JIA-2021-0994 苹果茎沟病毒与黄金蜜柚黄化斑驳花叶病相关性研究

doi:10.1016/S2095-3119(21)63823-6

摘要/Abstract

摘要：

苹果茎沟病毒 (apple stem grooving virus, ASGV) 是一种重要的潜隐类果树病毒，对柑橘、梨和苹果等多种果树的生产构成了严重的威胁。2018年，在中国南方广泛种植的黄金蜜柚 (Citrus grandis cv. Huangjinmiyou) 上观察到了严重的黄化、斑驳和花叶症状，推测其可能由病毒引起。取5株表现相关症状果树的叶片样品构建混库并送高通量测序分析，从其中鉴定到了3个ASGV变异体，通过RT-PCR和RACE技术验证了其基因组序列。序列分析显示，这3个变异体的基因组核苷酸序列一致性为81.03%–82.34%，其基因组结构与过往报道的侵染其它果树的变异体类似。基于病毒全基因组核苷酸序列和外壳蛋白氨基酸序列的系统发育分析显示，3个黄金蜜柚ASGV变异体分别与来自不同寄主和地区的ASGV变异体聚在一枝。重组分析显示，3个ASGV变异体可能来自于ASGV不同株系间的重组。在全国11个主要柑橘种植省份采集了507份黄金蜜柚样品进行RT-PCR检测发现，在每个省份表现上述相关症状的样品中，ASGV的检出率均在92.7%以上，而在40份没有症状的样品中，均未检测到ASGV。将其中6个省份的感病样品嫁接到ASGV的指示植物——Rusk枳橙上，新生的系统叶表现出典型的碎叶症状，进一步验证了黄金蜜柚中ASGV的侵染。进一步探究了病毒和症状与温度的关系，发现嫁接的感病黄金蜜柚样品在30°C–35°C条件下症状消失，同时RT-PCR也检测不到ASGV的存在。而后，再将其置于20°C–24°C的条件下一段时间后，黄金蜜柚症状恢复，且ASGV可以由RT-PCR检测到。本文揭示了黄金蜜柚黄化斑驳花叶病与ASGV侵染的相关性，并提示了该病害大面积流行的风险，为进一步的病害防控提供了相应参考。

Abstract: Although it is usually latent on citrus, apple, and pear, apple stem grooving virus (ASGV) poses a great risk to many sensitive cultivars. Since severe leaf yellow mottle mosaic (LYMM) symptoms have been observed on Huangjinmiyou (HJY) pummelos (Citrus grandis cv. Huangjinmiyou), a commercial variety that is widely cultivated in South China, high throughput sequencing (HTS) was used to find potential pathogens and only three divergent ASGV variants were identified. The three ASGV variants shared 81.03–82.34% genome-wide pairwise identities with each other, and were separately closest to other ASGV variants from different hosts and/or geographical regions, as indicated by viral phylogenies. However, these new variants may have developed from viral interstrain interactions, based on the results of recombination analysis. A large-scale survey using reverse transcription-PCR (RT-PCR) protocols designed for the three ASGV variants revealed a high incidence (92.7–100%) of ASGV in symptomatic HJY trees from 11 major citrus-producing regions in China. None of ASGV were detected in asymptomatic trees. Temperature treatments applied to the symptomatic HJY plants showed that ASGV is sensitive to high temperatures (30–35°C), at which not only the plants recovered, but also the viruses were not detected by RT-PCR, while at low temperatures (20–24°C), both the symptoms and viruses remained detectable. These data show that ASGV is associated with the LYMM disease prevalent on HJY in China, and this is the significant basis especially of taking appropriate measures timely to manage the disease.

Key words: citrus, pummelo , apple stem grooving virus , genome sequence , thermotherapy

. JIA-2021-0994 苹果茎沟病毒与黄金蜜柚黄化斑驳花叶病相关性研究[J]. Journal of Integrative Agriculture, 2022, 21(7): 2031-2041.

XUAN Zhi-you, ZHANG Song, LI Ping, YANG Fang-yun, CHEN Hong-ming, LIU Ke-hong, ZHOU Yan, LI Zhong-an, ZHOU Chang-yong, CAO Meng-ji. Apple stem grooving virus is associated with leaf yellow mottle mosaic disease on Citrus grandis cv. Huangjinmiyou in China[J]. Journal of Integrative Agriculture, 2022, 21(7): 2031-2041.

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