高温胁迫下火龙果转录组及热激蛋白响应分析

doi:10.3969/j.issn.1004-1524.20230843

浙江农业学报 ›› 2024, Vol. 36 ›› Issue (5): 1067-1075.DOI: 10.3969/j.issn.1004-1524.20230843

高温胁迫下火龙果转录组及热激蛋白响应分析

李俊成¹(), 党芸芝², 孙清明¹^,^*()

1.广东省农业科学院果树研究所/农业农村部南亚热带果树生物学与遗传资源利用重点实验室/广东省热带亚热带果树研究重点实验室,广东广州 510640
2.广东第二师范学院,广东广州 510303

收稿日期:2023-07-10 出版日期:2024-05-25 发布日期:2024-05-29
作者简介:李俊成(1988—),男,河南信阳人,博士,研究方向为火龙果遗传育种。E-mail:lijuncheng@gdaas.cn
通讯作者: * 孙清明,E-mail: sunqingming@gdaas.cn
基金资助:
国家自然科学基金(32102341);广州市科技计划(202201010527)

Transcriptome analysis and HSP response of pitaya (Selenicereus spp.) under heat stress

LI Juncheng¹(), DANG Yunzhi², SUN Qingming¹^,^*()

1. Institute of Fruit Tree Research, Guangdong Academy of Agricultural Sciences/Key Laboratory of South Subtropical Fruit Biology and Genetic Resource Utilization, Ministry of Agriculture and Rural Affairs/Guangdong Provincial Key Laboratory of Tropical and Subtropical Fruit Tree Research, Guangzhou 510640, China
2. Guangdong University of Education, Guangzhou 510303, China

Received:2023-07-10 Online:2024-05-25 Published:2024-05-29

摘要/Abstract

摘要：

为探究火龙果(Selenicereus spp.)高温胁迫后基因表达变化,研究火龙果高温胁迫响应的关键基因,挖掘耐高温基因资源。本研究通过RNA-seq对25 ℃及40 ℃处理24 h的火龙果枝条进行测序,从系统生物学角度对差异表达基因进行分析。结果表明,火龙果在遭遇40 ℃胁迫24 h后,与25 ℃胁迫相比,枝条中有2 408个基因的表达发生显著变化,其中基因包括165个转录因子,占基因总量的6.7%;表达量上调2~587倍或者下调2~59倍。热激蛋白(HSP)在胁迫后表达量上调尤为显著,差异表达基因中上调程度最高的10个基因均为HSP,可能具有重要的胁迫耐受作用。通过qPCR对不同胁迫时间样品该10个HSP表达量检测发现, HSP18.2,HSP21,HSP22,HSP23.5,HSP23.6在胁迫5 h后表达量即上调至50倍以上,尤其是HSP21,在胁迫5 h后表达量即上调391倍,在10 h时表达量达到最高水平,上调589倍,响应速度高于其他基因。HSP的快速响应可能和火龙果的高温耐受性有关,HSP21可以作为火龙果高温胁迫的信号基因。

关键词: 火龙果, 高温胁迫, 热激蛋白, 转录组

Abstract:

To investigate the changes in gene expression under high temperature stress of pitaya, we studied key genes that respond to such stress and screened heat-resistant gene resources. In this study, RNA-seq was used to sequence pitaya stems treated at 25 ℃ and 40 ℃ for 24 h, and differentially that under were analyzed from the perspective of systems biology. The results showed that after pitaya was subjected to 40 ℃ for 24 h, and 2 408 genes showed significant changes compared with expression relative to 25 ℃, including 165 transcription factors, accounting for 6.7% of the total number of genes; the expression level was up-regulated by 2 to 587 times or down-regulated by 2 to 59 times. Heat shock proteins (HSPs) were particularly up-regulated after stress, and the top 10 up-regulated genes in differentially expressed genes were all HSPs, which may have important stress tolerance effects. By qPCR detection of the expression levels of these 10 HSPs in samples at different stress time, it was found that HSP18.2, HSP21, HSP22, HSP23.5 and HSP23.6 were up-regulated by more than 50 times after stress for 5 h, especially HSP21, which was up-regulated by 391 times after stress for 5 h and reached the highest level after stress for 10 h with an up-regulation of 589 times. The rapid response of HSP may be related to the high temperature tolerance of pitaya. HSP21 can be used as a signal gene for high temperature stress of pitaya.

Key words: pitaya, heat stress, heat shock protein, transcriptome

中图分类号:

S667.9

李俊成, 党芸芝, 孙清明. 高温胁迫下火龙果转录组及热激蛋白响应分析[J]. 浙江农业学报, 2024, 36(5): 1067-1075.

LI Juncheng, DANG Yunzhi, SUN Qingming. Transcriptome analysis and HSP response of pitaya (Selenicereus spp.) under heat stress[J]. Acta Agriculturae Zhejiangensis, 2024, 36(5): 1067-1075.

图/表 9

参考文献 24

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基因名称 Gene ID	正向引物/反向引物 Forward primers/Reverse primers
HSP17.6II	TGTGAGGATGGAGAGGAGGATGG/GGAGCCACCTTCTGAGCAGTAAC
HSP23.6	AGATGGTTAGGGCTGACGATGATG/CTGGCTCACGCTCCGAGTTG
HSP21	CGAAGGCTCCTTGATGTCTCTCC/CCCGTTCCTCCTGGCATTGTC
HSP17.4	TCGGGAGAAGGAGGAGAAGACC/TCGGAACGGTCACAGTCAACAC
HSP17.8	AGCATAACCACCATCATCGGAAGG/AGCATACGAACCGCCCAAGTC
HSP18.2	GGAGGTCAAAGTGGAGGTGGAAG/GCATTCTCAGGGAGGCGGAAC
HSP22.0	TTGGACTCGGTCAAGGCTAAGC/CACTACCTTCACCACCTTCAATGC
HSP17.6C	CTCCCACTCCGCCCTCTCTC/TCCACCTTGACCTCCTCCTTCC
HSP17.6B	CCCTGATTCCCACATCTACTCTGC/GTCTTCCGCCTCCACCTTGATC
HSP23.5	AGATGGTTAGGGCTGACGATGATG/CTGGCTCACGCTCCGAGTTG

基因名称 Gene ID	正向引物/反向引物 Forward primers/Reverse primers
HSP17.6II	TGTGAGGATGGAGAGGAGGATGG/GGAGCCACCTTCTGAGCAGTAAC
HSP23.6	AGATGGTTAGGGCTGACGATGATG/CTGGCTCACGCTCCGAGTTG
HSP21	CGAAGGCTCCTTGATGTCTCTCC/CCCGTTCCTCCTGGCATTGTC
HSP17.4	TCGGGAGAAGGAGGAGAAGACC/TCGGAACGGTCACAGTCAACAC
HSP17.8	AGCATAACCACCATCATCGGAAGG/AGCATACGAACCGCCCAAGTC
HSP18.2	GGAGGTCAAAGTGGAGGTGGAAG/GCATTCTCAGGGAGGCGGAAC
HSP22.0	TTGGACTCGGTCAAGGCTAAGC/CACTACCTTCACCACCTTCAATGC
HSP17.6C	CTCCCACTCCGCCCTCTCTC/TCCACCTTGACCTCCTCCTTCC
HSP17.6B	CCCTGATTCCCACATCTACTCTGC/GTCTTCCGCCTCCACCTTGATC
HSP23.5	AGATGGTTAGGGCTGACGATGATG/CTGGCTCACGCTCCGAGTTG

基因名称 Gene ID	正向引物/反向引物 Forward primers/Reverse primers(5'—3')
HSFB2B	AGGAGGAGGAGGAGGATGAATGTC/TTGTTCTGGAGGCGTCTGTAGTG
HSFA3	CCTCAATCTCAGCAGCTCACTACC/GCACTCTTCTCCCTCCTCAACTTC
HSFA2	TGGGCTGGAAGAAGAGGTAGAGAG/AGTTGCCTGCTGCTGCTGTC
HSFB2A	AGCAGCAACAATAACAGCAGCAG/CTCAACCTCTCGTTCTCGTCCATC
HSF4	ACAGACGAAGAGACAGTGTGACG/TGACATTCCAACTCCGCCCTTG
HSFA7A	AGTGGGTAGATTCGGGTTTGATGG/TGCTCTTGTGTTCTGCTGTTGTTG
HSFA8	GCTGGATGGGGTGATGGTGAG/ACAATGGAGCCGGAGTTGATGAG

基因名称 Gene ID	正向引物/反向引物 Forward primers/Reverse primers(5'—3')
HSFB2B	AGGAGGAGGAGGAGGATGAATGTC/TTGTTCTGGAGGCGTCTGTAGTG
HSFA3	CCTCAATCTCAGCAGCTCACTACC/GCACTCTTCTCCCTCCTCAACTTC
HSFA2	TGGGCTGGAAGAAGAGGTAGAGAG/AGTTGCCTGCTGCTGCTGTC
HSFB2A	AGCAGCAACAATAACAGCAGCAG/CTCAACCTCTCGTTCTCGTCCATC
HSF4	ACAGACGAAGAGACAGTGTGACG/TGACATTCCAACTCCGCCCTTG
HSFA7A	AGTGGGTAGATTCGGGTTTGATGG/TGCTCTTGTGTTCTGCTGTTGTTG
HSFA8	GCTGGATGGGGTGATGGTGAG/ACAATGGAGCCGGAGTTGATGAG

样品编号 Sample ID	Reads数量 Reads number	GC含量 GC content/%	Q30/%	比对率 Mapping rate/%
25 ℃-1	35 028 564	45.27	96.78	81.35
25 ℃-2	36 523 020	45.65	96.89	78.29
25 ℃-3	30 664 466	45.47	96.74	80.98
40 ℃-1	31 077 180	44.84	96.79	79.60
40 ℃-2	31 374 155	44.25	96.73	77.03
40 ℃-3	33 091 371	44.74	96.71	76.99

高温胁迫下火龙果转录组及热激蛋白响应分析

Transcriptome analysis and HSP response of pitaya (Selenicereus spp.) under heat stress

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图/表 9

参考文献 24

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基因名称 Gene ID	上调倍数 Up-regulated	基因名称 Gene ID	下调倍数 Down-regulated
HSP22.0	587.071 4	Lipoprotein	-23.272 97
HSP21	558.786 2	GH9B8	-24.179 93
HSP17.6C	541.206 3	HISTONE 3.1	-24.574 07
HSP17.4	531.965 9	PPT2	-24.814 61
HSP18.2	500.420 2	IPMI1	-27.424 14
HSP23.5	404.966 4	ATLP-1	-31.941 21
HSP17.6B	392.680 5	FMO GS-OX3	-34.497 69
HSP17.8	267.216 9	ENODL15	-38.873 75
HSP23.6	215.171 0	HTA6	-39.878 57
HSP17.6II	177.829 3	DIR1-LIKE	-59.320 59

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