CMV通过影响效应因子MpC002的表达干预桃蚜种群增长的机制

doi:10.13560/j.cnki.biotech.bull.1985.2023-1057

生物技术通报 ›› 2024, Vol. 40 ›› Issue (4): 271-277.doi: 10.13560/j.cnki.biotech.bull.1985.2023-1057

CMV通过影响效应因子MpC002的表达干预桃蚜种群增长的机制

毛立杰¹^,²^,³(), 梁晓²^,³, 刘迎²^,³, 伍春玲²^,³, 韩晓燕¹, 陈青²^,³()

1.海南大学南繁学院（三亚南繁研究院），三亚 572025
2.中国热带农业科学院环境与植物保护研究所农业农村部热带作物有害生物综合治理重点实验室海南省热带农业有害生物监测与控制重点实验室海南省热带作物病虫害生物防治工程技术研究中心,海口 571101
3.中国热带农业科学院三亚研究院海南省南繁生物安全与分子育种重点实验室，三亚 572000

收稿日期:2023-11-10 出版日期:2024-04-26 发布日期:2024-04-30
通讯作者: 陈青，男，博士，研究员，研究方向：资源利用与虫害绿色防控；E-mail: chqingztq@163.com
作者简介:毛立杰，男，硕士研究生，研究方向：资源利用与虫害绿色防控；E-mail: maolijie9@163.com
基金资助:
海南省重大科技计划项目(ZDKJ202107-1);海南省自然科学基金青年项目(322QN366)

Mechanism of CMV Intervention in Peach Aphid Population Growth by Influencing the Expression of Effector MpC002

MAO Li-jie¹^,²^,³(), LIANG Xiao²^,³, LIU Ying²^,³, WU Chun-ling²^,³, HAN Xiao-yan¹, CHEN Qing²^,³()

1. School of Breeding and Multiplication（Sanya Institute of Breeding and Multiplication）,Hainan University, Sanya 572025
2. Chinese Academy of Tropical Agricultural Sciences/Key Laboratory of Integrated Pest Management on Tropical Crops, Ministry of Agriculture and Rural Affairs/Hainan Key Laboratory for Monitoring and Control of Tropical Agricultural Pests/Hainan Engineering Research Center for Biological Control of Tropical Crops Diseases and Insect Pests, Haikou 571101
3. Sanya Research Academy, Chinese Academy of Tropical Agriculture Sciences/Hainan Key Laboratory for Biosafety Monitoring and Molecular Breeding in Off-Season Reproduction Regions, Sanya 572000

Received:2023-11-10 Published:2024-04-26 Online:2024-04-30

摘要/Abstract

摘要：

【目的】桃蚜是CMV的重要传毒媒介，效应因子MpC002在桃蚜取食寄主过程中发挥关键作用，CMV和MpC002均存在于蚜虫唾液中，但两者如何相互作用从而利于病毒传播知之甚少。为探讨CMV影响MpC002表达干预桃蚜种群增长的机制。【方法】利用绝对定量的qPCR法建立CMV拷贝数检测的标准曲线方程y=-3.163 1x+39.763。【结果】桃蚜取食CMV感染辣椒10 s体内CMV含量最高，然后随着取食时间的延长CMV含量不断降低，其获毒过程符合非持久性传毒特征。取食CMV感染辣椒5 min-24 h的桃蚜MpC002表达量显著降低至取食前的37%-58%；取食CMV感染辣椒的3龄、4龄若蚜和1-4龄若蚜的平均发育历期分别为1.58 d、2.07 d和6.47 d，显著长于取食未处理辣椒的1.25 d、1.47 d和5.33 d，但1 龄和2 龄若蚜发育历期在CMV感染辣椒和未处理辣椒间无显著差异；平均产蚜期、总产蚜量和单日平均产蚜量分别为10.03 d、16.87头和1.67头，显著短于取食未处理辣椒桃蚜的14.27 d，39.73头和2.82头；净增殖率、内禀增长率、周限增长率分别为16.87、0.21、1.24，均显著低于取食未处理辣椒桃蚜的39.73、0.31、1.36，平均世代周期（13.02 d）和种群加倍时间（3.26 d）均显著长于取食未处理辣椒桃蚜的12.00 d和2.30 d。【结论】CMV能显著抑制桃蚜MpC002的表达，从而延长其发育历期和降低其繁殖，最终抑制其种群的增长。

关键词: CMV, 辣椒, 桃蚜, 效应因子MpC002, 种群增长

Abstract:

【Objective】Peach aphid（Myzus persicae）is an important vector of cucumber mosaic virus（CMV）, and the effector MpC002 plays a key role in the feeding process. CMV and MpC002 are both present in aphid saliva, but little is known about their interaction and how MpC002 facilitate virus transmission. Therefore, this study aims to preliminarily investigate the mechanism by which CMV influences the expression of MpC002 for interfering the population growth of GPA. 【Method】The CMV copy number was calculated by the standard curve y=-3.163 1x+39.763, via applying the absolute quantitative polymerase chain reaction（qPCR）. 【Result】The CMV content in the peach aphid fed with CMV-infected pepper reached the highest at 10 s, then the CMV content decreased gradually along with the feeding time, and its virulence acquisition process conformed to the characteristics of non-persistent transmission. In addition, the expressions of MpC002 in the peach aphids fed on CMV-infested peppers within 5 min-24 h significantly decreased to 37%-58%, compared with those before feeding. While feeding on CMV-infested peppers, the average developmental duration of third, fourth instar and the entire nymph stages were 1.58, 2.07 and 6.47 d, which were significantly longer than those fed on the untreated peppers（1.25 d, 1.47 d and 5.33 d）; in contrast, the developmental duration of the first and second instars did not show significantly difference between CMV-infested and untreated peppers. The average laying period, the total peach aphid number and the average peach aphid in a single day were 10.03 d, 16.87 and 1.67, which were significantly shorter than those of untreated peppers（14.27 d, 39.73 and 2.82）. In addition, analysis of GPA life table parameters revealed that the net growth rate, intrinsic rate of increase, and finite rate of increase were 16.87, 0.21, and 1.21, respectively, which were significantly lower than those of peach aphids feeding on untreated peppers（39.73, 0.31, and 1.36, respectively）, whereas the mean generation time（13.02 d）and the population doubling time（3.26 d）were both significantly longer than those of peach aphids feeding untreated peppers（12.00 d and 2.30 d）. 【Conclusion】CMV significantly inhibited the expressions of MpC002, and then prolongated the developmental duration and decreased the reproduction, which in turn adversely affected its population growth.

Key words: CMV, pepper, Myzus persicae, effector MpC002, population growth

毛立杰, 梁晓, 刘迎, 伍春玲, 韩晓燕, 陈青. CMV通过影响效应因子MpC002的表达干预桃蚜种群增长的机制[J]. 生物技术通报, 2024, 40(4): 271-277.

MAO Li-jie, LIANG Xiao, LIU Ying, WU Chun-ling, HAN Xiao-yan, CHEN Qing. Mechanism of CMV Intervention in Peach Aphid Population Growth by Influencing the Expression of Effector MpC002[J]. Biotechnology Bulletin, 2024, 40(4): 271-277.

图/表 5

图1 桃蚜取食CMV感染辣椒不同时间后的获毒量 A：CMV定量检测标准曲线； B：不同时间桃蚜体内病毒拷贝数，数据为平均数±标准误，“*”、“**”和“ns”分别表示同一个取食时间点内病毒拷贝数差异显著（P<0.05）、差异极显著（P<0.01）和无显著差异，下同

Fig. 1 CMV content in M. persicae after fed on CMV-infested pepper at different time A: Standard curve of CMV quantitative detection; B: the CMV copies in M. persicae at different times, data indicate mean value ± SE; *, ** and ns indicate significant（P<0.05）, extremely significant difference（P<0.01）and no significant difference differences in CMV copy number within the same feeding time point, respectively. The same below

图2 取食CMV感染辣椒、未处理辣椒后桃蚜效应因子MpC002基因相对表达量 A：取食CMV感染辣椒后MpC002基因相对表达量； B：取食未处理辣椒MpC002基因相对表达量

Fig. 2 Relative expressions of the effector MpC002 in M. persicae after feeding on CMV-infested and untreated pepper A: The transcription of MpC002 after feeding on CMV- infested pepper. B: The transcription of MpC002 after feeding on untreated pepper

图3 桃蚜取食CMV感染辣椒与未处理辣椒后的发育历期

Fig. 3 Developmental duration of M. persicae after feeding on CMV-infested or untreated peppers

图4 桃蚜取食CMV感染辣椒与未处理辣椒对桃蚜繁殖和寿命的影响 A：单雌平均产蚜期；B：单雌平均总产蚜量：C：单日平均产蚜量；D：平均成蚜寿命

Fig. 4 Influence on M. persicae reproduction and lifespan after feeding on CMV-infested and untreated peppers A: Average production period per female; B: average total production per female; C: average daily production; D: average lifespan of adult aphid

表1 桃蚜取食CMV感染辣椒与未处理辣椒后的生命表参数

Table 1 Life table parameters of aphids after feeding on CMV-infested and untreated peppers

处理 Treatment	净增殖率R₀ Net reproduction rate（R₀）	平均世代周期T Mean generation time（T）/d	内禀增长率r_m Intrinsic rate of increase（r_m）/（d^-1）	周限增长率λ Finite rate of increase（λ）/（d^-1）	种群加倍时间Dt Population doubling time（Dt）/d
未处理辣椒 Untreated peppers	39.73±2.08^**	12.00±0.24	0.31±0.01^**	1.36±0.01^**	2.30±0.06
CMV感染辣椒 CMV-infested peppers	16.87±0.98	13.02±0.40^*	0.21±0.00	1.24±0.01	3.26±0.07^**

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CMV通过影响效应因子MpC002的表达干预桃蚜种群增长的机制

Mechanism of CMV Intervention in Peach Aphid Population Growth by Influencing the Expression of Effector MpC002

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