葡萄糖-6-磷酸脱氢酶在紫花苜蓿干旱耐受性中的响应

doi:10.11686/cyxb2017308

草业学报 ›› 2018, Vol. 27 ›› Issue (2): 48-56.DOI: 10.11686/cyxb2017308

葡萄糖-6-磷酸脱氢酶在紫花苜蓿干旱耐受性中的响应

熊丽丽¹, 闫霜¹, 李萍^{2, *}, 杨国柱¹, 尹卫¹, 严晓霞¹, 张韩¹

1.青海大学生态环境工程学院,青海西宁810016;
2.青海大学省部共建三江源生态与高原农牧业国家重点实验室,青海西宁 810016

收稿日期:2017-07-18 修回日期:2017-10-18 出版日期:2018-02-20 发布日期:2018-02-20
通讯作者: liping051126@163.com
作者简介:熊丽丽(1993-),女,青海乐都人,在读硕士。E-mail: 843293453@qq.com
基金资助:
国家自然基金地区基金项目(31660063),青海省科技厅基础研究计划(2016-ZJ-930Q),青海大学省部共建三江源生态与高原农牧业国家重点实验室开放课题(2017-KF-05)和青海大学生态环境工程学院中青年科研基金项目(2016-T-4)资助

Role of glucose-6-phosphate dehydrogenase in the drought tolerance of alfalfa

XIONG Li-li¹, YAN Shuang¹, LI Ping^{2, *}, YANG Guo-zhu¹, YIN Wei¹, YAN Xiao-xia¹, ZHANG Han¹

1.College of Eco-Environmental Engineering, Qinghai University, Xining 810016, China;
2.State Key Laboratory of Plateau Ecology and Agriculture, Qinhai University, Xining 810016, China

Received:2017-07-18 Revised:2017-10-18 Online:2018-02-20 Published:2018-02-20

摘要/Abstract

摘要： 葡萄糖-6-磷酸脱氢酶(glucose-6-phosphate dehydrogenase,G6PDH,EC1.1.1.49)是磷酸戊糖途径的关键限速酶,参与NADPH和磷酸核糖的合成,在植物响应生物胁迫和非生物胁迫方面有重要作用。研究了葡萄糖-6-磷酸脱氢酶(G6PDH)在紫花苜蓿中对干旱胁迫的响应。首先采用不同浓度的PEG模拟干旱胁迫处理紫花苜蓿幼苗,通过测定不同PEG浓度下紫花苜蓿幼苗的株高、根长、干重、过氧化氢(H₂O₂)、丙二醛(MDA)含量的变化筛选胁迫浓度来确定干旱胁迫的条件。其次,检测胁迫浓度下紫花苜蓿幼苗中G6PDH的活性变化。第三,通过添加G6PDH的抑制剂Na₃PO₄,观察干旱胁迫下紫花苜蓿幼苗的生长情况并测定MDA、H₂O₂含量以及G6PDH活性的变化。最后,综合分析G6PDH对干旱胁迫的响应。结果显示,不同浓度PEG处理下,紫花苜蓿幼苗的生长受到限制,例如株高、根长、鲜重、干重都随PEG浓度的增加而逐渐减小,MDA和H₂O₂的含量随PEG浓度的增加而增加,其中以15% PEG处理的影响最大;同时,也发现随PEG浓度的增加紫花苜蓿幼苗中G6PDH的活性也较对照增加,15%PEG处理时达最高值;故15%的PEG处理为干旱胁迫模拟最佳条件。通过添加G6PDH的抑制剂Na₃PO₄后发现,干旱胁迫下紫花苜蓿幼苗的生长明显地受到抑制,其叶片中MDA、H₂O₂含量分别较干旱胁迫下的增长28.4%、19.9%,G6PDH的活性也明显降低了49.4%。以上结果初步说明G6PDH可能参与调节了干旱胁迫引起的氧化胁迫。

Abstract: Glucose-6-phosphate dehydrogenase (G6PDH, EC1.1.1.49) is the first and critical rate-limiting enzyme in the pentose phosphate pathway. It is involved in the synthesis of NADPH and ribose-5-phosphate, and plays an important role in plant responses to biotic and abiotic stresses. The aim of this study was to explore the role of G6PDH under drought stress in alfalfa. First, 7-day-old alfalfa seedlings were treated with different concentrations of polyethylene glycol (PEG), and then plant height and root length, fresh weight and dry weight, and the contents of hydrogen peroxide (H₂O₂) and malondialdehyde (MDA) were determined. These analyses were used to define the concentration of PEG causing drought stress to the seedlings. Then, the activity of G6PDH in alfalfa seedlings was determined under a range of stress-inducing PEG concentrations. The G6PDH inhibitor Na₃PO₄ was applied to alfalfa seedlings under drought stress, and changes in the indexes described above were determined. The results showed that PEG treatment significantly inhibited the growth of alfalfa seedlings. Plant height, root length, fresh weight, and dry weight decreased with increasing PEG concentrations. Compared with non-stressed seedlings, those subjected to PEG-induced drought stress (15% PEG) showed 28.4% and 19.9% higher contents of H₂O₂ and MDA, respectively, and 49.4% lower G6PDH activity. Application of Na₃PO₄ decreased the activity of G6PDH and caused the symptoms of drought stress to become more severe, suggesting that G6PDH is involved in the regulation of oxidative stress induced by PEG.

熊丽丽, 闫霜, 李萍, 杨国柱, 尹卫, 严晓霞, 张韩. 葡萄糖-6-磷酸脱氢酶在紫花苜蓿干旱耐受性中的响应[J]. 草业学报, 2018, 27(2): 48-56.

XIONG Li-li, YAN Shuang, LI Ping, YANG Guo-zhu, YIN Wei, YAN Xiao-xia, ZHANG Han. Role of glucose-6-phosphate dehydrogenase in the drought tolerance of alfalfa[J]. Acta Prataculturae Sinica, 2018, 27(2): 48-56.

参考文献

[1] Li Y, Sun H R, Ding N, et al. Alfalfa (Medicago sativa L.) root biomass. Acta Agrestia Sinica, 2011, (5): 872-879.
李扬, 孙洪仁, 丁宁, 等. 紫花苜蓿根系生物量. 草地学报, 2011, (5): 872-879.
[2] Lu S.Effects of drought stress on plant growth and physiological traits. Journal of Jiangsu Forestry Science & Technology, 2012, (4): 51-54.
鲁松. 干旱胁迫对植物生长及其生理的影响. 江苏林业科技, 2012, (4): 51-54.
[3] Zhou F.Effects of drought stress on plant growth and physiological traits. Beijing Agriculture, 2014, (3): 1.
周峰. 干旱胁迫对植物生长及其生理的影响. 北京农业, 2014, (3): 1.
[4] Jo B, Radhika D, Hancock J T, et al. ABA-induced NO generation and stomatal closure in Arabidopsis are dependent on H₂O₂ synthesis. Plant Journal, 2006, 45(1): 113-122.
[5] Yang X L, Zhu Y J.Advances of plant drought stress. Agricultural Engineening, 2012, (11): 44-45.
杨雪莲, 朱友娟. 植物干旱胁迫研究进展. 农业工程, 2012, (11): 44-45.
[6] Wang B F, Huang J P, Yang X L, et al. Advances on inhibition mechanism of crop photosynthesis by drought stress. Hubei Agricultural Sciences, 2014, (23): 5628-5632.
汪本福, 黄金鹏, 杨晓龙, 等. 干旱胁迫抑制作物光合作用机理研究进展. 湖北农业科学, 2014, (23): 5628-5632.
[7] Yu D Q, Tang H R, Zhang Y, et al. Research progress in glucose-6-phosphate dehydrogenase in higher plants. Chinese Journal of Biotechnology, 2012, (7): 800-812.
于定群, 汤浩茹, 张勇, 等. 高等植物葡萄糖-6-磷酸脱氢酶的研究进展. 生物工程学报, 2012, (7): 800-812.
[8] Kletzien R F, Harris P K W, Foellmi L A. Glucose-6-phosphate dehydrogenase, a “housekeeping” enzyme subject to tissue-specific regulation by hormones, nutrients, and oxidant stress. The FASEB Journal, 1994, 8: 174-181.
[9] Ghosh A K, Saini S, Das S, et al. Glucose-6-phosphate dehydrogenase and trypanothione reductase interaction protects Leishmania donovani from metalloid mediated oxidative stress. Free Radical Biology and Medicine, 2017, 106: 10-23.
[10] Valderrama R, Corpas F J, Carreras A, et al. The dehydrogenase-mediated recycling of NADPH is a key antioxidant system against salt-induced oxidative stress in olive plants. Plant Cell & Environment, 2006, 29(7): 1449-1459.
[11] Li J S.Studies on the Mechanism of Regulations of G6PDH, cGMP, H₂O₂ and Ca²+ in Plant Adaptation to Salt Tolerance. Lanzhou: Lanzhou University, 2011.
李积胜. G6PDH、cGMP、H₂O₂和Ca²+在植物耐盐适应中调节作用的机理研究. 兰州: 兰州大学, 2011.
[12] Stanton R C.Glucose-6-phosphate dehydrogenase, NADPH, and cell survival. Iubmb Life, 2012, 64(5): 362-369.
[13] Wang H H, Yang L D, Li Y, et al. Involvement of ABA- and H₂O₂- dependent cytosolic glucose-6-phosphate dehydrogenase in maintaining redox homeostasis in soybean roots under drought stress. Plant Physiology Biochemistry, 2016, 107: 126-136.
[14] Liu J, Wang X M, Hu Y F, et al. Glucose-6-phosphate dehydrogenase plays a pivotal role in tolerance to drought stress in soybean roots. Plant Cell Report, 2013, 32(3): 415-429.
[15] Zhao C Z, Wang X M, Wang X Y, et al. Glucose-6-phosphate dehydrogenase and alternative oxidase are involved in the cross tolerance of highland barley to salt stress and UV-B radiation. Journal of Plant Physiology, 2015, 181: 83-95.
[16] Cardi M, Castiglia D, Ferrara M, et al. The effects of salt stress cause a diversion of basal metabolism in barley roots: possible different roles for glucose-6-phosphate dehydrogenase isoforms. Plant Physiology Biochemistry, 2015, 86: 44-54.
[17] Hofmann N R.The GSK3-type kinase ASKα targets glucose-6-phosphate dehydrogenase to mediate oxidative stress responses in Arabidopsis. Plant Cell, 2012, 24(8): 3170.
[18] An X, Liu L Z, Hu P.Research in progress on glucose-6-phosphate dehydrogenase. Chinese Journal of Biologicals, 2011, (6): 745-748.
安选, 刘良忠, 胡鹏. 葡萄糖-6-磷酸脱氢酶的研究进展. 中国生物制品学杂志, 2011, (6): 745-748.
[19] Asai S, Yoshioka M, Nomura H, et al. A plastidic glucose-6-phosphate dehydrogenase is responsible for hypersensitive response cell death and reactive oxygen species production. Journal of General Plant Pathology, 2011, 77(3): 152-162.
[20] Hodges D M, Delong J M, Forney C F, et al. Improving the thiobarbituric acid-reactive-substances assay for estimating lipid peroxidation in plant tissues containing anthocyanin and other interfering compounds. Planta, 1999, 207: 604-711.
[21] Heath R L, Packer L.Photoperoxidation in isolated chloroplasts: I. Kinetics and stoichiometry of fatty acid peroxidation. Archives of Biochemistry and Biophysics, 1968, 125: 189-198.
[22] Bradford M M.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 1976, 72: 248-254.
[23] Esposito S, Carfagna S, Massaro G, et al. Glucose-6-phosphate dehydrogenase in barley roots: kinetic properties and localization of the isoforms. Planta, 2001, 212: 627-634.
[24] Liu Y G, Wu R R, Wan Q, et al. Glucose-6-phosphate dehydrogenase palys a pivotal role in nitric oxide-involved defense against oxidative stress under salt stress in red kidney bean roots. Plant and Cell Physioloy, 2007, 48(3): 511-522.
[25] Nie S H, Qi J C, Zhang H L, et al. Effect of drought stress simulated by PEG6000 on malondialdehyde content and activities of protective enzymes in barley seedings. Xingjiang Agricultural Sciences, 2011, (1): 11-17.
聂石辉, 齐军仓, 张海禄, 等. PEG6000模拟干旱胁迫对大麦幼苗丙二醛含量及保护酶活性的影响. 新疆农业科学, 2011, (1): 11-17.
[26] Wang H Z, Zhang L H, Jun M A, et al. Effects of water stress on reactive oxygen species generation and protection system in rice during grain-filling stage. Agricultural Sciences in China, 2010, 9(5): 633-641.
[27] Gong H, Chen G, Li F, et al. Involvement of G6PDH in heat stress tolerance in the calli from Przewalskia tangutica and Nicotiana tabacum. Biologia Plantarum, 2012, 56(3): 422-430.
[28] Dal Santo S, Stampfl H, Krasensky J, et al. Stress-induced GSK3 regulates the redox stress response by phosphorylating glucose-6-phosphate dehydrogenase in Arabidopsis. Plant Cell, 2012, 24(8): 3380-3392.

葡萄糖-6-磷酸脱氢酶在紫花苜蓿干旱耐受性中的响应

Role of glucose-6-phosphate dehydrogenase in the drought tolerance of alfalfa

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 0

编辑推荐

Metrics