Effects of High Temperature Stress on Gene Expression of Chemosensory Protein in Midgut and Fat Body of Bombyx mori

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

Abstract

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

CLC Number:

S881.24

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.

Figures/Tables 4

References 36

[1]	SALIN C, VERNON P, VANNIER G. Effects of temperature and humidity on transpiration in adults of the lesser mealworm,Alphitobius diaperinus (Coleop tera: Tenebrionidae)[J]. Journal of insect physiology, 1999, 45(10):907-914. doi: 10.1016/S0022-1910(99)00067-0 URL
[2]	HALLMAN G J, DENLINGER D L. Temperature sensitivity in insects and application in integrated pest management[M]. CRC press, 2019.
[3]	李庆荣, 肖阳, 吴福泉, 等. 高温多湿冲击对不同抗性家蚕品种中肠上皮细胞超微结构的影响[J]. 蚕业科学, 2013, 39(5):946-950.
[4]	魏淑花, 吴秀芝, 王颖, 等. 白纹雏蝗抗寒特性和高温耐受力及其机制[J]. 植物保护学报, 2021, 48(1):172-178.
[5]	OH S Y, KIM Y T. Temperature-sensitive paralytic behavior of shibire is altered in cAMP defective mutations of Drosophila[J]. Korean journal of genetics, 2003, 25(2):147-153.
[6]	FABRICE S, JEAN-FRANCOIS F. Temperature affects the ontogeny of sexually dimorphic cuticular hydrocarbons in Drosophila melanogaster[J]. The journal of experimental biology, 2002, 205:3241-3429. doi: 10.1242/jeb.205.20.3241 URL
[7]	张亚南, 樊建庭, 杨林, 等. 35℃下烟青虫夏滞育率、蛹重及代谢速率的变化[J]. 应用昆虫学报, 2014, 51(5):1274-1279.
[8]	CHRISTOPHE S, DAVID R, GUY V, et al. Critical thermal maximum and water loss in developmental stages of the lesser mealworm Alphitobius diaperinus[J]. Acta zoologica sinica, 2006, 52(1):79-86.
[9]	IWONA W. Temperature stress and insect immunity[J]. Journal of thermal biology, 2017, 68:96-103. doi: 10.1016/j.jtherbio.2016.12.002 URL
[10]	杨伟克, 唐芬芬, 刘增虎, 等. 高温和低温条件下琥珀蚕血淋巴SOD及CAT活性的变化[J]. 江苏农业科学, 2017, 45(1):153-155.
[11]	吴福泉, 陈列辉, 刘新涛, 等. 家蚕新品种粤蚕8号(越·春5×航诱7·研7)的育成[J]. 蚕业科学, 2009, 35(4):877-880.
[12]	NING X, XUN B, YAN Y L, et al. Molecular evidence of RNA editing in Bombyx chemosensory protein family[J]. PLOS one, 2014, 9(2):e86932. doi: 10.1371/journal.pone.0086932 URL
[13]	徐世才, 孙勇, 刘伟燕, 等. 桃小食心虫触角气味结合蛋白和化学感受蛋白基因的鉴定与进化分析[J/OL]. 基因组学与应用生物学:1-17. http://kns.cnki.net/kcms/detail/45.1369.Q.20210421.1221.002.html .
[14]	MCKENNA M P, HEKMAT-SCAFE D S, GAINES P, et al. Putative Drosophila pheromone-binding proteins expressed in a subregion of the olfactory system[J]. Jounal of biological chemistry, 1994, 269(23):16340-16347.
[15]	PIKIELNY C W, HASAN G, ROUYER F, et al. Members of a family of Drosophila putative odorant-binding proteins are expressed in different subsets of olfactory hairs[J]. Neuron, 1994, 12(1):35-49. doi: 10.1016/0896-6273(94)90150-3 URL
[16]	徐浩智, 游银伟, 张龙. 昆虫化学感受蛋白及其功能研究进展[J]. 农业生物技术学报, 2015, 23(1):118-125.
[17]	LI M Y, JIANG X Y, LIU X Y, et al. Genome-wide analysis of chemosensory protein genes in the small white butterfly Pieris rapae (Lepidoptera: Pieridae) [J]. Journal of Asia-Pacific Entomology, 2020(23):772-780.
[18]	QIAO H L, DENG P Y, LI D D, et al. Expression analysis and binding experiments of chemosensory proteins indicate multiple roles in bombyx mori[J]. Journal of insect physiology, 2013(59):667-675.
[19]	SABATIER L, JOUANGUY E, DOSTERT C, et al. Pherokine-2 and -3,two Drosophila molecules related to pheromone/odor-binding proteins induced by viral and bacterial infection[J]. European journal of biochemistry, 2003, 270(16) : 3398-3407. doi: 10.1046/j.1432-1033.2003.03725.x URL
[20]	GUO Z, CHENG Z Y, HUANG F, et al. Microarray analysis of global gene regulation in the Cry1Ab-resistant and Cry1Ab-susceptible strains of Diatraea saccharalis[J]. Pest manag science, 2012, 68(5) :718-730. doi: 10.1002/ps.v68.5 URL
[21]	GONG D P, ZHANG H J, ZHAO P, et al. Identification and expression pattern of the chemosensory protein gene family in the silkworm,Bombyx mori[J]. Insect biochemistry and molecular biology, 2007, 37:266-277. doi: 10.1016/j.ibmb.2006.11.012 URL
[22]	邓培渊, 李玉华, 袁丹丹, 等. 家蚕化学感受蛋BmCSP8表达谱及多克隆抗体制备[J]. 中国农学通报, 2014, 30(19):298-303.
[23]	LI Q R, XIAO Y, WU F Q, et al. Analysis of midgut gene expression profiles from different silkworm varieties after exposure to high temperature[J]. Gene, 2014, 549(1):85-96. doi: 10.1016/j.gene.2014.07.050 URL
[24]	李庆荣, 肖阳, 叶明强, 等. 高温高湿胁迫下家蚕不同耐受性品种的中肠组织数字基因表达谱分析[J]. 蚕业科学, 2015, 41(4):620-628.
[25]	李庆荣, 肖阳, 邢东旭, 等. 化学感受蛋白家族基因在家蚕5龄幼虫表皮中的表达分析[J]. 蚕业科学, 2017, 43(1):39-44.
[26]	LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCt method[J]. Methods, 2001, 25(4):402-408. doi: 10.1006/meth.2001.1262 URL
[27]	丁岩钦. 昆虫数学生态学[M]. 北京: 科学出版社, 1994: 318.
[28]	孙雪华, 夏江宝, 吴玉新, 等. 低温对美国白蛾越冬蛹的影响研究[J]. 应用昆虫学报, 2012, 49(5):1263-1267.
[29]	鲍晓文. 补充营养和短期高温对梨小食心虫生殖及成虫寿命的影响[D]. 杨凌:西北农林科技大学, 2010.
[30]	LI J X, MAO T T, WANG H, et al. The CncC/keap1 pathway is activated in high temperature-induced metamorphosis and mediates the expression of Cyp450 genes in silkworm, Bombyx mori[J]. Biochemical and biophysical research communications, 2019, 514:1045-1050. doi: 10.1016/j.bbrc.2019.05.052 URL
[31]	王晓云, 李双宇, 姚磊, 等. 大豆蚜Aghsp75基因对吡虫啉及温度胁迫的应激表达[J]. 中国生物防治学报, 2020, 36(6):913-919.
[32]	姚梦霜, 孟建玉, 杨昌利, 等. 草地贪夜蛾热激蛋白Hsc70基因克隆、表达谱及对不同环境胁迫的响应[J]. 植物保护学报, 2020, 47(4):797-806.
[33]	MIAO Z Q, TU Y Q, GUO P Y, et al. Antioxidant enzymes and heat shock protein genes from Liposcelis bostrychophila are involved in stress defense upon heat shock[J]. Insects, 2020, 11,839.doi: 10.3390/insects11120839. doi: 10.3390/insects11120839 URL
[34]	张虹. 高温胁迫下莲草直胸跳甲的代谢响应机制[D]. 北京:中国农业科学院, 2019.
[35]	WANG Y C, CHANG Y W, BAI J, et al. High temperature stress induces expression of CYP450 genes and contributes to insecticide tolerance in Liriomyza trifolii[J]. Pesticide biochemistry and physiology, 2021, 174:104826. doi: 10.1016/j.pestbp.2021.104826 URL
[36]	鲍忠赞, 张彩霞, 徐世清, 等. 应用基因表达系列分析(SAGE)技术研究高温处理前后家蚕的基因表达差异[J]. 蚕业科学, 2012, 38(3):456-467.

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