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    王杰, 贾月慧, 张克中, 崔金腾. 百合雌蕊中杂交亲和性与不亲和性差异表达基因分析[J]. 北京林业大学学报, 2017, 39(2): 82-91. DOI: 10.13332/j.1000-1522.20160292
    引用本文: 王杰, 贾月慧, 张克中, 崔金腾. 百合雌蕊中杂交亲和性与不亲和性差异表达基因分析[J]. 北京林业大学学报, 2017, 39(2): 82-91. DOI: 10.13332/j.1000-1522.20160292
    WANG Jie, JIA Yue-hui, ZHANG Ke-zhong, CUI Jin-teng. Analysis of different expressiongenes between cross-compatibility and cross-incompatibility within pistils of Lilium spp[J]. Journal of Beijing Forestry University, 2017, 39(2): 82-91. DOI: 10.13332/j.1000-1522.20160292
    Citation: WANG Jie, JIA Yue-hui, ZHANG Ke-zhong, CUI Jin-teng. Analysis of different expressiongenes between cross-compatibility and cross-incompatibility within pistils of Lilium spp[J]. Journal of Beijing Forestry University, 2017, 39(2): 82-91. DOI: 10.13332/j.1000-1522.20160292

    百合雌蕊中杂交亲和性与不亲和性差异表达基因分析

    Analysis of different expressiongenes between cross-compatibility and cross-incompatibility within pistils of Lilium spp

    • 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(clpB)、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: The hybridization of Lilium Oriental hybrids and wild lily of Sect. Sinomartagon Comber is the farmost genetic relationship and it's difficult to achieve. Researching the different expression genes in pistil of cross-compatibility and cross-incompatibility may explain the mechanism of causing orientated growth of pollen tube and the molecular mechanism of incompatible hybridization, and provide the basis for overcoming cross-incompatibility. We constructed a forward suppression subtractive hybridization (SSH) library by using the cross-incompatible Lilium Oriental hybrid pistils as the tester, and the cross-compatible ones as the driver. 180 positive clones were randomly screened out. Through sequencing, deleting the bad, removing the redundant and aligning the sequences, 113 ESTs were found to be homologous with known sequences in the bioinformatics databases. RT-PCR was performed to test the 10 selected different expression genes, which showed 1 gene was up-regulated, 7 genes were down-regulated and 2 genes were almost unchanged. Gene annotation and functional category analysis showed that the different expression genes mainly contained signal transduction(including PP2A-3, CDPK and small GTP-binding protein), transport-related(including clpB and CAT2), stress/defense(including ATPase and pyrophosphatase proton pump), protein fate related genes(including 60S ribosomal protein). It was speculated that the incompatibility in the section cross of Oriental hybrids ×Lilium pumilum was probably resulted from the abnormal signal communication between pistils and pollens, the deduced substance transport ability in pistils, the weaker stress resistance ability in pistils after incompatible hybridization.

       

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