ToCV和TYLCV侵染番茄对烟粉虱解毒酶的影响

doi:10.16420/j.issn.0513-353x.2023-0484

摘要/Abstract

摘要：

番茄病毒病已成为目前番茄生产最主要的病害之一，造成番茄的大量减产甚至绝收，其中番茄褪绿病毒（tomato chlorosis virus，ToCV）和番茄黄化曲叶病毒（tomato yellow leaf curl virus，TYLCV）危害最为严重，二者的复合侵染还会对烟粉虱的取食、获毒以及传毒等行为产生一系列影响，给番茄病毒病的防治造成了很大困难。为了明确在ToCV和TYLCV复合侵染条件下，烟粉虱取食带毒番茄对其体内解毒酶以及生理防御反应相关基因变化的影响，以便更好地对烟粉虱进行防治，通过酶活性测定和实时荧光定量PCR技术，研究烟粉虱取食ToCV、TYLCV单独侵染及ToCV + TYLCV复合侵染番茄48 h后其体内解毒酶变化，并以健康番茄作为对照。结果表明，除取食ToCV单独侵染番茄的烟粉虱谷胱甘肽S-转移酶（Glutathione S-transferase，GST）和UDP-葡萄糖醛酸转移酶（UDP-glucuronosy-ltransferase，UDPGT）活性较取食健康番茄烟粉虱下降，烟粉虱带毒后GST、羧酸酯酶（Carboxylesterase，CarE）、乙酰胆碱酯酶（Acetylcholinesterase，AchE）、细胞色素 P450（Cytochrome P450，CYP450）和UDP活性和基因表达量均较健康烟粉虱升高，且其中取食复合侵染番茄后CarE、AchE、P450、UDP酶活性和基因表达量变化均存在显著差异。研究结果表明，取食复合侵染番茄和TYLCV单独侵染番茄后，不同程度诱导了烟粉虱的解毒防御，从而有助于烟粉虱传播病毒。

关键词: 番茄, 烟粉虱, 番茄褪绿病毒, 番茄黄化曲叶病毒, 复合侵染, 解毒酶

Abstract:

Tomato virus disease has become one of the most important diseases in tomato production，resulting in a large reduction in tomato yield or even a failure of the harvest. Tomato chlorosis virus（ToCV） and tomato yellow leaf curl virus（TYLCV）cause severe losses in the field，and co-infection of ToCV and YLCV will also have a series of effects on the B. tabaci feeding，virus acquisition，and virus transmission，causing great difficulties in the prevention of tomato virus diseases. The changes of detoxification enzymes and gene related to physiological defense responses，after B. tabaci feeding on tomatoes co-infected with ToCV and TYLCV was clarified，to control B. tabaci effectively. In this study，the changes of detoxification enzymes in co-infected B. tabaci were studied by enzyme activity determination and real-time fluorescence quantitative PCR technology，and healthy tomatoes were used as the control. The results showed that expect the enzyme activity of glutathione S-transferase（GST）and UDP-glucuro- nosyltransferase（UDPGT）of B. tabaci fed on ToCV infected tomatoes were lower than those fed on healthy B. tabaci，viruliferous B. tabaci’s GST，carboxylesterase（CarE），acetylcholinesterase（AchE），cytochrome P450（CYP450）and UDP enzyme activities and gene expression levels were higher than those of healthy B. tabaci，and there were significant differences in CarE，AchE，P450，UDP enzyme activities and gene expression levels after B. tabaci feeding and infection. The results showed that the detoxification defense of B. tabaci was induced to different degrees after feeding on the co-infected tomato and TYLCV-infected tomato plant，thus helping B. tabaci to transmitt the virus.

Key words: tomato, Bemisia tabaci, ToCV, TYLCV, co-infection, detoxification enzymes

朱婕, 卢丁伊慧, 张德咏, 张战泓, 张卓, 史晓斌, 刘勇. ToCV和TYLCV侵染番茄对烟粉虱解毒酶的影响[J]. 园艺学报, 2024, 51(2): 411-422.

ZHU Jie, LU Dingyihui, ZHANG Deyong, ZHANG Zhanhong, ZHANG Zhuo, SHI Xiaobin, LIU Yong. Effect of ToCV and TYLCV Infection on Detoxification Enzymes of Bemisia tabaci[J]. Acta Horticulturae Sinica, 2024, 51(2): 411-422.

图/表 4

表1 试验中所用引物

Table 1 Primer list used in the experiment

目的 Purpose	引物名称 Primer name	引物序列（5′-3′） Primer sequence
ToCV RT-PCR	ToCV-3F	GGTTTGGATTTTGGTACTACATTCAGT
ToCV RT-PCR	ToCV-3R	AAACTGCCTGCATGAAAAGTCTC
TYLCV RT-PCR	TYLCV-F	GTTCACGGATTTCGTTGTATG
TYLCV RT-PCR	TYLCV-R	AGAGGGACTGGCAAAGCAACA
Bemisia tabaci 参考基因 Bemisia tabaci reference genes	Actin-F	CGCTGCCTCCACCTCATT
	Actin-R	ACCGCAAGATTCCATACCC
	EF-1α-F	AGCCTTGTGCCAATTTCCG
	EF-1α-R	CCTTCAGCATTACCGTCC
	CarE-F	CCCTCTCGAGCAGCAAAGAA
	CarE-R	GGCCTTCAAGAGAACGGACA
	GST-F	GAGCCGGTTCGTTTCATCCT
	GST-R	TCCCAAATGGTGTGGTTGGT
	AchE-F	CCACGAGGATTGCCTCTACC
	AchE-R	GAGAGCGGAACCATCCCAAA
	P450-F	CAACAATGCTGGTTCGGAGC
	P450-R	CCAAGTGAGCTGTGATGGGT
	UDP-F	CGGCGTAAATTTGCAGCGTA
	UDP-R	TGCAACCCGCCTATCAAACT

图1 烟粉虱不携带病毒（A）、携带TYLCV（B）、携带ToCV（C）、携带ToCV和TYLCV（D、E）检测结果 M：DNA marker；1 ~ 12：烟粉虱样品；N：阴性对照；P：阳性对照。

Fig. 1 The results of healthy（A），TYLCV-infected（B），ToCV-infected（C），ToCV and TYLCV-infected（D，E）Bemisia tabaci M：DNA marker；1-12：B. tabaci sample；N：Negative control；P：Positive control.

图2 取食健康和携带病毒（ToCV + TYLCV、ToCV、TYLCV）番茄48 h后的烟粉虱的解毒酶活性不同小写字母表示0.05水平差异显著。下同。

Fig. 2 Detoxification activity of Bemisia tabaci 48h after feeding on healthy and virus-carrying （ToCV + TYLCV，ToCV，TYLCV）tomatoes Different lowercase letters indicate significant difference at 0.05 level. The same below.

图3 取食健康和携带病毒（ToCV + TYLCV、ToCV、TYLCV）番茄48 h烟粉虱体内解毒酶相关基因表达量的变化

Fig. 3 Changes of detoxification enzyme related gene expression in healthy and virus-carrying tomatoes （ToCV + TYLCV，ToCV，TYLCV）for 48 h in Bemisia tabaci

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