Analysis of Different Resistant Gossypium hirsutum Inoculated with Verticillium dahliae Infection by Small RNA and Degradome Sequencing
Analysis of Different Resistant Gossypium hirsutum Inoculated with Verticillium dahliae Infection by Small RNA and Degradome Sequencing
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小RNA和降解组测序分析接种大丽黄萎病的不同抗性陆地棉
DOI：
10.1134/s1021443722040112
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发表时间：
2022-07
影响因子：
1.4
通讯作者：
陈勋基
中科院分区：
文献类型：
--
作者：
李晓荣;陈方圆;王剑;胡文冉;李波;杨洋;倪志勇;陈勋基
摘要本研究旨在鉴定不同抗病棉花品种中参与调控棉花黄萎病防御反应的miRNAs及其潜在靶点。构建了陆地棉(抗病品种‘中棉-49’和感病品种‘君棉-1号’)对照样品和病根的RNA文库和降解组文库。利用高通量小RNA测序技术对两个不同抗性的陆地棉品种在不同胁迫模式下的miRNAs进行了鉴定和分析。用变性DNA测序法检测切割后的靶基因。用实时定量逆转录-聚合酶链式反应(qRT-PCR)验证miRNAs，并检测与生长发育和抗性相关的靶基因。在所有文库中共鉴定出69个已知miRNAs和330个新miRNAs。通过比较对照样品和大丽叶霉应激库中miRNAs的表达水平，发现‘中棉49’有23个差异表达的miRNAs，‘军棉1号’有62个差异表达的miRNAs。此外，通过降解组分析，在615个目的基因中确定了615个降解点。生物信息学分析进一步表明，基因可能参与了棉花生长发育和抗性过程中的许多细胞和分子过程的调控。最后，qRT-PCR进一步确定了8个可能在棉花生长和抗性调控中发挥作用的目的基因。
Abstract This study aims to identify miRNAs and their potential targets involved in regulating the defense response of cotton verticillium wilt in different resistant Gossypium hirsutum varieties. RNA libraries and degradome libraries from control samples and infected roots of G. hirsutum (resistant cotton variety ‘Zhongmian-49’ and susceptible cotton variety ‘Junmian-1’) were constructed. High-throughput small RNA sequencing was used to identify and analyze miRNAs from two different resistant G. hirsutum varieties under different stress modes. Degradome sequencing was used to detect cleaved target genes. The real-time reverse transcription-PCR (qRT-PCR) was used to verify the miRNAs and detect target genes related to growth, development and resistance. A total of 69 known miRNAs and 330 novel miRNAs were identified in all libraries. By comparing the expression levels of miRNAs in control samples and V. dahliae stress libraries, we found that there are 23 differentially expressed miRNAs in ‘Zhongmian-49’, and 62 differentially expressed miRNAs in ‘Junmian-1’. In addition, 615 degradation sites were identified in 615 target genes by degradome analysis. Bioinformatics analysis further showed that genes might be involved in the regulation of many cellular and molecular processes in the growth, development and resistance process of cotton. Finally, qRT-PCR further confirmed 8 target genes with potential roles in cotton growth and resistance regulation.
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