草地贪夜蛾取食Cry1Ab和Cry1Fa蛋白后中肠转录组及ABC基因表达分析

doi:10.16380/j.kcxb.2022.09.006

昆虫学报 ›› 2022, Vol. 65 ›› Issue (9): 1127-1135.doi: 10.16380/j.kcxb.2022.09.006

草地贪夜蛾取食Cry1Ab和Cry1Fa蛋白后中肠转录组及ABC基因表达分析

黄晓丹^1,2, 静大鹏², 张天涛², 王振营^2,*

(1. 吉林农业大学生物防治研究所, 天敌昆虫应用技术工程研究中心, 长春 130118; 2. 中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193)

出版日期:2022-09-20 发布日期:2022-10-16

Analysis of the midgut transcriptome and ABC gene expression in Spodoptera frugiperda (Lepidoptera: Noctuidae) after feeding Cry1Ab and Cry1Fa proteins

HUANG Xiao-Dan^1,2, JING Da-Peng², ZHANG Tian-Tao², WANG Zhen-Ying^2,*

(1. Engineering Research Center of Natural Enemy Insects, Institute of Biological Control, Jilin Agricultural University, Changchun 130118, China; 2. State Key Laboratory of Biology of Plant Diseases and Insect Pest, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China)

Online:2022-09-20 Published:2022-10-16

摘要/Abstract

摘要： 【目的】为揭示草地贪夜蛾Spodoptera furgiperda幼虫取食Bt蛋白后与中肠上相关ATP结合盒转运子(ATP-binding cassette transporter, ABC)蛋白基因的表达量变化的关系。【方法】分别使用含活化晶体蛋白Cry1Ab (LC₇₀=240.2 μg/g)和Cry1Fa (LC₇₀=270.0 μg/g)蛋白的人工饲料饲喂草地贪夜蛾4龄幼虫48 h，利用高通量测序对中肠进行转录组测序并进行生物信息学分析，筛选处理后差异表达基因；利用RT-qPCR验证差异表达ABC基因的表达量。【结果】与饲喂正常人工饲料的对照相比，饲喂含240.2 μg/g Cry1Ab和270.0 μg/g Cry1Fa的人工饲料后草地贪夜蛾4龄幼虫中肠转录组中分别检测到1 305和1 202个差异表达基因。Cry1Ab和Cry1Fa处理组与对照组之间分别有994和912个差异表达基因被GO功能注释到生物学过程、分子功能和细胞组分三大类。在最终筛选到的9个差异表达的ABC家族基因中，Cry1Ab处理组与对照组之间有4个差异表达ABC基因，3个上调，1个下调； Cry1Fa处理组与对照组之间有5个差异表达ABC基因，2个上调，3个下调；Cry1Ab和Cry1Fa处理组与对照组之间有2个ABC基因(LOC118267200和LOC118267201)表达量均显著上调。RT-qPCR验证结果表明，与对照组相比，Cry1Ab处理组有3个ABC基因表达量极显著上调，2个ABC基因表达量下调；Cry1Fa处理组有5个ABC基因表达量上调，1个ABC基因表达量下调。【结论】Cry1Ab和Cry1Fa蛋白的摄入可以影响草地贪夜蛾幼虫中肠一些ABC家族基因的表达量变化，这些基因的表达量变化与昆虫抗性产生有关。经比对后发现，ABCC家族与ABCG8基因表达量变化显著。本研究为下一步明确草地贪夜蛾体内ABC转运蛋白在Bt蛋白杀虫机制中的作用，以及合理使用Bt蛋白防治草地贪夜蛾及延缓抗性提供了理论依据。

关键词: 草地贪夜蛾； Bt蛋白, ABC转运蛋白；转录组测序； Cry1Ab； Cry1Fa

Abstract: 【Aim】 To reveal the relationship between the feeding of Bt proteins by Spodoptera frugiperda larvae and the changes in the expression levels of related ABC (ATP-binding cassette transporter) genes in the midgut. 【Methods】 Artificial diets containing Cry1Ab (LC₇₀=240.2 μg/g) or Cry1Fa (LC₇₀=270.0 μg/g) activated crystal protein were used to feed the 4th instar larvae of S. frugiperda for 48 h, respectively. Transcriptome sequencing of midgut and bioinformatics analysis were used to screen differentially expressed genes in the midgut after treatment. RT-qPCR was used to verify the expression levels of the differentially expressed ABC genes. 【Results】 A total of 1 305 and 1 202 differentially expressed genes were detected in the midgut transcriptome of the 4th instar larvae of S. frugiperda fed with the artificial diet containing 240.2 μg/g Cry1Ab and 270.0 μg/g Cry1Fa, respectively, compared with those fed with the normal artificial diet (the control group). There were 994 and 912 differentially expressed genes between the Cry1Ab and Cry1Fa treatment groups and the control group, respectively, and were annotated by GO function into three categories biological process, molecular function and cell component. Among the nine differentially expressed ABC family genes screened, there were four differentially expressed ABC genes (three up-regulated and one down-regulated) between the Cry1Ab treatment group and the control group and five differentially expressed ABC genes (two up-regulated and three down-regulated) between the Cry1Fa treatment group
and the control group. The expression levels of two ABC genes (LOC118267200 and LOC118267201) in the Cry1Ab and Cry1Fa treatment groups were significantly up-regulated as compared to those in the control grpup. RT-qPCR validation results showed that the expression levels of three and two ABC genes in the Cry1Ab treatment group were extremely significantly up-regulated and down-regulated, respectively, and those of five ABC genes and one ABC gene in the Cry1Fa treatment group were up-regulated and down-regulated, respectively, as compared to those in the control grpup. 【Conclusion】 The intake of Cry1Ab and Cry1Fa proteins could affect the expression levels of certain ABC family genes in the midgut of S. frugiperda larvae, and the expression level changes of these genes are related to pest resistance. After comparison, we found that the expression levels of ABCC family and ABCG8 genes changed significantly. This study provides a theoretical basis for further clarifying the role of ABC transporter proteins in the insecticidal mechanism of Bt proteins in S. frugiperda and the rational use of Bt proteins for controlling S. frugiperda and delaying resistance.

Key words: Spodoptera frugiperda, Bt protein, ABC transporter protein, transcriptome sequencing, Cry1Ab, Cry1Fa

黄晓丹, 静大鹏, 张天涛, 王振营. 草地贪夜蛾取食Cry1Ab和Cry1Fa蛋白后中肠转录组及ABC基因表达分析[J]. 昆虫学报, 2022, 65(9): 1127-1135.

HUANG Xiao-Dan, JING Da-Peng, ZHANG Tian-Tao, WANG Zhen-Ying . Analysis of the midgut transcriptome and ABC gene expression in Spodoptera frugiperda (Lepidoptera: Noctuidae) after feeding Cry1Ab and Cry1Fa proteins [J]. Acta Entomologica Sinica, 2022, 65(9): 1127-1135.

草地贪夜蛾取食Cry1Ab和Cry1Fa蛋白后中肠转录组及ABC基因表达分析

Analysis of the midgut transcriptome and ABC gene expression in Spodoptera frugiperda (Lepidoptera: Noctuidae) after feeding Cry1Ab and Cry1Fa proteins

PDF (PC)

赞

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 0

Metrics

本文评价

推荐阅读 0