中国农业科学 ›› 2016, Vol. 49 ›› Issue (12): 2353-2367.doi: 10.3864/j.issn.0578-1752.2016.12.011

• 园艺 • 上一篇    下一篇

梨遗传连锁图谱的构建与比较分析

王磊1,2,王龙2,薛华柏2,李秀根2,李疆1

 
  

  1. 1新疆农业大学林学与园艺学院,乌鲁木齐 830052
    2中国农业科学院郑州果树研究所,郑州 450009
  • 收稿日期:2015-12-28 出版日期:2016-06-16 发布日期:2016-06-16
  • 通讯作者: 李秀根,Tel:13653825516;E-mail:lixiugen@caas.cn。李疆,Tel:13899823969;E-mail:lijiangxj@163.com
  • 作者简介:王磊,Tel:18538710151;E-mail:ewlei@163.com
  • 基金资助:
    国家“十二五”科技支撑计划(2013BAD02B01)、中国农业科学院创新工程(CAAS-ASTIP)

Construction of SSR Genetic Linkage Map and Comparison on Pears

WANG Lei1,2, WANG Long2, XUE Hua-bai2, LI Xiu-gen2, LI Jiang1   

  1. 1College of Horticulture & Forestry Sciences, Xinjiang Agricultural University, Urumqi 830052
    2Zhengzhou Fruit Research Institute, Chinese Academy of Agricultural Sciences, Zhengzhou 450009
  • Received:2015-12-28 Online:2016-06-16 Published:2016-06-16

摘要: 【目的】利用已公开发表的梨和苹果的SSR(Simple Sequence Repeat)引物以及从梨转录组开发的SSR引物构建本研究作图群体的遗传连锁图谱,为后期梨重要性状QTL定位和分子标记辅助选择等奠定基础。【方法】以西洋梨品种‘红茄’(Red Clapp Favorite)为母本,东方梨品种‘晚秀’(Mansoo)为父本,构建F1代作图群体。将所选用的SSR引物在亲本和4个子代个体进行PCR扩增,初步筛选出扩增结果符合JoinMap 4.0软件中“CP”作图模式要求的引物,随后在F1群体中检测,选用JoinMap 4.0软件对分离数据进行连锁分析,分别构建亲本的连锁图谱。以双亲图谱在各连锁群上的同源标记作为锚定位点,对双亲图谱进行整合。【结果】利用PCR技术对不同来源的共909对SSR引物(526对梨和283对苹果公开发表的SSR引物,从梨转录组开发的100对SSR引物)进行初步筛选后,发现来自苹果的SSR引物有效扩增片段的比例和多态性均较低,而来自梨和梨转录组开发的SSR引物相对较高。筛选出207对符合作图要求的SSR引物在群体中扩增,构建亲本的连锁图谱。母本图谱中的141个标记分布在17个连锁群上,总长度757.34 cM,标记间平均5.37 cM;父本图谱中的153个标记分布在19个连锁群上,总长度1 149.43 cM,标记间平均7.51 cM。【结论】对不同来源的SSR引物构建的双亲连锁图谱进行整合,最终得到一张由186个SSR标记,覆盖基因组长度1 125.33 cM的整合图谱。

关键词: 梨, SSR, 连锁图谱, 整合图谱

Abstract: 【Objective】The genetic linkage map was constructed based on the Simple Sequence Repeat primers of pears and apples that have been published and developed from the transcriptome of pears by our team, which will be the foundation for the important trait mapping of QTL and marker-assisted selection in the future.【Method】A F1 mapping population was built on a cross between a western pear (Red Clapp Favorite) as the female parent, and an oriental pear (Mansoo) as the male parent. The SSR primers which matched the "CP" mode in the JoinMap 4.0 software in parents and four individuals by PCR application were selected and then tested in the F1 group. The separation of data was analyzed by JoinMap 4.0 software, and the parent genetic map was constructed respectively. The genetic map of the parents was integrated based on anchor points which were homology marks in the parents’ map on each linkages.【Result】909 pairs of SSR primers from different sources (526 pairs from pears and 283 pairs from apples that have been published, and 100 pairs were developed from the pear transcriptome by our team) was preliminarily screened through a PCR technique. The results showed that amplified fragments and polymorphism of SSR primers from apples were lower, but that the SSR primers from pears and developed through the transcriptome of pears were higher. The parents’ genetic linkage map was constructed based on the 207 pairs of SSR primers which conformed to the drawing. A female genetic map was constructed by 141 markers which was distributed on 17 linkage groups and spanned 757.34 cM in genome with an average distance of 5.37 cM between markers; A male genetic map was constructed by 153 markers which was distributed on 19 linkage groups and spanned 1 149.43 cM in genome with an average distance of 7.51 cM between markers.【Conclusion】The integrated genetic linkage maps of parents were constructed by different sources of SSR molecular markers, which contained 186 SSR markers and finally covered the genome 1 125.33 cM.

Key words: pear, SSR, genetic linkage map, integrated the genetic map