高温胁迫对化学感受蛋白在家蚕中肠与脂肪体中基因表达的影响

doi:10.11924/j.issn.1000-6850.casb2021-0673

摘要/Abstract

摘要：

通过探讨高温胁迫对家蚕化学感受蛋白基因表达水平的影响,为家蚕高温耐受机理的阐明及耐高温品种选育奠定基础,本实验对BmCSPs基因序列进行聚类分析,并通过荧光定量PCR方法对高温胁迫不同时间后BmCSPs在中肠和脂肪体内的表达情况进行检测。实验结果表明,16个BmCSPs基因依据序列相似性主要聚为2个大类,其中BmCSP15在中肠、脂肪体中受高温胁迫的大部分时间点均出现表达上调,而BmCSP7、BmCSP9和BmCSP14在这两个组织中均出现表达下调,推测BmCSP15与家蚕对高温的耐受性密切相关,BmCSP7、BmCSP9和BmCSP14与高温敏感性相关,基因表达量检测结果与基因序列的聚类结果大致吻合。由此可见,在家蚕受高温胁迫前后BmCSPs基因发生了转录水平的变化,其参与家蚕对高温胁迫的应答,筛选出的差异表达基因可作为家蚕高温响应的候选基因。

关键词: 高温胁迫, 家蚕, 化学感受蛋白, 基因表达

Abstract:

To lay a foundation for clarifying the mechanism of high temperature tolerance of silkworm and breeding high temperature-resistant silkworm varieties, the effect of high temperature stress on the expression of chemosensory proteins (CSPs) in Bombyx mori was studied. In this research, cluster analysis of gene sequences of BmCSPs was carried out, and the expression of BmCSPs in midgut and fat body subjected to high temperature stress for different time was detected by fluorescence quantitative PCR. The results showed that the 16 BmCSPs genes were mainly clustered into two groups according to sequence similarity. Among them, the expression level of BmCSP15 was up-regulated in midgut and fat body at most time points under high temperature stress, while that of BmCSP7, BmCSP9 and BmCSP14 were down-regulated in these tissues. It was speculated that BmCSP15 was closely related to the tolerance to high temperature in silkworm, and BmCSP7, BmCSP9 and BmCSP14 might be correlated with the sensitivity to high temperature. The gene expression results were roughly consistent with the clustering results of gene sequences. From the results, it can be seen that the transcription levels of BmCSPs changed before and after high temperature stress, indicating that BmCSPs are involved in the response of silkworm to high temperature stress. The differentially expressed genes screened in the study can be used as candidate genes for high temperature response of silkworm.

Key words: high temperature stress, Bombyx mori, chemosensory protein, gene expression

中图分类号:

S881.24

肖阳, 李庆荣, 邢东旭, 杨琼. 高温胁迫对化学感受蛋白在家蚕中肠与脂肪体中基因表达的影响[J]. 中国农学通报, 2022, 38(6): 107-115.

XIAO Yang, LI Qingrong, XING Dongxu, YANG Qiong. Effects of High Temperature Stress on Gene Expression of Chemosensory Protein in Midgut and Fat Body of Bombyx mori[J]. Chinese Agricultural Science Bulletin, 2022, 38(6): 107-115.

图/表 4

参考文献 36

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基因及登录号	引物序列(5’-3’)	扩增产物大小/bp
BmCSP1(BGIBMGA004045)	F:TGTACTCGTGGCCGTACTTG R:AGTTCCTTCCCTTCGGGAGT	150
BmCSP2(BGIBMGA004067)	F:AAAAGTCGATTGCCAAGGCG R:TTGGTTCCAGCTTTCCGGTT	158
BmCSP3(NM_001043598.2)	F:CCCCTGACGCTAAGGAGTTG R:TTCGGGGTCGTATTTAGCGG	164
BmCSP4(NM_001043587.2)	F:GCGAGAAATGCACTGAAGCC R:TAGCGCGGTCTTCGTACTTC	144
BmCSP5(NM_001043597.1)	F:GAAAAGGGCCAATGTACCGC R:ATTTGGCCTTCAGCTCCTCC	169
BmCSP6(NM_001043935.2)	F:TGTCTTGCCGCTGATCTCTC R:GCAGTTGGTTGCAATGGTGT	168
BmCSP7(NM_001043603.1)	F:TCCTGAAGCTGTGGCAGAAG R:AAGAATTAGCGACGGCGGAT	176
BmCSP8(BGIBMGA004040)	F:TATCGGTCGTTGTTTGCCGA R:AATTCTGTTCCTTCGGCGGT	151
BmCSP9(BGIBMGA004035)	F:TCATCGCTCTGGCTGTAGTG R:AGCTGTGCACTTTCCCTTGT	155
BmCSP10(NM_001043599.1)	F:GATCCTCGAGTCCAACCGAC R:TTGTCAGCTCCCGACTTCTG	159
BmCSP11(BGIBMGA004044)	F:AAGGACCGTGTACCCCAGAT R:CTTCGGGATGGCGTTCGATA	141
BmCSP12(BGIBMGA004047)	F:AATCGTCCCGGTGCTCAAAT R:GGGGTGCATGGGTTTTTGTC	147
BmCSP13(NM_001043715.2)	F:GCCGTACTGGCTCAAGACAA R:TCGCACAATGGGTCTCCAAA	181
BmCSP14(NM_001043727.2)	F:GCAGCAGAGCTATCCCAGAA	180
BmCSP14(NM_001043727.2)	R:TTATTAGAGCACCGCGCACA	180
BmCSP15(NM_001098311.1 )	F:TTTGGTACAGAACCCGAGGC R:TCTTCCTGGCGAATCGCTTT	161
BmCSP16(BGIBMGA001469)	F:CCAGGTCACAGACACAGCTT R:TGAGCATTGAGGACAAGCCC	160
sw22934（DQ443290）	F:TCCAAAAATGGGCCATCGAA R:TGCTGGATTGCAGAAGGTTT	179

基因及登录号	引物序列(5’-3’)	扩增产物大小/bp
BmCSP1(BGIBMGA004045)	F:TGTACTCGTGGCCGTACTTG R:AGTTCCTTCCCTTCGGGAGT	150
BmCSP2(BGIBMGA004067)	F:AAAAGTCGATTGCCAAGGCG R:TTGGTTCCAGCTTTCCGGTT	158
BmCSP3(NM_001043598.2)	F:CCCCTGACGCTAAGGAGTTG R:TTCGGGGTCGTATTTAGCGG	164
BmCSP4(NM_001043587.2)	F:GCGAGAAATGCACTGAAGCC R:TAGCGCGGTCTTCGTACTTC	144
BmCSP5(NM_001043597.1)	F:GAAAAGGGCCAATGTACCGC R:ATTTGGCCTTCAGCTCCTCC	169
BmCSP6(NM_001043935.2)	F:TGTCTTGCCGCTGATCTCTC R:GCAGTTGGTTGCAATGGTGT	168
BmCSP7(NM_001043603.1)	F:TCCTGAAGCTGTGGCAGAAG R:AAGAATTAGCGACGGCGGAT	176
BmCSP8(BGIBMGA004040)	F:TATCGGTCGTTGTTTGCCGA R:AATTCTGTTCCTTCGGCGGT	151
BmCSP9(BGIBMGA004035)	F:TCATCGCTCTGGCTGTAGTG R:AGCTGTGCACTTTCCCTTGT	155
BmCSP10(NM_001043599.1)	F:GATCCTCGAGTCCAACCGAC R:TTGTCAGCTCCCGACTTCTG	159
BmCSP11(BGIBMGA004044)	F:AAGGACCGTGTACCCCAGAT R:CTTCGGGATGGCGTTCGATA	141
BmCSP12(BGIBMGA004047)	F:AATCGTCCCGGTGCTCAAAT R:GGGGTGCATGGGTTTTTGTC	147
BmCSP13(NM_001043715.2)	F:GCCGTACTGGCTCAAGACAA R:TCGCACAATGGGTCTCCAAA	181
BmCSP14(NM_001043727.2)	F:GCAGCAGAGCTATCCCAGAA	180
BmCSP14(NM_001043727.2)	R:TTATTAGAGCACCGCGCACA	180
BmCSP15(NM_001098311.1 )	F:TTTGGTACAGAACCCGAGGC R:TCTTCCTGGCGAATCGCTTT	161
BmCSP16(BGIBMGA001469)	F:CCAGGTCACAGACACAGCTT R:TGAGCATTGAGGACAAGCCC	160
sw22934（DQ443290）	F:TCCAAAAATGGGCCATCGAA R:TGCTGGATTGCAGAAGGTTT	179