百合雌蕊中杂交亲和性与不亲和性差异表达基因分析

发布时间：2018-04-17 21:51

本文选题：百合 + 杂交不亲和　；参考：《北京林业大学学报》2017年02期

【摘要】：东方百合与卷瓣组野生百合杂交(简称OS组合),属于亲缘关系最远的组间杂交,杂交障碍最大。挖掘雌蕊中杂交亲和性与不亲和性差异表达基因,有可能解释不亲和性杂交中引起花粉管定向生长异常的机理,进而阐明不亲和性杂交的分子机理,为实施克服杂交不亲和障碍技术措施提供依据。本研究采用抑制消减杂交技术(SSH技术),分别以不亲和性杂交和亲和性杂交的东方百合雌蕊为试验组(tester)和驱动组(driver),建立正向抑制消减杂交文库,随机挑选180个阳性克隆,经过测序、去劣、去冗余、序列比对,有113个EST与数据库中的已知序列具有同源性,最终获得10个差异表达基因。采用RT-PCR技术对10个差异表达基因进行验证,其中有1个基因在不亲和性杂交的雌蕊中表达上调、7个基因表达下调、2个基因无明显变化。对差异表达基因进行功能注释及分类,差异表达基因主要集中于信号转导(包括丝氨酸/苏氨酸蛋白磷酸酶PP2A-3、钙依赖蛋白激酶CDPK、小G结合蛋白)、抗逆防御(包括分子伴侣(clp B)、过氧化氢酶CAT2)、转运(包括质膜ATPase、焦磷酸酶质子泵)、蛋白命运(包括60S核糖体蛋白)等功能类别。据此推测,在东方百合×山丹组(系)间不亲合性杂交育种过程中,可能引起雌蕊与花粉管信号交流异常、物质转运能力减弱和雌蕊耐受逆境能力降低,从而导致杂交不亲和性。
[Abstract]:Hybrid between Oriental Lilium and Wild Lilium curl group (abbreviated as OS combination) is the most distant cross, and the cross obstacle is the biggest.Digging out the differentially expressed genes between incompatibility and incompatibility in pistil may explain the mechanism of pollen tube directional growth abnormality in incompatibility hybridization, and then elucidate the molecular mechanism of incompatibility hybridization.To provide the basis for the implementation of technical measures to overcome the barrier of cross incompatibility.In this study, the suppression subtractive hybridization (SSH) technique was used to establish a positive suppression subtractive hybridization library, and 180 positive clones were randomly selected.After sequencing, inferiority, redundancy removal, sequence alignment, 113 EST have homology with known sequences in the database, and finally 10 differentially expressed genes were obtained.Ten differentially expressed genes were verified by RT-PCR technique. Among them, 1 gene was up-regulated in pistil of incompatibility hybridization, 7 genes were down-regulated, and 2 genes had no obvious change.Functional annotation and classification of differentially expressed genes,Differentially expressed genes are mainly concentrated in signal transduction (including serine / threonine protein phosphatase PP2A-3, calcium-dependent protein kinase (CDPKK), small G-binding protein, anti-stress defense (including molecular chaperone, catalase CAT2, and transport).Plasma membrane Pase, pyrophosphatase proton pump, protein fate (including 60s ribosomal protein) and other functional categories.It is inferred that, in the process of incompatibility hybrids between Lilium oriental 脳 Shandan group (lines), it may lead to abnormal signal exchange between pistil and pollen tube, decrease of material transport ability and decrease of pistil tolerance to stress.This leads to cross incompatibility.
【作者单位】：北京农学院园林学院;城乡生态环境北京实验室;
【基金】：北京市科技提升计划项目(TJSHG201310020020) 北京林果业生态环境功能提升协同创新中心项目(PXM2016-014207-000038) 北京市属高等学校创新团队建设项目(IDHT20150503) 北京市自然科学基金项目(6122004)
【分类号】：S682.29

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