Endogenous hormone responses to nitric oxide in alfalfa seedlings under PEG stress

doi:10.11686/cyxb2020173

Abstract

Abstract:

This research studied the endogenous hormone responses to nitric oxide （NO） in leaves and roots of 50-day old alfalfa seedlings under PEG stress. Polyethylene glycol （PEG-6000） was used as the osmotic regulator to simulate drought stress. Exogenous spraying of NO releasing agent sodium nitroprusside （SNP） and NO scavenger carboxy-PTIO （cPTIO）， and liquid chromatography/mass spectrometry methods were used to study the response of four endogenous hormones （abscisic acid， ABA； indole-3-acetic acid， IAA； salicylic acid， SA； gibberellin， GA₃） to NO in the leaf and root systems of alfalfa seedlings under PEG stress. Treatments included in the experiment were： T₁， distilled water （CK）， T₂， PEG added； T₃， SNP+PEG added； and T₄， cPTIO+PEG added. It was found that： 1） The application of exogenous NO under PEG stress （T₃） can promote the NO content in the leaf and root of alfalfa， and the NO scavenger cPTIO （T₄） significantly reduced the NO content in leaves of alfalfa. 2） With increasing duration of PEG stress， the levels of ABA and SA in leaves and roots increased gradually， but the ABA and SA levels in the root system increased later than those in the leaves. On the 8th day of stress， the leaf ABA content in plants of T₃ was 6.68 times higher than the SA content， while the ABA content in plants of T₄ was 77.22% higher than the SA content， and the leaf ABA content in leaves of T₃ plants was significantly higher than that in roots. 3） NO rapidly induces an increase in the IAA and GA₃ concentrations in leaves and roots. On the 4th day of stress， the IAA levels in leaves under exogenous NO treatment were 1.65 times higher than in the cPTIO scavenger treatment. Meanwhile the IAA concentration in roots was 8.70% higher in T₄ than T₃， and the GA₃ content in leaves and roots of T₃ was 54. 49% and 84. 65% higher， respectively， than in T₄. GA₃ content reached its maximum on the second day of treatment and decreased thereafter. In summary， NO is here shown to regulate the growth and stress resistance of alfalfa plants by raising or lowering the metabolic levels of IAA， ABA， GA₃ and SA， the regulation of ABA and SA in alfalfa is particularly prominent.

Key words: Medicago sativa, PEG stress, nitric oxide, endogenous hormone

Xiao-fang ZHANG, Xiao-hong WEI, Fang LIU, Xue-mei ZHU. Endogenous hormone responses to nitric oxide in alfalfa seedlings under PEG stress[J]. Acta Prataculturae Sinica, 2021, 30(4): 160-169.

Figures/Tables 9

References 46

1	Yang P Z. Mechanism involved in drought/salt tolerance improvement in alfalfa due to symbiotic interaction with rhizobium.Xianyang： Northwest A & F University， 2012.
	杨培志. 紫花苜蓿根瘤菌共生对干旱及盐胁迫的响应机制研究. 咸阳：西北农林科技大学， 2012.
2	Zhang L Q， Zhang F Y， Hasi A. Research progress on alfalfa salt tolerance. Acta Prataculturae Sinica， 2012， 21（6）： 296-305.
	张立全，张凤英，哈斯阿古拉. 紫花苜蓿耐盐性研究进展. 草业学报， 2012， 21（6）： 296-305.
3	Sun J J， Alamusi， Zhao J M， et al. Analysis of amino acid composition and six native alfalfa cultivars. Scientia Agricultura Sinica， 2019， 52（13）： 2359-2367.
	孙娟娟，阿拉木斯，赵金梅，等. 6个紫花苜蓿品种氨基酸组成分析及营养价值评价. 中国农业科学， 2019， 52（13）： 2359-2367.
4	Yu L Q， Wang H， Zhang X J， et al. Morphological characteristics and variation of three species of Medicago. Grassland and Turf， 2008（3）： 29-33.
	于林清，汪慧，张旭婧，等. 3种苜蓿形态特征及变异分析. 草原与草坪， 2008（3）： 29-33.
5	Wendehenne D， Pugin A， Klessing D F. Nitric oxide： comparative synthesis and signaling in animal and plant cells. Trends in Plant Science， 2001， 6（4）： 177-183.
6	Zhao Y， Wei X H， Ji X Z， et al. Endogenous NO-mediated transcripts involved in photosynthesis and carbohydrate metabolism in alfalfa （Medicago sativa L.） seedlings under drought stress. Plant Physiology and Biochemistry， 2019， 141： 456-465.
7	Zhao Y， Yi Q， Wei X H， et al. Role of NO-mediated Ca²⁺ signaling in regulation of photosynthesis and resistance to osmotic stress in alfalfa seedlings. Acta Prataculturae Sinica， 2018， 27（5）： 130-140.
	赵颖，弋钦，魏小红，等. NO介导的Ca²⁺信号对渗透胁迫下紫花苜蓿幼苗光合特征及抗性的影响. 草业学报， 2018， 27（5）： 130-140.
8	Sanz L， Albertos P， Mateos I， et al. Nitric oxide （NO） and phytohormones crosstalk during early plant development. Journal of Experimental Botany， 2015， 66（10）： 2857-2868.
9	Campos R G， Osorio M P， Sánchez B R， et al. Plant hormone signaling in flowering： An epigenetic point of view. Journal of Plant Physiology， 2017， 214： 16-27.
10	Fang H， Wang C L， Liao W B， et al. NO is involved in ABA-regulated senescence of cut roses by maintaining water content and increasing antioxidant enzyme activities. Acta Horticulturae Sinica， 2019， 46（5）： 901-909.
	方华，王春蕾，廖伟彪，等. 一氧化氮通过调控水分和抗氧化酶活性参与脱落酸延缓切花月季衰老. 园艺学报， 2019， 46（5）： 901-909.
11	Zhang S M， Wang L X， Chen Y H. Research progress on the interaction between nitric oxide and phytohormones. Journal of Southern Agricultural， 2011， 42（3）： 261-266.
	张盛敏，王罗霞，陈银华. 一氧化氮与植物激素之间相互作用研究进展. 南方农业学报， 2011， 42（3）： 261-266.
12	Shan C J， Dai H F， Sun H L， et al. Study on nitric oxide involved in regulation of drought tolerance of maize seedling roots by salicylic acid. Jiangsu Agricultural Sciences， 2016， 44（8）： 133-135.
	单长卷，代海芳，孙海丽，等. 一氧化氮参与水杨酸对玉米幼苗根系抗旱性的调控. 江苏农业科学， 2016， 44（8）： 133-135.
13	Zhong D L， Ding M， Tang F B， et al. Determination of four endogenous phytohormones in bamboo shoots by liquid chromatography-tandem mass spectrometry. Chinese Journal of Analytical Chemistry， 2013， 41（11）： 1739-1743.
	钟冬莲，丁明，汤富彬，等. 高效液相色谱-串联质谱同时测定毛竹笋中4种内源性植物激素. 分析化学， 2013， 41（11）： 1739-1743.
14	Deng W H， Zhao X R， Zhang J Q， et al. Determination of plant hormones in plant tissues by UPLC-MS/MS. Journal of Beijing Forestry University， 2019， 41（8）： 154-160.
	邓文红，赵欣蕊，张俊琦，等. UPLC-MS/MS测定植物组织中植物激素的含量. 北京林业大学学报， 2019， 41（8）： 154-160.
15	Zhang L G， Deng X P. Advances in studies on physiology and biochemistry of wheat drought resistance. Agricultural Research in the Arid Areas， 2000， 18（3）： 87-92.
	张林刚，邓西平. 小麦抗旱性生理生化研究进展. 干旱地区农业研究， 2000， 18（3）： 87-92.
16	Zhao X， Wang Y L， Wang Y J， et al. Effects of exogenous Ca²⁺on stomatal movement and plasma membrane K⁺ channels of Vicia faba guard cell under salt stress. Acta Agronomica Sinica， 2008， 34（11）： 1970-1976.
	赵翔，汪延良，王亚静，等. 盐胁迫条件下外源Ca²⁺对蚕豆气孔运动及质膜K⁺通道的调控. 作物学报， 2008， 34（11）： 1970-1976.
17	Shao R X， Li L L， Zheng H F， et al. Effects of exogenous nitric oxide on photosynthesis of maize seedlings under drought stress. Scientia Agricultura Sinica， 2016， 49（2）： 251-259.
	邵瑞鑫，李蕾蕾，郑会芳，等. 外源一氧化氮对干旱胁迫下玉米幼苗光合作用的影响. 中国农业科学， 2016， 49（2）： 251-259.
18	Zhao L Q， Liu Y L， Sun B T， et al. Effect of nitric oxide on ions absorption in excised root tips of wheat seedlings under drought stress. Acta Agronomica Sinica， 2009， 35（3）： 530-534.
	赵立群，刘玉良，孙宝腾，等. 干旱胁迫下一氧化氮对小麦离体根尖离子吸收的影响. 作物学报， 2009， 35（3）： 530-534.
19	Zhou Y Q， Bu Q Y. Research on changing of endogenous hormones of fruit trees under drought stress. Northern Fruits， 2011（3）： 1-4.
	周晏起，卜庆雁. 干旱胁迫下果树内源激素变化规律研究进展. 北方果树， 2011（3）： 1-4.
20	Ruan H H， Shen W B， Xu L L. Nitric oxide is involved in the abscisic acid induced proline accumulation in wheat seedling leaves under salt stress. Acta Botanica Sinica， 2004， 46（11）： 1307-1315.
21	García M C， Lamattina L. Nitric oxide induces stomatal closure and enhances the adaptive plant responses against drought stress. Plant Physiology， 2001， 126（3）： 1196-1204.
22	Desikan R， Cheung M K， Bright J， et al. ABA， hydrogen peroxide and nitric oxide signalling in stomatal guard cells. Journal of Experimental Botany， 2004， 55： 205-212.
23	Chang L， Xue J P. Advances in study of polar auxin transport. Journal of Biology， 2008， 25（6）： 9-13.
	常莉，薛建平. 生长素极性运输研究进展. 生物学杂志， 2008， 25（6）： 9-13.
24	Saini S， Sharma I， Kaur N， et al. Auxin： A master regulator in plant root development. Plant Cell Reports， 2013， 32（6）： 741-757.
25	Shi H， Chen L， Ye T， et al. Modulation of auxin content in Arabidopsis confers improved drought stress resistance. Plant Physiology and Biochemistry， 2014， 82： 209-217.
26	Shen H W， Zhang S， Li J B， et al. Effect of exogenous nitric oxide on plant hormones and enzyme content of Acanthopanax senticosus seeds. Chinese Journal of Experimental Traditional Medical Formulae， 2019， 25（3）： 137-142.
	沈宏伟，张爽，李佳宾，等. 不同浓度的外源NO对刺五加种子激素及酶含量变化的影响. 中国实验方剂学杂志， 2019， 25（3）： 137-142.
27	Peto A， Lehotai N， Lozano J J， et al. Involvement of nitric oxide and auxin in signal transduction of copper induced morphological responses in Arabidopsis seedlings. Annals of Botany， 2011， 108： 449-457.
28	Chen W W， Yang J L， Qin C. Nitric oxide acts downstream of auxin to trigger root ferric-chelate reductase activity in response to iron deficiency in Arabidopsis. Plant Physiology， 2010， 154（2）： 810-819.
29	Yan Z P. Effect of drought stress on IAA oxidase of wheat. Inner Mongolia Agricultural Science and Technology， 2005（4）： 16-17.
	闫志佩. 干旱胁迫对小麦吲哚乙酸氧化酶的影响. 内蒙古农业科技， 2005（4）： 16-17.
30	Pagnussat G C， Simontacchi M， Puntarulo S， et al. Nitric oxide is required for root organogenesis. Plant Physiology， 2002， 129（3）： 954-956.
31	Correa A N， Graziano M， Lamattina L. Nitric oxide plays a central role in determining lateral root development in tomato. Planta， 2004， 218（6）： 900-905.
32	Meng X J， Di K， Ding G H. Progress of study on the physiological role of salicylic acid in plant. Chinese Agricultural Science Bulletin， 2010， 26（15）： 207-214.
	孟雪娇，邸昆，丁国华. 水杨酸在植物体内的生理作用研究进展. 中国农学通报， 2010， 26（15）： 207-214.
33	Wang D H， Li X X， Su Z K， et al. The role of salicylic acid in response of two rice cultivars to chilling stress. Biologia Plantarum， 2009， 53（3）： 545-552.
34	Mahdavian K， Kalantai K M， Ghorbanli M， et al. The effects of salicylic acid on pigment contents in ultraviolet radiation stressed pepper plants. Biologia Plantarum， 2008， 52（1）： 170-172.
35	Gautam S， Singh P K. Salicylic acid-induced salinity tolerance in corn grown under NaCl stress. Acta Physiologiae Plantarum， 2009， 31（6）： 1185-1190.
36	Sun D Z， Yang H S， Peng J， et al. Effects of exogenous salicylic acid and nitric oxide on growth， photosynthesis， and ion distribution in tomato seedlings under NaCl stress. Acta Ecologica Sinica， 2014， 34（13）： 3519-3528.
	孙德智，杨恒山，彭靖，等. 外源SA和NO对NaCl胁迫下番茄幼苗生长、光合及离子分布的影响. 生态学报， 2014， 34（13）： 3519-3528.
37	Shan C J， Zhang F Y. Effects of salicylic acid on antioxidant properties of maize “Xindan 29” seedling roots under drought stress. Jiangsu Agricultural Sciences， 2015， 43（2）： 102-104.
	单长卷，张飞扬. 水杨酸对干旱下新单29玉米幼苗根系抗氧化特性的影响. 江苏农业科学， 2015， 43（2）： 102-104.
38	Azooz M M， Youssef M M. Evaluation of heat shock and salicylic acid treatments as inducers of drought stress tolerance in Hassawi wheat. American Journal of Plant Physiology， 2010， 5（2）： 56-70.
39	Loutfy N， El T M A， Hassanen A M， et al. Changes in the water status and osmotic solute contents in response to drought and salicylic acid treatments in four different cultivars of wheat （Triticum aestivum）. Journal of Plant Research， 2012， 125（1）： 173-184.
40	Klessig D F， Durner J， Noad R， et al. Nitric oxide and salicylic acid signaling in plant defense. Proceedings of the National Academy of Sciences， 2000， 97（16）： 8849-8855.
41	Zottini M， Costa A， De Michele R， et al. Salicylic acid activates nitric oxide synthesis in Arabidopsis. Journal of Experimental Botany， 2007， 58（6）： 1397-1405.
42	Gémes K， Poór P， Horváth E， et al. Cross-talk between salicylic acid and NaCl-generated reactive oxygen species and nitric oxide in tomato during acclimation to high salinity. Physiologia Plantarum， 2011， 142（2）： 179-192.
43	Liu X， Zhang S Q， Lou C H. Nitric oxide is involved in the regulation of stomatal movement of broad bean by salicylic acid. Chinese Science Bulletin， 2003， 48（1）： 60-63.
	刘新，张蜀秋，娄成后. 一氧化氮参与水杨酸对蚕豆气孔运动的调控. 科学通报， 2003， 48（1）： 60-63.
44	Wang D， Pan Y， Zhao X， et al. Genome-wide temporal-spatial gene expression profiling of drought responsiveness in rice. BMC Genomics， 2011， 12： 149-163.
45	Bai T H， Yin R， Li C Y， et al. Comparative analysis of endogenous hormones in leaves and roots of two contrasting malus species in response to hypoxia stress. Journal of Plant Growth Regulation， 2011， 30（2）： 119-127.
46	Beligni M V， Fath A， Bethke P C， et al. Nitric oxide acts as an antioxidant and delays programmed cell death in barley aleurone layers. Plant Physiology， 2002， 129（4）： 1642-1650.

激素 Plant hormone	保留时间 Retention time (s)	母离子 Mother ion (m·z^-1)	子离子 Child ion (m·z^-1)	碎裂电压 Fragmentor voltage (V)	碰撞能量 Collision energy (eV)
脱落酸ABA	2.547	263.1	153.1	90	10
生长素IAA	3.035	174.1	129.9	80	13
水杨酸SA	3.389	136.9	93.0	70	22
赤霉素GA₃	4.010	345.1	239.0	75	9

激素 Plant hormone	保留时间 Retention time (s)	母离子 Mother ion (m·z^-1)	子离子 Child ion (m·z^-1)	碎裂电压 Fragmentor voltage (V)	碰撞能量 Collision energy (eV)
脱落酸ABA	2.547	263.1	153.1	90	10
生长素IAA	3.035	174.1	129.9	80	13
水杨酸SA	3.389	136.9	93.0	70	22
赤霉素GA₃	4.010	345.1	239.0	75	9

植物激素 Plant hormone	线性方程 Linear equation	相关系数 Correlation coefficient (r)	线性范围 Linear range (μg·L^-1)
脱落酸ABA	C=0.1204X+0.5550	0.9997	1~100
生长素IAA	C=0.6629X+0.9960	0.9990	1~100
水杨酸SA	C=0.0557X-0.0379	0.9995	1~100
赤霉素GA₃	C=0.0624X+0.7897	0.9997	1~100

植物激素 Plant hormone	线性方程 Linear equation	相关系数 Correlation coefficient (r)	线性范围 Linear range (μg·L^-1)
脱落酸ABA	C=0.1204X+0.5550	0.9997	1~100
生长素IAA	C=0.6629X+0.9960	0.9990	1~100
水杨酸SA	C=0.0557X-0.0379	0.9995	1~100
赤霉素GA₃	C=0.0624X+0.7897	0.9997	1~100

[1]	Xin MA, Zhu-zhu LUO, Yao-quan ZHANG, Jia-he LIU, Yi-ning NIU, Li-qun CAI. Distribution characteristics and ecological function predictions of soil bacterial communities in rainfed alfalfa fields on the Loess Plateau [J]. Acta Prataculturae Sinica, 2021, 30(3): 54-67.
[2]	Dong LI, Hong-tao SHEN, Yan-fang WANG, Yue-hua WANG, Li-jun WANG, Shi-min ZHAO, Ling LIU. Effects of exogenous melatonin on photosynthetic carbon assimilation and endogenous hormones in tobacco seedlings under drought stress [J]. Acta Prataculturae Sinica, 2021, 30(1): 130-139.
[3]	LIU Yan, YU Mei-ling, ZHANG Ran, NIU Kui-ju, LI Yu-zhu, ZHANG Jin-qing, MA Hui-ling. The relationship between endogenous hormone content and apomixis rate of wild Kentucky bluegrass in Gansu Province [J]. Acta Prataculturae Sinica, 2020, 29(7): 99-111.
[4]	ZHAO Ying, WEI Xiao-hong, LI Tao-tao. Effects of exogenous nitric oxide on seed germination and seedling growth of Chenopodium quinoa under complex saline-alkali stress [J]. Acta Prataculturae Sinica, 2020, 29(4): 92-101.
[5]	BAO Gen-sheng, SONG Mei-ling, WANG Yu-qin, LIU Jing, WANG Hong-sheng. Interactive effects of different densities of Pedicularis kansuensis parasitism and Epichloё endophyte infection on the endogenous hormone levels and alkaloid contents of Stipa purpurea [J]. Acta Prataculturae Sinica, 2020, 29(4): 147-156.
[6]	YI Kun, ZHAO Yi-hang, HU Yao, LIU Jia-xue, HE Tao-tao, LI Xu, SONG Peng, CUI Guo-wen, YIN Xiu-jie. Effect of GA₃ and 6-BA on rhizome segment growth and endogenous hormone content of Caucasian clover [J]. Acta Prataculturae Sinica, 2020, 29(2): 22-30.
[7]	WANG Ning, FU Ya-jun, YUAN Mei-li, LIU Zheng-yang, ZHANG Ming-xin, MI Yin-fa. Effectiveness of exogenous GA₃ for dormancy breaking in invasive Aegilops tauschii, and effect on germination physiology [J]. Acta Prataculturae Sinica, 2020, 29(2): 73-81.
[8]	XIANG Hong-tao, QI De-qiang, LI Wan, ZHENG Dian-feng, WANG Yue-xi, WANG Tong-tong, WANG Li-zhi, ZENG Xian-nan, YANG Chun-jie, ZHOU Hang, ZHAO Hai-dong. Effect of exogenous ABA on the endogenous hormone levels and physiology of chilling resistance in the leaf sheath of rice at the flowering stage under low temperature stress [J]. Acta Prataculturae Sinica, 2019, 28(4): 81-94.
[9]	ZHAO Ying, YI Qin, WEI Xiao-hong, XIN Xia-qing, HAN Ting, YUE Kai, WANG Fang-lin. Role of NO-mediated Ca²⁺ signaling in regulation of photosynthesis and resistance to osmotic stress in alfalfa seedlings [J]. Acta Prataculturae Sinica, 2018, 27(5): 130-140.
[10]	XIN Xia-qing, WEI Xiao-hong, HAN Ting, YUE Kai, ZHAO Ying. Research on exogenous NO and reverse regulation to the dynamic changes of antioxidant enzymes and isoenzymes in alfalfa seed germination under PEG stress [J]. Acta Prataculturae Sinica, 2018, 27(10): 105-112.
[11]	LIU Jian-Xin, WANG Jin-Cheng, LIU Xiu-Li. Effect of exogenous nitric oxide on active oxygen metabolism and mineral contents in oat seedlings under lanthanum stress [J]. Acta Prataculturae Sinica, 2017, 26(5): 135-143.
[12]	WANG Ri-Ming, XIONG Xing-Yao. Effect of temperature stress on growth and metabolism in perennial ryegrass [J]. Acta Prataculturae Sinica, 2016, 25(8): 81-90.
[13]	CHANG Qing-Shan, ZHANG Li-Xia, YANG Wei, ZHOU Shan-Shan, HUANG Qing-Zhe, LV Feng-Juan, HUANG Yue, GE Shu-Hui, ZHANG Tian-Meng. Effects of exogenous nitric oxide on antioxidant activity and photosynthetic characteristics of Prunella vulgaris seedlings under NaCl stres [J]. Acta Prataculturae Sinica, 2016, 25(7): 121-130.
[14]	YANG Yong, LOU Yan-Hong, YANG Zhi-Jian, XIANG Zuo-Xiang, XU Qing-Guo, HU Long-Xing. Effect of low temperature on phytohormones and carbohydrates metabolism in Bermuda grass [J]. Acta Prataculturae Sinica, 2016, 25(2): 205-215.
[15]	WU Xu-Hong, LV Cheng-Min, FENG Jing-Min. Protective effect of exogenous nitric oxide against oxidative damage in pumpkin seedlings under chilling stress [J]. Acta Prataculturae Sinica, 2016, 25(12): 161-169.