利用DDRT-PCR鉴定芸芥自交不亲和系与自交亲和系的差异表达cDNA

doi:10.7668/hbnxb.2016.03.007

摘要/Abstract

摘要： 芸芥与芸薹属植物具有亲缘关系,也是芸薹属植物的重要育种资源。自交亲和性是芸芥的一种重要变异性状。为了探明芸芥自交亲和基因的表达器官,并分离与自交亲和性有关的cDNA片段,以芸芥的一对近等基因系,自交亲和系(SC)与自交不亲和系(SI)为试材,利用差异显示RT-PCR技术分别对开花前和开花后的叶片、花药和柱头进行鉴定。结果表明,不论开花前还是开花后,芸芥自交亲和系与自交不亲和系的叶片或花药间的扩增带型是一样的,但在近等基因系的柱头间得到了差异表达条带。这证明芸芥自交亲和基因不是组成型表达,而是组织特异性表达,柱头是其唯一的表达器官。在开花前的自交亲和系中共得到了2条cDNA片段,一条小于100 bp,另一条为750~1000 bp,在开花后的自交亲和系中得到了1条300 bp的cDNA片段。据分析这3条cDNA片段可能与芸芥的自交亲和性密切相关。在开花前的自交不亲和系中共得到了2条cDNA片段,一条为500 bp,另一条为750 bp,同时在开花后的自交不亲和系中得到了1条500~600 bp的cDNA片段。这些cDNA片段可以用来区分芸芥的自交亲和系与自交不亲和系,还可用于克隆自交亲和基因。

关键词: 芸芥, 自交亲和系, 自交不亲和系, DDRT-PCR

Abstract: Yunjie is closely related to important vegetables and oil seed crops,and can be considered a genetic resource for all Brassiceae crops.Self compatibility is an important variant traits to Yunjie.This study aimed to probe the expression organ of self-compatible gene and isolate cDNA fragments related to self-compatibility of Yunjie.Leaf,anther and stigma from self-incompatible (SI) and self-compatible (SC) lines of Yunjie were examined by DDRT-PCR.The results showed that the amplification pattern of leaves and anthers were the same in SI and SC lines.However,different bands were obtained in stigmas of SI and SC lines before and after flowering.SC gene of E.sativa was not showing constitutive expression but showed tissue-specific expression in stigma.Only two fragments were amplified in SC lines before flowering,the one was less than 100 bp,the other was 750-1 000 bp.A 300 bp fragment was found unique to self-compatible line after flowering.These three cDNA fragments might be closely related to the self-compatibility in Yunjie.In addition,two cDNA fragments were generated in SI line before flowering,one was 500 bp,the other was 750 bp.One band of 500-600 bp was identified in SI line after flowering.These cDNA fragments could efficiently distinguish self-compatible and self-incompatible lines,and could also be useful for cloning self-compatible gene of Yunjie.

Key words: Yunjie, Self-compatibility, Self-incompatibility, DDRT-PCR

中图分类号:

范惠玲, 白生文, 李华清, 孙万仓. 利用DDRT-PCR鉴定芸芥自交不亲和系与自交亲和系的差异表达cDNA[J]. 华北农学报, 2016, 31(3): 44-50. doi: 10.7668/hbnxb.2016.03.007.

FAN Huiling, BAI Shengwen, LI Huaqing, SUN Wancang. Identification of cDNA Fragments in Self-incompatible and Self-compatible Lines of Eruca sativa Mill.by DDRT-PCR[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2016, 31(3): 44-50. doi: 10.7668/hbnxb.2016.03.007.

http://www.hbnxb.net/CN/Y2016/V31/I3/44

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