两类新疆狗牙根抗旱基因型抗氧化酶保护系统及其基因表达差异分析

doi:10.11686/cyxb2021178

摘要/Abstract

摘要：

新疆狗牙根具发达的根状茎和细长的匍匐茎，在耐旱性方面表现出广泛的遗传变异特性，对抗旱型C138和不抗旱型C32狗牙根两个基因型进行自然干旱7 d后复水3 d处理，明确2个基因型在干旱胁迫及复水下抗氧化酶系统生理响应及基因表达水平上的差异。结果表明：干旱胁迫降低了狗牙根草坪表观质量、叶片相对含水量和叶绿素含量，电导率及丙二醛（malondialdehyde， MDA）含量升高，且C32较C138下降或升高更为明显。随干旱时间延长，狗牙根体内超氧阴离子自由基（O₂^·-）、过氧化氢（H₂O₂）、MDA含量均显著上升，且C32较C138升高更显著，而超氧化物歧化酶（superoxide dismutase， SOD）、过氧化物酶（peroxidase， POD）和过氧化氢酶（catalase， CAT）活性呈先升后降趋势，抗坏血酸过氧化物酶（ascorbate peroxidase， APX）活性呈增加趋势。抗旱型C138对比不抗旱型C32表现为SOD、POD、APX、DHAR和GPX等抗氧化基因高水平的表达，而不抗旱型C32较C138表现出高的脂质过氧化和低的抗氧化酶活性及基因表达水平；在复水过程中，POD活性在提高抗氧化能力及清除活性氧（reactive oxygen species， ROS）过程中起着至关重要的作用。干旱胁迫诱导抗氧化酶及其相关基因上调表达，增强了细胞内的抗氧化防御能力，有助于狗牙根维持细胞膜稳定性，延缓叶片的脱水，从而提高了狗牙根的抗旱能力。

关键词: 干旱胁迫, 狗牙根, 抗氧化系统, 基因表达

Abstract:

Xinjiang bermudagrass ecotypes have well-developed rhizomes and slender stolons， and exhibit extensive genetic variation in drought tolerance. In this study drought tolerant （C138） and drought-sensitive （C32） bermudagrass genotypes were exposed to natural drought for 7 days followed by 3 days of rehydration treatment and the differences in physiological response and gene expression levels of the antioxidant enzyme systems under drought stress and rehydration were evaluated. It was found that drought stress reduced the apparent quality of bermudagrass lawn， relative leaf water content and chlorophyll content， increased electrical conductivity and malondialdehyde （MDA） content， and these responses were expressed more significantly in C32 than in C138. With increased duration of drought exposure， the levels of O₂^·-， H₂O₂， and MDA in bermudagrass leaves increased significantly， and C32 increased more significantly than C138， while activities of superoxide dismutase （SOD）， peroxidase （POD） and catalase （CAT） initially increased and then decreased， and ascorbate peroxidase （APX） activities increased. Drought-resistant C138 and non-drought-resistant C32 showed high-levels of expression of antioxidant genes such as SOD， POD， APX， DHAR and GPX， while non-drought-resistant C32 showed the highest lipid peroxidation and the lowest antioxidant enzyme activity and gene expression levels. In the process of rehydration， POD activity plays a vital role in improving antioxidant capacity and removing reactive oxygen species. Drought stress induces the up-regulation of the expression of antioxidant enzymes and related genes， enhances the antioxidant defense in cells， helps bermudagrass maintain cell membrane stability， delays the dehydration of leaves， and improves the bermudagrass drought resistance ability.

Key words: drought stress, bermudagrass, antioxidant system, gene expression

曾令霜, 李培英, 孙宗玖, 孙晓梵. 两类新疆狗牙根抗旱基因型抗氧化酶保护系统及其基因表达差异分析[J]. 草业学报, 2022, 31(7): 122-132.

Ling-shuang ZENG, Pei-ying LI, Zong-jiu SUN, Xiao-fan SUN. Analysis of antioxidant enzyme protection systems and gene expression differences in two Xinjiang bermudagrass genotypes with contrasting drought resistance[J]. Acta Prataculturae Sinica, 2022, 31(7): 122-132.

图/表 5

表 1 实时荧光定量引物信息

Table 1 Real-time fluorescence quantitative primer information

基因 Gene	正/反向 Forward/reverse	引物序列 Primer sequences （5'-3'）	扩增产物大小 Amplication product size （bp）
Cu/Zn-SOD	Forward	AGCGAGCGACTTACAATGGC	208
	Reverse	AACACTGAAGGCGTGGCTGA	208
2-Cys POD	Forward	TTTGATCCCTGATCAGGGCA	185
	Reverse	GCCTGAAGAGTCCTCATGGT	185
GPX 4	Forward	TAAGTGAAGCCAGTATCAGC	230
	Reverse	AGGAAAGCAGATGAGAAGAC	230
DHAR	Forward	GCAATGTGCGGGATGAAGAT	260
	Reverse	GCTCCAATGAAACCTGGGCT	260
CAT	Forward	CCATGAGATCAAGGCCATCT	103
	Reverse	ATCTTACATGCTCGGCTTGG	103
Cyt-APX	Forward	GGTTCAATTAGATACGAGGAAGAGT	129
	Reverse	GGCAAGCTGATGAAGGTCTG	129
内参基因 Actin	Forward	GCTCAACCCCAAGGCTAAC	186
	Reverse	AGAGCGTATCCCTCGTAGATG	186

图1 在干旱胁迫和非胁迫条件下，两种不同的耐旱狗牙根基因型的表型（A）、土壤水分（B）和草坪质量（C）的变化竖线表示在采样日之间比较处理之间的最小显著差异值（P<0.05），C138-CK、C32-CK表示C138、C32材料正常浇水处理，C138-D、C32-D表示C138、C32材料干旱-复水处理，Re-2 d表示复水第2天，下同。Vertical bars indicate least significant difference values （P<0.05） for the comparison among treatments at the sampling day. C138-CK and C32-CK indicate normal watering treatment for C138 and C32 materials， and C138-D and C32-D indicate drought-rewatering treatment for C138 and C32 materials， Re-2 means the second day of rewatering. The same below.

Fig.1 Change in the phenotypes （A）， soil water content （B） and turf quality （C） of two different drought tolerant bermudagrass genotypes under stressed and non-stressed conditions

图2 干旱胁迫下狗牙根膜透性及氧化产物的变化

Fig.2 Changes of bermudagrass membrane permeability and oxidation products under drought stress

图3 干旱胁迫下狗牙根抗氧化酶活性的变化

Fig.3 Changes of antioxidant enzyme activities of bermudagrass under drought stress

图4 干旱胁迫下狗牙根抗氧化酶基因的表达变化1表示C138； 2表示C32； Re表示复水。*表示同一天数不同基因型存在显著差异（P<0.05）。 1 means C138； 2 means C32； Re means rewatering. * indicates that there are significant different drought tolerant bermudagrass at the same day （P<0.05）.

Fig.4 Changes of antioxidant enzyme gene expression in bermudagrass under drought stress

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