低温胁迫对3份葛种质萌发过程中脯氨酸和抗坏血酸-谷胱甘肽循环代谢的影响

doi:10.3969/j.issn.1004-1524.20230863

浙江农业学报 ›› 2024, Vol. 36 ›› Issue (7): 1558-1568.DOI: 10.3969/j.issn.1004-1524.20230863

低温胁迫对3份葛种质萌发过程中脯氨酸和抗坏血酸-谷胱甘肽循环代谢的影响

曾洪学(), 屈兴红^*()

浙江同济科技职业学院水利工程学院,浙江杭州 311231

收稿日期:2023-07-11 出版日期:2024-07-25 发布日期:2024-08-05
作者简介:曾洪学(1974—),男,江西乐安人,硕士,副教授,主要从事植物生理和植物生态及设施农业方面的研究工作。E-mail: zenghongxue@zjtongji.edu.cn
通讯作者: *屈兴红,E-mail: quxinghong@zjtongji.edu.cn
基金资助:
浙江省水利厅科研项目

Effect of low-temperature stress on proline metabolism and ascorbic acid-glutathione cycle during germination of three Pueraria lobata germplasm lines

ZENG Hongxue(), QU Xinghong^*()

School of Hydraulic Engineering, Zhejiang Tongji Vocational College of Science and Technology, Hangzhou 311231, China

Received:2023-07-11 Online:2024-07-25 Published:2024-08-05

摘要/Abstract

摘要：

为探究不同温度处理对葛种子萌发时脯氨酸代谢和抗坏血酸-谷胱甘肽(AsA-GSH)循环的影响,以来自澳大利亚(AUS),以及中国江苏(JS)和湖南(HN)的3份葛种质为研究对象,通过纸上发芽试验,研究不同处理(CK,25 ℃;LT,15 ℃;LTI1,10 ℃;LTI2,0.1% H₂O₂浸种+10 ℃)下3份葛种质脯氨酸含量、脯氨酸代谢关键酶活性、AsA-GSH循环代谢中非酶抗氧化剂含量和酶抗氧化剂活性的变化。结果表明,与CK相比,低温胁迫下葛的丙二醛(MDA)、过氧化氢(H₂O₂)含量升高,脯氨酸脱氢酶(ProDH)活性降低,添加H₂O₂促使脯氨酸、(还原型)抗坏血酸(AsA)、氧化型抗坏血酸(DHA)、(还原型)谷胱甘肽(GSH)、氧化型谷胱甘肽(GSSG)含量增加,Δ¹-吡咯林-5-羧酸合成酶(P5CS)、鸟氨酸转氨酶(OAT)、抗坏血酸过氧化物酶(APX)和谷胱甘肽还原酶(GR)活性增强,提高葛藤的耐低温性能。结合隶属函数分析,判定3份葛种质的耐低温性从强到弱依次为HN>AUS>JS。

关键词: 葛, 低温胁迫, 脯氨酸代谢

Abstract:

To investigate the effects of different temperature on proline metabolism and ascorbic acid-glutathione (AsA-GSH) circulatory system during germination of Pueraria lobata seeds, three lines of P. lobata collected from Australian (AUS), and Jiangsu Province (JS), Hunan Province (HN) in China, were subjected to germination tests by using the paper interval method to study the changes in proline content, key enzymes activities of proline metabolism, and contents of non-enzymatic antioxidants and activities of enzymatic antioxidants in the metabolism of AsA-GSH cycle under different treatments (CK, 25 ℃; LT, 15 ℃; LTI1, 10 ℃; LTI2, seed soaking by 0.1% H₂O₂+10 ℃). The results showed that compared to the CK, low temperature stress increased contents of malondialdehyde (MDA), hydrogen peroxide (H₂O₂) and decreased proline dehydrogenase (ProDH) activity. Application of H₂O₂ increased the contents of proline, ascorbic acid (AsA), dehydro ascorbic acid (DHA), glutathione (GSH), glutathione disulfide (GSSG), and activities of Δ¹-pyrroline-5-carboxylic acid synthase (P5CS), ornithine aminotransferase (OAT), ascorbate peroxidase (APX) and glutathione reductase (GR), hence improved the low-temperature resistance of P. lobata. Based on the membership function analysis, the three P. lobata germplasm lines were ranked as HN>AUS>JS in order of cold-tolerance.

Key words: Pueraria lobata, low temperature stress, proline metabolism

中图分类号:

曾洪学, 屈兴红. 低温胁迫对3份葛种质萌发过程中脯氨酸和抗坏血酸-谷胱甘肽循环代谢的影响[J]. 浙江农业学报, 2024, 36(7): 1558-1568.

ZENG Hongxue, QU Xinghong. Effect of low-temperature stress on proline metabolism and ascorbic acid-glutathione cycle during germination of three Pueraria lobata germplasm lines[J]. Acta Agriculturae Zhejiangensis, 2024, 36(7): 1558-1568.

图/表 7

图1 低温胁迫对3份葛种质丙二醛(MDA)和过氧化氢(H2O2)含量的影响柱上无相同大写字母的表示同一处理下不同种质间差异显著(P<0.05),无相同小写字母的表示同一种质不同处理间差异显著(P<0.05)。下同。

Fig.1 Effect of low temperature stress on malondialdehyde (MDA) and hydrogen peroxide (H2O2) contents of three P. lobata germplasms Bars marked without the same uppercase letters indicate significant (P<0.05) difference within germplasms under the same treatment, while bars marked without the same lowercase letters indicate significant (P<0.05) difference within treatments for the same germplasm. The same as below.

图2 低温胁迫对3份葛种质脯氨酸含量和脯氨酸代谢关键酶活性的影响

Fig.2 Effects of low temperature stress on proline content and key enzymes activity for proline metabolism of three P. lobata germplasms P5CS, Pyrroline-5-carboxylate synthase; OAT, Ornithine aminotransferase; ProDH, Proline dehydrogenase.

图3 低温胁迫对3份葛种质(还原型)抗坏血酸(AsA)、氧化型抗坏血酸(DHA)含量的影响 AsA/DHA表征AsA与DHA含量之比。

Fig.3 Effects of low temperature stress on ascorbic acid (AsA), dehydroascorbic acid (DHA) contents of three P. lobata germplasms AsA/DHA represents the ratio of AsA content to DHA content.

图4 低温胁迫对3份葛种质(还原型)谷胱甘肽(GSH)、氧化型谷胱甘肽(GSSG)含量的影响 GSH/GSSG表征GSH与GSSG含量之比。

Fig.4 Effects of low temperature stress on glutathione (GSH), glutathione disulfide (GSSG) contents of three P. lobata germplasms GSH/GSSG represents the ratio of GSH content to GSSG content.

图5 低温胁迫对3份葛种质抗坏血酸过氧化物酶(APX)和谷胱甘肽还原酶(GR)的影响

Fig.5 Effects of low temperature stress on ascorbate peroxidase (APX) and glutathione reductase (GR) activities of three P. lobata germplasms

表1 低温胁迫下3份葛种质生理生化特性的相关性分析

Table 1 Correlation analysis of physiological and biochemical characteristics of three P. lobata germplasms under low temperature stress

指标 Indices	指标间的相关系数Correlation coefficient within indices
指标 Indices	H₂O₂	MDA	Pro	P5CS	ProDH	OAT	GSSG	GSH	AsA	DHA	APX	GR	AsA/DHA
MDA	0.836^**
Pro	0.428^**	0.669^**
P5CS	0.693^**	0.724^**	0.365^*
ProDH	-0.460^**	-0.541^**	-0.787^**	-0.273
OAT	0.735^**	0.897^**	0.634^**	0.755^**	-0.491^**
GSSG	0.647^**	0.797^**	0.265	0.648^**	-0.124	0.810^**
GSH	0.672^**	0.802^**	0.470^**	0.696^**	-0.256	0.895^**	0.865^**
AsA	0.759^**	0.816^**	0.328	0.769^**	-0.173	0.825^**	0.876^**	0.931^**
DHA	0.806^**	0.894^**	0.524^**	0.649^**	-0.388^*	0.928^**	0.853^**	0.889^**	0.843^**
APX	0.604^**	0.765^**	0.628^**	0.693^**	-0.411^*	0.912^**	0.716^**	0.898^**	0.777^**	0.833^**
GR	0.604^**	0.765^**	0.628^**	0.693^**	-0.411^*	0.913^**	0.715^**	0.899^**	0.777^**	0.828^**	0.998^**
AsA/DHA	-0.326	-0.403^*	-0.476^**	-0.054	0.465^**	-0.483^**	-0.240	-0.244	-0.041	-0.569^**	-0.398^*	-0.390^*
GSH/GSSG	-0.211	-0.291	0.238	-0.159	-0.143	-0.160	-0.590^**	-0.106	-0.248	-0.259	0.034	0.040	0.066

表2 低温胁迫下3份葛种质生理生化指标的隶属度

Table 2 Membership degree among all physiological and biochemical indices of three P. lobata germplasms under low temperature stress

指标 Indices	不同种质的隶属度Member degree of germplasms
指标 Indices	AUS	HN	JS
H₂O₂	0.534	0.502	0.525
MDA	0.469	0.603	0.408
Pro	0.494	0.487	0.421
P5CS	0.527	0.510	0.474
ProDH	0.517	0.532	0.461
OAT	0.462	0.475	0.573
GSSG	0.479	0.482	0.523
GSH	0.566	0.527	0.507
AsA	0.572	0.533	0.468
DHA	0.474	0.462	0.485
APX	0.506	0.483	0.502
GR	0.506	0.483	0.502
AsA/DHA	0.429	0.569	0.524
GSH/GSSG	0.520	0.545	0.461
平均值Average	0.504	0.514	0.488

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低温胁迫对3份葛种质萌发过程中脯氨酸和抗坏血酸-谷胱甘肽循环代谢的影响

Effect of low-temperature stress on proline metabolism and ascorbic acid-glutathione cycle during germination of three Pueraria lobata germplasm lines

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