枇杷RAPD扩增产物的不同电泳检测及其序列特征分析

›› 2010, Vol. 22 ›› Issue (5): 570-575.

枇杷RAPD扩增产物的不同电泳检测及其序列特征分析

张晓莹¹,钱剑林²,王化坤²,宋长年¹,张彦苹¹,房经贵¹

¹南京农业大学园艺学院,江苏南京210095； ²江苏省太湖常绿果树中心,江苏苏州 215107

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-09-25 发布日期:2010-09-25

Analysis of different electrophoresis and sequence characteristics of RAPD amplification products of loquat

ZHANG Xiao-ying;QIAN Jian-lin;WANG Hua-kun;SONG Chang-nian;ZHANG Yan-ping;FANG Jing-gui

¹College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China; ² Evergreen Tree Center of Taihu Lake in Jiangsu Province, Suzhou 215107, China

Received:1900-01-01 Revised:1900-01-01 Online:2010-09-25 Published:2010-09-25

摘要/Abstract

摘要： 为了更好地将RAPD技术应用于枇杷品种资源的鉴定以及遗传基础的研究,在选用11个碱基引物以及严格筛选PCR退火温度的最新RAPD技术优化的基础上,以16个枇杷品种为试验材料,对琼脂糖凝胶以及聚丙烯酰胺凝胶(PAGE)电泳检测的RAPD PCR扩增产物效果进行比较。结果表明，聚丙烯酰胺凝胶(PAGE)电泳检测出的总谱带数以及多态性谱带数均高于琼脂糖凝胶。根据两种电泳系统获得的标记信息进行枇杷品种遗传聚类分析,结果表明PAGE电泳的分析结果与枇杷的分类情况更为一致。随机对挑选的38个片段进行克隆与测序，发现37个片段都是对应引物的RAPD扩增产物，其中有6条是编码蛋白的基因片段,说明RAPD不仅扩增基因组上的非编码蛋白序列,而且可以扩增编码蛋白的基因片段,是能够揭示枇杷品种间基因组信息异同的理想的DNA标记技术。

关键词: 枇杷, RAPD, 琼脂糖凝胶电泳, PAGE

Abstract: In order to utilize RAPD technology in identification of loquat cultivars and genetic analysis, the RAPD fingerprints of 16 loquat cultivars from agarose and polyacrylamide gel electrophoreses were compared, which were carried out with latest optimization of RAPD technique in choosing 11 nt primers and strict screening PCR annealing temperature. The results showed that more bands and more polymorphic bands could be detected by the polyacrylamide gel electrophoresis (PAGE) than the agarose gel electrophoresis. Two polygenetic trees derived from the fingerprint data collected from two electrophoreses were constructed, showing that the result from PAGE electrophoresis was more consistent with the regular classification of these loquat cultivars by phenotypic traits. The results of cloning and sequencing of 38 PCR amplified fragments showed that 37 fragments were RAPD amplified products generated by the corresponding primers, among which 6 sequences were segments of protein-encoding genes, suggesting RAPD could amplify both protein-encoding and non-protein-encoding gene sequences, and could reveal the genetic variability of different loquat cultivars.

Key words: loquat, RAPD, agarose gel electrophoresis, PAGE

张晓莹;钱剑林;王化坤;宋长年;张彦苹;房经贵. 枇杷RAPD扩增产物的不同电泳检测及其序列特征分析[J]. , 2010, 22(5): 570-575.

ZHANG Xiao-ying;QIAN Jian-lin;WANG Hua-kun;SONG Chang-nian;ZHANG Yan-ping;FANG Jing-gui. Analysis of different electrophoresis and sequence characteristics of RAPD amplification products of loquat[J]. , 2010, 22(5): 570-575.

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