云南山茶在干旱-复水过程中抗氧化酶活性变化及关键基因差异表达分析

doi:10.3969/j.issn.1004-1524.20221626

浙江农业学报 ›› 2023, Vol. 35 ›› Issue (11): 2611-2620.DOI: 10.3969/j.issn.1004-1524.20221626

云南山茶在干旱-复水过程中抗氧化酶活性变化及关键基因差异表达分析

林先玉¹(), 李紫倩¹, 柏松¹, 罗军¹, 屈燕¹^,²^,^*()

1.西南林业大学园林园艺学院,云南昆明 650224
2.国家林业和草原局西南风景园林工程技术研究中心,云南昆明 650224

收稿日期:2022-12-16 出版日期:2023-11-25 发布日期:2023-12-04
作者简介:林先玉(1995—),女,四川凉山人,硕士研究生,主要从事园林植物资源开发与利用研究。E-mail:1159688949@qq.com
通讯作者: * 屈燕,E-mail:quyan@swfu.edu.cn
基金资助:
“十三五”国家重点研发计划(2019YFD1001005);云南省万人计划青年拔尖人才项目(YNWR-QNBJ-2019-211)

Changes of antioxidant enzyme activity and differential expression of key genes in Camellia reticulata during drought-rehydration process

LIN Xianyu¹(), LI Ziqian¹, BAI Song¹, LUO Jun¹, QU Yan¹^,²^,^*()

1. School of Landscape Architecture, Southwest Forestry University, Kunming 650224, China
2. Southwest Landscape Architecture Engineering Technology Research Center, National Forestry and Grassland Administration, Kunming 650224, China

Received:2022-12-16 Online:2023-11-25 Published:2023-12-04

摘要/Abstract

摘要：

为研究在干旱胁迫及复水过程中云南山茶抗氧化酶活性变化及关键基因的表达差异,本文以半年生云南山茶幼苗为材料,利用聚乙二醇(PEG-6000)对其进行干旱胁迫,测定云南山茶叶片过氧化物酶(POD)和超氧化物歧化酶(SOD)活性,并将对照组和重度干旱组的叶片进行转录组测序。结果表明:在胁迫期间,各处理组SOD活性持续上升,重度干旱胁迫下POD活性先上升后下降;在复水期间,POD、SOD活性都呈下降趋势,除轻度干旱胁迫组外其余两组POD和SOD活性均未能恢复到胁迫之前的水平。通过对关键基因差异表达分析发现,在重度干旱胁迫及复水过程中,33个抗氧化酶相关基因表达量与抗氧化酶活性有极显著相关性(P<0.01),19个显著相关(P<0.05)。POD相关基因均富集于苯丙烷类生物合成通路中,SOD相关基因主要富集于过氧化物酶体通路中,均为下调表达。在胁迫开始后相关基因表达量均发生了显著性变化,说明抗氧化酶相关基因在干旱胁迫及复水过程中参与了调控,积极响应干旱胁迫。

关键词: 云南山茶, 干旱胁迫, 复水, 抗氧化酶, 基因表达

Abstract:

To study the changes of antioxidant enzyme activity and the expression of key genes in Camellia reticulata during drought stress and rehydration process, this paper used semi-annual Camellia reticulata seedlings to determine the peroxidase (POD) and superoxide dismutase (SOD) activities of Camellia reticulata leaves under drought stress using polyethylene glycol (PEG-6000), and transcriptome sequencing was performed on leaves of the control group and the severely drought stress group. The results showed that during the stress period, the SOD activity of each treatment group showed a continuous upward trend, and the POD activity firstly increased and then decreased under severe drought stress. During the rehydration period, the activities of POD and SOD showed a downward trend, except for the mild drought stress group, the POD and SOD activities of the other two groups could not recover to the level before stress. Through the analysis of differential expression of key genes, it was found that the expression of 33 antioxidant enzyme-related genes was very significantly correlated with antioxidant enzyme activity (P<0.01) and 19 antioxidant enzyme-related genes showed significant correlation (P<0.05) during severe drought stress and rehydration. POD-related genes were enriched in the phenylpropane biosynthesis pathway, and SOD-related genes were mainly enriched in the peroxisome pathway, all of which were down-regulated. The relevant expression levels changed significantly after the start of stress, indicating that antioxidant enzyme-related genes participated in the regulation of drought stress and rehydration, and actively responded to drought stress.

Key words: Camellia reticulata, drought stress, rehydration, antioxidant enzymes, gene expression

中图分类号:

S685.14

林先玉, 李紫倩, 柏松, 罗军, 屈燕. 云南山茶在干旱-复水过程中抗氧化酶活性变化及关键基因差异表达分析[J]. 浙江农业学报, 2023, 35(11): 2611-2620.

LIN Xianyu, LI Ziqian, BAI Song, LUO Jun, QU Yan. Changes of antioxidant enzyme activity and differential expression of key genes in Camellia reticulata during drought-rehydration process[J]. Acta Agriculturae Zhejiangensis, 2023, 35(11): 2611-2620.

图/表 9

参考文献 23

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基因ID及编号 Gene ID and number	相关性 Correlation	基因ID及编号 Gene ID and number	相关性 Correlation
Cluster-23907.121858 CrPOD1	-0.714^**	Cluster-23907.193947 CrPOD27	-0.778^**
Cluster-23907.205799 CrPOD2	-0.834^**	Cluster-23907.191538 CrPOD28	0.850^**
Cluster-23907.131313 CrPOD3	-0.705^*	Cluster-23907.186821 CrPOD29	0.816^**
Cluster-23907.133854 CrPOD4	-0.799^**	Cluster-23907.130118 CrPOD30	0.676^*
Cluster-23907.133898 CrPOD5	-0.800^**	Cluster-23907.135693 CrPOD31	0.769^**
Cluster-23907.215532 CrPOD6	-0.689^*	Cluster-23907.226143 CrPOD32	0.605^*
Cluster-23907.216128 CrPOD7	-0.629^*	Cluster-23907.135700 CrPOD33	0.621^*
Cluster-23907.136199 CrPOD8	-0.749^**	Cluster-23907.229292 CrPOD34	0.843^**
Cluster-23907.136924 CrPOD9	-0.839^**	Cluster-23907.172436 CrPOD35	0.765^**
Cluster-23907.141902 CrPOD10	-0.606^*	Cluster-23907.247053 CrPOD36	0.639^*
Cluster-23907.144090 CrPOD11	-0.687^*	Cluster-23907.26403 CrPOD37	0.805^**
Cluster-23907.145111 CrPOD12	-0.805^**	Cluster-23907.177551 CrPOD38	0.746^**
Cluster-23907.149843 CrPOD13	0.686^*	Cluster-23907.86700 CrPOD39	0.688^*
Cluster-23907.157313 CrPOD14	-0.887^**	Cluster-23907.89908 CrPOD40	0.675^*
Cluster-23907.159841 CrPOD15	-0.683^*	Cluster-23907.97564 CrPOD41	0.738^**
Cluster-23907.162853 CrPOD16	-0.848^**	Cluster-23907.185111 CrPOD42	0.626^*
Cluster-23907.169530 CrPOD17	-0.814^**	Cluster-23907.154253 CrSOD1	-0.918^**
Cluster-23907.169635 CrPOD18	-0.888^**	Cluster-23907.154259 CrSOD2	-0.631^*
Cluster-23907.169636 CrPOD19	-0.686^*	Cluster-23907.154756 CrSOD3	-0.837^**
Cluster-23907.227562 CrPOD20	-0.874^**	Cluster-23907.156212 CrSOD4	-0.762^**
Cluster-23907.233931 CrPOD21	-0.768^**	Cluster-23907.158564 CrSOD5	-0.864^**
Cluster-23907.177905 CrPOD22	-0.665^*	Cluster-23907.170549 CrSOD6	-0.847^**
Cluster-23907.179414 CrPOD23	-0.785^**	Cluster-23907.183528 CrSOD7	-0.789^**
Cluster-23907.180565 CrPOD24	-0.732^**	Cluster-23907.187659 CrSOD8	-0.843^**
Cluster-23907.183161 CrPOD25	-0.695^*	Cluster-23907.204201 CrSOD9	-0.808^**
Cluster-23907.80759 CrPOD26	-0.706^*	Cluster-23907.210282 CrSOD10	-0.742^**

基因ID及编号 Gene ID and number	相关性 Correlation	基因ID及编号 Gene ID and number	相关性 Correlation
Cluster-23907.121858 CrPOD1	-0.714^**	Cluster-23907.193947 CrPOD27	-0.778^**
Cluster-23907.205799 CrPOD2	-0.834^**	Cluster-23907.191538 CrPOD28	0.850^**
Cluster-23907.131313 CrPOD3	-0.705^*	Cluster-23907.186821 CrPOD29	0.816^**
Cluster-23907.133854 CrPOD4	-0.799^**	Cluster-23907.130118 CrPOD30	0.676^*
Cluster-23907.133898 CrPOD5	-0.800^**	Cluster-23907.135693 CrPOD31	0.769^**
Cluster-23907.215532 CrPOD6	-0.689^*	Cluster-23907.226143 CrPOD32	0.605^*
Cluster-23907.216128 CrPOD7	-0.629^*	Cluster-23907.135700 CrPOD33	0.621^*
Cluster-23907.136199 CrPOD8	-0.749^**	Cluster-23907.229292 CrPOD34	0.843^**
Cluster-23907.136924 CrPOD9	-0.839^**	Cluster-23907.172436 CrPOD35	0.765^**
Cluster-23907.141902 CrPOD10	-0.606^*	Cluster-23907.247053 CrPOD36	0.639^*
Cluster-23907.144090 CrPOD11	-0.687^*	Cluster-23907.26403 CrPOD37	0.805^**
Cluster-23907.145111 CrPOD12	-0.805^**	Cluster-23907.177551 CrPOD38	0.746^**
Cluster-23907.149843 CrPOD13	0.686^*	Cluster-23907.86700 CrPOD39	0.688^*
Cluster-23907.157313 CrPOD14	-0.887^**	Cluster-23907.89908 CrPOD40	0.675^*
Cluster-23907.159841 CrPOD15	-0.683^*	Cluster-23907.97564 CrPOD41	0.738^**
Cluster-23907.162853 CrPOD16	-0.848^**	Cluster-23907.185111 CrPOD42	0.626^*
Cluster-23907.169530 CrPOD17	-0.814^**	Cluster-23907.154253 CrSOD1	-0.918^**
Cluster-23907.169635 CrPOD18	-0.888^**	Cluster-23907.154259 CrSOD2	-0.631^*
Cluster-23907.169636 CrPOD19	-0.686^*	Cluster-23907.154756 CrSOD3	-0.837^**
Cluster-23907.227562 CrPOD20	-0.874^**	Cluster-23907.156212 CrSOD4	-0.762^**
Cluster-23907.233931 CrPOD21	-0.768^**	Cluster-23907.158564 CrSOD5	-0.864^**
Cluster-23907.177905 CrPOD22	-0.665^*	Cluster-23907.170549 CrSOD6	-0.847^**
Cluster-23907.179414 CrPOD23	-0.785^**	Cluster-23907.183528 CrSOD7	-0.789^**
Cluster-23907.180565 CrPOD24	-0.732^**	Cluster-23907.187659 CrSOD8	-0.843^**
Cluster-23907.183161 CrPOD25	-0.695^*	Cluster-23907.204201 CrSOD9	-0.808^**
Cluster-23907.80759 CrPOD26	-0.706^*	Cluster-23907.210282 CrSOD10	-0.742^**

基因ID Gene ID	引物序列 Primer sequence(5'-3')
Cluster-23907.105087	F:GGTGAGAAGCGACCAATGGA
	R:TGGGTGGCTTGTGTTCCATT
Cluster-23907.21994	F:CGGGAAGCTGAGATGGAAGG
	R:ATCAAATCCCCGACGTTGGT
Cluster-23907.22854	F:TCTCCTTGGTGTAGCTCCGA
	R:ACATTCATTCCGCCTCAGCA
Cluster-23907.128010	F:GCTTGAACGAGCCTTTCACC
	R:TCACGAAGTTTGCGGAGGAA
Cluster-23907.131185	F:CCATCAACTCTGGCGTCACT
	R:TTCAGAGCCTTGCCGAGAAG
Cluster-23907.35810	F:CGTTCTCTCCGTTGCCTTCT
	R:CTCCGAGAGTGTTCCAGAGC
Cluster-23907.43736	F:GCCGGAGTTTCCTTTGAGGA
	R:AGAGAACCAGCTTCGTTGGG
Cluster-23907.45879	F:TGGCGTGGAAGTTCATCCTC
	R:ACCGAATTTCTCCTGCCCTG
Cluster-23907.42603	F:GGGAACAATGAGGGCTCACA
	R:GAGGTGCCAGAGGATGATGG

基因ID Gene ID	引物序列 Primer sequence(5'-3')
Cluster-23907.105087	F:GGTGAGAAGCGACCAATGGA
	R:TGGGTGGCTTGTGTTCCATT
Cluster-23907.21994	F:CGGGAAGCTGAGATGGAAGG
	R:ATCAAATCCCCGACGTTGGT
Cluster-23907.22854	F:TCTCCTTGGTGTAGCTCCGA
	R:ACATTCATTCCGCCTCAGCA
Cluster-23907.128010	F:GCTTGAACGAGCCTTTCACC
	R:TCACGAAGTTTGCGGAGGAA
Cluster-23907.131185	F:CCATCAACTCTGGCGTCACT
	R:TTCAGAGCCTTGCCGAGAAG
Cluster-23907.35810	F:CGTTCTCTCCGTTGCCTTCT
	R:CTCCGAGAGTGTTCCAGAGC
Cluster-23907.43736	F:GCCGGAGTTTCCTTTGAGGA
	R:AGAGAACCAGCTTCGTTGGG
Cluster-23907.45879	F:TGGCGTGGAAGTTCATCCTC
	R:ACCGAATTTCTCCTGCCCTG
Cluster-23907.42603	F:GGGAACAATGAGGGCTCACA
	R:GAGGTGCCAGAGGATGATGG

云南山茶在干旱-复水过程中抗氧化酶活性变化及关键基因差异表达分析

Changes of antioxidant enzyme activity and differential expression of key genes in Camellia reticulata during drought-rehydration process

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