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作 者:郑翠芳[1] 朱晓东[1] 方丽娜[1] 陈友铃[1] 吴文珊[1]
机构地区:[1]福建师范大学生命科学学院,福建福州350108
出 处:《热带作物学报》2009年第12期1740-1745,共6页Chinese Journal of Tropical Crops
基 金:福建省科技厅重点资助项目(No.2008I0014);福建师范大学生物学国家级实验教学示范中心学生创新性研究项目(No.2009ls006);福建省教育厅科技计划项目(No.JB09030)
摘 要:爱玉子雌、雄异株,其性别以及品系在结果前难以通过枝叶等形态特征进行辨别。作者将序列相关扩增多态性(SRAP)标记应用于爱玉子性别及品系鉴别研究,建立了完整的PCR反应体系,扩增效果好,结果稳定可靠,可重复性强。在153对SRAP引物中,只有引物me5-em14在爱玉子雌株上扩增出约220bp的特异条带,将其命名为FAme5-em14-220,该标记与雄株没有关联,可作为爱玉子性别鉴别的SRAP标记。从153对引物中筛选出8对多态性好、分辨率高的引物组合,在24个爱玉子雌性品系和16个雄性品系的DNA样品中扩增出171条多态性带,并获得爱玉子雌、雄各品系独特的核酸指纹,其中em6-me6引物与me3-em2、me1-em8、me1-em9中的任何一对引物进行组合,都能鉴别24个爱玉子雌性品系;me3-em2与me9-em142对引物组合也能鉴别24个爱玉子雌性品系;me1-em8单独一对引物,以及me1-em9与me3-em2(或me6-em6)2对引物组合能鉴别16个爱玉子雄性品系,并获得2个雄性品系的特异性片段,即片段me1-em8-350bp为大洋57特有,me1-em8-240bp为大洋241特有。研究结果对爱玉子种质资源遗传多样性的分析、品种和品系的鉴定、以及种苗质量仲裁检验等具有重要的理论意义和实践价值。Jelly fig(Ficus awkeotsang Makino) is dioecious, and its sex and cultivars are difficult to be distinguished from their morphological characteristics of the branches and leaves without fruit. In this context sequence-related amplified polymorphism(SRAP) was then used for identification of plant sex and cultivars. An efficient PCR reaction system was developed for detecting the SRAP and it was very reliable, effective and reproducible. Of 153 pairs of SRAP primers only me5-em14 primer was successful to amplify a specific band of 220 bp from a female plant, named FA me5-em14-220. This marker was not associated with male plant and can be used as SRAP marker for sex identification of F. awkeotsang. Eight pairs of primers with high polymorphism and quality were selected from 153 pairs of primers, and 171 polymorphic bands were amplified from the DNA samples of 24 female and 16 male cultivars of F. awkeotsang. Specific fingerprints of each cultivar of F. awkeotsang had been obtained. Twenty four female cultivars of F. awkeotsang were effectively identified by using any combination of em6-me6 with me3-em2, me1-em8 or me1-em9, or combination of me3-em2 with me9-em14; 16 male cultivars of F. awkeotsang were identified with a single pair of primers me1-em8, or combinations of me1-em9 with me3-em2 or me6-em6.Two SRAP markers, me1-em8-350 bp and me1-em8-240 bp, related to male cultivars were obtained. The fragment me1-em8-350 bp is unique to cultivar Dayang 57, and the fragment me1-em8-240 bp to Dayang 241. DNA fingerprinting data of F. awkeotsang cultivars are important basis for validation, patent protection and utilization of varieties or clones. This study provides important theoretical guidance and practical value for identification of genetic diversity, evaluation of varieties or clones and quality control of planting materials.
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