Quantification and ecological study of ‘Candidatus Liberibacter asiaticus’ in citrus hosts,rootstocks and the Asian citrus psyllid

The use of proper management strategies for citrus huanglongbing (HLB), caused by ‘Candidatus Liberibacter asiaticus’ (Las) and transmitted by Asian citrus psyllid (ACP) (Diaphorina citri), is a priority issue. HLB control is based on healthy seedlings, tolerant rootstock cultivars and reduction of ACP populations. Here, dynamic populations of Las in different citrus hosts and each instar of ACP were studied, together with the seasonal growth and distribution of Las in different tissues, using conventional and TaqMan real‐time PCR. Different levels of susceptibility/tolerance to HLB were seen, resulting in different degrees of symptom severity and growth effects on hosts or rootstocks. Troyer citrange, Swingle citrumelo and wood apple were highly tolerant among 11 rootstock cultivars. Regarding distribution and seasonal analysis of Las, mature and old leaves contained high concentrations in cool temperatures in autumn and spring. Las was detected earlier through psyllid transmission than through graft inoculation, and the amounts of Las (AOL) varied in different hosts. Thus, different AOL (10⁴–10⁷ copy numbers μL^?1) and Las‐carrying percentages (LCP; 40–53.3%) were observed in each citrus cultivar and on psyllids, respectively. Furthermore, both AOL and LCP were lower in nymphs than in adult psyllids, whereas the LCP of psyllids were not affected by increasing the acquisition‐access time. The present study has significant implications for disease ecology. The combination of early detection, use of suitable rootstocks and constraint of psyllid populations could achieve better management of HLB disease.     

48%盖灌能EC对柑橘黄龙病树伐桩处理试验

在柑橘黄龙病重灾区,病树清理量大,病树伐桩处理困难,目前普遍推行的处理方法费工费时,处理措施难以到位,导致病树伐桩萌芽率高,成为新的传染源。本文利用48%盖灌能EC使用柴油为溶剂对柑橘黄龙病树伐桩进行处理。结果表明,5个处理试验对抑制病树伐桩萌芽均有很好的防效,270 d防效全南县果园达到98.7%,其他都达到100%;同时具有很好的催腐效果。该处理方法操作简单易行,省时省工,可以节省大量的人力和财力,是一种经济、安全、有效的柑橘黄龙病树伐桩处理方法。     

柑橘黄龙病蒸汽快速热处理升温特性及田间防治效果

为了深入了解柑橘黄龙病湿热蒸汽快速热处理时韧皮部的升温特性,针对柑橘黄龙病细菌位于韧皮部的特点,构建了室内和田间试验装置,主要包含加热罩、湿热蒸汽发生装置、树体表面和韧皮部温度采集装置。通过比较热处理前后韧皮部温度变化,作物恢复程度差异和黄龙病病菌浓度变化,分析获取湿热蒸汽快速热处理合理参数组合。室内试验选用1 a生柑橘树,研究了树体表面温度为55~65℃,蒸汽输送压力为0.015和0.03 MPa,不保温和保温处理30 s对于韧皮部升温和作物恢复的影响。试验结果:树体表面温度为65℃及以上时,作物都因热损而死亡,采用60℃作为热处理温度阈值,有利于韧皮部温度提高。较高的蒸汽输送压力(如0.03 MPa),虽有利于作物所在环境温度的快速升高,但其作用于韧皮部的传热时间短,韧皮部温度并没有得到有效提高,而加热后的30 s保温处理普遍有利于韧皮部温度提高。田间试验选用9 a生柑橘树,研究了树体表面温度为60℃,蒸汽输送压力为0.03 MPa,保温时间30 s的组合参数对于黄龙病病菌浓度和作物恢复的影响。试验结果:4棵染病柑橘树经过热处理,其中2棵恢复到未染病状态,2棵病菌浓度降低。2棵未经过热处理的染病柑橘树,其病菌浓度没有变化。室内和田间试验结果表明:湿热蒸汽快速热处理能有效促进染病柑橘树的生命力恢复,明显降低病菌浓度;对于柑橘黄龙病田间湿热蒸汽快速热处理,不能只关注树体表面温度的快速提高,而应关注韧皮部温度的有效提高;热处理应考虑直接蒸汽加热和关闭蒸汽后保温的综合影响,为了增加韧皮部传热,应尽量选取较高升温上限和较低输送压力。研究结果为柑橘黄龙病田间热处理防治提供参考。     

柑桔木虱与烟草的关系研究

本文对柑桔木虱与烟草的关系进行了研究。结果显示：木虱在烟草上存活的时间不会超过36h;对烟草嫩叶的趋性强于“默科特”桔橙嫩叶;带菌木虱刺吸烟草后,可把黄龙病病菌传给烟草,但感病烟草不表现黄龙病病症。     

柑橘黄龙病碘—淀粉法快速检测技术优化

为使一种柑橘黄龙病碘—淀粉法快速检测技术更好地应用于大田检测,以砂糖橘为试验材料,研究无需冷冻和缩短叶片脱色时间的技术.结果表明,磨掉叶片正反面表面蜡质层后,直接脱色,可以省去冷冻处理步骤,且不影响检测结果;检测结果与常规PCR检出结果的一致率为92.5%;同时,叶片脱色时间缩短7.17~8.47 h.优化后,冬季样品叶绿素脱除所需时间均值为5.0 h,其他季节叶片脱色时间均值为3.7 h.     

Distribution,biology, ecology and control of the psyllid Diaphorina citri Kuwayama,a major pest of citrus: A status report for China

Abstract

The Asiatic citrus psyllid Diaphorina citri Kuwayama (Hemiptera: Psyllidae) is a major pest of citrus in China. Its status derives, not from the damage it causes, but from its role as the only known vector in China of huanglongbing, a phloem-limited bacterial disease of international importance. The disease can devastate orchards within a few years of planting. It also poses a major threat to endangered indigenous citrus germplasm in Asia and Australasia. The distribution, biology, ecology and control of the psyllid in China are reviewed in these contexts. Constraints and challenges related to control of the vector in China are discussed.     

Effect of Candidatus Liberibacter asiaticus infection on susceptibility of Asian citrus psyllid, Diaphorina citri, to selected insecticides

BACKGROUND: In the present investigation, the effect of Candidatus Liberibacter asiaticus (Las), a bacterium considered to be responsible for causing huanglongbing (HLB) disease in citrus, on the physiology of its vector, the Asian citrus psyllid (ACP) Diaphorina citri Kuwayama, was determined. Specifically, the effects of Las infection on the susceptibility of ACP to selected insecticides were determined. Furthermore, total protein content and general esterase activity were quantified in Las‐infected and uninfected ACP to gain insight into the possible mechanism(s) responsible for altered susceptibility to insecticides owing to Las infection. RESULTS: LC₅₀ values were significantly lower in Las‐infected than in uninfected ACP adults for chlorpyrifos and spinetoram. Furthermore, there was a general trend towards lower LC₅₀ values for three other insecticides for Las‐infected ACP; however, the differences were not statistically significant. Total protein content (µg mL^?1) was significantly lower in Las‐infected (23.5 ± 1.3 in head + thorax; 27.7 ± 1.9 in abdomen) than in uninfected (29.7 ± 2.1 in head + thorax; 35.0 ± 2.3 in abdomen) ACP. Likewise, general esterase enzyme activity (nmol min^?1 mg^?1 protein) was significantly lower in Las‐infected (111.6 ± 4.5 in head + thorax; 109.5 ± 3.7 in abdomen) than in uninfected (135.9 ± 7.5 in head + thorax; 206.1 ± 23.7 in abdomen) ACP. CONCLUSION: Susceptibility of ACP to selected insecticides from five major chemistries was greater in Las‐infected than in uninfected ACP. The lower total protein content and reduced general esterase activity in Las‐infected than in uninfected ACP may partly explain the observed higher insecticide susceptibility of Las‐infected ACP. Copyright © 2010 Society of Chemical Industry     

基于光谱信息的柑橘黄龙病无损检测及分级模型构建

柑橘黄龙病(Huanglongbing,HLB)是世界柑橘生产上最具毁灭性的病害,给果农和相关产业造成了巨大的损失.以柑橘叶片为载体,利用高光谱图像技术采集柑橘叶片表面的高光谱图像,用ENVI4.7进行图像处理,提取感兴趣区域(Region of Intest,ROI),统计感兴趣区域平均光谱数据,并进行相关植被植物的运算,最后通过PLS-DA(Partial Least Squares Discrimination Analysis)判别法进行鉴别并分类.结果表明:基于平均光谱值和植被指数的PLS-DA判别模型都能对健康、缺锌和HLB叶片进行鉴别.其中基于平均光谱值的PLS-DA模型鉴别健康柑橘叶片样品的灵敏度为100％,特异度为100％,准确度为100％;鉴别缺锌柑橘叶片样品的灵敏度为80.6％,特异度为91.7％,准确度为88.9％;鉴别HLB叶片的灵敏度为89.3％,特异度为88.3％,准确度为88.9％.基于植被指数的PLS-DA判别模型鉴别健康柑橘叶片样品的灵敏度为100％,特异度为100％,准确度为100％;鉴别缺锌柑橘叶片样品灵敏度为92.5％,特异度为89.3％,准确度为90.1％;鉴别HLB叶片的灵敏度为86.4％,特异度为95.3％,准确度为90.1％.识别正确率较高,说明利用高光谱进行柑橘黄龙病病情分类是可行的.     

种子传播黄龙病可能性研究

[目的]探讨带菌种子能否传播黄龙病,为黄龙病防控策略的制定提供科学依据.[方法]采用单管双引物对TaqMan探针实时荧光定量PCR(STDP-qPCR)对从感染黄龙病沙田柚树上收集的种子进行黄龙病菌亚洲种(Candidatus Liberibacter asi-aticus,Las)检测,观察播种的沙田柚种子萌发生长情况,并用STDP-qPCR对萌发的幼苗进行Las检测.[结果]两份种子样品均能检测到Las.播种的108颗种子共萌发91颗(84.3％),部分沙田柚苗在生长初期表现叶片畸形或黄化症状,但随后恢复正常.由种子萌发而来的沙田柚苗经30、60、90、180和360d生长后取样检测,均未检测出Las.[结论]黄龙病通过带菌种子进行传播缺乏证据.