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寻找非传毒粉虱:亚洲柑橘木虱同型雌系获得和传播黄龙病菌“亚洲韧皮杆菌”的天然变异。

The quest for a non-vector psyllid: Natural variation in acquisition and transmission of the huanglongbing pathogen 'Candidatus Liberibacter asiaticus' by Asian citrus psyllid isofemale lines.

机构信息

USDA-ARS, US Horticultural Research Laboratory, Fort Pierce, Florida, United States of America.

University of Florida, Lake Alfred & Fort Pierce, Florida, United States of America.

出版信息

PLoS One. 2018 Apr 13;13(4):e0195804. doi: 10.1371/journal.pone.0195804. eCollection 2018.

Abstract

Genetic variability in insect vectors is valuable to study vector competence determinants and to select non-vector populations that may help reduce the spread of vector-borne pathogens. We collected and tested vector competency of 15 isofemale lines of Asian citrus psyllid, Diaphorina citri, vector of 'Candidatus Liberibacter asiaticus' (CLas). CLas is associated with huanglongbing (citrus greening), the most serious citrus disease worldwide. D. citri adults were collected from orange jasmine (Murraya paniculata) hedges in Florida, and individual pairs (females and males) were caged on healthy Murraya plants for egg laying. The progeny from each pair that tested CLas-negative by qPCR were maintained on Murraya plants and considered an isofemale line. Six acquisition tests on D. citri adults that were reared as nymphs on CLas-infected citrus, from various generations of each line, were conducted to assess their acquisition rates (percentage of qPCR-positive adults). Three lines with mean acquisition rates of 28 to 32%, were classified as 'good' acquirers and three other lines were classified as 'poor' acquirers, with only 5 to 8% acquisition rates. All lines were further tested for their ability to inoculate CLas by confining CLas-exposed psyllids for one week onto healthy citrus leaves (6-10 adults/leaf/week), and testing the leaves for CLas by qPCR. Mean inoculation rates were 19 to 28% for the three good acquirer lines and 0 to 3% for the three poor acquirer lines. Statistical analyses indicated positive correlations between CLas acquisition and inoculation rates, as well as between CLas titer in the psyllids and CLas acquisition or inoculation rates. Phenotypic and molecular characterization of one of the good and one of the poor acquirer lines revealed differences between them in color morphs and hemocyanin expression, but not the composition of bacterial endosymbionts. Understanding the genetic architecture of CLas transmission will enable the development of new tools for combating this devastating citrus disease.

摘要

昆虫媒介的遗传变异性对于研究媒介能力决定因素和选择非媒介种群很有价值,这些种群可能有助于减少媒介传播病原体的传播。我们收集并测试了 15 条亚洲柑橘木虱(Diaphorina citri)的同型雌线,亚洲柑橘木虱是“亚洲韧皮杆菌(CLas)”的媒介。CLas 与黄龙病(柑橘绿化)有关,这是世界上最严重的柑橘病害。佛罗里达州的橙子茉莉(Murraya paniculata)树篱中采集了成年柑橘木虱,并将每对个体(雌性和雄性)关在健康的 Murraya 植物上产卵。每对经 qPCR 检测为 CLas 阴性的后代都在 Murraya 植物上饲养,并被视为同型雌线。对来自不同代的各种同型雌线的幼虫进行了 6 次成虫获取试验,以评估它们的获取率(qPCR 阳性成虫的百分比)。有 3 条线的平均获取率为 28%至 32%,被归类为“良好”获取者,另外 3 条线被归类为“较差”获取者,获取率仅为 5%至 8%。所有的线都进一步通过将暴露于 CLas 的木虱限制在健康柑橘叶片上一周(每叶/周 6-10 只成虫)并通过 qPCR 测试叶片中的 CLas 来测试它们的接种能力。对于三个良好获取者线,平均接种率为 19%至 28%,而对于三个较差获取者线,平均接种率为 0%至 3%。统计分析表明,CLas 的获取和接种率之间以及木虱中 CLas 的滴度与 CLas 的获取或接种率之间存在正相关关系。对一个良好获取者线和一个较差获取者线的表型和分子特征进行了研究,发现它们在颜色形态和血蓝蛋白表达方面存在差异,但细菌共生体的组成没有差异。了解 CLas 传播的遗传结构将使开发新的工具来对抗这种毁灭性的柑橘病害成为可能。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/311e/5898736/1c5c062e1deb/pone.0195804.g001.jpg

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