柑橘木虱两地理种群的内共生菌群落组成及传菌能力

柑橘黄龙病（Huanglongbing, HLB）是经柑橘木虱Diaphorina citri传播的最主要柑橘病害之一, 危害严重时能对柑橘产业造成毁灭性的破坏。为了鉴定福建和海南2个地理种群柑橘木虱的内共生菌群落组成, 本研究对16S rRNA部分保守序列进行PCR扩增, 并利用特异性引物对不同内共生菌进行了感染率检测; 另外, 还通过人工接虫的方法, 探索柑橘木虱成虫在带黄龙病菌蕉柑Citrus reticulata cv. Tankan上的获菌能力, 以及带菌柑橘木虱成虫对黄岩蜜橘C. reticulata cv. Subcompressa的传菌能力。研究发现, 这2个地理种群的柑橘木虱含有相同的内共生菌组成, 包括α-Proteobacteria, Wolbachia spp., γ-Proteobacteria, mycetocyte symbionts, β-Proteobacteria, Oxalobacter和β-Proteobacteria, Herbaspirillum, 而且这2个地理种群柑橘木虱的4种内共生菌的携带率均在95%以上。柑橘木虱成虫在带菌蕉柑上饲菌28 d后, 带菌率可达到82%, 而带菌柑橘木虱成虫在黄岩蜜橘上传菌75 d后, 可导致橘树整体带菌。本研究为柑橘木虱的进一步研究和防虫治病途径提供了一些理论依据。     

黄龙病病菌在柑橘木虱体内的分布及感染动态

【目的】柑橘木虱Diaphorina citri Kuwayama是传播黄龙病病菌唯一的自然媒介昆虫,本文研究了黄龙病病菌在柑橘木虱各个发育阶段体内的分布以及感染动态,为有效防控柑橘木虱提供科学数据。【方法】利用常规PCR检测柑橘木虱若虫体内的黄龙病病菌,利用q RT-PCR和荧光原位杂交技术分别检测黄龙病病菌在柑橘木虱不同发育虫态体内的含量与分布形式。【结果】常规PCR可以在3-5龄若虫以及成虫体内检测到黄龙病病菌,而q RT-PCR除了3-5龄若虫外,还可以在2龄若虫体内检测到黄龙病病菌。卵和各龄期若虫中黄龙病病菌的含量是随着龄期的变大而不断增多。产卵盛期的成虫木虱体内黄龙病病菌的含量最高,显著高于产卵前期以及产卵后期。荧光原位杂交技术可以检测到4龄、5龄若虫及成虫体内的黄龙病病菌分布形态,病菌在若虫主要分布在U型含菌体内,而在雌雄成虫体内均是散布型分布。【结论】柑橘黄龙病在柑橘木虱的2-5龄若虫及成虫中都有感染,但其含量与分布形式因发育阶段不同而有显著差异。     

亚洲柑橘木虱成虫和5龄若虫在感染黄龙病的柑橘上的取食行为及获菌效率比较

【目的】亚洲柑橘木虱Diaphorina citri是柑橘的毁灭性病害——黄龙病亚洲种‘Candidatus Liberibacter asiaticus’(‘C las’)的主要传播媒介。本研究的目的是明确木虱成虫和5龄若虫的取食行为、获菌效率是否有差异,以及寄主感染黄龙病是否对5龄若虫取食产生影响。【方法】利用直流型刺吸电位仪(DC-EPG Giga-4)记录柑橘木虱成虫和5龄若虫在携带黄龙病的酸橘Citrus reticulata cv.Sunki嫩梢上10 h的取食行为,用qPCR单头检测其获得黄龙病病原菌的效率,并比较5龄若虫在感病和健康植株嫩梢上的取食行为。【结果】柑橘木虱成虫与5龄若虫在感染黄龙病的酸橘上的取食行为有显著差异。5龄若虫比成虫更快地开始在韧皮部和木质部进行吸食,口针在韧皮部的总过程以及吸食时间显著长于成虫。此外,5龄若虫和成虫在EPG测定(同时饲菌)10 h后获菌率分别为37.5%和20.0%,若虫明显高于成虫。寄主植物感染黄龙病对5龄若虫的取食行为有一定的影响,表现在感病植株上的刺探次数、唾液分泌次数和韧皮部吸食次数都显著少于健康植株,而两者分泌唾液和韧皮部吸食时间没有显著差异。另外,在感病植株上首次韧皮部取食出现时间较在健康植株上要早。【结论】在同等时间下,柑橘木虱5龄若虫比成虫在感染黄龙病的酸橘上的取食能力更强、取食量更大、获菌率更高,原因可能是若虫需要更多的营养物质供其生长发育所致。寄主感染黄龙病对木虱5龄若虫的取食有利,使其更快地开始取食,而且更改取食位点次数变少,推测可能与黄龙病菌破坏了植物的防御有关。     

藿香蓟和假臭草对柑橘木虱种群发展的影响

果园生草是病虫生态控制中的重要手段之一,藿香蓟可为柑橘害螨的自然天敌捕食螨提供栖息和繁殖场所,在橘园内被大量保留和引种。本文测试了重要柑橘害虫柑橘木虱成虫对藿香蓟的选择偏好、栖居于藿香蓟上的存活时长,以及藿香蓟存在条件下农药对木虱成虫防治效果的影响。结果显示:在无柑橘时,木虱成虫对藿香蓟和假臭草的选择比率分别为31.52%和16.22%,两者差异极显著,而在有柑橘同时存在的条件下,木虱成虫对藿香蓟和假臭草也分别有1.24%和0.83%的选择比率,但两者差异不显著;而木虱在藿香蓟上存活最长达49 d,在假臭草上仅12 d;在柑橘枝条上喷施1.8%阿维菌素乳油90 mg/L 11 d后,若无藿香蓟同时存在,木虱成虫死亡率为79.49%;若有藿香蓟同时存在,木虱成虫死亡率仅为42.76%,两者差异极显著。结果表明,藿香蓟对农药防治柑橘木虱的效果存在一定的干扰作用,对木虱种群的发展有较大的影响。     

亚洲柑橘木虱雌成虫对砂糖橘幼嫩部位的产卵选择性研究

【目的】明确亚洲柑橘木虱Diaphorina citri Kuwayama雌成虫的产卵过程及其对柑橘嫩芽产卵的偏好性。【方法】利用手机和微距镜头观察记录柑橘木虱雌成虫的产卵过程,并结合田间调查与室内试验明确了亚洲柑橘木虱雌成虫对砂糖橘嫩叶和不同发育状态嫩芽的偏好性。【结果】亚洲柑橘木虱雌成虫产卵前,头部前倾,尾部上翘,触角不断抖动,一段时间后,雌成虫爬行到合适的位点开始产卵,或爬至其他部位,继续上述行为,直至找到合适的位置产卵。产卵时,触角静止不动,腹部收缩带动产卵器向腹部弯曲,肛上板与下生殖板张开,产卵器插入产卵位点产卵。产卵结束后,雌成虫迅速抽出产卵器,肛上板与下生殖板在抽离过程中缓慢闭合。田间调查发现,亚洲柑橘木虱雌成虫偏向于未展开和半展开的柑橘嫩芽上产卵。室内试验发现,当同时供给新叶和多种发育状态的砂糖橘嫩芽时,亚洲柑橘木虱产卵偏好于未展开和半展开的砂糖橘嫩芽,且不会选择在砂糖橘新叶上产卵。当仅供给一种发育状态的砂糖橘嫩芽或新叶时,亚洲柑橘木虱雌成虫在新叶和嫩芽上都能产卵,但在新叶上的产卵量明显比嫩芽上的产卵量小。【结论】明确了亚洲柑橘木虱雌成虫的产卵过程。亚洲柑橘木虱雌成虫产卵具有明显趋嫩性,对于砂糖橘,雌成虫偏向于0-10 mm的嫩芽上产卵。     

柑橘木虱成虫趋光行为反应

柑橘木虱Diaphorina citri Kuwayama是柑橘黄龙病(huanglongbing,HLB)的重要传播媒介。为了利用灯光诱控技术防治柑橘木虱,本实验于室内条件下研究柑橘木虱对波长为360 nm、400 nm、440 nm、480 nm、520 nm、560 nm和600 nm的LED光源和不同光照强度趋光行为反应。结果表明:柑橘木虱对7种单色光都有正趋向性。其中雌雄混合存在时对400 nm的紫光趋向性最强,其次是560 nm的绿光;单独处理时,雌成虫对400 nm的紫光趋性最强,其次是520 nm的绿光,雄成虫则是对520 nm的绿光趋性最强,其次是400 nm的紫光。在200μw/cm 2到1000μw/cm 2的光照强度范围内,随着光照强度的增大,柑橘木虱雄成虫趋光行为逐渐增强,在光照强度为1000μw/cm 2时趋光行为最强,但雌成虫趋光行为变化不明显。该研究表明:柑橘木虱雌雄成虫具有明显的正趋光性,且对光谱和光强的反应存在差异。这一结果可为柑橘木虱田间的灯光诱控提供实验依据。     

柑橘木虱蜜露排泄机制:进展与展望

半翅目昆虫柑橘木虱Diaphorina citri是柑橘类果树重要害虫,主要以高渗透压的植物韧皮部汁液为食,是柑橘毁灭性病害——柑橘黄龙病(HLB)的主要传播媒介。柑橘木虱取食韧皮部汁液时自身进化出一套完整的渗透调节机制调节其体内渗透压,将体内过量摄入的糖类转化成长链寡糖并以蜜露形式排出体外。本文从柑橘木虱蜜露排泄行为、蜜露组成成分以及影响蜜露排泄的多个因素进行了论述,同时综述了可能参与柑橘木虱蜜露排泄行为的渗透调节基因。研究表明,柑橘木虱雌雄成虫及若虫在蜜露排泄行为上存在显著差异,且排泄的蜜露在颜色、纹路及组成成分方面均有不同;寄主植物、杀虫剂、病原微生物及天敌化合物均会影响柑橘木虱蜜露排泄行为。分子机制探究发现,α-葡萄糖苷水解酶、水通道蛋白及糖基转移酶基因等关键渗透调节基因可能参与调控柑橘木虱蜜露排泄行为。本文可为未来有关柑橘木虱蜜露排泄行为方面的研究以及为研制柑橘木虱防治新型药剂开发新靶标提供参考。     

不同种类寄主对柑橘黄龙病菌侵染能力及虫媒适应性的影响

柑橘木虱Diaphorina citri是柑橘黄龙病菌“Candidatus Liberibacter asiaticus”的主要媒介昆虫，探明不同植物种类寄主对柑橘黄龙病菌侵染能力及柑橘木虱适应性的影响，在设计黄龙病防控策略及寄主抗性研究中具有参考价值。本文选取蕉柑Citrus reticulata、蜜柚Citrus grandis、黄皮Clausena lansium和九里香Murraya exotica为研究材料，对比其田间植株黄龙病发病率和柑橘木虱数量及带菌情况，并在实验室条件下比较柑橘木虱对不同寄主的适应性（包括发育历期、存活率、携菌能力和传菌能力）。结果发现：（1）田间蕉柑和蜜柚的感染率最高，分别为47%和37%；而黄皮和九里香的带菌率较低，分别为18%和4%，且菌浓度均较轻微；（2）田间九里香上柑橘木虱卵、若虫及成虫数量均最多，蕉柑和黄皮其次，蜜柚最少，但蕉柑和蜜柚上柑橘木虱的携菌率和携菌量最高；（3）4种寄主上柑橘木虱卵和1~3龄若虫的发育历期没有显著性差异，柑橘木虱4龄、5龄若虫和总若虫历期在九里香上最短，而在蜜柚上最长；4种寄主上4龄、5龄若虫存活率没有显著性差异，但蜜柚上木虱卵的孵化率、1~3龄及总若虫存活率最低。（4）蕉柑和蜜柚上带菌木虱的携菌能力及传菌能力最高，其次为黄皮，九里香最低。该研究证明了除了寄主植物本身抗病性差异外，虫媒对寄主植物的适应性差异也是造成田间黄龙病发病差异的重要因素。     

利用虫生真菌生物防治柑橘木虱的研究进展

柑橘木虱是柑橘黄龙病的重要传播虫媒。目前对柑橘黄龙病的防治尚缺有效的药剂和抗病品种,加强对柑橘木虱的防治,对控制柑橘黄龙病的蔓延具有重要意义。目前防治柑橘木虱多采用化学防治,杀虫剂的频繁使用造成了农药残留、环境污染、生物多样性被破坏和害虫产生抗药性等诸多问题,生物防治以其高效、低毒、低残留、不易产生抗药性等优点逐渐受到重视。昆虫病原真菌能侵入昆虫寄主体内,导致昆虫发病死亡,具有良好的病害流行潜力及生产应用便利性,利用昆虫病原真菌防治柑橘木虱具有广阔的发展空间。本文总结了用于柑橘木虱生物防治的虫生真菌种类,重点介绍了国内外利用球孢白僵菌、玫烟色棒束孢、淡紫紫孢菌、宛氏拟青霉、蜡蚧菌等虫生真菌在防治柑橘木虱中的应用,并对虫生真菌防治柑橘木虱的发展前景进行了展望,以期为柑橘黄龙病的防控提供借鉴。     

防治果园周边柑橘木虱控制黄龙病效果研究

在新种柑橘园中栽种柑橘无毒苗,定期监测和防治果园内柑橘木虱Diaphorina citri Kuwayama,防治时期为各新梢萌芽期,重点防治期为早夏梢期和早秋梢期;与此同时,调查柑橘园周围柑橘木虱寄主植物,每年2月下旬、5月上旬、8月下旬和11月下旬定期喷药,清除柑橘园周围寄主植物上的柑橘木虱。结果表明,果园内柑橘木虱和黄龙病均得到有效的控制,果园内柑橘木虱种群数量控制在很低的水平,最高为225头/公顷,最低为0;黄龙病发病率也很低,2008年为8‰,实施该技术后,发病率逐年下降,2012年降低到01‰。     

A new Huanglongbing Species, "Candidatus Liberibacter psyllaurous," found to infect tomato and potato, is vectored by the psyllid Bactericera cockerelli (Sulc)

A new huanglongbing (HLB) "Candidatus Liberibacter" species is genetically characterized, and the bacterium is designated "Candidatus Liberibacter psyllaurous." This bacterium infects the psyllid Bactericera cockerelli and its solanaceous host plants potato and tomato, potentially resulting in "psyllid yellowing." Host plant-dependent HLB transmission and variation in psyllid infection frequencies are found.     

Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), and huanglongbing disease do not exist in the Stapleton Station area of the Northern Territory of Australia

Abstract  A series of specimens of the Asian citrus psyllid, Diaphorina citri , collected from the Northern Territory (NT) in 1915 was recently rediscovered in the Natural History Museum, London. Surveys were conducted in 2002 on suitable hosts in the locality of the 1915 collections to see if the infestation had persisted. These failed to detect either D. citri or the bacterium that it transmits and that causes huanglongbing disease in citrus. It is presumed that D. citri was eradicated fortuitously by the removal of all citrus plants above latitude 19°S during an eradication program for citrus canker in the NT from 1916 until 1922.     

Transmission parameters for Candidatus liberibacter asiaticus by Asian citrus psyllid (Hemiptera: Psyllidae)

The purpose of this investigation was to evaluate acquisition and inoculation (together, transmission) efficiency of Candidatus Liberibacter asiaticus (Las), the pathogen associated with citrus huanglongbing (HLB) by the Asian citrus psyllid, Diaphorina citri (Kuwayama) (Hemiptera: Psyllidae). In laboratory studies, nymphs reared on Las infected plants were more likely to acquire the bacterium than adults. Acquisition by nymphs ranged from 60 to 100%, whereas acquisition by adults only reached 40% after 5 wk of feeding on Las-infected plants. Similar rates of pathogen acquisition by psyllids after nymphal and adult feeding were observed in the field. Transmission of Las from parent to offspring (transovarial) occurred at a rate of 2-6%. One year after psyllid inoculations, successful transmission by individual D. citri ranged from 4 to 10%, whereas groups of 100 or more D. citri transmitted the pathogen at a rate of approximately 88%. In addition, the proportion of Las-positive adult psyllids, determined using quantitative real-time polymerase chain reaction, decreased over time when held on healthy plants. Due to the low rate of pathogen acquisition and long time period required for successful inoculation by adult D. citri, experiments designed to determine the latent period required for replication and successful inoculation of Las by D. citri did not result in Las-infected plants after >1 yr of incubation after inoculation. Collectively, these results indicate that adult D. citri which acquire the HLB pathogen as adults are poor vectors of the pathogen compared with adults that acquired the pathogen as nymphs.     

A New Huanglongbing Species, “Candidatus Liberibacter psyllaurous,” Found To Infect Tomato and Potato, Is Vectored by the Psyllid Bactericera cockerelli (Sulc)

A new huanglongbing (HLB) “Candidatus Liberibacter” species is genetically characterized, and the bacterium is designated “Candidatus Liberibacter psyllaurous.” This bacterium infects the psyllid Bactericera cockerelli and its solanaceous host plants potato and tomato, potentially resulting in “psyllid yellowing.” Host plant-dependent HLB transmission and variation in psyllid infection frequencies are found.     

Effects of the fungus Isaria fumosorosea (Hypocreales: Cordycipitaceae) on reduced feeding and mortality of the Asian citrus psyllid,Diaphorina citri (Hemiptera: Psyllidae)

The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is an important citrus pest primarily because it transmits bacteria putatively responsible for huanglongbing, a serious disease of citrus. We present a study on the effects of blastospore and conidial formulations of Isaria fumosorosea Wize on feeding rates and mortality of adult psyllids in laboratory bioassays. Information on quantities of honeydew droplets was used to make inferences on feeding rates. Psyllids treated with the blastospore formulation of I. fumosorosea produced fewer honeydew droplets compared to the conidial treatment and control beginning within the first 24 h after treatment. The highest daily mean number of droplets thereafter never exceeded 2.4 drops compared to 4 and 8 for the conidial treatment and control, respectively. The mean number (±SEM) of honeydew droplets produced per psyllid per day over 7 days was significantly higher in the control (5.5±0.5) compared to the blastospore treatment; however, there were no significant differences between the treatments. Psyllids treated with the conidial formulation of the pathogen showed no significant reduction in feeding activity until 4 days after treatment. One and 2 day's post-exposure, mortality of psyllids in the blastospore treatment ranged from 8 to 25% compared to 0% in the conidial and control treatments. By 7 days post-exposure, psyllid mortality reached 100% under both fungal treatments compared to none in the controls. This study documented that adult psyllids infected by I. fumosorosea (PFR 97) produce less honeydew than healthy psyllids and suggests that they may feed less, which could potentially reduce the spread of huanglongbing.     

Enhanced proliferation and efficient transmission of Candidatus Liberibacter asiaticus by adult Diaphorina citri after acquisition feeding in the nymphal stage

We carried out a quantitative detection of Candidatus Liberibacter asiaticus, the bacterium associated with the disease of huanglongbing, in the vector psyllid Diaphorina citri by using a TaqMan real‐time PCR assay. The concentration of the bacterium was monitored at 5‐day intervals for a period of 20 days after psyllids were exposed as fifth instar nymphs or adults to a Ca. L. asiaticus‐infected plant for an acquisition access period of 24 h. When adults fed on Ca. L. asiaticus‐infected plant, the concentration of the bacterium did not increase significantly and the pathogen was not transmitted to any citrus seedlings. In contrast, when psyllids fed on infected plant as nymphs, the concentration of the pathogen significantly increased by 25‐, 360‐ and 130‐fold from the initial acquisition day to 10, 15 and 20 days, respectively. Additionally, the pathogen was successfully transmitted to 67% of citrus seedlings by emerging adults. Our data suggested that multiplication of the bacterium into the psyllids is essential for an efficient transmission and show that it is difficult for adults to transmit the pathogen unless they acquire it as nymphs.     

Combined effects of drought stress and psyllid herbivory on the invasive weed Scotch broom,Cytisus scoparius

Increases in the frequency and duration of droughts under global climate change could have implications for plant–insect interactions, and could either increase or decrease the effects of weed biological control agents. In this study, we used greenhouse and field experiments to examine the impacts of drought stress on the abundance and impacts of the adventive psyllid Arytainilla spartiophila Forster (Hemiptera: Psyllidae, Arytaininae) on its target weed Scotch broom, Cytisus scoparius (L.) Link (Fabaceae), in California, USA. The psyllid impacted plant growth in both field and greenhouse experiments, whereas drought stress affected plant growth in the greenhouse only, suggesting that other factors besides water availability may be more limiting for plants in the field. Effects on psyllid survival were consistent with the plant vigor hypothesis, which predicts that herbivores will perform better on vigorously growing plants; psyllid numbers were lower on drought-stressed plants in the greenhouse and were correlated with plant growth in both the greenhouse and the field. In the greenhouse, the combined effects of the psyllid and drought stress were additive, indicating that the effects of the psyllid were consistent across unstressed and drought-stressed plants. Although the psyllid is unlikely to control Scotch broom on its own, results suggest that it may work in conjunction with drought stress to suppress Scotch broom.     

Secondary hosts of the Asian citrus psyllid,Diaphorina citri Kuwayama: Survivorship and preference

The Asian citrus psyllid, Diaphorina citri Kuwayama, is the most economically important and difficult to manage citrus pest in Florida due to its role as vector of huanglongbing or citrus greening disease. Spread of the disease is a function of dispersal patterns and thus influenced by availability of suitable hosts. Young leaves of citrus or related plants are required for reproduction, but in their absence, secondary hosts may provide needed resources to enhance survival. Therefore, survivorship on and preference for three abundant weed species was investigated. The suitability of potential secondary host plants Bidens alba (L.), DC, Ludwigia octovalvis (Jacqu.) P. H. Raven, and Eupatorium capillifolium (Lam.) Small was compared to a reproductive host, Murraya paniculata (L.) Jack, in no‐choice tests by assessing survivorship of D. citri adults confined to these plants in cages. Preference was evaluated by choice tests where D. citri adults were released into cages containing all three secondary hosts alone or with M. paniculata. Both B. alba and E. capillifolium increased D. citri survivorship by twofold compared to starvation conditions with only water available. Choice trials revealed no difference in initial selection between true and secondary hosts; however, the true host was favoured over time. This result suggested that hosts were selected initially by sight, and only later by taste and/or smell. While secondary hosts are unable to support reproduction or long‐term survival, these findings establish the ability of D. citri to use secondary hosts that are ubiquitous in Florida citrus groves as temporary reservoirs for food and moisture when ideal host conditions are scarce or absent.     

Assessment of Variation in Feeding Behavior by Color Morph in the Asian citrus Psyllid (Diaphorina citri)

Journal of Insect Behavior - The Asian citrus psyllid (ACP) Diaphorina citri (Hemiptera:Liviidae), vector of huanglongbing disease, displays a high degree of color polyphenism. In the adult stage,...