Mechanism of Tolerance of Protein Phosphatase AhPDCP37 in Peanut to Drought

doi:10.13560/j.cnki.biotech.bull.1985.2024-0721

Abstract

Abstract:

Objective To investigate the mechanism of tolerance of the peanut protein phosphatase AhPDCP37 to drought under dehydration stress, and to provide a research basis for biotechnological means of modifying new plant varieties. Method The phenotype and leaf stomatal opening of transgenic Arabidopsis plants overexpressing AhPDCP37 were observed under drought conditions, and physiological and biochemical characteristics as well as changes in the expression of genes related to the response to drought stress and those related to the ABA signaling pathway were measured. Result The malondialdehyde content in Arabidopsis overexpressing AhPDCP37 was less up-regulated under drought stress compared with the wild type, whereas superoxide dismutase and peroxidase activities showed a significant up-regulation of expression under drought treatment. Meanwhile, leaf stomatal openings showed a decreasing trend in all groups under drought stress, but the decrease in the overexpression of AhPDCP37 treatment group was significantly lower than that in the wild-type treatment group. This indicates that AhPDCP37 enhanced the survival ability of plants under drought conditions by decreasing stomatal opening and increasing the content of cellular antioxidant enzymes. Under drought conditions, the expressions of positively regulated genes (WDR55, APA1) and genes related to ABA signaling pathway (NCED3, ABF3, and RD29A) were significantly up-regulated in the Arabidopsis treatment group overexpressing AhPDCP37 compared with the wild-type treatment group, and the expressions of negatively regulated genes of drought stress response WRKY70 significantly decreased compared with the wild-type treatment group. Conclusion AhPDCP37 improves tolerance in plants to drought under drought stress conditions mainly through its involvement in the regulation of stomatal movement and antioxidant enzyme activities.

Key words: AhPDCP37, drought stress, ABA, antioxidant enzyme, stomatal opening

QIAN Qi, WANG Zeng-hui, SUN Rong-hua, LUO Ying-zhi, SU Liang-chen. Mechanism of Tolerance of Protein Phosphatase AhPDCP37 in Peanut to Drought[J]. Biotechnology Bulletin, 2025, 41(3): 98-103.

Figures/Tables 6

Table 1 Primer list of drought tolerance-related genes in Arabidopsis

引物名称Primer name	引物序列Primer sequence （5′‒3′）
Actin-F	GTAACATTGTGCTCAGTGGTGG
Actin-R	AACGACCTTAATCTTCATGCTGC
ABF3-F	TTCAATGATGGGAAACAATACC
ABF3-R	GACTAATCGTCCGAGGCAAG
NCED3-F	ATCCGTAACGACCCTTCA
NCED3-R	GCAGTATTCTCGGCTATTTT
RD29A-F	CTTGTCGACGAGAAGCAAAGAA
RD29A-R	TCTTGATGGAGAATTCGTGTCC
WRKY70-F	ACACCAACGCAGAAACTCCC
WRKY70-R	CCGCCACCTCCAAACACCAT
WDR55-F	TAAAGTTCGTGCCCATAA
WDR55-R	CTCCTGAAGCGATAGTTG
APA1-F	ACAGTTGGCGATTTAGTT
APA1-R	TATGTATGTTTGCCCTTG

Fig. 1 Drought tolerance analysis of AhPDCP37-overexpressing strainsWT: Wild-type Arabidopsis; OE4, OE6, OE7: AhPDCP37 overexpressing Arabidopsis

Fig. 2 Changes of stomatal opening (A) and stomatal phenotypic (B) in Arabidopsis leaves overexpressing AhPDCP37 under drought stressDifferent uppercase letters indicate significant differences between strains under the same culture conditions, and different lowercase letters indicate significant differences between strains under different culture conditions (P<0.05). The same below

Fig. 3 Antioxidant capacity of AhPDCP37 overexpressing Arabidopsis under drought stressA: MDA content in AhPDCP37 overexpressing Arabidopsis under drought stress. B: Changes in POD activity in AhPDCP37 overexpressing Arabidopsis under drought stress. C: Changes in SOD activity in AhPDCP37 overexpressing Arabidopsis under drought stress

Fig. 4 Effect of drought stress on the expression of WRKY70, WDR55 and APA1 in AhPDCP37 overexpressing Arabidopsis

Fig. 5 Effect of drought stress on the expression of NCED3, ABF3 and RD29A in AhPDCP37 overexpressing Arabidopsis

References 21

1	Chieb M, Gachomo EW. The role of plant growth promoting rhizobacteria in plant drought stress responses [J]. BMC Plant Biol, 2023, 23(1): 407.
2	Zeng RE, Chen TT, Wang XY, et al. Physiological and expressional regulation on photosynthesis, starch and sucrose metabolism response to waterlogging stress in peanut [J]. Front Plant Sci, 2021, 12: 601771.
3	Katam R, Sakata K, Suravajhala P, et al. Comparative leaf proteomics of drought-tolerant and-susceptible peanut in response to water stress [J]. J Proteomics, 2016, 143: 209-226.
4	Banavath JN, Chakradhar T, Pandit V, et al. Stress inducible overexpression of AtHDG11 leads to improved drought and salt stress tolerance in peanut (Arachis hypogaea L.) [J]. Front Chem, 2018, 6: 34.
5	Zhang H, Tang YY, Yue YL, et al. Advances in the evolution research and genetic breeding of peanut [J]. Gene, 2024, 916: 148425.
6	Jung C, Nguyen NH, Cheong JJ. Transcriptional regulation of protein phosphatase 2C genes to modulate abscisic acid signaling [J]. Int J Mol Sci, 2020, 21(24): 9517.
7	Guo YZ, Shi YB, Wang YL, et al. The clade F PP2C phosphatase ZmPP84 negatively regulates drought tolerance by repressing stomatal closure in maize [J]. New Phytol, 2023, 237(5): 1728-1744.
8	Li J, Liu XY, Ahmad N, et al. CePP2C19 confers tolerance to drought by regulating the ABA sensitivity in Cyperus esculentus [J]. BMC Plant Biol, 2023, 23(1): 524.
9	Huang Y, Yang RQ, Luo HL, et al. Arabidopsis protein phosphatase PIA1 impairs plant drought tolerance by serving as a common negative regulator in ABA signaling pathway [J]. Plants (Basel), 2023, 12(14): 2716.
10	Zhai ZK, Ao QQ, Yang LQ, et al. Rapeseed PP2C37 interacts with PYR/PYL abscisic acid receptors and negatively regulates drought tolerance [J]. J Agric Food Chem, 2024, 72(22): 12445-12458.
11	胡凤, 彭莉萍, 周涛, 等. 组蛋白乙酰化影响的蛋白磷酸酶基因 AhPDCP37 在花生叶片中的转录特征分析 [J]. 分子植物育种, 2022, 20(12): 3847-3854.
	Hu F, Peng LP, Zhou T, et al. Transcriptional characteristics of a protein phosphatase gene AhPDCP37 affected by histone acetylation in peanut leaves [J]. Mol Plant Breed, 2022, 20(12): 3847-3854.
12	可静, 李进, 吕海英, 等. 不同条件下黑果枸杞叶片气孔开度和超微结构的变化 [J]. 干旱区研究, 2017, 34(6): 1362-1370.
	Ke J, Li J, Lv HY, et al. Change of stomatal aperture and ultrastructure on Lycium ruthenicum Murr. leaves under different conditions [J]. Arid Zone Res, 2017, 34(6): 1362-1370.
13	Lin QF, Wang S, Dao YH, et al. Arabidopsis thaliana trehalose-6-phosphate phosphatase gene TPPI enhances drought tolerance by regulating stomatal apertures [J]. J Exp Bot, 2020, 71(14): 4285-4297.
14	Zhang GB, Zhang ZY, Luo SL, et al. Genome-wide identification and expression analysis of the cucumber PP2C gene family [J]. BMC Genomics, 2022, 23(1): 563.
15	Choudhury FK, Rivero RM, Blumwald E, et al. Reactive oxygen species, abiotic stress and stress combination [J]. Plant J, 2017, 90(5): 856-867.
16	Xiang LE, Jian DQ, Zhang FY, et al. The cold-induced transcription factor bHLH112 promotes artemisinin biosynthesis indirectly via ERF1 in Artemisia annua [J]. J Exp Bot, 2019, 70(18): 4835-4848.
17	王艺喆, 苏良辰. 花生种子在低温胁迫下的萌发抑制生理研究 [J]. 种子, 2023, 42(2): 28-33.
	Wang YZ, Su LC. Study on germination inhibition physiology of peanut seeds under low temperature stress [J]. Seed, 2023, 42(2): 28-33.
18	Li J, Besseau S, Törönen P, et al. Defense-related transcription factors WRKY70 and WRKY54 modulate osmotic stress tolerance by regulating stomatal aperture in Arabidopsis [J]. New Phytol, 2013, 200(2): 457-472.
19	Chen JN, Nolan TM, Ye HX, et al. Arabidopsis WRKY46, WRKY54, and WRKY70 transcription factors are involved in brassinosteroid-regulated plant growth and drought responses [J]. Plant Cell, 2017, 29(6): 1425-1439.
20	Sebastián D, Fernando FD, Raúl DG, et al. Overexpression of Arabidopsis aspartic protease APA1 gene confers drought tolerance [J]. Plant Sci, 2020, 292: 110406.
21	Park SR, Hwang J, Kim M. The Arabidopsis WDR55 is positively involved in ABA-mediated drought tolerance response [J]. Plant Biotechnol Rep, 2020, 14(4): 407-418.

[1]	ZHANG Wen-fei, YANG Fei, LIU Xu-xia. Theoretical Proof， International Comparison and Chinese Approach for Gene Edited Food Labeling System [J]. Biotechnology Bulletin, 2025, 41(3): 25-34.
[2]	LIU Jie, WANG Fei, TAO Ting, ZHANG Yu-jing, CHEN Hao-ting, ZHANG Rui-xing, SHI Yu, ZHANG Yi. Overexpression of SlWRKY41 Improves the Tolerance of Tomato Seedlings to Drought [J]. Biotechnology Bulletin, 2025, 41(2): 107-118.
[3]	KONG Qing-yang, ZHANG Xiao-long, LI Na, ZHANG Chen-jie, ZHANG Xue-yun, YU Chao, ZHANG Qi-xiang, LUO Le. Identification and Expression Analysis of GRAS Transcription Factor Family in Rosa persica [J]. Biotechnology Bulletin, 2025, 41(1): 210-220.
[4]	YUAN Liu-jiao, HUANG Wen-lin, CHEN Chong-zhi, LIANG Min, HUANG Zi-qi, CHEN Xue-xue, CHEN Ri-Meng, WANG Li-yun. Effects of Salt Stress on Physiological Characteristics, Ultrastructure and Medicinal Components of Pogostemon cablin Leaves [J]. Biotechnology Bulletin, 2025, 41(1): 230-239.
[5]	LI Yi-jun, YANG Xiao-bei, XIA Lin, LUO Zhao-peng, XU Xin, YANG Jun, NING Qian-ji, WU Ming-zhu. Cloning and Functional Analysis of NtPRR37 Gene in Nicotiana tabacum L. [J]. Biotechnology Bulletin, 2024, 40(8): 221-231.
[6]	HAN Kai, ZHOU Yong-shun, ZHANG Kai-yue, WANG Lu, GAO Jian-feng, CHEN Fu-long. Evaluation of Drought Resistance of Three Chlorella Strains [J]. Biotechnology Bulletin, 2024, 40(8): 244-254.
[7]	DU Zhong-yang, YANG Ze, LIANG Meng-jing, LIU Yi-zhen, CUI Hong-li, SHI Da-ming, XUE Jin-ai, SUN Yan, ZHANG Chun-hui, JI Chun-li, LI Run-zhi. Effect of Nano-selenium（SeNPs）in Alleviating Lead Stress and Promoting Growth of Tobacco Seedlings [J]. Biotechnology Bulletin, 2024, 40(7): 183-196.
[8]	WEN Jie, DU Yuan-xin, WU An-bo, YANG Guang-rong, LU Min, AN Hua-ming, NAN Hong. Identification and Expression Pattern Analysis of Rosa roxburghii SOD Gene Family [J]. Biotechnology Bulletin, 2024, 40(5): 153-166.
[9]	LI Jing-yan, ZHOU Jia-jing, YUAN Yuan, SU Xiao-yi, QIAO Wen-hui, XUE Yan-lei, LI Guo-jing, WANG Rui-gang. Response of Arabidopsis AtiPGAM2 Gene to Abiotic Stress [J]. Biotechnology Bulletin, 2024, 40(5): 215-224.
[10]	GUO Chun, SONG Gui-mei, YAN Yan, DI Peng, WANG Ying-ping. Genome Wide Identification and Expression Analysis of the bZIP Gene Family in Panax quinquefolius [J]. Biotechnology Bulletin, 2024, 40(4): 167-178.
[11]	SHAO Chang-xuan, ZHANG Shao-hua, DENG Hao-ran, YU Wei-kang, ZHU Yong-jie, SHAN An-shan. Database-aided Design of Antimicrobial Peptides [J]. Biotechnology Bulletin, 2024, 40(12): 12-19.
[12]	HE Wen-chuang, XU Qiang, QIAN Qian, SHANG Lian-guang. Development and Prospects of Rice Pan-genomics: Important Tools and Applications [J]. Biotechnology Bulletin, 2024, 40(10): 9-18.
[13]	WANG Zi-ying, LONG Chen-jie, FAN Zhao-yu, ZHANG Lei. Screening of OsCRK5-interacted Proteins in Rice Using Yeast Two-hybrid System [J]. Biotechnology Bulletin, 2023, 39(9): 117-125.
[14]	LIU Wen-jin, MA Rui, LIU Sheng-yan, YANG Jiang-wei, ZHANG Ning, SI Huai-jun. Cloning of StCIPK11 Gene and Analysis of Its Response to Drought Stress in Solanum tuberosum [J]. Biotechnology Bulletin, 2023, 39(9): 147-155.
[15]	DING Kai-xin, WANG Li-chun, TIAN Guo-kui, WANG Hai-yan, LI Feng-yun, PAN Yang, PANG Ze, SHAN Ying. Research Progress in Uniconazole Alleviating Plant Drought Damage [J]. Biotechnology Bulletin, 2023, 39(6): 1-11.