Effects of High Temperature Stress on Antioxidant Enzyme Activity and Gene Expression in Larvae of Silkworm Varieties with Different Tolerance

doi:10.11924/j.issn.1000-6850.casb2021-1144

Abstract

Abstract:

The effects of high temperature environment on antioxidant enzyme activity and gene expression in hemolymph of silkworm were explored, which may provide reference for revealing the physiological and molecular adaptation mechanism of silkworm to high temperature environmental stress. The tolerant variety to high temperature 932G and sensitive variety HY were taken as the studied objects, and the changes of antioxidant enzyme activities and enzyme gene expression levels in the hemolymph of the 5th instar larvae under high temperature stress were measured by real-time fluorescence quantitative PCR. The results showed that under 35℃ high temperature stress, the change trend of relative activities of SOD, CAT and GST in tolerant variety and sensitive variety were similar. The change ranges were all small in the early stage of stress, then increased significantly in the middle and late stages, and decreased significantly in the final stage of stress. The overall change trend was first increase then decrease. The relative expression levels of enzyme genes increased in varying degrees in the middle and late stages of stress, and decreased in the final stage of stress, which were basically consistent with the change trend of enzyme activities, but the change ranges among the varieties were different. The relative expression levels of BmSOD and most of BmGSTs genes increased significantly in tolerant variety, while that of BmCAT increased significantly in sensitive variety. In conclusion, the high temperature response trend of antioxidant enzymes SOD, CAT and GST in silkworm hemolymph are relatively consistent, and there are only differences in the change ranges among the varieties.

Key words: high temperature stress, tolerance, antioxidant enzyme activity, gene expression, Bombyx mori

CLC Number:

S881.24

XIAO Yang, LI Qingrong, XING Dongxu, YANG Qiong. Effects of High Temperature Stress on Antioxidant Enzyme Activity and Gene Expression in Larvae of Silkworm Varieties with Different Tolerance[J]. Chinese Agricultural Science Bulletin, 2022, 38(35): 111-118.

Figures/Tables 4

References 31

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基因	正向引物序列(5’-3’)	正向引物序列(5’-3’)
BmSOD	CAGCTGGAGCTCATTTCAAC	TAAAGTGCGACCAATGATGC
BmCAT	GACAGGGAGCGTATTCCAGA	CCACTCTCTCCACCAACTGT
BmGST 1(GSTe1)	CCTGAAGGCCTGGCCAAAAT	TAGGCAAGAGCGCATCCAAA
BmGST 2(GSTs1)	GGAAAGCTGACATGGGGTGA	AAGCCTTCACTTTGGGCTGT
BmGST 4(GSTz1)	CACTGGATCACAAGGGGCTT	CAGTTCACGGTCTATGCGGA
BmGST 5(GSTe2)	CGCAGATATCCAGGAAGCGT	ATCGACTGGCAACACACAGT
BmGST 6(GSTo2)	ACTGCCGCAAGATCCTTTGA	AGAACACAGTGCCTCGGTTT
BmGST 11(GSTe5)	TTGCCGGTGATGAGTTCTCC	ATCGGCAGTTGAGAGGAACG
BmGST 12(GSTe6)	TGATCAAGAACATCGAGGATGC	TCTGTTTATCCATTTGCTTTTCTCA
BmGST 13(GSTo4)	CGTGGCACAAGATTTCTCGG	GTATTGACTGACGGCGGGAT
BmGST d1	CTCAGCACACAATCCCGACT	GTCGATAATGGCACGTTGGC
BmGST d2	TGGTGAACAAGTACGCCAAAG	ACCTTCTCGTTCTTGGCCTTG
BmGST d3	CGCTTTGGACCTACAGCTCA	ACTGCTACTGTCTCCTCCGT
sw22934	TCCAAAAATGGGCCATCGAA	TGCTGGATTGCAGAAGGTTT

基因	正向引物序列(5’-3’)	正向引物序列(5’-3’)
BmSOD	CAGCTGGAGCTCATTTCAAC	TAAAGTGCGACCAATGATGC
BmCAT	GACAGGGAGCGTATTCCAGA	CCACTCTCTCCACCAACTGT
BmGST 1(GSTe1)	CCTGAAGGCCTGGCCAAAAT	TAGGCAAGAGCGCATCCAAA
BmGST 2(GSTs1)	GGAAAGCTGACATGGGGTGA	AAGCCTTCACTTTGGGCTGT
BmGST 4(GSTz1)	CACTGGATCACAAGGGGCTT	CAGTTCACGGTCTATGCGGA
BmGST 5(GSTe2)	CGCAGATATCCAGGAAGCGT	ATCGACTGGCAACACACAGT
BmGST 6(GSTo2)	ACTGCCGCAAGATCCTTTGA	AGAACACAGTGCCTCGGTTT
BmGST 11(GSTe5)	TTGCCGGTGATGAGTTCTCC	ATCGGCAGTTGAGAGGAACG
BmGST 12(GSTe6)	TGATCAAGAACATCGAGGATGC	TCTGTTTATCCATTTGCTTTTCTCA
BmGST 13(GSTo4)	CGTGGCACAAGATTTCTCGG	GTATTGACTGACGGCGGGAT
BmGST d1	CTCAGCACACAATCCCGACT	GTCGATAATGGCACGTTGGC
BmGST d2	TGGTGAACAAGTACGCCAAAG	ACCTTCTCGTTCTTGGCCTTG
BmGST d3	CGCTTTGGACCTACAGCTCA	ACTGCTACTGTCTCCTCCGT
sw22934	TCCAAAAATGGGCCATCGAA	TGCTGGATTGCAGAAGGTTT

基因	品种	高温胁迫时间/h
基因	品种	1	6	12	18	24	36	48
BmSOD	932G	1.04±0.04^C	0.76±0.09^D*	0.55±0.06^E*	1.00±0.09^C*	1.79±0.32^B*	5.54±0.48^A*	5.24±0.51^A*
BmSOD	HY	0.91±0.10^c	0.24±0.02^d	0.15±0.03^e	0.14±0.02^e	1.06±0.10^c	2.59±0.36^b	3.63±0.41^a
BmCAT	932G	0.93±0.05^B	0.82±0.09^B	0.24±0.09^C*	0.25±0.04^C*	0.32±0.04^C*	0.35±0.03^C*	1.10±0.09^A
BmCAT	HY	0.95±0.11^b	0.87±0.02^b	0.47±0.15^c	0.37±0.05^c	0.84±0.08^b	1.13±0.12^a	1.35±0.12^a
BmGST 1	932G	0.97±0.05^D	0.76±0.09^E	0.55±0.09^F*	0.40±0.04^G*	5.61±0.38^C*	9.19±0.82^B*	11.82±0.88^A*
BmGST 1	HY	1.31±0.11^f	1.22±0.02^f	1.89±0.15^e	2.71±0.28^d	22.20±3.05^c	35.19±2.28^a	28.17±1.96^b
BmGST 2	932G	0.93±0.03^D	1.60±0.12^C	2.80±0.23^B*	7.29±0.50^A*	6.84±0.59^A	4.16±0.43^B*	3.45±0.48^B*
BmGST 2	HY	1.04±0.28^e	1.36±0.15^de	1.54±0.16^d	1.82±0.11^c	6.96±0.10^b	11.05±0.82^a	7.33±0.49b
BmGST 4	932G	0.86±0.12^F	1.35±0.11^E*	2.56±0.28^D*	4.09±0.38^C*	5.26±0.45^B*	7.83±0.63^A*	7.65±0.32^A*
BmGST 4	HY	1.16±0.21^bc	0.98±0.07^c	1.28±0.13^b	1.32±0.15^b	1.52±0.14^ab	1.83±0.24^a	1.34±0.10^b
BmGST 5	932G	1.17±0.09^G	7.64±0.11^F*	24.08±0.90^E*	82.23±1.62^C*	105.73±2.82^A*	99.00±1.37^B*	76.20±1.08^D*
BmGST 5	HY	1.09±0.05^g	2.30±0.13^f	3.84±0.24^e	8.67±0.38^d	31.45±0.46^c	67.67±0.95^b	70.16±1.04^a
BmGST 6	932G	1.01±0.05^EF*	1.05±0.04^E*	0.94±0.05^F*	3.47±0.06^D*	6.82±0.09^B*	7.29±0.11^A*	4.11±0.05^C*
BmGST 6	HY	1.17±0.04^d	0.82±0.06^e	0.57±0.04^g	0.68±0.03^f	1.92±0.06^b	2.41±0.08^a	1.60±0.10^c
BmGST 7	932G	1.07±0.05^G*	4.24±0.06^F*	23.90±0.91^E*	96.56±1.42^D*	278.08±8.63^C*	497.83±12.59^A*	401.51±10.27^B*
BmGST 7	HY	0.86±0.05^g	6.04±0.12^f	27.31±1.01^e	75.33±1.58^d	102.96±3.65^c	156.52±5.02^a	138.28±8.48^b
BmGST 8	932G	0.93±0.02^D*	0.73±0.12^E	1.25±0.28^F*	1.41±0.29^G*	31.04±0.87^C*	68.38±1.54^A*	57.56±1.98^B*
BmGST 8	HY	0.99±0.02^d	0.75±0.03^f	0.86±0.02^e	0.88±0.04^e	14.46±0.66^c	34.22±1.05^a	28.29±0.83^b
BmGST 9	932G	1.06±0.05^F	1.16±0.06^F*	2.22±0.03^D*	1.53±0.02^E*	44.26±0.95^C*	137.00±4.83^A*	116.53±3.60^B*
BmGST 9	HY	0.97±0.04^d	0.83±0.02^e	0.78±0.05^e	0.56±0.03^f	26.62±0.73^c	37.36±0.77^a	32.37±1.28^b
BmGST 10	932G	1.03±0.06^G	2.35±0.27^F*	5.48±0.32^E*	11.62±0.36^D*	30.27±0.80^C*	39.54±1.02^A*	34.82±0.54^B*
BmGST 10	HY	1.01±0.03^d	0.88±0.01^e	0.32±0.02^f	0.16±0.01^g	12.78±0.45^c	48.92±1.03^a	25.26±0.86^b
BmGST 11	932G	1.02±0.02^D*	0.72±0.01^E	0.36±0.02^F*	0.12±0.01^G*	47.83±0.94^C*	169.53±4.37^A*	132.69±3.82^B*
BmGST 11	HY	1.03±0.02^e	0.72±0.02^f	0.66±0.01^g	1.31±0.04^d	17.74±0.65^c	61.25±2.72^a	47.08±1.59^b
BmGST 12	932G	0.92±0.03^F*	0.86±0.01^F*	1.44±0.04^E*	2.29±0.13^D*	5.84±0.20^A*	5.01±0.14^B*	3.47±0.09^C*
BmGST 12	HY	1.02±0.04^g	1.37±0.05^f	3.45±0.04^c	6.35±0.15^a	4.33±0.11^b	1.68±0.03^e	2.28±0.05^d
BmGST 13	932G	0.93±0.02^G	1.88±0.06^F	2.50±0.08^E*	7.37±0.18^D*	562.35±10.33^C*	3583.07±42.80^A*	1022.04±35.27^B*
BmGST 13	HY	0.98±0.03^g	1.64±0.02^f	2.27±0.09^e	5.89±0.16^d	103.36±1.08^c	247.71±4.64^a	198.18±5.29^b
BmGST d1	932G	0.97±0.05^F	1.36±0.06^D*	1.15±0.08^E*	0.83±0.02^G*	4.94±0.10^C*	25.36±0.62^B*	28.27±0.58^A*
BmGST d1	HY	1.03±0.03^g	0.89±0.02^f	2.33±0.05^e	4.46±0.08^d	9.52±0.17^c	33.41±0.88^a	22.73±0.92^b
BmGST d2	932G	1.02±0.01^F	1.11±0.05^E*	0.92±0.03^G*	2.48±0.07^D*	6.44±0.13^C*	29.77±0.46^A*	26.02±0.59^B*
BmGST d2	HY	1.02±0.02^c	0.78±0.01^d	0.21±0.01^f	0.03±0.00^g	0.57±0.01^e	1.40±0.03^b	1.86±0.05^a
BmGST d3	932G	0.98±0.02^D	1.03±0.01^C	0.91±0.02^E*	0.55±0.02^F*	1.03±0.04^CD*	2.05±0.05^B*	2.28±0.04^A
BmGST d3	HY	0.98±0.01d	0.52±0.02e	0.13±0.01g	0.26±0.02f	1.67±0.03c	3.39±0.24a	2.08±0.16b

基因	品种	高温胁迫时间/h
基因	品种	1	6	12	18	24	36	48
BmSOD	932G	1.04±0.04^C	0.76±0.09^D*	0.55±0.06^E*	1.00±0.09^C*	1.79±0.32^B*	5.54±0.48^A*	5.24±0.51^A*
BmSOD	HY	0.91±0.10^c	0.24±0.02^d	0.15±0.03^e	0.14±0.02^e	1.06±0.10^c	2.59±0.36^b	3.63±0.41^a
BmCAT	932G	0.93±0.05^B	0.82±0.09^B	0.24±0.09^C*	0.25±0.04^C*	0.32±0.04^C*	0.35±0.03^C*	1.10±0.09^A
BmCAT	HY	0.95±0.11^b	0.87±0.02^b	0.47±0.15^c	0.37±0.05^c	0.84±0.08^b	1.13±0.12^a	1.35±0.12^a
BmGST 1	932G	0.97±0.05^D	0.76±0.09^E	0.55±0.09^F*	0.40±0.04^G*	5.61±0.38^C*	9.19±0.82^B*	11.82±0.88^A*
BmGST 1	HY	1.31±0.11^f	1.22±0.02^f	1.89±0.15^e	2.71±0.28^d	22.20±3.05^c	35.19±2.28^a	28.17±1.96^b
BmGST 2	932G	0.93±0.03^D	1.60±0.12^C	2.80±0.23^B*	7.29±0.50^A*	6.84±0.59^A	4.16±0.43^B*	3.45±0.48^B*
BmGST 2	HY	1.04±0.28^e	1.36±0.15^de	1.54±0.16^d	1.82±0.11^c	6.96±0.10^b	11.05±0.82^a	7.33±0.49b
BmGST 4	932G	0.86±0.12^F	1.35±0.11^E*	2.56±0.28^D*	4.09±0.38^C*	5.26±0.45^B*	7.83±0.63^A*	7.65±0.32^A*
BmGST 4	HY	1.16±0.21^bc	0.98±0.07^c	1.28±0.13^b	1.32±0.15^b	1.52±0.14^ab	1.83±0.24^a	1.34±0.10^b
BmGST 5	932G	1.17±0.09^G	7.64±0.11^F*	24.08±0.90^E*	82.23±1.62^C*	105.73±2.82^A*	99.00±1.37^B*	76.20±1.08^D*
BmGST 5	HY	1.09±0.05^g	2.30±0.13^f	3.84±0.24^e	8.67±0.38^d	31.45±0.46^c	67.67±0.95^b	70.16±1.04^a
BmGST 6	932G	1.01±0.05^EF*	1.05±0.04^E*	0.94±0.05^F*	3.47±0.06^D*	6.82±0.09^B*	7.29±0.11^A*	4.11±0.05^C*
BmGST 6	HY	1.17±0.04^d	0.82±0.06^e	0.57±0.04^g	0.68±0.03^f	1.92±0.06^b	2.41±0.08^a	1.60±0.10^c
BmGST 7	932G	1.07±0.05^G*	4.24±0.06^F*	23.90±0.91^E*	96.56±1.42^D*	278.08±8.63^C*	497.83±12.59^A*	401.51±10.27^B*
BmGST 7	HY	0.86±0.05^g	6.04±0.12^f	27.31±1.01^e	75.33±1.58^d	102.96±3.65^c	156.52±5.02^a	138.28±8.48^b
BmGST 8	932G	0.93±0.02^D*	0.73±0.12^E	1.25±0.28^F*	1.41±0.29^G*	31.04±0.87^C*	68.38±1.54^A*	57.56±1.98^B*
BmGST 8	HY	0.99±0.02^d	0.75±0.03^f	0.86±0.02^e	0.88±0.04^e	14.46±0.66^c	34.22±1.05^a	28.29±0.83^b
BmGST 9	932G	1.06±0.05^F	1.16±0.06^F*	2.22±0.03^D*	1.53±0.02^E*	44.26±0.95^C*	137.00±4.83^A*	116.53±3.60^B*
BmGST 9	HY	0.97±0.04^d	0.83±0.02^e	0.78±0.05^e	0.56±0.03^f	26.62±0.73^c	37.36±0.77^a	32.37±1.28^b
BmGST 10	932G	1.03±0.06^G	2.35±0.27^F*	5.48±0.32^E*	11.62±0.36^D*	30.27±0.80^C*	39.54±1.02^A*	34.82±0.54^B*
BmGST 10	HY	1.01±0.03^d	0.88±0.01^e	0.32±0.02^f	0.16±0.01^g	12.78±0.45^c	48.92±1.03^a	25.26±0.86^b
BmGST 11	932G	1.02±0.02^D*	0.72±0.01^E	0.36±0.02^F*	0.12±0.01^G*	47.83±0.94^C*	169.53±4.37^A*	132.69±3.82^B*
BmGST 11	HY	1.03±0.02^e	0.72±0.02^f	0.66±0.01^g	1.31±0.04^d	17.74±0.65^c	61.25±2.72^a	47.08±1.59^b
BmGST 12	932G	0.92±0.03^F*	0.86±0.01^F*	1.44±0.04^E*	2.29±0.13^D*	5.84±0.20^A*	5.01±0.14^B*	3.47±0.09^C*
BmGST 12	HY	1.02±0.04^g	1.37±0.05^f	3.45±0.04^c	6.35±0.15^a	4.33±0.11^b	1.68±0.03^e	2.28±0.05^d
BmGST 13	932G	0.93±0.02^G	1.88±0.06^F	2.50±0.08^E*	7.37±0.18^D*	562.35±10.33^C*	3583.07±42.80^A*	1022.04±35.27^B*
BmGST 13	HY	0.98±0.03^g	1.64±0.02^f	2.27±0.09^e	5.89±0.16^d	103.36±1.08^c	247.71±4.64^a	198.18±5.29^b
BmGST d1	932G	0.97±0.05^F	1.36±0.06^D*	1.15±0.08^E*	0.83±0.02^G*	4.94±0.10^C*	25.36±0.62^B*	28.27±0.58^A*
BmGST d1	HY	1.03±0.03^g	0.89±0.02^f	2.33±0.05^e	4.46±0.08^d	9.52±0.17^c	33.41±0.88^a	22.73±0.92^b
BmGST d2	932G	1.02±0.01^F	1.11±0.05^E*	0.92±0.03^G*	2.48±0.07^D*	6.44±0.13^C*	29.77±0.46^A*	26.02±0.59^B*
BmGST d2	HY	1.02±0.02^c	0.78±0.01^d	0.21±0.01^f	0.03±0.00^g	0.57±0.01^e	1.40±0.03^b	1.86±0.05^a
BmGST d3	932G	0.98±0.02^D	1.03±0.01^C	0.91±0.02^E*	0.55±0.02^F*	1.03±0.04^CD*	2.05±0.05^B*	2.28±0.04^A
BmGST d3	HY	0.98±0.01d	0.52±0.02e	0.13±0.01g	0.26±0.02f	1.67±0.03c	3.39±0.24a	2.08±0.16b