Ectopic Express of EgrNAC1 Enhances Cold Tolerance and Sensitivity to Drought and Salt in Arabidopsis thaliana

doi:10.11869/j.issn.100-8551.2021.03.0567

Journal of Nuclear Agricultural Sciences ›› 2021, Vol. 35 ›› Issue (3): 567-575.DOI: 10.11869/j.issn.100-8551.2021.03.0567

• Induced Mutations for Plant Breeding·Agricultural Biotechnology • Previous Articles Next Articles

Ectopic Express of EgrNAC1 Enhances Cold Tolerance and Sensitivity to Drought and Salt in Arabidopsis thaliana

CONG Qing, NI Xiaoxiang, CHENG Longjun^*

Zhejiang Agriculture and Forestry University/State Key Laboratory of Subtropical Silviculture, Hangzhou, Zhejiang 311300

Received:2019-10-25 Online:2021-03-10 Published:2021-01-19

异源表达EgrNAC1提高拟南芥抗寒性和对干旱、高盐的敏感性

从青, 倪晓祥, 程龙军^*

浙江农林大学/亚热带森林培育国家重点实验室,浙江杭州 311300

通讯作者: ^*程龙军,男,副教授,主要从事巨桉基因组学与分子遗传学研究。E-mail:ljcheng@zju.edu.cn
作者简介:从青,女,主要从事巨桉抗逆分子机理研究。E-mail:348617225@qq.com
基金资助:
国家自然科学基金(31270657),浙江省科技厅林木新品种选育重大科技专项(2016C02056-9)

Abstract

Abstract: Eucalyptus is a very important economic tree species in southern China. However, most cultivated Eucalyptus is sensitive to abiotic stresses such as low temperature, drought and salinity, which limit its planting region and the improvement of cultivation benefits. EgrNAC1 (Eucgr.I00058) is a NAC-like transcription factor gene in Eucalyptus grandis which is induced by low temperature, drought and salt treatment. To further explore its function in abiotic stress response, homozygous transgenic lines that overexpressed EgrNAC1 were obtained by ectopic expression in Arabidopsis thaliana. And, subjected to low temperature(-6℃), drought, high salt(300 mmol·L^-1 NaCl)and ABA(0.5 μmol·L^-1) treatment, the responses of transgenic lines to these abiotic stress were analyzed. The results showed that overexpression of EgrNAC1 in Arabidopsis could increase the tolerance of plants to low temperature. After 12 hours treatment at -6℃ and then 5 days of recovering, the survival rate of the transgenic lines of EgrNAC1-OE1 and EgrNAC1-OE8 were 88.9% and 81.5% respectively, while only 29.6% for wild type. Further analysis showed the expression levels of three CBF pathway related genes AtCBF1, AtCBF2 and AtRD29A were significantly up-regulated in the EgrNAC1 over-expression lines under low temperature. On the other hand, sensitivity to drought and salinity in transgenic plants were increased and ABA sensitivity showed a decrease compared to wild type These results implied that EgrNAC1 possibly act as a positive regulator in the low temperature response through regulating the expression of genes invoved in ICE-CBF pathway in Eucalyptus grandis, thereby improving the resistance of plants to low temperature stress. However, under drought and high salt stress, EgrNAC1 may play a negative regulatory role, and ABA signal transduction possibly be involved in this regulation process. This study provides valuable information for elucidating of the function of EgrNAC1 and benefits for molecular-assisted breeding of abiotic stress-resistance in Eucalyptus.

Key words: Eucalyptus grandis, EgrNAC1, low temperature, drought, salt stress

摘要： 桉树是我国南方重要用材树种,但其栽培种对低温、干旱和高盐等非生物逆境抗性弱,限制了其栽培范围的扩大和栽培效益的提高。EgrNAC1(Eucgr.I00058)是巨桉中受低温、干旱和高盐诱导表达的NAC类转录因子。为进一步研究EgrNAC1的功能,通过异源转化拟南芥,获得了超表达EgrNAC1的转基因纯合株系,并对其进行低温(-6℃)、干旱、高盐(NaCl 300 mmol·L^-1 )和脱落酸(0.5 μmol·L^-1)处理,分析转基因株系对非生物逆境的响应。结果表明,超表达EgrNAC1能提高拟南芥植株对低温的抗性。-6℃处理12 h,恢复5 d后,转基因株系EgrNAC1-OE1和EgrNAC1-OE8的存活率分别达到88.9%和81.5%,而野生型仅有29.6%。低温处理下,转基因株系中3个低温信号传导(CBF)途径相关的基因,AtCBF1、AtCBF2和AtRD29A的表达量相对于野生型显著上调;超表达EgrNAC1提高了转基因植株对干旱和高盐的敏感程度。同时,转基因植株相对于野生型表现出对ABA敏感性的下降。综上,EgrNAC1在巨桉低温响应中可能作为一个正向调控因子,通过参与调控CBF途径中低温响应相关基因的表达,提高植物在低温逆境下的适应性。但在干旱和高盐逆境下,EgrNAC1可能发挥负调控作用,且这种作用可能与ABA的影响有关。本研究结果为深入了解桉树EgrNAC1的功能以及开展桉树抗逆分子辅助育种提供了一定的理论参考。

关键词: 巨桉, EgrNAC1, 低温, 干旱, 高盐

CONG Qing, NI Xiaoxiang, CHENG Longjun. Ectopic Express of EgrNAC1 Enhances Cold Tolerance and Sensitivity to Drought and Salt in Arabidopsis thaliana[J]. Journal of Nuclear Agricultural Sciences, 2021, 35(3): 567-575.

从青, 倪晓祥, 程龙军. 异源表达EgrNAC1提高拟南芥抗寒性和对干旱、高盐的敏感性[J]. 核农学报, 2021, 35(3): 567-575.

References

[1] Zhu J K.Abiotic stress signaling and responses in plants[J]. Cell 2016, 167(2): 313-324
[2] Anjum S A, Xie X Y, Wang L C, Saleem M F, Lei W.Morphological, physiological and biochemical responses of plants to drought stress[J]. African Journal of Agricultural Research, 2011, 6(9): 2026-2032
[3] Nakashima K, Ito Y, Yamaguchi-Shinozaki K.Transcriptional regulatory networks in response to abiotic stresses in Arabidopsis and grasses[J]. Plant Physiology, 2009, 149(1): 88-95
[4] Singh K B, Foley R C, Oñate-Sánchez L.Transcription factors in plant defense and stress responses[J]. Current Opinion in Plant Biology, 2002, 5(5): 430-436
[5] Harfouche A, Meilan R, Altman A.Molecular and physiological responses to abiotic stress in forest trees and their relevance to tree improvement[J]. Tree Physiology, 2014, 34(11): 1181-1198
[6] Olsen A, Ernst H A, Leggio L, Skriver K.NAC transcription factors: Structurally distinct, functionally diverse[J]. Trends in Plant Science, 2005, 10(2): 79-87
[7] Puranik S, Sahu P, Srivastava P S, Prasad M.NAC proteins: Regulation and role in stress tolerance[J]. Trends in Plant Science, 2012, 17(6): 369-381
[8] Souer E, Houwelingen A V, Kloos D, Mol J, Koes R.The no apical meristem gene of Petunia is required for pattern formation in embryos and flowers and is expressed at meristem and primordia boundaries[J]. Cell, 1996, 85(2): 159-170
[9] Petricka J J, Winter C M, Benfey P N.Control of Arabidopsis root development[J]. Annual Review of Plant Biology, 2012, 63(1): 563
[10] Hao Y J, Wei W, Song Q X, Chen H W, Zhang Y Q, Wang F, Zou H F, Lei G, Tian A G, Zhang W K.Soybean NAC transcription factors promote abiotic stress tolerance and lateral root formation in transgenic plants[J]. Plant Journal, 2011, 68(2): 302-313
[11] Zimmermann R, Werr W.Pattern formation in the monocot embryo as revealed by NAM and CUC3 orthologues from Zea mays L[J]. Plant Molecular Biology, 2005, 58(5): 669-685
[12] Negi S, Tak H, Ganapathi T R.Native vascular related NAC transcription factors are efficient regulator of multiple classes of secondary wall associated genes in banana[J]. Plant Science, 2017, 265(1): 70-86
[13] Krizek B A, Fletcher J C.Molecular mechanisms of flower development: An armchair guide[J]. Nature Reviews Genetics. 2005, 6(9): 688-698
[14] Guo Y F, Gan S S.AtNAP, a NAC family transcription factor, has an important role in leaf senescence[J]. Plant Journal for Cell and Molecular Biology, 2006, 46(4): 601-612
[15] Nakashima K, Takasaki H, Mizoi J, Shinozaki K, Yamaguchi-Shinozaki K.NAC transcription factors in plant abiotic stress responses[J]. BBA-Gene Regulatory Mechanisms, 2012, 1819(2): 97-103
[16] Hu H H, Dai M Q, Yao J L, Xiao B Z, Li X H, Zhang Q F, Xiong L Z.Overexpressing a NAM, ATAF, and CUC (NAC) transcription factor enhances drought resistance and salt tolerance in rice[J]. Proceedings of the National Academy of Sciences of the United States of America, 2006, 103(35): 12987-12992
[17] Liu Y M, Yu X W, Liu S S, Peng H, Mijiti A, Wang Z, Zhang H, Ma H.A chickpea NAC-type transcription factor, CarNAC6, confers enhanced dehydration tolerance in Arabidopsis[J]. Plant Molecular Biology Reporter, 2017, 35(1): 83-96
[18] Lu X, Zhang X F, Duan H, Lian C L, Liu C, Yin W L, Xia X L.Three stress-responsive NAC transcription factors from Populus euphratica differentially regulate salt and drought tolerance in transgenic plants[J]. Physiologia Plantarum, 2018, 162:77-79
[19] 孙丽娟, 王晓荣, 倪晓祥, 程龙军. 巨桉非生物逆境响应基因EgrNAC1的基因结构和表达分析[J]. 林业科学, 2017, 53(10): 60-69
[20] Steponkus P L, Uemura M, Joseph R A, Gilmour S J, Thomashow M F.Mode of action of the COR15a gene on the freezing tolerance of Arabidopsis thaliana[J]. Proceedings of the National Academy of Sciences of the United States of America, 1998, 95(24): 14570-14575
[21] 阎依超, 万春雁, 古咸彬, 郭成宝, 陈月红, 高志红. 低温胁迫下转RdreB1BI草莓逆境相关基因表达及抗性评价[J]. 核农学报, 2018, 32(11): 61-72
[22] Liu X, Hong L, Li X Y, Yao Y, Hu B, Li L.Improved drought and salt tolerance in transgenic Arabidopsis overexpressing a NAC Transcriptional Factor from Arachis hypogaea[J]. Bioscience, Biotechnology, and Biochemistry, 2011, 75(3): 443-450
[23] 刘静妍, 施怡婷, 杨淑华. CBF:平衡植物低温应答与生长发育的关键[J]. 植物学报, 2017(6): 12-21
[24] Livak K J, Schmittgen T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2^-ΔΔCt method[J]. Methods, 2000, 25(4): 402-408
[25] 曹云飞, 张海娜, 肖凯. CBF转录因子介导的植物低温信号转导研究进展[J]. 棉花学报, 2007, 19(4): 304-311
[26] 童超. ABA生理功能与信号转导相关综述[J]. 科技资讯, 2008(10): 44-45
[27] Lu P L, Chen N Z, An R, Su Z, Qi B S, Ren F, Chen J, Wang X C.A novel drought-inducible gene, ATAF1, encodes a NAC family protein that negatively regulates the expression of stress-responsive genes in Arabidopsis[J]. Plant Molecular Biology, 2007, 63(2): 289-305
[28] Han X M, Feng Z Q, Xing D D, Yang Q, Wang R G, Qi L W, Li G J.Two NAC transcription factors from Caragana intermedia altered salt tolerance of the transgenic Arabidopsis[J]. BMC Plant Biology, 2015, 15(1): 1-12
[29] Tran L-S P, Nakashima K, Sakuma Y, Simpson S D, Fujita Y, Maruyama K, Fujita M, Seki M, Shinozaki K, Yamaguchi-Shinozaki K. Isolation and functional analysis of Arabidopsis stress-inducible NAC transcription factors that bind to a drought-responsive cis-element in the early responsive to dehydration stress 1 promoter[J]. The Plant Cell, 2004, 16(9): 2481-2498
[30] Liu G, Li X, Jin S, Liu X, Zhu L, Nie Y, Zhang X.Overexpression of rice NAC gene SNAC1 improves drought and salt tolerance by enhancing root development and reducing transpiration rate in transgenic cotton[J]. PLoS One, 2014, 9(1): e86895
[31] Yang X W, Wang X Y, Ji L, Yi Z L, Fu C X, Ran J C, Hu R B, Zhou G K.Overexpression of a Miscanthus lutarioriparius NAC gene MlNAC5 confers enhanced drought and cold tolerance in Arabidopsis[J]. Plant Cell Reports, 2015, 34(6): 943-958
[32] Jin H X, Huang F, Cheng H, Song H N, Yu D Y.Overexpression of the GmNAC2 gene, an NAC transcription factor, reduces abiotic stress tolerance in tobacco[J]. Plant Molecular Biology Reporter, 2013, 31(2): 435-442
[33] Huang L, Hong Y B, Zhang H J, Li D Y, Song F M.Rice NAC transcription factor ONAC095 plays opposite roles in drought and cold stress tolerance[J]. BMC Plant Biology, 2016, 16(1): 203
[34] You J, Zong W, Li X K, Ning J, Hu H h, Li X H, Xiao J H, Xiong L Z. The SNAC1-targeted gene OsSRO1c modulates stomatal closure and oxidative stress tolerance by regulating hydrogen peroxide in rice[J]. Journal of Experimental Botany, 2013, 64(2): 569-583
[35] Huang Q J, Wang Y, Li B, Chang J L, Chen M J, Li K X, Yang G X, He G Y.TaNAC29, a NAC transcription factor from wheat, enhances salt and drought tolerance in transgenic Arabidopsis[J]. BMC Plant Biology, 2015, 15(1): 1-15
[36] Shi Y, Ding Y, Yang S.Molecular regulation of CBF signaling in cold acclimation[J]. Trends in Plant Science, 2018, 23(7): 623-637
[37] Viswanathan C, Zhu J H, Zhu J K.Cold stress regulation of gene expression in plants[J]. Trends in Plant Science, 2007, 12(10): 444-451
[38] Shan W, Kuang J F, Jin L W, Chen J Y.Banana fruit NAC transcription factor MaNAC1 is a direct target of MaICE1 and involved in cold stress through interacting with MaCBF1[J]. Plant Cell and Environment, 2014, 37(9): 2116-2127
[39] Qu Y T, Mei D, Zhang Z Q, Dong J L, Tao W.Overexpression of the Medicago falcata NAC transcription factor MfNAC3 enhances cold tolerance in Medicago truncatula[J]. Environmental and Experimental Botany, 2016, 129: 67-76

Metrics

Comments

创刊：1987年
主管部门：农业农村部
主办单位：中国原子能农学会与中国农业科学院农产品加工研究所（前中国农业科学院原子能利用研究所）
ISSN：1000-8551

访问统计
总访问：
今日访问：
当前在线：

Ectopic Express of EgrNAC1 Enhances Cold Tolerance and Sensitivity to Drought and Salt in Arabidopsis thaliana

异源表达EgrNAC1提高拟南芥抗寒性和对干旱、高盐的敏感性

RichHTML

PDF

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 0

Recommended Articles

Metrics

Comments