Effect of Exogenous ATP on Cold Tolerance of Brassica campestris Seedlings

doi:10.7525/j.issn.1673-5102.2021.02.018

Abstract

Abstract:

The effects of exogenous ATP on the cold tolerance of rape seedlings were studied with “Longyou 7”Brassica campestris L. as experimental material. Compared with low temperature stress alone， MDA content and content of rapeseed seedlings were significantly decreased after exogenous ATP pretreatment and low temperature stress， while chlorophyll content and antioxidant enzyme activities（SOD， POD， CAT， APX） were observed. The expressions of RBOHD， RBOHF， CPK4 and CPK5 were increased. Compared with exogenous ATP+ low temperature， MDA content was significantly increased under EGTA+exogenous ATP+ low temperature treatment. Total chlorophyll content， T-AOC enzyme activity， RBOHD and RBOHF gene expression were all increased. Significantly decreased， MDA content was increased significantly under DMTU+exogenous ATP+low temperature treatment， total chlorophyll content， Ca²⁺-ATPase activity， CPK4， CPK5 gene expression were decreased， indicating that exogenous ATP affected cold tolerance of rape seedlings by Ca²⁺and H₂O₂-dependent mechanisms.

Key words: Brassica campestris L., exogenous ATP, low temperature stress, calcium, antioxidant enzymes

CLC Number:

S565.4

Xin-Dan HU, Yao LI, Xiao-Hua ZHANG, Juan-Hong LIANG, Teng-Guo ZHANG. Effect of Exogenous ATP on Cold Tolerance of Brassica campestris Seedlings[J]. Bulletin of Botanical Research, 2021, 41(2): 302-311.

Figures/Tables 7

Table 1

Fig.1

Fig.2

Fig.3

Fig.4

Fig.5

Fig.6

References 49

1	Sun L Y，Li X N，Wang Z S，et al.Cold priming induced tolerance to subsequent low temperature stress is enhanced by melatonin application during recovery in wheat［J］.Molecules，2018，23（5）：1091.
2	Yadav S K.Cold stress tolerance mechanisms in plants.A review［J］.Agronomy for Sustainable Development，2010，30（3）：515-527.
3	Liu Y F，Han X R，Zhan X M，et al.Regulation of calcium on peanut photosynthesis under low night temperature stress［J］.Journal of Integrative Agriculture，2013，12（12）：2172-2178.
4	Choudhury F K，Rivero R M，Blumwald E，et al.Reactive oxygen species，abiotic stress and stress combination［J］.The Plant Journal，2017，90（5）：856-867.
5	Huang S，Van Aken O，Schwarzländer M，et al.The roles of mitochondrial reactive oxygen species in cellular signaling and stress response in plants［J］.Plant Physiology，2016，171（3）：1551-1559.
6	Sun W C，Wu J Y，Fang Y，et al.Growth and development characteristics of winter rapeseed northern-extended from the cold and arid regions in China［J］.Acta Agronomica Sinica，2010，36（12）：2124-2134.
7	Kavi Kishor P B，Sreenivasulu N.Is proline accumulation per se correlated with stress tolerance or is proline homeostasis a more critical issue？［J］.Plant，Cell & Environment，2014，37（2）：300-311.
8	Saijo Y，Kinoshita N，Ishiyama K，et al.A Ca²⁺-dependent protein kinase that endows rice plants with cold- and salt-stress tolerance functions in vascular bundles［J］.Plant and Cell Physiology，2001，42（11）：1228-1233.
9	Khakh B S，Burnstock G.The double life of ATP［J］.Scientific American，2009，301（6）：84-92.
10	Lew R R，Dearnaley J D W.Extracellular nucleotide effects on the electrical properties of growing Arabidopsis thaliana root hairs［J］.Plant Science，2000，153（1）：1-6.
11	Tang W Q，Brady S R，Sun Y，et al.Extracellular ATP inhibits root gravitropism at concentrations that inhibit polar auxin transport［J］.Plant Physiology，2003，131（1）：147-154.
12	Cho S H，Choi J，Stacey G.Molecular mechanism of plant recognition of extracellular ATP［M］.//Atassi M.Protein Reviews.Singapore：Springer，2017：233-253.
13	Sun J，Zhang X，Deng S R，et al.Extracellular ATP signaling is mediated by H₂O₂ and cytosolic Ca²⁺ in the salt response of Populus euphratica cells［J］.PLoS One，2012，7（12）：e53136.
14	Deng S R，Sun J，Zhao R，et al.Populus euphratica APYRASE2 enhances cold tolerance by modulating vesicular trafficking and extracellular ATP in Arabidopsis plants［J］.Plant Physiology，2015，169（1）：530-548.
15	Huo K，Li X，He Y F，et al.Exogenous ATP enhance signal response of suspension cells of transgenic rice（Oryza sativa L.） expressing maize C₄-pepc encoded phosphoenolpyruvate carboxylase under PEG treatment［J］.Plant Growth Regulation，2017，82（1）：55-67.
16	Feng H Q，Jiao Q S，Sun K，et al.Extracellular ATP affects chlorophyll fluorescence of kidney bean（Phaseolus vulgaris） leaves through Ca²⁺ and H₂O₂-dependent mechanism［J］.Photosynthetica，2015，53（2）：201-206.
17	Tonón C，Terrile M C，Iglesias M J，et al.Extracellular ATP，nitric oxide and superoxide act coordinately to regulate hypocotyl growth in etiolated Arabidopsis seedlings［J］.Journal of Plant Physiology，2010，167（6）：540-546.
18	Clark G，Torres J，Finlayson S，et al.Apyrase（Nucleoside Triphosphate-Diphosphohydrolase） and extracellular nucleotides regulate cotton fiber elongation in cultured ovules［J］.Plant Physiology，2010，152（2）：1073-1083.
19	孙苑玲，邓澍荣，邓佳音，等.胞外ATP对胡杨细胞囊泡运输的影响［J］.北京林业大学学报，2016，38（7）：33-39.
	Sun Y L，Deng S R，Deng J Y，et al.Effect of eATP on vesicular trafficking in Populus euphratica cells［J］.Journal of Beijing Forestry University，2016，38（7）：33-39.
20	Zhou J H，Fan C Y，Liu K，et al.Extracellular ATP is involved in the initiation of pollen germination and tube growth in Picea meyeri［J］.Trees，2015，29（2）：563-574.
21	Hao L H，Wang W X，Chen C，et al.Extracellular ATP promotes stomatal opening of Arabidopsis thaliana through heterotrimeric G protein α subunit and reactive oxygen species［J］.Molecular Plant，2012，5（4）：852-864.
22	Kim S Y，Sivaguru M，Stacey G.Extracellular ATP in plants.Visualization，localization，and analysis of physiological significance in growth and signaling［J］.Plant Physiology，2006，142（3）：984-992.
23	Sun J，Zhang C L，Deng S R，et al.An ATP signalling pathway in plant cells：extracellular ATP triggers programmed cell death in Populus euphratica［J］.Plant，Cell and Environment，2012，35（5）：893-916.
24	吴锦程，陈宇，吴毕莎，等.钙处理对低温胁迫下枇杷幼苗Ca²⁺-ATPase活性和膜脂过氧化水平的影响［J］.西北农林科技大学学报：自然科学版，2016，44（2）：121-128.
	Wu J C，Chen Y，Wu B S，et al.Effects of calcium on Ca²⁺-ATPase activity and lipid peroxidation level of loquat seedling under low temperature stress［J］.Journal of Northwest A&F University：Natural Science Edition，2016，44（2）：121-128.
25	Sun T P.Gibberellin-GID1-DELLA：a pivotal regulatory module for plant growth and development［J］.Plant Physiology，2010，154（2） 567-570.
26	及利，韩姣，王芳，等.干旱胁迫对不同土壤基质下核桃楸幼苗的生理特性的影响［J］.植物研究，2019，39（5）：722-732.
	Ji L，Han J，Wang F，et al.Effects of drought stress on photosynthetic and physiological characteristics of Juglans mandshurica seedling in different soil substrates［J］.Bulletin of Botanical Research，2019，39（5）：722-732.
27	Demmig-Adams B，Adams W W.Xanthophyll cycle and light stress in nature：uniform response to excess direct sunlight among higher plant species［J］.Planta，1996，198（3）：460-470.
28	Heath R L，Packer L.Photoperoxidation in isolated chloroplasts：I.Kinetics and stoichiometry of fatty acid peroxidation［J］.Archives of Biochemistry & Biophysics，1968，125（1）：189-198.
29	Bates L S，Waldren R P，Teare I D.Rapid determination of free proline for water-stress studies［J］.Plant & Soil，1973，39（1）：205-207.
30	郝建军，康宗利，于洋.植物生理学实验技术［M］.北京：化学工业出版社，2007.
	Hao J J，Kang Z L，Yu Y.Plant physiology experiment technique［M］.Beijing：Chemical Industry Press，2007.
31	Frahry G，Schopfer P.NADH-stimulated，cyanide-resistant superoxide production in maize coleoptiles analyzed with a tetrazolium-based assay［J］.Planta，2001，212（2）：175-183.
32	Dhindsa R S，Matowe W.Drought tolerance in two mosses：correlated with enzymatic defence against lipid peroxidation［J］.Journal of Experimental Botany，1981，32（1）：79-91.
33	Rao M V，Paliyath G，Ormrod D P，et al.Influence of salicylic acid on H₂O₂ production，oxidative stress，and H₂O₂-metabolizing enzymes（salicylic acid-mediated oxidative damage requires H₂O₂）［J］.Plant Physiology，1997，115（1）：137-149.
34	乔新荣，张继英.植物谷胱甘肽过氧化物酶（GPX）研究进展［J］.生物技术通报，2016，32（9）：7-13.
	Qiao X R，Zhang J Y.Research progress on GPX in plants［J］.Biotechnology Bulletin，2016，32（9）：7-13.
35	池春玉，丁国华，连永权，等.低温胁迫对三种冷季型草坪草脯氨酸含量及膜透性的影响［J］.中国农学通报，2007，23（1）：101-104.
	Chi C Y，Ding G H，Lian Y Q，et al.Effect of low temperature stress on proline content and membrane permeability in three kind cold-season turfgrass［J］.Chinese Agricultural Science Bulletin，2007，23（1）：101-104.
36	Albornoz K，Cantwell M I，Zhang L，et al.Integrative analysis of postharvest chilling injury in cherry tomato fruit reveals contrapuntal spatio-temporal responses to ripening and cold stress［J］.Scientific Reports，2019，9（1）：2795.
37	项洪涛，梁晓艳，郑殿峰，等.花期低温对水稻叶鞘生理及结实率的影响［J］.干旱地区农业研究，2019，37（2）：114-122.
	Xiang H T，Liang X Y，Zheng D F，et al.Effects of low temperature stress during flowering stage on physiological characteristics of rice leaf sheath and seed-setting rate［J］.Agricultural Research in the Arid Areas，2019，37（2）：114-122.
38	张牡丹.几种外源物缓解环境胁迫对植物生理影响的研究［D］.兰州：西北师范大学，2019.
	Zhang M D.Studies on the effects of several exogenous substances on plant physiology by mitigating environmental stress［D］.Lanzhou：Northwest Normal University，2019.
39	李亮，董春娟，尚庆茂.内源水杨酸参与黄瓜叶片光合系统对低温胁迫的响应［J］.园艺学报，2013，40（3）：487-497.
	Li L，Dong C J，Shang Q M.Role of endogenous salicylic acid in responding of cucumber leaf photosynthetic systems to low temperature stress［J］.Acta Horticulturae Sinica，2013，40（3）：487-497.
40	王春萍，雷开荣，李正国，等.低温胁迫对水稻幼苗不同叶龄叶片叶绿素荧光特性的影响［J］.植物资源与环境学报，2012，21（3）：38-43.
	Wang C P，Lei K R，Li Z G，et al.Effects of chilling stress on chlorophyll fluorescence characteristics of seedling leaves with different leaf ages of Oryza sativa［J］.Journal of Plant Resources and Environment，2012，21（3）：38-43.
41	Jiao Q S，Feng H Q，Tian W Y，et al.Extracellular ATP functions in alleviating the decrease of PSII photochemistry caused by the infection of Xanthomonas campestris pv.phaseoli［J］.Plant Pathology，2016，65（5）：819-825.
42	Baxter A，Mittler R，Suzuki N.ROS as key players in plant stress signalling［J］.Journal of Experimental Botany，2014，65（5）：1229-1240.
43	Sathiyaraj G，Srinivasan S，Kim Y J，et al.Acclimation of hydrogen peroxide enhances salt tolerance by activating defense-related proteins in Panax ginseng C.A.Meyer［J］.Molecular Biology Reports，2014，41（6）：3761-3771.
44	张腾国，李瑶，刁志宏，等.白菜型油菜RbohC和RbohF基因克隆与表达分析［J］.西北植物学报，2018，38（10）：1767-1776.
	Zhang T G，Li Y，Diao Z H，et al.Cloning and expression analysis of RbohC and RbohF genes in Brassica rapa L.［J］.Acta Botanica Boreali-Occidentalia Sinica，2018，38（10）：1767-1776.
45	Liu Y K，Zhou J.MAPping kinase regulation of ICE1 in freezing tolerance［J］.Trends in Plant Science，2018，23（2）：91-93.
46	Almadanim M C，Alexandre B M，Rosa M T G，et al.Rice calcium-dependent protein kinase OsCPK17 targets plasma membrane intrinsic protein and sucrose-phosphate synthase and is required for a proper cold stress response［J］.Plant，Cell & Environment，2017，40（7）：1197-1213.
47	Jeter C R，Tang W Q，Henaff E，et al.Evidence of a novel cell signaling role for extracellular adenosine triphosphates and diphosphates in Arabidopsis［J］.The Plant Cell，2004，16（10）：2652-2664.
48	Demidchik V，Shabala S N，Davies J M.Spatial variation in H₂O₂ response of Arabidopsis thaliana root epidermal Ca²⁺ flux and plasma membrane Ca²⁺ channels［J］.The Plant Journal，2007，49（3）：377-386.
49	Costa A，Drago I，Behera S，et al.H₂O₂ in plant peroxisomes：an in vivo analysis uncovers a Ca²⁺-dependent scavenging system［J］.The Plant Journal，2010，62（5）：760-772.

基因 Gene	基因号 Genbank number	引物 Primer	碱基序列 Sequence（5’to 3’）
Actin	XM_013830703.2	Actin-F	TGTGCCAATCTACGAGGGTTT
Actin	XM_013830703.2	Actin-R	TTTCCCGCTCTGCTGTTGT
BrRbohD	XM_009153345.2	RbohD-F	CACTAGCTCGCCGTAATAACATCTC
BrRbohD	XM_009153345.2	RbohD-R	CGTCCATCTTCGTCTTTGTCC
BrRbohF	XM_009114548.2	RbohF-F	CAGAGCCGATGAAACAACAAAG
BrRbohF	XM_009114548.2	RbohF-R	GCCAAGACCAACCAATAAGAGAA
BrCPK4	XM_009135901.2	CPK4-F	CAATTCGGGACGACCTATCT
BrCPK4	XM_009135901.2	CPK4-R	GGTGCATGATCTGAATCTCG
BrCPK5	XM_018653206.1	CPK5-F	ACTTTGATTCGGATCCTTGG
BrCPK5	XM_018653206.1	CPK5-R	ACACCGTTCTCACAGATCCA

[1]	Senyao LIU, Fenglin JIA, Qing GUO, Gaofeng FAN, Boru ZHOU, Tingbo JIANG. Response Analysis of Transcription Factor PsnbHLH162 Gene in Populus simonii × P. nigra under Salt Stress and Low Temperature Stress [J]. Bulletin of Botanical Research, 2023, 43(2): 300-310.
[2]	Dongmei HUANG, Ying CHEN, Lu BAI, Di’an NI, Yiyang XU, Zhiguo ZHANG, Qiaoping QIN. Transcriptome Analysis of Hemerocallis fulva Leaves Respond to Low Temperature Stress [J]. Bulletin of Botanical Research, 2022, 42(3): 424-436.
[3]	Lin LU, Shang-Yu YANG, Wei-Dong LIU, Li-Ming LU. To Explore the Transcription Factor in Response to Low Temperature Stress in Nicotiana alata by Transcriptome Sequencing [J]. Bulletin of Botanical Research, 2021, 41(1): 119-129.
[4]	CHEN Long-Tao, YANG Bo-Wen, WANG Ying-Nan, FAN Ming-Shou. Effect of Calcium treat on the Development and Fruit Setting of Hazwlnut Tree [J]. Bulletin of Botanical Research, 2018, 38(4): 597-603.
[5]	HE Wen-Ping, WANG Xin-Xia, ZHANG Xu-Qiang, YANG Peng-Jun, QIU Yi-Zhi, YANG Ning. Exogenous ABA Affected on Antioxidant Enzymes and Osmotic Adjustment in Choraspora bungeana Under Low Temperature Stress [J]. Bulletin of Botanical Research, 2016, 36(6): 870-877.
[6]	LI Yan-Hong;JIANG Yong;WANG Wen-Jie;ZHANG Bao-You*. Effect of Organic and Inorganic Carbon on Extracellular Enzyme Activity of Acid Phosphatase and Proteases in Three Kinds of Fungal Hyphae [J]. Bulletin of Botanical Research, 2013, 33(4): 404-409.
[7]	LI Mei-Lan;LI De-Wen;YU Jing-Hua;ZU Yuan-Gang. Effect of Exogenous Nitric Oxide on Photosynthetic Pigment and Antioxidant Enzymes in Taxus chinensis* var. marei Seedlings [J]. Bulletin of Botanical Research, 2013, 33(1): 39-44.
[8]	YOU Jia;ZHANG Jin;WANG Wen-Rui;LU Jin;JIA Peng-Xiang;MIAO Jian-Shun;YANG Ying-Li. Changes of Physiological Properties in Limonium aureum*(Linn.) Hill Seedlings under Salt Stress [J]. Bulletin of Botanical Research, 2013, 33(1): 45-50.
[9]	SHEN Lei;GUO Shui-Liang;YANG Wu;CAO Tong;Janice M. Glime. Physiological Responses of Hypnum fertile* Sendtn.(Musci:Hypnaceae) to Short-term Extreme Temperature Stress [J]. Bulletin of Botanical Research, 2011, 31(1): 40-48.
[10]	FAN Qing;LÜXiu-Jun;YANG Liu;YANG Ying-Li. Effects of Cd Stress on the Seed Germination,Seedling Growth and Activities of Antioxidant Enzymes in Petunia hybrida* [J]. Bulletin of Botanical Research, 2010, 30(6): 685-691.
[11]	XU Na;DONG Xiao-Hong;GUAN Yang;WANG Jing. The Effect of Low Temperature Stress on Antioxidant Enzyme Activities of Buxus sempervirens* L. Under Different Light Conditions [J]. Bulletin of Botanical Research, 2007, 27(5): 574-577.
[12]	NIE Qing-Juan;MENG Zhao;LIANG Hai-Yong;MENG Qing-Rui;LI Yan-Hui*. Effect on Low Temperature Stress on Lipid Peroxidation and Activity of Cell Defense Enzymes of Four Evergreen Broadleaved Tree Species [J]. Bulletin of Botanical Research, 2007, 27(5): 578-581.
[13]	YAN Yue-Hong, HE Zu-Xia, SHEA She-Sang, HUANG Zhong-Liang, XING Fu-Wu. A survey on the invasive plants in Kap O Islands, Northeastern Hong Kong [J]. Bulletin of Botanical Research, 2005, 25(2): 242-248.
[14]	WU Jian-Hui, YANG Ling, SUN Guo-Rong. Generation of activated oxygen and change of cell defense enzyme activity in leaves of maize seedling under the stress of low temperature [J]. Bulletin of Botanical Research, 2004, 24(4): 456-459.
[15]	LIU Jin-Wen, SHA Wei, WANG Yan-Jun. The physiological effect of the different phenological period Ledum palustre subsp. decumdens on low temperature stress [J]. Bulletin of Botanical Research, 2004, 24(2): 197-200.