柑桔黄龙病及其控防对策

柑桔黄龙病是柑桔生产中危害最重、威胁最大的一种毁灭性病害。文章在分别对柑桔黄龙病的病名、危害与分布、病原的认识过程、诊断技术、病原侵染来源、病原脱除等进行综述的前提下,提出严格实施检疫、重病园成片更新、应用无病毒苗木、及时挖除病株、大面积联防联治、严格防治柑桔木虱等柑桔黄龙病的防控对策。     

柑桔青霉病的发生及生物防治研究进展(英文)

柑桔青霉病是柑桔采后贮运的一种重要病害,是影响柑桔长期贮藏的主要因素。文章首先介绍了柑桔青霉病的病原、危害症状及浸染循环,然后综述了近年来柑桔青霉病的生物防治技术研究进展,旨在为柑桔青霉病的防治提供参考。     

柑桔根结线虫病研究初报

柑桔根结线虫病是江西省新发现的一种病害,经室外调查、室内研究确定,病原线虫为南方根结线虫(Meloidogyne incognita)和爪哇根结线虫(M.javanica).本文对病害的症状、病原、发病条件和综合防治进行了探讨。     

柑桔黄龙病的防治措施

柑桔黄龙病是一种毁灭性的柑桔类病害，其病原是一种病毒性的类质菌体。此病主要通过带病的苗木、接穗、柑桔木虱等传播。该病的症状表现为：树冠顶部梢黄化或斑驳，叶脉肿胀，吉质脆硬，导致果实品味酸，无经济价植，后期烂根，1～2年后全株枯死。该病给果农带来较大的风险。柑桔黄龙病的防治可用以下措施     

快速检测柑桔黄龙病病原的研究

应用ＰＣＲ技术对柑桔黄龙病病原ＤＮＡ进行体外扩增,可建立一套快速,准确,有效的检测该病原的方法,研究结果表明：ＰＣＲ技术对柑桔黄龙病病原的检测具有很强的特异性,只有感染了黄龙病病原的样品,ＰＣＲ才呈阳性反应。文章报道了应用ＰＣＲ技术能检测已带病但尚未症状的柑桔黄龙病病株,该检测技术可以对柑桔黄龙病进行早期诊断,及是地控制带病苗木的传播,这对选育无病苗木及病害的综合治理具有很高的应用价值。     

柑桔黄龙病防控技术研究与应用

根据柑桔黄龙病病原PCR检测结果,分析其病原检出率不高与检测方法、检测部位等相关,提供了"红鼻果"作为芦柑黄龙病田间诊断的依据。通过田间调查,分析总结了至2000年前永春县柑桔黄龙病长期未蔓延危害的原因在于果园管理均衡,施药次数较多,柑桔木虱没有大量生存繁衍的空间;而近年柑桔黄龙病蔓延危害的主要原因是"三农"状况发生变化,劳力转移,失管果园多,病源多、柑桔木虱大量危害,病害迅速传播。论述了近年永春县柑桔黄龙病防控技术的实践应用与取得的成效;指出柑桔规模化连片种植,分散管理,难以统一落实防控措施是黄龙病防控工作的难点。探讨了黄龙病产区发展柑桔生产的措施。     

鄂西南山区黄芪主要病害及综合防治技术

通过对鄂西南山区黄芪[Astragalus membranceus(Fisch)Bunge]病害发生情况的调查发现黄芪的主要病害有根腐病、紫纹羽病、白粉病、白绢病4种,在实验室对该4种病害的病原进行了分离培养,并对致病性进行了初步鉴定;通过防治试验总结出一套以农业防治为主的综合防治技术。     

柑桔青枯病的发生与防控技术

柑桔青枯病是我国上世纪60年代所出现的一种南方桔区常见病害,它的危害性较大,能够让柑桔植株迅速死亡。本文从福建三明市三元区莘口镇柑桔区的青枯病发病病原谈起,重点探讨了它的青枯病症状与病程,基于阶段性分析方法给出了当地柑桔青枯病的防控技术措施。     

柑桔微芽嫁接苗黄龙病的早期检测

柑桔微芽嫁接组织培养试管苗采用微量黄龙病病原DNA提取法,获得的DNA用作PCR检测的模板,可快速检测柑桔黄龙病病毒.为工厂化生产无病毒苗的选育及病害防治提供早期诊断依据.     

鲟鱼病害研究进展

鲟鱼病害大体可分为病原性疾病和非病原性疾病两大类。针对国内外鲟鱼病害的临床症状、病原体、致病机制以及防治方法等研究情况进行概述,以期为鲟鱼病害的诊断与防治提供参考。     

中国柑橘黄龙病研究30年

综述了我国自1978年以来有关柑橘黄龙病的研究概况,包括柑橘黄龙病在我国的分布与危害、黄龙病寄主、病原研究、病害诊断与检测、传播途径与病害流行、防控技术等.     

柑橘黄龙病菌对沙田柚田间性状和果实品质的影响

【目的】明确柑橘黄龙病菌对沙田柚Citrus maxima (Burm.) Merr. cv. Shatian Yu田间性状和果实品质的影响。【方法】结合田间调查和室内试验系统研究沙田柚感染黄龙病菌后叶片症状、果实内外品质和风味感官品质的变化,及病菌浓度和树龄对病程发展的影响。【结果】沙田柚嫁接病芽后,黄龙病菌浓度保持在较低水平(Ct28)时,植株叶片的黄龙病症状表现不明显,老叶轻微斑驳黄化,新叶轻微均匀黄化,抽梢正常。与健康植株(Ct35)相比,低黄龙病菌浓度(28Ct32)的植株,叶片症状、果实产量和各品质指标均无显著变化;随着发病程度的加深,高黄龙病菌浓度(Ct26)的柚树叶片出现典型斑驳黄化症状,单株柚果总产量和总结果数显著降低,果实变小变轻,着色不均匀,可食率、出汁率、可溶性固形物含量、甜味度、果肉饱满度和综合风味显著下降,酸味度和异味度反而显著提高,失去食用价值。【结论】沙田柚黄龙病病程进展较慢,但随着发病程度加深,其经济价值受到严重影响。本研究可为评价柑橘黄龙病对沙田柚品质的影响提供科学依据,为解析柚类植物的黄龙病耐性机理提供理论支撑。     

巢式PCR检测油茶根腐病菌的研究

为建立油茶Camellia oleifera根腐病菌层生镰刀菌Fusarium proliferatum的巢式聚合酶链式反应（PCR）快速检测体系．扩增层生镰刀菌核糖体DNA基因间隔序列（ITS）区并测定其序列,比较该序列与GenBank中近似种的ITS序列差异．设计了特异性引物GF1和GR2,利用该对引物与ITS1和ITS4进行巢式PCR扩增检测层生镰刀菌。结果显示,巢式PCR对层生镰刀菌的检测灵敏度可达100ag基因组DNA,比常规扩增方法提高了1万倍。利用设计的GF1／GR2特异性引物与ITS区通用引物进行巢式PCR扩增．可灵敏地扩增出油茶根腐病菌层生镰刀菌DNA。     

Citrus disease recognition based on weighted scalable vocabulary tree

Citrus Huanglongbing (HLB) is a destructive disease in citrus production that causes huge economic damage to citrus producers and related industries in the world. Early and accurate detection of HLB is a critical management step to control the spread of this disease. However, existing HLB detection methods cannot be widely adopted in citrus production due to long-time and high-cost detection period in specific laboratory environments. In view of this, a fast-response and low-cost computer vision technique is investigated for diagnosing HLB in citrus leaves. Specifically, the Gaussian mixture density (GMD) is performed to extract the leaf object from the citrus image, followed by the feature extraction and recognition of the existence of HLB in the leaf based on scalable vocabulary tree. A citrus leaf image dataset is constructed, and the experimental results show that the proposed HLB recognition method with GMD object extraction performs 95–100 % accuracy within 1 s.     

Molecular Detection of Verticillium albo-atrum by PCR Based on Its Sequences

We developed one species-specific PCR assays for rapid and accurate detection of the pathogenic fungi Verticillium albo-atrum in diseased plant tissues and soil. Based on differences in internal transcribed spacer （ITS） sequences of Verticillium spp., a pair of species-specific primers, Vaal/Vaa2, was synthesized. After screening 17 isolates of V. alboatrum, 121 isolates from the Ascomycota, B asidiomycota, Deuteromycota, and Oomycota, the Vaal/Vaa2 primers amplified only a single PCR band of approximately 330 bp from V. albo-atrum. The detection sensitivity with primers Vaal/Vaa2 was 10 fg of genomic DNA. Using ITS1/ITS4 as the first-round primers, combined with Vaa1/Vaa2, the nested PCR procedures were developed, and the detection sensitivity increased 1 000-fold to 10 ag. The detection sensitivity for the soil pathogens was 100-conidiag^-1 soil. The PCR-based methods developed here could simplify both plant disease diagnosis and pathogen monitoring as well as guide plant disease management.     

超分支滚环扩增法检测小麦矮腥黑穗菌

 【目的】建立小麦矮腥黑穗病菌（Tilletia controversa Kühn,TCK）的超分支滚环扩增（hyper- branched rolling cycle amplification,HRCA）检测体系,为小麦矮腥黑穗病的鉴定以及早期诊断提供了一种新的稳定、可靠的检测技术。【方法】锁式探针包含一个公共连接序列和在探针两端与靶DNA序列互补的2个序列。根据TCK的差异序列设计锁式探针两端序列,以此为基础建立了TCK超分支滚环扩增反应体系。以优化的HRCA反应条件为基础,确定检测体系的特异性和灵敏度。比较HRCA体系和常规PCR体系的性能,并利用这2种体系对来自中国出入检验检疫局截获的51个小麦样品进行了验证检测。【结果】HRCA体系能专一地检出小麦矮腥黑穗菌的DNA靶带,而TCT、TCL等近源种及健康小麦样品都不能扩增。HRCA检测TCK靶序列质粒DNA的下限为1 fg?μl-1,检测基因组DNA的下限为10 pg?μl-1,检测灵敏度比常规PCR检测体系高10倍。HRCA检测体系具有很好的特异性、灵敏度和准确性,更适合于TCK的检测及鉴定。【结论】稳定、灵敏、特异的小麦矮腥黑穗菌的HRCA检测体系的建立,为小麦矮腥黑穗病的早期诊断及其近源种的多靶标检测提供了同步检测技术。     

Development of a sensitive and reliable droplet digital PCR assay for the detection of ‘Candidatus Liberibacter asiaticus'

Citrus Huanglongbing(HLB, yellow shoot disease) is one of the most serious citrus diseases worldwide. To better improve the detection sensitivity, a droplet digital PCR(ddPCR) assay was developed for the rapid detection of ‘Candidatus Liberibacter asiaticus'(Las), the putative causal agent of HLB. The detection of sensitivity comparison using positive plasmid indicated that dd PCR was superior to quantitative PCR(qPCR) for detecting and quantifying Las at low concentrations. The Las detection of 40 field samples also showed that six of 13 asymptomatic samples(46.15%) with high Ct value(35) were positive by dd PCR. This methodology showed great potential for early HLB infection diagnosis.     

膜吸附法结合可视化环介导等温扩增技术检测柑橘黄龙病菌

【背景】柑橘黄龙病（Huanglongbing,HLB）是由候选韧皮部杆菌亚洲种（‘Candidatus Liberibacter asiaticus’,CLas）侵染引起的一种柑橘病害。对于该病害的防治主要采取综合措施,包括实施检疫、种植无病苗木、及时挖除病树和集中大面积联防联控柑橘木虱等。前3种方法都需要依靠准确的柑橘黄龙病诊断技术。【目的】利用环介导等温扩增技术（loop-mediated isothermal amplification, LAMP）,结合膜吸附法DNA快速提取和Gelgreen荧光染料可视化,建立黄龙病田间核酸快速检测方法。【方法】以黄龙病菌β-操纵子与原噬菌体DNA聚合酶基因为模板设计LAMP特异性引物,包括外引物F3/B3、内引物FIP/BIP、环引物LoopF/LoopB和茎引物StemF/StemB。通过对环引物和茎引物设定不同的用量组合,对LAMP引物体系进行优化,确定合适的引物浓度。用优化后的LAMP引物体系对188份田间柑橘叶片进行检测,并构建受试者工作特征（receiver operating characteristic,ROC）曲线分析实时荧光LAMP（qLAMP）在CLas检测上的准确性。对qLAMP预混反应液进行两步的常温干燥,分别设定不同的存放条件,评估LAMP常温干燥试剂的稳定性。用本研究建立的CLas可视化LAMP快速检测方法,对田间71个柑橘叶片样品和35个柑橘果实样品进行检测,与实时荧光定量PCR（qPCR）比较两者的符合率。【结果】在LAMP体系中加入环引物、茎引物或增加其浓度都能促进反应速率的提升,并且同时加入终浓度均为1.6 μmol·L^-1的环引物和茎引物能进一步提高LAMP反应速率。LAMP预混液通过两步法干燥在不同温度存放1—4周均能保持反应活性基本不变,表明制备的两步法干燥LAMP试剂检测性能较好,在低温和常温环境下的稳定性尚可,仅35℃高温存放会略微增加LAMP试剂的反应时间。使用孔径0.1 μm尼龙膜代替纤维素滤纸作为核酸吸附材料能提升CLas快速诊断技术的灵敏度。结合DNA快速提取和可视化LAMP建立的CLas快速核酸诊断方法最低能检测到浓度为10² copies/μL的重组质粒样品,总体准确率高。经配对卡方检验,该方法的诊断结果与qPCR无显著差异。可视化LAMP快速检测相较常规检测有着更低的成本和耗时,而且可视化LAMP快速检测无需离心机和PCR仪等昂贵的仪器,仅需一台65℃恒温设备即可。【结论】建立的CLas快速核酸检测方法成本低,30 min即可观察到检测结果,操作简便,准确性高,可替代qPCR在田间进行黄龙病的快速鉴定。     

Spectral difference analysis and airborne imaging classification for citrus greening infected trees

Citrus greening, also called Huanglongbing (HLB), became a devastating disease spread through citrus groves in Florida, since it was first found in 2005. Multispectral (MS) and hyperspectral (HS) airborne images of citrus groves in Florida were acquired to detect citrus greening infected trees in 2007 and 2010. Ground truthing including field and indoor spectral measurement, infection status along with GPS coordinates was conducted for both healthy and infected trees. Ground spectral measurements showed that healthy canopy had higher reflectance in the visible range, and lower reflectance in the near-infrared (NIR) range than HLB infected canopy. Red edge position (REP) also showed notable difference between healthy and HLB canopy. But the difference in the NIR range and REP were comparably more sensitive to the environment or the background noise. Accuracy for separating HLB and healthy samples reached more than 90% when a simple REP threshold method was implemented in the ground reflectance datasets, regardless of field or indoor measurement; but it did not work well with the HS images because of its low spatial resolution. Support vector machine (SVM) was able to provide a fast, easy and adoptable way to build a mask for tree canopy. High positioning error of the ground truth in the 2007 HS image led to validation accuracy of less than 50% for most of classification methods. In the 2010 image from Southern Gardens (SG) grove, with better ground truth records, higher classification accuracies (about 90% in training sets, more than 60% in validation sets for most of the methods) were achieved. Disease density maps were also generated from the classification results of each method; most of them were able to identify the severely infected areas. Simpler classification methods such as minimum distance (MinDist) and Mahalanobis distance (MahaDist) showed more stable and balanced detection accuracy between the training and validation sets in the 2010 images. Their similar infection trend with ground scouted maps showed a promising future to manage HLB disease with airborne spectral imaging.