Salt Stress Response and Functional Indentification of HbICEs from Rubber Tree

doi:10.3969/j.issn.1000-2561.2022.11.003

Abstract

Abstract:

ICE (inducer of CBF expression), the MYC (myelocytomatosis)-like bHLH (basic helix-loop-helix) transcription factor, plays an important role in regulating the responses of plants to abiotic stresses such as low temperature, drought, and high salt. In this study, the expression patterns of rubber tree ICE genes were analyzed by qPCR. The expression of five HbICEs were up-regulated in rubber tree seedlings upon salt stress, HbICE2, HbICE3, HbICE4 and HbICE5 were significantly up-regulated under salt stress for 2 h. At 4 h of salt stress, the expression of HbICE4 and HbICE5 reached the maximum with the most up-regulated level of HbICE4. Under salt stress for 4-24 h, HbICE3 was significantly down regulated. The expression of HbICE1 had no obvious change before 4 h of salt stress, but increased significantly after 8 h. Therefore, all five HbICE genes may participate in the salt stress response of rubber tree. Overexpression of HbICEs, especially HbICE4, led to an enhanced salt-tolerance of transgenic Arabidopsis based on survival rate, average fresh and dry weight per plant. Under salt stress, all wild-type Arabidopsis wilted. The survival rate of HbICE4 overexpressing Arabidopsis was 96.29%, the average fresh weight per plant was 0.03 g, and the average dry weight per plant was 0.0035 g, the three growth parameters of HbICE4 overexpressing plants were the highest among the transformed strains of rubber tree HbICE genes. Compared with the physiological indexes before salt stress, the relative conductivity of WT and HbICEs transgenic plants increased significantly under salt stress, and the chlorophyll content and relative water content decreased significantly. Among them, relative conductivity increase of HbICE4 transformed Arabidopsis was the smallest, and the decrease of SPAD value (chlorophyll content) and relative water content was the smallest. Therefore, the enhanced salt-tolerance was associated with lower relative conductivity, a physiological parameter for evaluating membrane system stability, as well as higher chlorophyll content and relative water content in comparison to those of WT. Salt stress leads to the damage of plant cell membrane system, resulting in the decrease of water content, the destruction of chloroplasts and the degradation of chlorophyll. The membrane system of plants overexpressing HbICE4 was slightly damaged under salt stress, delaying the decrease of water content and the degradation of chlorophyll. Rubber tree HbICEs can not only improve the cold and drought tolerance of transgenic plants, but also improve the salt-tolerance of transgenic plants. This study would deepen the understanding of the biological function of rubber tree HbICEs, and provide candidate target genes for improving salt-tolerance of crops.

Key words: Hevea brasiliensis Muell. Arg., inducer of CBF expression, salt tolerance, functional identification

CLC Number:

S794.1

LI Yan, YANG Shuguang, SHI Minjing, TIAN Weimin. Salt Stress Response and Functional Indentification of HbICEs from Rubber Tree[J]. Chinese Journal of Tropical Crops, 2022, 43(11): 2199-2206.

Figures/Tables 3

Fig. 1 Expression analysis of five HbICE family genes in leaves of ‘93-114’ under NaCl stress ** and * indicate extremely significant difference (P<0.01) and significant difference (P<0.05), respectively.

Fig. 2 Effect of overexpression of HbICEs on salt tolerance of Arabidopsis seedlings Different capital letters and lowercase letters indicate extremely significant difference (P<0.01) and significant difference (P<0.05), respectively.

Fig. 3 Determination of physiological indexes of wild-type Arabidopsis and transgenic lines under NaCl stress Different capital letters and lowercase letters indicate extremely significant difference (P<0.01) and significant difference (P<0.05), respectively.

References 31

[1]	VERMA R K, KUMAR V V S, YADAV S K, KUMAR T S, RAO M V, CHINNUSAMY V. Overexpression of Arabidopsis ICE1 enhances yield and multiple abiotic stress tolerance in indica rice[J]. Plant Signaling and Behavior, 2020, 15(11): 1814547. DOI URL
[2]	BADAWI M, REDDY Y V, AGHARBAOUI Z, TOMINAGAY, DANYLUK J, SARHAN F, HOUDE M. Structure and functional analysis of wheat ICE (inducer of CBF expression) genes[J]. Plant Cell Physiology, 2008, 49: 1237-1249. DOI URL
[3]	CHINNUSAMY V. ICE1: a regulator of cold-induced transcriptome and freezing tolerance in Arabidopsis[J]. Genes Development, 2003, 17: 1043-1054. DOI URL
[4]	NAKAMURA J, TAKASHI YUASA T, HUONG TT, HARANO K, IWAYA-INOUE M. Rice homologs of inducer of CBF expression (OsICE) are involved in cold acclimation[J]. Plant Biotechnology, 2011, 28: 303-309. DOI URL
[5]	LI Y, QUAN C, YANG S, WU S, SHI M, WANG J, TIAN W M. Functional identification of ICE transcription factors in rubber tree[J]. Forests, 2022, 13(1): 52. DOI URL
[6]	TOLEDO-ORTIZ G, HUQ E, QUAIL P H. The Arabidopsis basic/helix-loop-helix transcription factor family[J]. Plant Cell, 2003, 15: 1749-1770. DOI URL
[7]	NIU Y, FIGUEROA P, BROWSE J. Characterization of JAZ interacting bHLH transcription factors that regulate jasmonate responses in Arabidopsis[J]. Journal of Experimental Botany, 2011, 62(6): 2143-2154. DOI URL
[8]	MIURA K, NOZAWA R. Overexpression of SIZ1 enhances tolerance to cold and salt stresses and attenuates response to abscisic acid in Arabidopsis thaliana[J]. Plant Biotechnology, 2014, 31(2): 167-172. DOI URL
[9]	HU Y, JIANG Y, HAN X, WANG H, PAN J, YU D. Jasmonate regulates leaf senescence and tolerance to cold stress: crosstalk with other phytohormones[J]. Journal Experimental Botany, 2017, 68(6): 1361-1369. DOI URL
[10]	HU Y, JIANG L, WANG F, YU D. Jasmonate regulates the inducerof cbf expression-c-repeat binding factor/dre bindingfactor1 cascade and freezing tolerance in Arabidopsis[J]. The Plant Cell, 2013, 25(8): 2907-2924. DOI URL
[11]	FENG H L, MA N N, MENG X, ZHANG S, WANG J R, CHAI S, MENG Q W. A novel tomato MYC-type ICE1-like transcription factor, SlICE1a, confers cold, osmotic and salt tolerance in transgenic tobacco[J]. Plant Physiology Biochemistry, 2013, 73: 309-320. DOI URL
[12]	GUO J, REN Y, TANG Z, SHI W, ZHOU M. Characterization and expression profiling of the ICE-CBF-COR genes in wheat[J]. Peer Journal, 2019, 7: e8190. DOI URL
[13]	ZUO Z F, KANG H G, PARK M Y, JEONG H, SUN H J, SONG P S, LEE H Y. Zoysia japonica MYC type transcription factor ZjICE1 regulates cold tolerance in transgenic Arabidopsis[J]. Plant Science, 2019, 289: 110254. DOI URL
[14]	DANIEL G Z, JONATHAN T V, DANIEL C, THOMASHOW M F. Cold induction of Arabidopsis CBF genes involves multiple ICE (inducer of CBF expression) promoter elements and a cold-regulatory circuit that is desensitized by low temperature[J]. Plant Physiology, 2003, 133(2): 910-918. DOI URL
[15]	杜忠杰, 林电, 许杰, 林建明. 海南省橡胶园土壤养分状况研究[J]. 广东农业科学, 2011, 38(11): 73-77.
	DU Z J, LIN D, XU J, LIN J M. The nutrient status of rubber plantation soil in Hainan province[J]. Guangdong Agricultural Sciences, 2011, 38(11): 73-77. (in Chinese)
[16]	王旺年, 葛均筑, 杨海昌, 阴法庭, 黄太利, 蒯婕, 王晶, 汪波, 周广生, 傅廷栋. 大田作物在不同盐碱地的饲料价值评价[J]. 作物学报, 2022, 48(6): 1451-1462. DOI
	WANG W N, GE J Z, YANG H C, YIN F T, HUANG T L, KUAI J, WANG J, WANG B, ZHOU G S, FU T D. Adaptation of feed crops to saline-alkali soil stress and effect of improving saline-alkali soil[J]. Acta Agronomica Sinica, 2022, 48(6): 1451-1462. (in Chinese) DOI URL
[17]	KOTULA L, GARCIA C P, ZORB C, COLMER T D, FLOWERS T J. Improving crop salt tolerance using transgenic approaches: an update and physiological analysis[J]. Plant Cell Environment. 2020, 43(12): 2932-2956. DOI URL
[18]	李言, 余文才, 陆庆志, 杨署光, 田维敏. 非生物胁迫对橡胶树热激转录因子家族成员表达的影响[J]. 热带作物学报, 2021, 42(8): 2119-2125.
	LI Y, YU W C, LU Q Z, YANG S G, TIAN W M. Effects of abiotic stresses on the expression of heat shock transcription factor (HSF) family members in rubber tree (Hevea brasiliensis Muell. Arg.)[J]. Chinese Journal of Tropical Crops, 2021, 42(8): 2119-2125. (in Chinese)
[19]	LI Y, YU W, CHEN Y, YANG S G, WU S H, CHAO J Q, WANG X L, TIAN W M. Genome-wide identification and characterization of heat-shock transcription factors in rubber tree[J]. Forests, 2019, 10(12): 1157. DOI URL
[20]	LI F, GUO S, ZHAO Y, CHEN D, CHONG K, XU Y. Overexpression of a homopeptide repeat-containing bHLH protein gene (OrbHLH001) from Dongxiang wild rice confers freezing and salt tolerance in transgenic Arabidopsis[J]. Plant Cell Reports, 2010, 29: 977-986. DOI URL
[21]	ZHOU J, LI F, WANG J L, MA Y, CHONG K, XU Y Y. Basic helix-loop-helix transcription factor from wild rice (OrbHLH2) improves tolerance tosalt-and osmotic stress in Arabidopsis[J]. Journal of Plant Physiology, 2009, 166(12): 1296-1306. DOI URL
[22]	王昕嘉, 李昆志. 植物bHLH转录因子参与非生物胁迫信号通路研究进展[J]. 生命科学, 2015, 27(2): 208-216.
	WANG X J, LI K Z. Progress of plant bHLH transcription factors involved in abiotic stress signaling pathways[J]. Chinese Bulletin of Life Sciences, 2015, 27(2): 208-216. (in Chinese)
[23]	CHEN Y, LI F, MA Y, CHONG K, XU Y. Overexpression of OrbHLH001, a putative helix-loop-helix transcription factor, causes increased expression of AKT1 and maintains ionic balance under salt stress in rice[J]. Journal of Plant Physiology, 2013, 170: 93-100. DOI URL
[24]	ZHANG T, MO J, ZHOU K, CHANG Y, LIU Z. Overexpression of Brassica campestris BcICE1 gene increases abiotic stress tolerance in tobacco[J]. Plant Physiology and Biochemistry, 2018, 132: 515-523. DOI URL
[25]	潘凌云, 马家冀, 李建民, 尹兵兵, 付畅. 植物盐胁迫应答转录因子的研究进展[J]. 生物工程学报, 2022, 38(1): 50-65.
	PAN L Y, MA J J, LI J M, YIN B B, FU C. Advances of salt stress-responsive transcription factors in plants[J]. Chinese Journal of Biotechnology, 2022, 38(1): 50-65. (in Chinese)
[26]	李光道, 白生才, 张秀志, 李军, 唐学清, 王宗明, 陈国斌. 植物抗盐性研究综述[J]. 甘肃农业科技, 2011(2): 29-33.
	LI G D, BAI S C, ZHANG X Z, LI J, TANG X Q, WANG Z M, CHEN G B. Review on salt resistance of plants[J]. Gansu Agricultural Science and Technology, 2011(2): 29-33. (in Chinese)
[27]	PARDO J M. Biotechnology of water and salinity stress tolerance[J]. Current Opinion in Biotechnology, 2010, 21(2): 185-196. DOI PMID
[28]	QIN H, LI Y, HUANG R. Advances and challenges in the breeding of salt-tolerant rice[J]. International Journal of Molecular Sciences, 2020, 21(21): 8385. DOI URL
[29]	HOU Z, LI Y, CHENG Y, LI W, LI T, DU H, KONG F, DONG L, ZHENG D, FENG N, LIU B, CHENG Q. Genome-wide analysis of DREB genes identifies a novel salt tolerance gene in wild soybean (Glycine soja)[J]. Frontiers in Plant Science, 2022, 13: 821647. DOI URL
[30]	TU T Q, VACIAXA P, LO T T M, NGUYEN N H, PHAM N T T, NGUYEN Q H, DO P T, NGUYEN L T N, NGUYEN Y T H, CHU M H. GmDREB6, a soybean transcription factor, notably affects the transcription of the NtP5CS and NtCLC genes in transgenic tobacco under salt stress conditions[J]. Saudi Journal of Biological Sciences, 2021, 28(12): 7175-7181. DOI URL
[31]	SHAH N, ANWAR S, XU J, HOU Z, SALAH A, KHAM S, GONG J, SHANG Z, QIAN L, ZHANG C. The response of transgenic Brassica species to salt stress: a review[J]. Biotechnol Lett, 2018, 40(8): 1159-1165. DOI URL