATTGCTCCCAACCATGCTTCACTCAAGCCCCATGGGGAGGCA

[0436] ACAAACGCAGTGGCTTTGGTCGAGAACTCGGGGAATGGTATATTTTGCATTTTAAACTAC

[0437] GTCTTAAATAAGTTTGGCTATATAAAAAAACGAAAATTTAGGGTCATATTTTGAAAATTGT

[0438] TCTAGAAACTACTCCATGAAGCACTTGTTTGGGAGTTACAACATAGGTTTATGTTGAAAC

[0439] AGAGCATTGAAACACGATTTTCAAATAAAATCATTAGCTGAAGAATGTAAACATTGGATG

[0440] AGCAGATTTACTGAGTGTGTGTTTGATTTGTTGGATGAACAGGGGACTTGATAATTATCT

[0441] GACTGTAAAGCAGGTTACTGAGTATGTTTCCAATGAACCATGGGGATGGTACAAGTCTC

[0442] CTTCTAAATTGTAGAAGAAGACAACACGAGCCTCTCTTGCGTCAAAAAGTTGCAGAACC

[0443] GTCCAGAAAAATAAGCACTGCGACATCTATACTTCGTGTAGTTGGAAGAAACTCTTGCT

[0444] CAAGTGTGTAGTGCTTCGCCCTGAGTTTCCTGGTTAGTTCAGTTTAGGATGTGTCTAAGA

[0445] ACTCGATGAATCACTTGAAACAGTAGACTGTAACTCAATAGAAAATGCAGTTGTTGGTA

[0446] GCCTCAAAAAGGATTGTGATCTGAGATTTGTTTGAGATCTTTTACTTTCTTTTCAGTCTTC

[0447] TCTATTACTCAAATCTTGGCTTTGAAAAGATGAGAATAAACAAGGATTTCTTCTGTGGTA

[0448] AAATATTGTAGATTCTGAGATATCCTTAGAATTGAGATGCTTTTTCCATGAAAATTCATCA

[0449] AACTTGGCTGAAAAGGTACGACATCTGTCTGGAATCTCATGATAATCGCATCTTAATCCA

[0450] GAAGCAACGTTGTTCATGGCTGAGTTCTTCACCTAAACCTATATTATAATCAACCACTATA

[0451] TCATTATAAAATCTGATAACACTTGATTACAATACAATATTCATCCTCATTGCCCCTTTTTC

[0452] CTAAGAAGAAATTTACCCTTTAGACATCATCAGAGATCTGTTAACCAAATCATTCCAATTA

[0453] TTGGTGGTAAAGTTGATAATTACATTCTTCAAGATGAGCAAAGATCCAGTCAAGATAATT

[0454] ACAATATTCATCTTCTCCTTCATTCCCTCTTCTCAGACACAATGATTTCTTCCCTCATCATT

[0455] CTTTATTCAAGAAACGATGCTGAAAACTCACAAGATAATCATCTAATGCAACGAATTAAG

[0456] ACAGCCCAAATACTCAAAAACAAAAAGGAAAATCAACAGATGCTTAATCTTACAGGCA

[0457] AAGTTTTGTTGAGCAGTATGAAGAAGACCCTGATGCGGTATGAAAACTTGTCGTATTATG

[0458] TACAGTTCTTGGTGGGGGTGTGATAAAATCATGGGATCGGTAAAATTGATGCGAGGAATG

[0459] ATCCTGGATCCTTCCTTCAACCGACAAATGCACAAGGTTGAGTCAAATTGGCATTGACTT

[0460] AAGTGGGCCAACACAATCCTTGACGATGGGCAACCCAATATCACTAACTCTGCCATTAAT

[0461] TTCCACTGGTTGGCGTCCACAAATCTCTCTTTTTGCACAAACGCTGCCTCCAAAAGTGG

[0462] AGAGAGAGAGGGTAACAGAGAGTTGGAAAGTTGATTGTTCTGGCTGGAATTTGGCCAC

[0463] ATATCGTAAGAATCTGGCGAAGTACTGCATTAATTCAAATCCCGAATCCCCACGGCTTGT

[0464] ATTTCACAACGTTTCGTTTCGCTGTTCTTTGTTTGTTTGTGTTTCTCCCTGAATTCAGAGA

[0465] TTCTGTTCACAATACCATTTCTGGGTTGGGCATATTTACCTAAATTATCAATTTACTTGGAT

[0466] TTCCCATATCACTTTCTTGTTGTTGAGATCAGTCGGCGGCGGTATTGGGTTTTAGGGATCT

[0467] GGGTTTACTTCGATCAGCGATGGGGAGCGCAGAGGAGAAGGTTGTTGCTGTGATCATGG

[0468] TCGGCGGCCCCACTAAAGGTATATGAATTCCTAACTTCTTCCCATTCTGTT

[0469] Example 2 Verification of Indel molecular marker related to the sweet potato flavor trait in parents

[0470] For the mutation of the insertion of betaine aldehyde dehydrogenase gene (4597bp large fragment), the amplification primers were designed at the upstream and downstream positions of the insertion site, including the upstream primer (TTGTTAATCGCCTTGCAGG, SEQ ID NO: 4) and the downstream primer (CGGAGAAGCTAAATAGCCG, SEQ ID NO: 5).

[0471] The DNA of the two parents in Example 1 was extracted, and PCR amplification was performed using the above amplification primers, wherein the reaction system of PCR amplification was as follows: ddH2O 18μL, 2×Phanta Max Buffer 25μL, dNTPMix (10mM each) 1μL, upstream primer (10μM) 2μL, downstream primer (10μM) 2μL, Phanta Max Super-Fidelity DNA Polymerase 1μL, template DNA 1μL. The reaction program was as follows: 95℃ 3min; 95℃, 15sec, 53℃, 15sec, 72℃, 5min, 34 cycles; 72℃ 5min.

[0472] After the PCR amplification reaction was completed, the amplified fragments were sequenced, and the results showed that the amplification product of the sweet potato flavor pumpkin resource was 5137bp, and the amplification product of the non-sweet potato flavor pumpkin resource was 538bp (part of the BADH fragment, since the primers were designed at both ends of the insertion site for amplification, the amplification product of the sweet potato flavor pumpkin resource was slightly different from the size of the inserted fragment).

[0473] Example 3 Further verification of the Indel molecular marker related to the sweet potato flavor trait of pumpkin

[0474] Using the method of Example 2, 10 sweet potato flavor pumpkin varieties (No. 22108, 22118, Hei3, Huananda1, Mu2, S18-1, Xiang1hao, Za2-3, Za8-1, Za9-2, created resources of Guangdong Academy of Agricultural Sciences Vegetable Research Institute) and 30 non-sweet potato flavor pumpkin varieties (commercial commodity varieties) were subjected to PCR amplification and gel electrophoresis detection.

[0475] Results are shown in Table 1 Figure 3 As shown in Table 1, the size of the amplified product is about 500 bp in 30 non-earthy pumpkin varieties, and the size of the amplified product is about 5100 bp in 10 earthy pumpkin varieties. The genotype is 100% consistent with the earthy phenotype.

[0476] Each technical feature of the above-described embodiments can be combined arbitrarily, and for the sake of brevity, each technical feature in the above-described embodiments is not described in all possible combinations, however, as long as the combination of technical features does not exist, it should be considered as the scope of the description.

[0477] The above-described embodiments only express several embodiments of the present application, which are described in detail and in detail, but should not be construed as limiting the scope of the patent. It should be noted that for ordinary skilled in the art, without departing from the concept of the present application, a number of modifications and improvements can be made, which are within the scope of the present application. Therefore, the scope of protection of the present application patent should be subject to the appended claims.

Claims

1. An Indel molecular marker associated with the taro aroma trait of pumpkin fruit, characterized in that, The sequence of the Indel molecular marker is shown in SEQ ID NO:1 or SEQ ID NO:3, wherein SEQ ID NO:3 is SEQ ID NO:1 with the sequence shown in SEQ ID NO:2 inserted.

2. The application of the reagent for detecting the Indel molecular marker described in claim 1 in detecting whether pumpkin fruits have a taro aroma or in screening taro-flavored pumpkin varieties.

3. The application according to claim 2, characterized in that, The reagents include a forward primer with the sequence shown in SEQ ID NO:4 and a reverse primer with the sequence shown in SEQ ID NO:

5.

4. A method for screening taro-flavored pumpkin varieties, characterized in that, Includes the following steps: Using the DNA of the pumpkin to be tested as a template, PCR amplification was performed using the forward primer with the sequence shown in SEQ ID NO:4 and the reverse primer with the sequence shown in SEQ ID NO:

5. If the size of the amplification product is 5137 bp, the pumpkin to be tested is a taro-flavored pumpkin variety; if the size of the amplification product is 538 bp, the pumpkin to be tested is a non-taro-flavored pumpkin variety.

5. A method for detecting whether pumpkin fruit has a taro aroma, characterized in that, Includes the following steps: Using the DNA of the pumpkin to be tested as a template, PCR amplification was performed using the forward primer with the sequence shown in SEQ ID NO:4 and the reverse primer with the sequence shown in SEQ ID NO:

5. If the size of the amplification product is 5137bp, the pumpkin fruit to be tested has a taro aroma; if the size of the amplification product is 538bp, the pumpkin fruit to be tested does not have a taro aroma.

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