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小麦-簇毛麦异附加二体染色体系种质SN6306中与抗白粉病相关基因的转录组分析

Transcriptome analysis of genes related to resistance against powdery mildew in wheat-Thinopyrum alien addition disomic line germplasm SN6306.

作者信息

Li Quanquan, Niu Zubiao, Bao Yinguang, Tian Qiuju, Wang Honggang, Kong Lingrang, Feng Deshun

机构信息

State Key Laboratory of Crop Biology, Shandong Key Laboratory of Crop Biology, College of Agronomy, Shandong Agricultural University, Tai'an 271018, PR China.

State Key Laboratory of Crop Biology, Shandong Key Laboratory of Crop Biology, College of Agronomy, Shandong Agricultural University, Tai'an 271018, PR China.

出版信息

Gene. 2016 Sep 15;590(1):5-17. doi: 10.1016/j.gene.2016.06.005. Epub 2016 Jun 2.

Abstract

Wheat powdery mildew, which is mainly caused by Blumeria graminis f. sp. tritici (Bgt), seriously damages wheat production. The wheat-Thinopyrum intermedium alien addition disomic line germplasm SN6306, being one of the important sources of genes for wheat resistance, is highly resistant to Bgt E09 and to many other powdery mildew physiological races. However, knowledge on the resistance mechanism of SN6306 remains limited. Our study employed high-throughput RNA sequencing based on next-generation sequencing technology (Illumina) to obtain an overview of the transcriptome characteristics of SN6306 and its parent wheat Yannong 15 (YN15) during Bgt infection. The sequencing generated 104,773 unigenes, 9909 of which showed varied expression levels. Among the 9909 unigenes, 1678 unigenes showed 0 reads in YN15. The expression levels in Bgt-inoculated SN6306 and YN15 of exactly 39 unigenes that showed 0 or considerably low reads in YN15 were validated to identify the genes involved in Bgt resistance. Among the 39 unigenes, 12 unigenes were upregulated in SN6306 by 3-45 times. These unigenes mainly encoded kinase, synthase, proteases, and signal transduction proteins, which may play an important role in the resistance against Bgt. To confirm whether the unigenes that showed 0 reads in YN15 are really unique to SN6306, 8 unigenes were cloned and sequenced. Results showed that the selected unigenes are more similar to SN6306 and Th. intermedium than to the wheat cultivar YN15. The sequencing results further confirmed that the unigenes showing 0 reads in YN15 are unique to SN6306 and are most likely derived from Th. intermedium (Host) Nevski. Thus, the genes from Th. intermedium most probably conferred the resistance of SN6306 to Bgt.

摘要

小麦白粉病主要由小麦白粉菌(Blumeria graminis f. sp. tritici,Bgt)引起,严重损害小麦生产。小麦-中间偃麦草异附加二体染色体系种质SN6306是小麦抗性基因的重要来源之一,对Bgt E09及许多其他白粉病生理小种具有高度抗性。然而,关于SN6306抗性机制的了解仍然有限。我们的研究采用基于下一代测序技术(Illumina)的高通量RNA测序,以全面了解SN6306及其亲本小麦扬麦15(YN15)在Bgt侵染过程中的转录组特征。测序产生了104,773个单基因,其中9909个表现出不同的表达水平。在这9909个单基因中,有1678个单基因在YN15中 reads 数为0。对在YN15中reads数为0或相当低的39个单基因在接种Bgt的SN6306和YN15中的表达水平进行验证,以鉴定参与Bgt抗性的基因。在这39个单基因中,有12个单基因在SN6306中上调了3至45倍。这些单基因主要编码激酶、合酶、蛋白酶和信号转导蛋白,它们可能在抗Bgt中发挥重要作用。为了确认在YN15中reads数为0的单基因是否真的是SN6306特有的,克隆并测序了8个单基因。结果表明,所选单基因与SN6306和中间偃麦草的相似性高于与小麦品种YN15的相似性。测序结果进一步证实,在YN15中reads数为0的单基因是SN6306特有的,最有可能来自中间偃麦草(寄主)涅夫斯基。因此,来自中间偃麦草的基因很可能赋予了SN6306对Bgt的抗性。

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