The role of HaFT-9， a 14-3-3 protein from Haloxylon ammodendron， in the cross regulation of high temperature and drought stress

doi:10.11686/cyxb2024403

Abstract

Abstract:

Previous research has demonstrated that Haloxylon ammodendron seedlings acquire resistance to stressors like high temperatures and drought through adaptive stress conditioning during their growth. Additionally， the expression of the 14-3-3 protein gene HaFT-9 is significantly upregulated in response to high temperature and drought stress， suggesting a potential role in mediating crosstalk between these stress pathways. This study further investigates the role of HaFT-9 in the cross-regulation of high temperature and drought stress signals. Pre-treatment of H. ammodendron seedlings with heat stress， followed by a subsequent drought stress challenge， significantly increased HaFT-9 expression during drought stress. Comparative experiments demonstrated that Arabidopsis thaliana lines overexpressing HaFT-9 showed higher survival rates and reduced cellular mortality under sequential heat and drought stress compared to wild-type controls. Notably， the lines overexpressing HaFT-9 exhibited enhanced expression of several key stress-responsive genes， including P5CS1， P5CS2， CAT2， CHLI1， HSP21， HsfA2 and BI-1， in response to subsequent drought stress.In addition， these lines showed significantly enhanced catalase （CAT） activity， increased proline and chlorophyll levels， while the content of malondialdehyde （MDA） was significantly reduced. Moreover， staining with diaminobenzidine （DAB） and nitroblue tetrazolium （NBT） in overexpressing lines was lighter. In conclusion， HaFT-9 overexpression markedly improves drought tolerance in A. thaliana， underscoring its critical regulatory role in modulating the interplay between high temperature and drought stress responses. This study provides valuable insights into the molecular mechanisms underlying stress tolerance in H. ammodendron seedlings and provides data relevant to the conservation of its germplasm resources.

Key words: Haloxylon ammodendron, high temperature, drought, cross regulation, HaFT-9

Yuan-yuan LIU, Xu WANG, Qi WEI, Li-juan CHE, Meng YUAN, Bo WANG. The role of HaFT-9， a 14-3-3 protein from Haloxylon ammodendron， in the cross regulation of high temperature and drought stress[J]. Acta Prataculturae Sinica, 2025, 34(9): 134-146.

Figures/Tables 7

Table 1 Primers used in this study

基因 Gene	正向引物Forward primer （5′→3′）	反向引物 Reverse primer （5′→3′）
Ha18SrRNA	CTCTGCCCGTTGCTCTGATGAT	CCTTGGATGTGGTAGCCGTTTC
HaFT-9	TACTTTGGGTGAAGAGTCCTACA	TTTGATCCTGCATGTCTGATGT
P5CS1	GATACGGATATGGCAAAGCG	CCAAGTCCAAATCGGAAACC
P5CS2	GTTAAGCGTATCGTCGTCAAGGTT	CCTAAACGTCCAAGAGCCAATCT
CAT2	ACATTCGTTGACCCTTTTCAC	GGCCACACACGATAACAAC
CDS1	CTGCATCTCTACTGGACCTC	CCACATCCAACTCTCGAGC
CHLI1	TGTTGATGGGTTGAGAGGAG	AACGGTTGCAACATCATCTG
DREB2a	CAACAGCAGGATTCGCTATCTG	ACATCGTCGCCATTTAGGTCA
HSP21	TGGACGTCTCTCCTTTCGGATTGT	TGCACGAATCTCTGACACTCCACT
HsfA2	GAGATTTTCCGGCGTGATTT	ACATCGGACCTGAAAGAGAG
BI-1	ACATTCGTTGACCCTTTTCAC	GGCCACACACGATAACAAC
At18SrRNA	AAACGGCTACCACATCCAAG	CCTCCAATGGATCCTCGTTA

Fig.1 The expression of the HaFT-9 gene under the heat stress priming-the secondary drought stress treatment

Fig.2 The effects of the heat stress priming-the secondary drought stress treatment on the growth of Arabidopsis

Fig.3 The effects of the heat stress priming-the secondary drought stress treatment on the survival rate and cell death rate in Arabidopsis

Fig.4 The expression of relevant genes under the heat stress priming-the secondary drought stress treatment in Arabidopsis

Fig.5 Diaminobenzidine （DAB） and nitroblue tetrazolium （NBT） staining of Arabidopsis under the heat stress priming-the secondary drought stress treatment

Fig.6 Antioxidant enzyme activity， proline content， and chlorophyll content under the heat stress priming-the secondary drought stress treatment in Arabidopsis

References 31

[1]	Paul A L， Denison F C， Schultz E R， et al. 14-3-3 phosphoprotein interaction networks-does isoform diversity present functional interaction specification？ Frontiers in Plant Science， 2012， 3： 190.
[2]	Li X D， Wu J H， Sun F， et al. Enhanced tolerance of Arabidopsis over expressing Fa14-3-3C from tall fescue （Festuca arundinacea） to low-nitrogen stress. Acta Prataculturae Sinica， 2017， 26（9）： 104-112.
	李小冬，吴佳海，孙方，等. 过量表达Fa14-3-3C促进拟南芥对低氮胁迫耐受性的研究. 草业学报， 2017， 26（9）： 104-112.
[3]	Zhou H P， Lin H X， Chen S， et al. Inhibition of the Arabidopsis salt overly sensitive pathway by 14-3-3 proteins. Plant Cell， 2014， 26（3）： 1166-1182.
[4]	Hilker M， Schmülling T. Stress priming， memory， and signalling in plants. Plant Cell and Environment， 2019， 42（3）： 753-761.
[5]	Liang X H， Ai F F， Zhong T X， et al. Cross adaptation under drought and low temperature stress in perennial ryegrass. Acta Prataculturae Sinica， 2016， 25（1）： 163-170.
	梁小红，艾非凡，钟天秀，等. 多年生黑麦草对干旱-低温交叉适应的生理响应. 草业学报， 2016， 25（1）： 163-170.
[6]	Liu H P， Able A J， Able J A. Priming crops for the future： rewiring stress memory. Trends in Plant Science， 2022， 27（7）： 699-716.
[7]	Gallusci P， Agius D R， Moschou P N， et al. Deep inside the epigenetic memories of stressed plants. Trends in Plant Science， 2023， 28（2）： 142-153.
[8]	Lu Y， Lei J Q， Zeng F J， et al. Effects of salt treatments on the growth and ecophysiological characteristics of Haloxylon ammodendron. Acta Prataculturae Sinica， 2014， 23（3）： 152-159.
	鲁艳，雷加强，曾凡江，等. NaCl处理对梭梭生长及生理生态特征的影响. 草业学报， 2014， 23（3）： 152-159.
[9]	Ma H， Zhang H， Ma L， et al. None-watering and tube-protecting planting technique for Haloxylon ammodendron under desert and its extension. Scientia Sinica （Vitae）， 2014， 44（3）： 248-256.
	麻浩，张桦，马林，等. 无灌溉管件防护梭梭荒漠造林新技术及其示范推广. 中国科学（生命科学）， 2014， 44（3）： 248-256.
[10]	Pan R， Ren W J， Liu S S， et al. Ectopic over-expression of HaFT-1， a 14-3-3 protein from Haloxylon ammodendron， enhances acquired thermotolerance in transgenic Arabidopsis. Plant Molecular Biology， 2023， 112（4/5）： 261-277.
[11]	Cao Y H， Ren W， Gao H J， et al. HaASR2 from Haloxylon ammodendron confers drought and salt tolerance in plants. Plant Science， 2023， 328： 111572.
[12]	Wang J. Expression patterns and functional analysis of 14-3-3 genes related to stress resistance of Haloxylon ammodendron. Urumqi： Xinjiang Agricultural University， 2017.
	王娇. 与抗逆相关的梭梭14-3-3蛋白基因的表达模式和功能分析. 乌鲁木齐：新疆农业大学， 2017.
[13]	Liu S S， Wang B， Zong X F， et al. Cloning and characterization analysis of HaFT-9 gene in 14-3-3 protein in Haloxylon ammodendron. Xinjiang Agricultural Sciences， 2019， 56（6）： 1083-1094.
	刘栓栓，王波，宗兴风，等. 梭梭14-3-3蛋白基因HaFT-9的克隆及特性分析. 新疆农业科学， 2019， 56（6）： 1083-1094.
[14]	Wei Q. Functional analysis of heat stress memory key genes HaHsfA2a and HaFT-9 under abiotic stress in Haloxylon ammodendron. Urumqi： Xinjiang Agricultural University， 2023.
	魏琪. 梭梭高温胁迫记忆关键基因HaHsfA2a与HaFT-9在非生物胁迫下的功能分析. 乌鲁木齐：新疆农业大学， 2023.
[15]	Montero-Barrientos M， Hermosa R， Cardoza R E， et al. Transgenic expression of the Trichoderma harzianum hsp70 gene increases Arabidopsis resistance to heat and other abiotic stresses. Journal of Plant Physiology， 2010， 167（8）： 659-665.
[16]	Livak K J， Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2^-∆∆CT method. Methods， 2001， 25（4）： 402-408.
[17]	Xu M， Liu J， Ayiguli A. Comparative methods on chlorophyll extraction in plant physiology experiment teaching. Experiment Science and Technology， 2018， 16（4）： 129-133.
	徐敏，刘君，阿衣古力·阿布都瓦依提. 植物生理实验教学中叶绿素提取方法比较. 实验科学与技术， 2018， 16（4）： 129-133.
[18]	Li W Y， Chen Y F， He H， et al. Effects of high temperature exercise on growth and physiological and biochemical characters of Taxus wallichiana var. mairei under drought stress. Shandong Agricultural Sciences， 2023， 55（9）： 57-63.
	李文杨，陈亚飞，何辉，等. 高温锻炼对干旱胁迫下南方红豆杉生长和生理生化指标的影响. 山东农业科学， 2023， 55（9）： 57-63.
[19]	Fan Z H， Zhu Y Q， Kuang W， et al. The 14-3-3 protein GRF8 modulates salt stress tolerance in apple via the WRKY18-SOS pathway. Plant Physiology， 2024， 194（3）： 1906-1922.
[20]	Ren Y R， Yang Y Y， Zhang R， et al. MdGRF11， an apple 14-3-3 protein， acts as a positive regulator of drought and salt tolerance. Plant Science， 2019， 288： 110219.
[21]	Sirichandra C， Davanture M， Turk B E， et al. The Arabidopsis ABA-activated kinase OST1 phosphorylates the bZIP transcription factor ABF3 and creates a 14-3-3 binding site involved in its turnover. PLoS One， 2010， 5（11）： e13935.
[22]	Jiang W， Tong T， Li W， et al. Molecular evolution of plant 14-3-3 proteins and function of Hv14-3-3A in stomatal regulation and drought tolerance. Plant Cell Physiology， 2023， 63（12）： 1857-1872.
[23]	Wiese A J， Steinbachová L， Timofejeva L， et al. Arabidopsis bZIP18 and bZIP52 accumulate in nuclei following heat stress where they regulate the expression of a similar set of genes. International Journal of Molecular Sciences， 2021， 22（2）： 530.
[24]	Liu J Z， Feng L L， Gu X T， et al. An H3K27me3 demethylase-HSFA2 regulatory loop orchestrates transgenerational thermomemory in Arabidopsis. Cell Research， 2019， 29（5）： 379-390.
[25]	Kim J M， To T K， Seki M. An epigenetic integrator： new insights into genome regulation， environmental stress responses and developmental controls by histone deacetylase 6. Plant and Cell Physiology， 2012， 53（5）： 794-800.
[26]	Liu M Y， Guo L Z， Yue Y S， et al. Physiological and antioxidant enzyme gene expression differences between female and male Buchloe dactyloides plants under drought stress. Acta Prataculturae Sinica， 2023， 32（10）： 93-103.
	刘牧野，郭丽珠，岳跃森，等. 干旱胁迫下不同性别野牛草生理及抗氧化酶基因表达差异. 草业学报， 2023， 32（10）： 93-103.
[27]	Zhang J， Cheng K， Wang Y C. Analysis of the calcium-dependent protein kinase RtCDPK16 response to abiotic stress in Reaumuria trigyna. Acta Prataculturae Sinica， 2023， 32（2）： 97-109.
	张洁，程凯，王迎春. 长叶红砂钙依赖蛋白激酶RtCDPK16的非生物胁迫应答分析. 草业学报， 2023， 32（2）： 97-109.
[28]	Liu J P， Sun X J， Liao W C， et al. Involvement of OsGF14b adaptation in the drought resistance of rice plants. Rice， 2019， 12（1）： 82.
[29]	Lukaszewicz M， Matysiak-Kata I， Aksamit A， et al. 14-3-3 protein regulation of the antioxidant capacity of transgenic potato tubers. Plant Science， 2002， 163（1）： 125-130.
[30]	Wang L Y， Tian Y C， Shi W， et al. The miR396-GRFs module mediates the prevention of photo-oxidative damage by brassinosteroids during seedling de-etiolation in Arabidopsis. Plant Cell， 2020， 32（8）： 2525-2542.
[31]	Liu X， Challabathula D K， Quan W L， et al. Transcriptional and metabolic changes in the desiccation tolerant plant Craterostigma plantagineum during recurrent exposures to dehydration. Planta， 2019， 249（4）： 1017-1035.