出 处:《植物遗传资源学报》2021年第3期860-872,共13页Journal of Plant Genetic Resources
基 金:国家重点研发计划(2018YFD1000107);中央高校基本业务费(KYZZ202011,KYZZ2021002);农业部农作物种质资源保护项目(19200363);“一带一路”创新合作项目:园艺作物分子育种技术的海外应用合作开发(BZ2019012)。
摘 要:自交不亲和是植物避免近亲繁殖保持优良种性的重要途径,梅是典型的配子体(GSI)自交不亲和植物。本研究采用等位基因特异性PCR(AS-PCR,allele-specific PCR)和序列测定等技术,分别以梅S-RNase基因与SFB基因核苷酸序列保守区设计引物Pru-C2和PCE-R、SFB-C1F和Pm-Vb,然后以11个品种的基因组DNA为模板进行AS-PCR扩增和PCR产物测序。经过序列对比分析明确了11个梅品种的S-RNase基因型与SFB基因型,分别为软条红梅(S14S13)(F-box2/SFB14)、小叶猪肝(S3S33)(SFB51/SFB53)、中红(S3S24)(SFB2/SFB24)、青佳915(S3S15)(SFB2/SFB43)、美林红(S1S37)(SFB1/SFB24)、赤凤红梅(S14S38)(SFB12/SFB49)、大龙梅(S20S30)(SFB7/SFB41)、红光梅(S3S33)(SFB24/SFB55)、南农丰羽(S3S5)(SFB31/SFB43)、南农丰娇(S3S6)(SFB2/SFB47)、南农龙霞(S15S37)(SFB41/SFB43)。对获得的S-RNase基因与SFB基因序列进一步分析发现,除了已知的S-RNase基因与SFB基因外,还鉴定到S37-RNase、S38-RNase 2个未登录的新S-RNase基因与SFB47、SFB49、SFB51、SFB53、SFB555个未登录的新SFB基因,并将其序列登录在Gen Bank数据库,其登录号分别为MT950061、MT950062,以及MW186463、MW186465、MW186467、MW186469、MW186471。研究结果为梅生产栽培、授粉树的配置和提高育种效率提供了依据。Self-incompatibility is an important way for plants to avoid inbreeding and thus to maintain excellent species. Japanese apricot is a kind of typical gametophyte self-incompatibility(GSI)species. In this study,AS-PCR(Allele Specific PCR)technique and sequencing were used to analyze and identify S-RNase genotypes and SFB genotypes of 11 cultivars. First,primer pairs Pru-C2 and PCE-R,SFB-C1 F and Pm-Vb were designed according to the conservative region of the S-RNase gene and SFB gene of Japanese apricot. Then,the genomic DNA was used as a template for AS-PCR amplification and PCR products sequenced directly. After sequences comparison and analysis,the S-RNase genotype and SFB genotype of 11 Japanese apricot cultivars were Ruantiaohongmei of S13 S14 and F-box2/SFB14,Xiaoyezhugan of S3 S33 and SFB51/SFB53,Zhonghong of S3 S24 and SFB2/SFB24,Qingjia915 of S3 S15 and SFB2/SFB43,Meilinhong of S1 S37 and SFB1/SFB24,Chifenghongmei of S14 S38 and SFB12/SFB49,Dalongmei of S20 S30 and SFB7/SFB41,Hongguangmei of S3 S33 and SFB24/SFB55,Nannongfengyu of S3 S5 and SFB31/SFB43,Nannongfengjiao of S3 S6 and SFB2/SFB47,Nannonglongxia of S15 S37 and SFB41/SFB43. Further analysis of the obtained S-RNase gene and SFB gene sequence showed that in addition to the known S-RNase gene and SFB gene,and two unregistered S-RNase genes of S37-RNase and S38-RNase,and five unregistered new SFB genes SFB47,SFB49,SFB51,SFB53 and SFB55 were also identified. Their sequences have been registered in the GenBank database,the accession numbers were MT950061 and MT950062,MW186463,MW186465,MW186467,MW186469 and MW186471,respectively. The research results provide a basis for the allocation of pollination trees for Japanese apricot production and cultivation and the improvement of breeding efficiency.
关 键 词:梅 S-RNase基因型 SFB基因 AS-PCR 新基因
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