花生蛋白磷酸酶AhPDCP37的抗旱性功能研究

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

摘要/Abstract

摘要：

目的探究花生蛋白磷酸酶（protein phosphatase）AhPDCP37在干旱胁迫中的功能，为生物技术手段改造植物新品种提供研究基础。方法观察干旱条件下过表达AhPDCP37拟南芥的表型及叶片气孔开度等情况，测定生理生化特征及干旱胁迫反应相关基因和ABA信号通路相关基因的表达变化。结果与野生型相比，过表达AhPDCP37拟南芥中的丙二醛含量在干旱胁迫下小幅上调，而超氧化物歧化酶和过氧化物酶活性在干旱处理下则呈显著上调表达的变化趋势。同时，叶片气孔开度在干旱胁迫下的各个组中均呈下降趋势，但是在过表达AhPDCP37处理组中下降的幅度较野生型处理组更为显著。表明干旱条件下AhPDCP37通过降低气孔开度和提高细胞抗氧化酶的含量增强植物的逆境生存能力。干旱条件下，过表达AhPDCP37拟南芥处理组中的干旱胁迫响应正调控基因（WDR55、APA1）和ABA信号通路相关基因（NCED3、ABF3、RD29A）的表达较野生型处理组明显上调表达，干旱胁迫响应负调控基因WRKY70的表达量较野生型处理组明显下降。结论干旱胁迫条件下AhPDCP37主要通过参与调节气孔运动与抗氧化酶活性来提高植物的耐旱性。

关键词: AhPDCP37, 干旱胁迫, ABA, 抗氧化酶, 气孔开度

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

钱祺, 王增辉, 孙荣华, 罗英智, 苏良辰. 花生蛋白磷酸酶AhPDCP37的抗旱性功能研究[J]. 生物技术通报, 2025, 41(3): 98-103.

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.

图/表 6

表1 拟南芥抗旱相关基因引物表

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

图1 AhPDCP37过表达株系抗旱分析WT：野生型拟南芥；OE4、OE6、OE7：AhPDCP37过表达型拟南芥

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

图2 干旱胁迫下AhPDCP37过表达型拟南芥叶片气孔开度（A）和气孔表型（B）变化不同大写字母表示相同培养条件下不同株系具有显著性差异，不同小写字母表示相同株系不同培养条件下具有显著性差异（P<0.05），下同

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

图3 干旱胁迫下AhPDCP37过表达型拟南芥的抗氧化能力A：干旱胁迫下AhPDCP37过表达型拟南芥中MDA含量；B：干旱胁迫下AhPDCP37过表达型拟南芥中POD活性的变化；C：干旱胁迫下AhPDCP37过表达型拟南芥中SOD活性的变化

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

图4 干旱胁迫对AhPDCP37过表达型拟南芥中WRKY70、 WDR55、 APA1表达的影响

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

图5 干旱胁迫对AhPDCP37过表达型拟南芥中NCED3、 ABF3、 RD29A表达的影响

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

参考文献 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.

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