苜蓿体内H2S信号与Ca2+调节气孔运动的作用机制

doi:10.7525/j.issn.1673-5102.2023.02.013

摘要/Abstract

摘要：

为探究H₂S信号在苜蓿（Medicago sativa）体内调节气孔运动的作用，及在此过程中H₂S与Ca²⁺的关系，以蒺藜苜蓿（Medicago truncatula）的野生型和钙离子转运体突变体为试验材料，分别从转录水平、细胞水平和生理水平开展研究。采用qRT-PCR比较相关基因的表达量变化、荧光探针显示体内Ca²⁺含量、电极法测定H₂S含量、光学显微镜观察和测量气孔孔径等。结果表明：蒺藜苜蓿突变体NF3011和NF2734体内H₂S的含量与野生型相比极显著降低（P<0.01）；H₂S信号在一定程度上抑制钙离子转运体编码基因MTR_6g027580的表达；外源生理浓度H₂S熏蒸可诱导蒺藜苜蓿气孔关闭，与Ca²⁺通道阻断剂LaCl₃联合处理对野生型气孔运动未产生影响，而在突变体中的结果截然相反；利用荧光探针测定保卫细胞内的Ca²⁺含量，所得结果与气孔孔径的变化规律完全一致。综上所述，H₂S信号促进叶片保卫细胞内Ca²⁺的含量增加，最终表现为植物气孔孔径变小，在此过程中胞内Ca²⁺含量变化主要通过Ca²⁺转运体进行，少部分依赖Ca²⁺离子通道。该研究结果不仅在理论上丰富了H₂S信号的作用机制，更具应用于苜蓿生产实践并推广于其他作物的潜力。

关键词: H₂S信号, Ca²⁺, 气孔运动, 蒺藜苜蓿

Abstract:

To explore the role of H₂S signal in regulating stomatal movement in alfalfa and the relationship between H₂S and Ca²⁺ during this process， wild type and the calcium transporter mutants of Medicago truncatula were used as experimental materials， and the transcriptional level， cellular level and physiological level were studied respectively， and the expression levels of related genes were compared by qRT-PCR， the content of Ca²⁺in vivo was detected by fluorescent probe， the content of H₂S was measured by electrode method， and the stomatal aperture was observed by microscope respectively. The results showed that compared with the wild type， the content of H₂S in the mutant NF3011 and NF2734 decreased significantly； the H₂S signal inhibited the expression of calcium transporter encoding gene MTR_6g027580， the exogenous physiological concentration of H₂S fumigation could induce stomatal closure of Medicago truncatula， and the combination of treatment with LaCl₃， Ca²⁺ channel blocker， had no effect on stomatal movement of wild type， but opposite in mutant； the concentration of Ca²⁺ in guard cells was measured by fluorescence probe， which were consistent with the change law of stomatal aperture. In conclusion， the content of Ca²⁺ in guard cells of leaves is induced by H₂S， whose stomatal aperture become smaller at the same time. In this process， the change of intracellular Ca²⁺ concentration mainly depend on Ca²⁺ transporter， and small partly depend on Ca²⁺ channel. The results of this study not only enrich the mechanism of H₂S signaling in theory， but also have the potential to be applied to alfalfa production and other crops.

Key words: H₂S signal, Ca²⁺ transporter, stomatal movement, Medicago truncatula

中图分类号:

Q945.78

郝雪峰, 亢春霞, 裴雁曦, 金竹萍. 苜蓿体内H₂S信号与Ca²⁺调节气孔运动的作用机制[J]. 植物研究, 2023, 43(2): 281-287.

Xuefeng HAO, Chunxia KANG, Yanxi PEI, Zhuping JIN. The Mechanism of H₂S Signal and Ca²⁺ Regulating Stomatal Movement in Medicago sativa[J]. Bulletin of Botanical Research, 2023, 43(2): 281-287.

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参考文献 36

1	WILSON L G， BRESSAN R A， FILNER P.Light-dependent emission of hydrogen sulfide from plants［J］.Plant Physiology，1978，61（2）：184-189.
2	裴雁曦.植物中的气体信号分子硫化氢：无香而立，其臭如兰［J］.中国生物化学与分子生物学报，2016，32（7）：721-733.
	PEI Y X.Gasotransmitter hydrogen sulfide in plants：stinking to high heaven，but refreshing to fine life［J］.Chinese Journal of Biochemistry and Molecular Biology，2016，32（7）：721-733.
3	AROCA A， GOTOR C， BASSHAM D C，et al.Hydrogen sulfide：from a toxic molecule to a key molecule of cell life［J］.Antioxidants，2020，9（7）：621.
4	WANG C L， DENG Y Z， LIU Z S，et al.Hydrogen sulfide in plants：crosstalk with other signal molecules in response to abiotic stresses［J］.International Journal of Molecular Sciences，2021，22（21）：12068.
5	CHEN Z， HUANG Y W， YANG W J，et al.The hydrogen sulfide signal enhances seed germination tolerance to high temperatures by retaining nuclear COP1 for HY5 degradation［J］.Plant Science，2019，285：34-43.
6	LI H， GHOTO K， WEI M Y，et al.Unraveling hydrogen sulfide-promoted lateral root development and growth in mangrove plant Kandelia obovata：insight into regulatory mechanism by TMT-based quantitative proteomic approaches［J］.Tree Physiology，2021，41（9）：1749-1766.
7	LI H， CHEN H Y， CHEN L L，et al.The role of hydrogen sulfide in plant roots during development and in response to abiotic stress［J］.International Journal of Molecular Sciences，2022，23（3）：1024.
8	MA X L， ZHANG L P， PEI Z Y，et al.Hydrogen sulfide promotes flowering in heading Chinese cabbage by S-sulfhydration of BraFLCs［J］.Horticulture Research，2021，8：19.
9	LIU D， XU S， HU H L，et al.Endogenous hydrogen sulfide homeostasis is responsible for the alleviation of senescence of postharvest daylily flower via increasing antioxidant capacity and maintained energy status［J］.Journal of Agricultural and Food Chemistry，2017，65（4）：718-726.
10	HU K D， ZHANG X Y， YAO G F，et al.A nuclear-localized cysteine desulfhydrase plays a role in fruit ripening in tomato［J］.Horticulture Research，2020，7（1）：211.
11	LIU Z Q， LI Y W， CAO C Y，et al.The role of H₂S in low temperature-induced cucurbitacin C increases in cucumber［J］.Plant Molecular Biology，2019，99（6）：535-544.
12	CHEN X D， CHEN Q， ZHANG X M，et al.Hydrogen sulfide mediates nicotine biosynthesis in tobacco（Nicotiana tabacum） under high temperature conditions［J］.Plant Physiology and Biochemistry，2016，104：174-179.
13	WANG L J， MU X J， CHEN X，et al.Hydrogen sulfide attenuates intracellular oxidative stress via repressing glycolate oxidase activities in Arabidopsis thaliana ［J］.BMC Plant Biology，2022，22（1）：98.
14	FANG H H， LIU Z Q， LONG Y P，et al.The Ca²⁺/calmodulin2-binding transcription factor TGA3 elevates LCD expression and H₂S production to bolster Cr⁶⁺ tolerance in Arabidopsis ［J］.The Plant Journal，2017，91（6）：1038-1050.
15	FANG H H， JING T， LIU Z Q，et al.Hydrogen sulfide interacts with calcium signaling to enhance the chromium tolerance in Setaria italica ［J］.Cell Calcium，2014，56（6）：472-481.
16	HE H Y， LI Y Q， HE L F.The central role of hydrogen sulfide in plant responses to toxic metal stress［J］.Ecotoxicology and Environmental Safety，2018，157：403-408.
17	JIN Z P， SHEN J J， QIAO Z J，et al.Hydrogen sulfide improves drought resistance in Arabidopsis thaliana ［J］.Biochemical and Biophysical Research Communications，2011，414（3）：481-486.
18	ZHOU H， CHEN Y， ZHAI F C，et al.Hydrogen sulfide promotes rice drought tolerance via reestablishing redox homeostasis and activation of ABA biosynthesis and signaling［J］.Plant Physiology and Biochemistry，2020，155：213-220.
19	HAO X F， JIN Z P， WANG Z Q，et al.Hydrogen sulfide mediates DNA methylation to enhance osmotic stress tolerance in Setaria italica L.［J］.Plant and Soil，2020，453（1/2）：355-370.
20	WEI M Y， LIU J Y， LI H，et al.Proteomic analysis reveals the protective role of exogenous hydrogen sulfide against salt stress in rice seedlings［J］.Nitric Oxide，2021，111/112：14-30.
21	ZHANG X W， FU X， LIU F J，et al.Hydrogen sulfide improves the cold stress resistance through the CsARF5-CsDREB3 module in cucumber［J］.International Journal of Molecular Sciences，2021，22（24）：13229.
22	XUAN L J， WU H J， LI J，et al.Hydrogen sulfide reduces cell death through regulating autophagy during submergence in Arabidopsis ［J］.Plant Cell Reports，2022，41（7）：1531-1548.
23	VOJTOVIČ D，LUHOVÁL， PETŘIVALSKÝ M.Something smells bad to plant pathogens：production of hydrogen sulfide in plants and its role in plant defence responses［J］.Journal of Advanced Research，2021，27：199-209.
24	GARCÍA-MATA C， LAMATTINA L.Hydrogen sulphide，a novel gasotransmitter involved in guard cell signalling［J］.New Phytologist，2010，188（4）：977-984.
25	LIU H， XUE S W.Interplay between hydrogen sulfide and other signaling molecules in the regulation of guard cell signaling and abiotic/biotic stress response［J］.Plant Communications，2021，2（3）：100179.
26	ZHU J K.Salt and drought stress signal transduction in plants［J］.Annual Review of Plant Biology，2002，53：247-273.
27	JIN Z P， XUE S W， LUO Y N，et al.Hydrogen sulfide interacting with abscisic acid in stomatal regulation responses to drought stress in Arabidopsis ［J］.Plant Physiology and Biochemistry，2013，62：41-46.
28	LI Z G， GONG M， XIE H，et al.Hydrogen sulfide donor sodium hydrosulfide-induced heat tolerance in tobacco （Nicotiana tabacum L.） suspension cultured cells and involvement of Ca²⁺ and calmodulin［J］.Plant Science，2012，185/186：185-189.
29	GONG C M， SHI C， DING X T，et al.Hydrogen sulfide induces Ca²⁺ signal in guard cells by regulating reactive oxygen species accumulation［J］.Plant Signaling & Behavior，2020，15（11）：1805228.
30	FRESQUEZ A M， WHITE C.Extracellular cysteines C226 and C232 mediate hydrogen sulfide-dependent inhibition of Orai3-mediated store-operated calcium entry［J］.American Journal of Physiology，2022，322（1）：C38-C48.
31	ZHANG C M， SHI S L， LIU Z，et al.Drought tolerance in alfalfa （Medicago sativa L.） varieties is associated with enhanced antioxidative protection and declined lipid peroxidation［J］.Journal of Plant Physiology，2019，232：226-240.
32	李辉.蒺藜苜蓿节间缩短控制基因COSTIN的克隆及功能研究［D］.太原：山西大学，2018.
	LI H.Identification and characterization of the COMPACT STALK INTERNODES（COSTIN） gene required for plant height in model legume Medicago truncatula ［D］.Taiyuan：Shanxi University，2018.
33	JIN Z P， WANG Z Q， MA Q X，et al.Hydrogen sulfide mediates ion fluxes inducing stomatal closure in response to drought stress in Arabidopsis thaliana ［J］.Plant and Soil，2017，419（1/2）：141-152.
34	郑远，陈兆进.植物细胞器钙信号研究进展［J］.植物生理学报，2015，51（8）：1195-1203.
	ZHENG Y， CHEN Z J.Organellar calcium signaling in plants［J］.Plant Physiology Journal，2015，51（8）：1195-1203.
35	刘建新，欧晓彬，王金成.胞质Ca²⁺参与外源H₂S促进盐碱胁迫下裸燕麦种子萌发［J］.植物研究，2020，40（1）：58-65.
	LIU J X， OU X B， WANG J C.Cytosolic Ca²⁺ involved in seed germination of naked oat enhanced by exogenous H₂S under saline-alkali stress［J］.Bulletin of Botanical Research，2020，40（1）：58-65.
36	刘小龙，李霞，钱宝云，等.植物体内钙信号及其在调节干旱胁迫中的作用［J］.西北植物学报，2014，34（9）：1927-1936.
	LIU X L， LI X， QIAN B Y，et al.Ca²⁺ signal transduction and its regulation role under drought stress in plant［J］.Acta Botanica Boreali-Occidentalia Sinica，2014，34（9）：1927-1936.

处理方式 Treatment	PBS /mL	NaHS /μL	LaCl₃ /μL
1	4	—	—
2	4	25	—
3	4	—	40
4	4	25	40

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[2]	刘建新, 欧晓彬, 王金成. 胞质Ca²⁺参与外源H₂S促进盐碱胁迫下裸燕麦种子萌发[J]. 植物研究, 2020, 40(1): 58-65.
[3]	马泓思1；潘亚婕1；王艳1；尹静1*；詹亚光1,2；赵微1；张梦岩1；梁　甜1. Ca²⁺在介导MeJA诱导白桦悬浮培养三萜合成中的作用[J]. 植物研究, 2015, 35(1): 117-126.
[4]	钱宝云;李霞*. 植物气孔运动调节的新进展[J]. 植物研究, 2013, 33(1): 120-128.
[5]	陈平波;李霞. 浓度NO对高表达转玉米C₄型pepc*水稻光合的促进[J]. 植物研究, 2012, 32(4): 402-409.
[6]	宰学明;钦　佩*;吴国荣;王　光;闫道良. 高温胁迫对花生幼苗光合速率、叶绿素含量、叶绿体Ca²⁺-ATPase、Mg²⁺-ATPase及Ca²⁺分布的影响[J]. 植物研究, 2007, 27(4): 416-420.
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苜蓿体内H₂S信号与Ca²⁺调节气孔运动的作用机制

The Mechanism of H₂S Signal and Ca²⁺ Regulating Stomatal Movement in Medicago sativa

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