茉莉酸甲酯对猕猴桃果实抗葡萄座腔菌过程中能量代谢和膜脂代谢的影响

doi:10.3864/j.issn.0578-1752.2024.07.013

中国农业科学 ›› 2024, Vol. 57 ›› Issue (7): 1377-1393.doi: 10.3864/j.issn.0578-1752.2024.07.013

茉莉酸甲酯对猕猴桃果实抗葡萄座腔菌过程中能量代谢和膜脂代谢的影响

肖刘华(), 康乃慧(), 李树成, 郑致远, 罗绕绕, 陈金印, 陈明, 向妙莲()

江西农业大学农学院/江西省果蔬采后处理关键技术与质量安全协同创新中心/江西省果蔬保鲜与无损检测重点实验室，南昌 330045

收稿日期:2023-09-13 接受日期:2024-01-30 出版日期:2024-04-01 发布日期:2024-04-09
通信作者:
向妙莲，E-mail：mlxiang@jxau.edu.cn
联系方式: 肖刘华，E-mail：liuhua1846@163.com。康乃慧，E-mail：knh08256458@163.com。肖刘华和康乃慧为同等贡献作者。
基金资助:
国家自然科学基金(32160399); 江西省自然科学基金(20224BAB205031); 江西省研究生创新专项基金(YC2022-s399); 江西省研究生创新专项基金(YC2023-B135)

Effect of Methyl Jasmonate on Energy Metabolism and Membrane Lipid Metabolism During Resistance to Botryosphaeria dothidea in Kiwifruit

XIAO LiuHua(), KANG NaiHui(), LI ShuCheng, ZHENG ZhiYuan, LUO RaoRao, CHEN JinYin, CHEN Ming, XIANG MiaoLian()

College of Agronomy, Jiangxi Agricultural University/Collaborative Innovation Center of Postharvest Key Technology and Quality Safety of Fruits and Vegetables in Jiangxi Province/Jiangxi Key Laboratory for Postharvest Technology and Non-Destructive Testing of Fruits & Vegetables, Nanchang 330045

Received:2023-09-13 Accepted:2024-01-30 Published:2024-04-01 Online:2024-04-09

摘要/Abstract

摘要：

【背景】由葡萄座腔菌（Botryosphaeria dothidea）引起的软腐病是猕猴桃果实贮藏期的重要病害，对猕猴桃产业造成了严重的经济损失。茉莉酸甲酯（MeJA）是一类广泛存在于植物体内的生物信号分子，前期研究发现MeJA能够有效诱导猕猴桃果实抵御B. dothidea的侵染。【目的】分析MeJA对果实能量代谢和膜脂代谢的影响，深入解析MeJA介导的猕猴桃果实抗病机理。【方法】以‘红阳’猕猴桃（Actinidia chinensis cv. Hongyang）为试验材料，分3组进行试验：接种组（Inoculation），果实不经过MeJA处理，但接种B. dothidea；MeJA+接种组（MeJA+inoculation），果实经过0.1 mmol·L^-1 MeJA熏蒸处理24 h并接种B. dothidea；对照组（Control），不作任何处理，即未经MeJA处理且不接种B. dothidea。所有果实于培养箱（（20±1）℃、相对湿度90%—95%）中贮藏8 d。利用2,7-二氯二氢荧光素乙酰乙酸（H₂DCFDA）染色分析果实活性氧（ROS）积累情况，测定丙二醛（MDA）含量和相对电导率，分析磷脂酶D（PLD）、脂肪酶（LPS）、脂氧合酶（LOX）、琥珀酸脱氢酶（SDH）、苹果酸脱氢酶（MDH）、H⁺-ATP酶（H⁺-ATPase）、Ca²⁺-ATP酶（Ca²⁺-ATPase）和细胞色素C氧化酶（CCO）活性及其相应基因的表达，利用高效液相色谱仪（HPLC）检测三磷酸腺苷（ATP）、二磷酸腺苷（ADP）、单磷酸腺苷（AMP）、草酰乙酸和延胡索酸的含量，并对接种组和MeJA+接种组猕猴桃果实膜脂代谢和能量代谢指标进行相关性分析。【结果】与接种组相比，MeJA处理抑制了接种B. dothidea果实中活性氧（ROS）的过量累积，减缓了膜脂过氧化程度，并有效降低了PLD、LPS和LOX活性及AcLOXs、AcPLD和AcLPS的表达，但提高了SDH、MDH、H⁺-ATPase、Ca²⁺-ATPase和CCO的活性及AcSDH、AcMDH、AcH⁺-ATPase、AcCa²⁺-ATPase和AcCCO转录水平，促进了ATP、ADP、草酰乙酸和延胡索酸的产生，延缓了果实能荷的下降，保障了果实抵御病原菌所需的能量。相关性分析表明，猕猴桃果实膜脂代谢与ROS的积累呈正相关，而与能荷呈负相关。【结论】MeJA诱导猕猴桃果实对B. dothidea的抗性与其调控果实能量水平和减缓ROS参与的膜脂代谢进程有关。

关键词: 茉莉酸甲酯, 猕猴桃果实, 葡萄座腔菌, 诱导抗性, 能量代谢, 膜脂代谢

Abstract:

【Objective】Soft rot caused by Botryosphaeria dothidea is one of important diseases during kiwifruit storage, which causes serious economic losses for kiwifruit industry. Methyl jasmonate (MeJA) is a kind of biological signal molecule widely existing in plants, which had been found that it could effectively induce kiwifruit’s resistance to B. dothidea in previous studies. In order to further investigate the mechanism of MeJA-mediated resistance to disease in postharvest kiwifruit, the effects of MeJA on energy metabolism and membrane lipid metabolism were analyzed in this study. 【Method】 Hongyang kiwifruit (Actinidia chinensis cv. Hongyang) were used as experiment material and divided into three groups as follows: inoculation group, which the fruits were inoculated with B. dothidea without MeJA treatment; MeJA+inoculation group, which the fruits were fumigated with 0.1 mmol·L^-1 MeJA for 24 h then inoculated with B. dothidea; control group, the fruits without MeJA treatment or inoculation with B. dothidea. All fruits were stored in an incubator ((20±1) ℃, 90%-95% relative humidity) for 8 d. The reactive oxygen species (ROS) accumulation was analyzed by 2, 7-dichlorodihydrofluorescein acetoacetate (H₂DCFDA) method, malondialdehyde (MDA) content and relative conductivity were also measured. Meanwhile, phospholipase D (PLD), lipase (LPS), lipoxygenase (LOX), succinate dehydrogenase (SDH), malate dehydrogenase (MDH), H⁺-ATPase, Ca²⁺-ATPase and cytochrome C oxidase (CCO) activities and the related genes expression were analyzed. The content of Adenosine triphosphate (ATP), Adenosine diphosphate (ADP), Adenosine monophosphate (AMP), oxaloacetic acid and fenugreek acid were determined by high performance liquid chromatography (HPLC). Then the correlation between membrane lipid metabolism and energy metabolism of kiwifruit in inoculation group and MeJA+ inoculation group was analyzed. 【Result】Compared with the inoculated group, MeJA treatment could inhibit the excessive accumulation of ROS in inoculated B. dothidea fruits, slowed down the degree of membrane lipid peroxidation, and effectively decreased PLD, LPS, and LOX activities and the expression levels of AcLOXs, AcPLD and AcLPS. However, MeJA increased the activities of SDH, MDH, H⁺-ATPase, Ca²⁺-ATPase and CCO and the transcription levels of AcSDH, AcMDH, AcH⁺-ATPase, AcCa²⁺-ATPase and AcCCO, promoted the production of ATP, ADP, oxaloacetic acid and fumaric acid, delayed the decline of fruit energy charge then ensured the energy required for fruit resistance to pathogens. In addition, correlation analysis showed that kiwifruit membrane lipid metabolism was positively correlated with ROS accumulation, but negatively correlated with energy charge. 【Conclusion】The above results indicated that MeJA-induced resistance to B. dothidea in kiwifruit was related to its regulation of fruit energy levels and mitigation of ROS-involved membrane lipid metabolism.

Key words: methyl jasmonate, kiwifruit, Botryosphaeria dothidea, induced resistance, energy metabolism, membrane lipid metabolism

肖刘华, 康乃慧, 李树成, 郑致远, 罗绕绕, 陈金印, 陈明, 向妙莲. 茉莉酸甲酯对猕猴桃果实抗葡萄座腔菌过程中能量代谢和膜脂代谢的影响[J]. 中国农业科学, 2024, 57(7): 1377-1393.

XIAO LiuHua, KANG NaiHui, LI ShuCheng, ZHENG ZhiYuan, LUO RaoRao, CHEN JinYin, CHEN Ming, XIANG MiaoLian. Effect of Methyl Jasmonate on Energy Metabolism and Membrane Lipid Metabolism During Resistance to Botryosphaeria dothidea in Kiwifruit[J]. Scientia Agricultura Sinica, 2024, 57(7): 1377-1393.

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基因 Gene	登录号 Accession number	序列 Primer sequence (5′-3′)
AcActin	Ach05g107181	F: GCTTACAGAGGCACCACTCAACC R: CCGGAATCCAGCACAATACCAG
AcPLD	Actinidia10522	F: CCCGACTATGATGTGGAGCC R: GACCCATCCCTGTTCGTCTG
AcLPS	Actinidia27609	F: GTCCGATAAACCGACCGTCT R: GTCGTACCTCTTTCCGCCTC
AcLOX1	DQ497792.1	F: TCGGGTCAGAGATCAAAAGCA R: GAAGCCTACGAGACAATCCATCAT
AcLOX2	DQ497797.1	F: TGCCTTACACACTGCTATTTCC R: AACGACATCCCAAATGAAAAG
AcLOX3	DQ497795.1	F: GTTAGTGCCACGGGCTACGT R: CATACATGTTCTCGACGATAGAGTTGA
AcLOX4	DQ497793.1	F: GAGAAGGGAGTGGTTTACGTGTGT R: AACGGAGTATTAAATTGTGCTTGTTG
AcLOX5	DQ497796.1	F: AGTGAATGTGCCATACACTTTGCT R: ACGAAACCGGAAACCCTTAGA
AcLOX6	DQ497794.1	F: GGGTTGTACCCTATGAGCTTTTGA R: ATGCGTAAACACACTCGCTTCTC
AcH⁺-ATPase	Actinidia38808	F: TCGGAAAGGATGCTTTGGCT R: CCATCCATACCAGCACCTCG
AcCa²⁺-ATPase	Actinidia16431	F: GAGGAACCTCTGTGGGGTTG R: GTCATCACTTTGCGCACCTG
AcSDH	Actinidia19180	F: TGAATCGACTGGGGGACGAT R: TGTGCGAGCGAGTAGGAAAT
AcMDH	Actinidia12555	F: AATCCAATCGTCCCTCAGGC R: GCGATATCGTAGAGGGCGAG
AcCCO	Actinidia22368	F: GGAGGAGAGCGTCGAATCAG R: AAGTCGAGTGTTGGGACGAC

茉莉酸甲酯对猕猴桃果实抗葡萄座腔菌过程中能量代谢和膜脂代谢的影响

Effect of Methyl Jasmonate on Energy Metabolism and Membrane Lipid Metabolism During Resistance to Botryosphaeria dothidea in Kiwifruit

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