Transcriptome analysis of small RNAs in response to Bt insecticidal protein Cry1Ca in Asiatic rice borer Chilo suppressalis.
Transcriptome analysis of small RNAs in response to Bt insecticidal protein Cry1Ca in Asiatic rice borer Chilo suppressalis.
复制标题
DOI：
10.13802/j.cnki.zwbhxb.2021.2021893
复制
发表时间：
2021-01-01
期刊：
影响因子：
--
通讯作者：
Zhang Wei
中科院分区：
文献类型：
--
作者：
Li Zhenzhen;Yao Zhuotian;Zhang Wei
为有效防治二化螟，采用高通量Illumina Solexa测序技术对Bt杀虫蛋白Cry 1Ca处理组和未处理对照组中肠小RNA转录组进行测序，并进行比较分析。通过实时荧光定量PCR验证测序结果。结果表明，Cry 1Ca对C.二化螟为0.061 μ g/g。大多数microRNA序列长度为22 nt。在小RNA数据库中，治疗组和对照组中注释序列的比例都很低，分别仅为7.2%和7.9%。经Cry 1Ca处理后，miRNA、snoRNA、snRNA、rRNA和皮尔纳的数量减少。与对照组相比，治疗组有358个miRNAs表达上调，747个miRNAs表达下调。选择具有最高转录水平的25个已知miRNA用于进一步分析。Cry 1Ca处理组与对照组相比，25种miRNAs的转录水平有显著性差异，其中3种miRNAs的转录水平显著上调，22种miRNAs的转录水平显著下调。在新的miRNAs中，23个上调，43个下调。KEGG分析表明Cry 1Ca诱导的miRNAs参与了丝裂原活化蛋白激酶途径。该通路包含17个靶基因，其中11个受多个miRNAs调控。调控每个靶基因的miRNAs数量在59 ~ 665个之间，相对表达量在0.01 ~ 23289倍之间。这些结果表明，miRNAs在调控C. suppressalis.实时荧光定量PCR结果与测序结果一致，表明测序结果准确。
In order to effectively control Asiatic rice borer Chilo suppressalis, the midgut small RNA transcriptome was sequenced by using high-throughput Illumina Solexa sequencing technology, which was compared and analyzed between the group fed with Bt insecticidal protein Cry1Ca and the control group without Cry1Ca treatment. The sequencing results were verified by quantitative real-time PCR. The results showed that the 30% lethal concentration (LC30) of Cry1Ca against C. suppressalis was 0.061 [mu]g/g. Most microRNA sequences were 22 nt in length. In the small RNA database, the proportions of annotated sequences were very low in both treatment and control groups, only 7.2% and 7.9%, respectively. After treated with Cry1Ca, the numbers of miRNA, snoRNA, snRNA, rRNA and piRNA were decreased. Compared with the control, 358 miRNAs were up-regulated and 747 miRNAs were down-regulated in the treatment group. Twenty-five known miRNAs with the highest transcriptional level were selected for further analysis. The transcriptional levels of these 25 miRNAs were significantly different between the Cry1Ca-treated group and the control, among which, three miRNAs were significantly up-regulated and 22 were significantly down-regulated. Among the novel miRNAs, 23 were upregulated and 43 were down-regulated. KEGG analysis indicated that miRNAs induced by Cry1Ca were involved in the mitogen-activated protein kinase pathway. This pathway contained 17 target genes, of which 11 were regulated by multiple miRNAs. The numbers of miRNAs regulating each target gene ranged from 59 to 665, and the relative expression levels of miRNAs varied from 0.01 times to 23 289 times. These results suggested that miRNAs played an important role in regulation of defense against Bt insecticidal protein Cry1Ca in C. suppressalis. The results of quantitative real-time PCR were consistent with the sequencing results, suggesting an accurate sequencing result.