云南柠檬园橘臀纹粉蚧的发生及防治初探

对德宏柠檬产区柠檬园树体冠层和土壤耕作层橘臀纹粉蚧的发生情况及防治措施进行了研究,结果表明：橘臀纹粉蚧每年7至10月在柠檬树地上冠层部分形成一个为害期,4月和6月在地下耕作层根部形成两个为害高峰,于橘臀纹粉蚧低龄期采用10%吡虫啉可湿性粉剂2000倍液进行灌根,防治效果最好。     

Spiroplasma citri Spiralin Acts In Vitro as a Lectin Binding to Glycoproteins from Its Insect Vector Circulifer haematoceps

ABSTRACT In order to understand the molecular mechanisms underlying transmission of Spiroplasma citri by the leafhopper Circulifer haematoceps, we screened leafhopper proteins as putative S. citri-binding molecules using a spiroplasma overlay assay of protein blots (Far-western assay). Insect proteins were separated by one- or two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis, blotted, and probed with S. citri proteins. In this in vitro assay, we found that spiroplasma proteins exhibited affinity for seven leafhopper proteins. The interactions between S. citri proteins and insect proteins with molecular masses of 50 and 60 kDa were found to be sugar sensitive. These insect proteins were identified as high mannose N-glycoproteins, which support an interaction of glycoprotein-lectin type with S. citri proteins. Lectin detection in S. citri has revealed only one protein of 24 kDa. Using a leafhopper protein overlay assay on an S. citri protein blot, one spiroplasma protein with a similar molecular mass of 24 kDa was shown to display an insect protein-binding capacity. This protein was identified as the spiralin, which is the most abundant membrane protein of S. citri. Far-western experiments performed with purified spiralin and insect glycoproteins confirmed the binding of spiralin to the insect glycoproteins of 50 and 60 kDa. Thus, the spiralin could play a key role in the transmission of S. citri by mediating spiroplasma adherence to epithelial cells of insect vector gut or salivary gland.     

Detection of Spiroplasma citri in plants and insect hosts by ELISA

Enzyme-linked immunosorbent assay (ELISA) was shown to be a sensitive method for the detection of Spiroplasma citri in plants and insect hosts. S. citri was detected in Vinca rosea less than 1 week after infection by grafting, and reached a peak litre of up to 10⁹ spiroplasmas per gram of tissue in infected shoots and root tips. Multiplication of S. citri in the leafhopper Euscelidius variegatus was also monitored by ELISA. S. citri could be detected in a single insect, showing that this technique is suitable for screening insect populations in the field for potential vectors of Spiroplasma diseases. A method is described for raising a pathogen-specific antiserum from V. rosea infected with S. citri which reacted with cultured S. citri and also with S. citri in plant tissue.     

Characterization of phenotypically distinct strains of Xanthomonas axonopodis pv. citri from Southwest Asia

Strains of Xanthomonas axonopodis pv. citri were isolated from Mexican lime (Citrus aurantifolia) trees in several countries in southwest Asia. These strains produced typical erumpent bacterial canker lesions on Mexican lime but not on grapefruit (C. paradisi). Lesions on grapefruit were watersoaked and blister-like in contrast to the typical erumpent lesions seen after artificial inoculation with all described pathotypes of X. axonopodis pv. citri. This group of strains hydrolysed gelatin and casein and grew in the presence of 3% NaCl as is typical of X. axonopodis pv. citri pathotype A. RFLP analyses and DNA probe hybridization assays also gave results consistent with X. axonopodis pv. citri pathotype A. Metabolic fingerprints prepared with the Biolog® system showed similarities as well as differences to X. axonopodis pv. citri pathotype A. In spite of the physiological and genetic similarities to pathotype A of X. axonopodis pv. citri, these strains had no or very little affinity for polyclonal antiserum prepared against any of the reference strains of X. axonopodis pv. citri and also did not react with monoclonal antibody A1, an antibody that detects all strains of pathotype A of X. axonopodis pv. citri. These strains were also insensitive to bacteriophage Cp3 like X. axonopodis pv. citri pathotype A and unlike X. axonopodis pv. citri pathotype B. We conclude that these strains, designated Xcc-A*, represent a variant of X. axonopodis pv. citri pathotype-A with pathogenicity limited to C. aurantifolia. The existence of extensive genotypic and phenotypic variation within pathotype A of X. axonopodis pv. citri was unexpected and further complicates the systematics of this species.     

一株侵染柑橘木虱的球孢白僵菌的分离及鉴定

从柑橘木虱虫体分离得到一株具有强致病性的虫生真菌GZMS-28,本文测定了该菌株对柑橘木虱的致病性,并分析了其rDNA ITS基因序列。试验结果表明,该菌株对柑橘木虱具有较强的致病性,浓度为1.0×108个/mL的分生孢子悬浮液处理柑橘木虱成虫7d后,其校正死亡率达到95.7%。菌株GZMS-28的培养特征与白僵菌属球孢白僵菌较为一致,其rDNA ITS序列与GenBank中白僵菌属多个球孢白僵菌菌株的对应序列相似性达到99%以上,因此将菌株GZMS-28鉴定为球孢白僵菌Beauveria bassiana。     

7种挥发性化合物对柑橘木虱引诱效果的评价

柑橘黄龙病对全球的柑橘产业造成了严重的经济损失,其传播媒介主要为柑橘木虱。在前期研究中利用柑橘木虱对4种芸香科植物的取食偏好结合代谢组学分析筛选出一批可能对柑橘木虱具有引诱效果的挥发性化合物,本研究鉴定了邻氨基苯甲酸甲酯、癸酸、正辛醇、左旋香芹酮、2-环己烯酮、反式-1,2-环己二醇、吡嗪等7种挥发性化合物在0、0.1%、1%、10%4个浓度和10、20、30、40、50、60 min 6个持续时间下对柑橘木虱的引诱效果,并比较了其中5种脂溶性挥发物在各自最优浓度下的引诱效果。试验结果表明,7种挥发物对柑橘木虱都有引诱效果,但是引诱效果与挥发物浓度及持续时间有关。吡嗪、左旋香芹酮、正辛醇、2-环己烯酮、癸酸、邻氨基苯甲酸甲酯和反式-1,2-环己二醇的最佳引诱浓度分别为0.1%、1%、1%、1%、10%、10%、10%。六臂嗅觉仪试验结果显示5种脂溶性挥发物中1%左旋香芹酮的引诱效果高于10%癸酸、10%邻氨基苯甲酸甲酯、1%2-环己烯酮、1%正辛醇及对照(矿物油),但与其他4种挥发物没有显著差异。研究结果可为柑橘木虱引诱剂的开发提供参考,促进柑橘黄龙病的绿色防控。     

乙唑螨腈与螺螨酯混用对柑橘全爪螨的田间防治效果评价

柑橘全爪螨 Panonychus citri 是一种重要的世界性害螨, 已对多种常用杀螨剂产生了不同程度的抗性?为有效控制该害螨的危害, 本研究于2017年-2018年通过田间小区试验评价了30%乙唑螨腈·螺螨酯SC?30%乙唑螨腈SC?240 g/L螺螨酯SC?1.8%阿维菌素EC及43%联苯肼酯SC对柑橘全爪螨的防治效果及其对柑橘树的安全性?研究结果表明:试验浓度下, 叶面喷施1次上述药剂均对柑橘树安全?30%乙唑螨腈·螺螨酯SC及30%乙唑螨腈SC能有效控制柑橘全爪螨危害, 速效性好, 持效期长达30 d, 药后1~30 d的防治效果分别为76.55%~100%和81.21%~98.30%?240 g/L螺螨酯SC和43%联苯肼酯SC对柑橘全爪螨的控制作用较好, 速效性一般而持效期可达30 d, 药后1~30 d的防治效果分别为69.15%~91.55%和64.63%~88.46%?1.8%阿维菌素EC难以控制柑橘全爪螨危害, 速效性较差, 持效期约15 d; 药后1~30 d对该害螨的防治效果为57.08%~83.39%?综上所述, 在柑橘生产实践中, 为有效控制柑橘全爪螨为害, 应于害螨初发期叶面喷施1次30%乙唑螨腈·螺螨酯SC 75 mg/kg或30%乙唑螨腈SC 100 mg/kg?     

壳聚糖和2株芽孢杆菌对柑橘溃疡病菌的杀菌效果评价

柑橘溃疡病是重要的柑橘细菌性病害,为探明壳聚糖和枯草芽孢杆菌对柑橘溃疡病的防治潜力,本文用平板菌落计数法评价了3种粘度壳聚糖对柑橘溃疡病菌的杀菌效果,以改良的纸碟法测定2株枯草芽孢杆菌(Bacillus subtilis)发酵液对柑橘溃疡病菌的抑菌效果。综合处理0.5 h、3 h和6 h的结果显示,3种粘度壳聚糖对柑橘溃疡病的杀菌强弱依次为:粘度12粘度100粘度20。枯草芽孢杆菌菌株BS 101和54-6在YPG培养基中有氧发酵3 d产生发酵液原液的抑菌圈直径分别为3.45 cm和3.21 cm,比0.1 g/mL氨苄青霉素的抑菌圈分别大45%和35%。可见,发酵液中存在对溃疡病菌高度有效的成分。因此,认为粘度12的壳聚糖和芽孢杆菌菌株BS 101和54-6具有防治柑橘溃疡病的潜力。     

Green Fluorescent Detection of Fungal Colonization and Endopolygalacturonase Gene Expression in the Interaction of Alternaria citri with Citrus

ABSTRACT Alternaria citri, a postharvest pathogen, produces endopolygalacturonase (endoPG) and causes black rot on citrus fruit. We previously described that an endoPG-disrupted mutant of Alternaria citri was significantly reduced in its ability to macerate plant tissue and cause black rot symptoms on citrus. In order to investigate colonization of citrus fruit tissues by Alternaria citri, pTEFEGFP carrying a green fluorescent protein (GFP) gene was introduced into wild-type Alternaria citri and its endoPG-disrupted mutant (M60). Green fluorescence was observed in spores, germ tubes, appressoria, and infection hyphae of transformants G1 (derived from wild type) and GM4 (derived from M60). Hyphae of G1 but not GM4 vertically penetrated the peel, but the hyphae of both G1 and GM4 spread equally in the juice sac area of citrus fruit. Green fluorescence of Alternaria citri transformant EPG7 carrying a GFP gene under control of the endoPG gene promoter of Alternaria citri was induced by pectin in the peel during the infection stage, but repressed completely in the juice sac area, likely by carbon catabolite repression by sugars in the juice.     

泰和县郊柑桔木虱发生初步调查

柑桔木虱为柑桔黄龙病的传播介体。本文对泰和县郊柑桔木虱的发生情况进行了初步调查，发现泰和县郊柑桔术虱发生已较严重。     

Insertion sequence- and tandem repeat-based genotyping techniques for Xanthomonas citri pv. mangiferaeindicae

Molecular fingerprinting techniques that have the potential to identify or subtype bacteria at the strain level are needed for improving diagnosis and understanding of the epidemiology of pathogens such as Xanthomonas citri pv. mangiferaeindicae, which causes mango bacterial canker disease. We developed a ligation-mediated polymerase chain reaction targeting the IS1595 insertion sequence as a means to differentiate pv. mangiferaeindicae from the closely related pv. anacardii (responsible for cashew bacterial spot), which has the potential to infect mango but not to cause significant disease. This technique produced weakly polymorphic fingerprints composed of ≈70 amplified fragments per strain for a worldwide collection of X. citri pv. mangiferaeindicae but produced no or very weak amplification for pv. anacardii strains. Together, 12 tandem repeat markers were able to subtype X. citri pv. mangiferaeindicae at the strain level, distinguishing 231 haplotypes from a worldwide collection of 299 strains. Multilocus variable number of tandem repeats analysis (MLVA), IS1595-ligation-mediated polymerase chain reaction, and amplified fragment length polymorphism showed differences in discriminatory power and were congruent in describing the diversity of this strain collection, suggesting low levels of recombination. The potential of the MLVA scheme for molecular epidemiology studies of X. citri pv. mangiferaeindicae is discussed.     

益害比及花粉存在对江原钝绥螨和柑橘全爪螨种群动态的影响

研究了在以不同益害比释放江原钝绥螨的情况下,益害比及花粉存在对江原钝绥螨捕食效果的影响.结果表明,当柑橘全爪螨平均每片叶低于5头时,按益害比1:10和1:20的比例释放江原钝绥螨,当平均每片叶柑橘全爪螨多于5头时,按益害比1:5释放江原钝绥螨,可将柑橘全爪螨数量控制在经济阈值以下.花粉的存在能减缓猎物密度下降对江原钝绥...     

柑桔溃疡病菌对杀菌剂拌种灵抗逆性生理研究

在离体条件下对柑桔溃疡病菌 Xanthomonas citri胞外产物的分泌量、胞外蛋白酶活性、拌种灵的作用动态进行了测定 ,并对病原菌细胞的超微结构进行了电镜观察。结果表明 ,在一定浓度拌种灵的选择压力下 ,柑桔溃疡病菌胞外产物及胞外多糖的分泌量增多 ,胞外蛋白酶的活性在拌种灵浓度低于 10 μg/m L时亦增高 ,但胞外蛋白的产量受到抑制。接触药剂的菌体细胞原生质浓缩。胞外产物对拌种灵具有拮抗作用 ,从而对菌体细胞产生保护功效。     

克菌丹·啶氧菌酯对柑橘沙皮病的防治效果评价

为筛选防治柑橘沙皮病的高效低毒药剂,连续两年进行了新型复配药剂50％克菌丹·啶氧菌酯水分散粒剂防治柑橘沙皮病的田间药效试验,对该药剂防治柑橘沙皮病的效果及安全性进行了评价.两年的试验结果表明:50％克菌丹·啶氧菌酯水分散粒剂有效成分用量625mg/kg对柑橘沙皮病具有较好的防治效果,2018年药剂处理两次后对柑橘叶片和...     

5种寄主上的尼氏真绥螨对柑橘红蜘蛛的捕食能力差异研究

室内测定了5种寄主上的尼氏真绥螨对柑橘红蜘蛛的日捕食量和持续捕食能力。结果表明,柑橘上的尼氏真绥螨成螨对柑橘红蜘蛛成螨的日均捕食量最大,显著高于紫苏;而梧桐、薄荷和茶叶3种寄主上的尼氏真绥螨对柑橘红蜘蛛雌成螨的捕食力与柑橘上的无显著差异。对柑橘红蜘蛛若螨的捕食能力以薄荷和梧桐上的显著为高,柑橘和茶叶上的次之,紫苏上的最低;而5种寄主上的尼氏真绥螨对柑橘红蜘蛛幼螨的日均捕食量之间无显著差异。对柑橘红蜘蛛成螨的持续捕食能力结果显示,各处理连续3天的日均捕食量基本呈现略微降低的趋势,但3天之间均无显著差异。上述结果证明不同寄主上的尼氏真绥对柑橘红蜘蛛的捕食能力存在差异,且对柑橘红蜘蛛成螨的捕食能力较为稳定。     

R44型直升机喷雾防治柑橘木虱的效果初报

大区域内实施柑橘木虱统防统治是控制柑橘黄龙病的关键措施,也是难点。本试验研究R44型直升机施药防治柑橘木虱的可行性和效果。在连片柑橘园,利用R44型直升机喷洒30%噻虫嗪悬浮剂防治柑橘木虱,施药3、7和10d后,柑橘木虱的虫口减退率分别为84.2%、84.8%和85.0%,防治效果分别为85.3%、87.7%和89.6%;对果树无损伤。试验结果表明,利用R44直升机施药进行柑橘木虱大面积防治可行有效,是一种省工、快速、高效的施药方式,值得大面积推广应用。     

Transcriptomic analysis of changes in Citrus × microcarpa gene expression post Xanthomonas citri subsp. citri infection

European Journal of Plant Pathology - Citrus canker (CC) is an economically important bacterial disease caused by Xanthomonas citri subsp. citri (Xcc). There is variability in resistance to CC...     

基于MaxEnt模型的柑橘木虱在西南地区潜在分布模拟

本研究应用生态位软件MaxEnt和地理信息系统软件ArcGIS预测柑橘木虱在西南地区的潜在地理分布范围,筛选影响柑橘木虱分布的关键变量。结果表明柑橘木虱在西南地区高适生区主要位于西藏东南部、四川中东部、重庆大部、贵州大部和云南中北部,总面积为52.03万km 2,占西南地区总面积的23.89%。影响柑橘木虱分布的关键变量为最冷月最低温度、最冷季度平均温度、最湿季度降水量、最暖季度降水量、最暖季度平均温度和最暖月最高温度。本研究不仅模拟了柑橘木虱在西南地区的潜在地理分布,更为该虫的分布与环境变量之间关系的研究提供了理论参考。     

The Effect of Irrigation Practices on the Spatio-temporal Increase of Asiatic Citrus Canker in Simulated Nursery Plots in Reunion Island

Asiatic citrus canker is a potentially severe disease of several citrus species and cultivars in many tropical and subtropical areas. In such areas, infected nursery plants constitute an important source of primary inoculum for newly established citrus groves. The influence of overhead, drip, and mist irrigation systems on the development of Asiatic citrus canker was studied in simulated, Mexican-lime nurseries in Reunion Island. Overhead irrigation exacerbated the increase of disease incidence and severity caused by a streptomycin-resistant strain of Xanthomonas axonopodis pv. citri. The temporal development of Asiatic citrus canker for overhead irrigated nursery plots was best described by an exponential model, because disease incidence in these plots did not come close to an asymptote during the experimental period. This can be explained by the continuous production of new growth, susceptible to infection by Xanthomonas axonopodis pv. citri, and splash dispersal of Xanthomonas axonopodis pv. citri associated with overhead irrigation. Based on spatial correlation and spatio-temporal analyses, aggregated disease patterns were found irrespective of the irrigation system. In overhead-irrigated plots, the spread of Xanthomonas axonopodis pv. citri lacked directionnality. Rainstorms of short duration and high intensity were apparently associated with disease increase in drip-irrigated plots. There is a need to improve cultivation practices in Reunion Island citrus nurseries to minimize Asiatic citrus canker incidence in nurseries and to minimize the introduction of Xanthomonas axonopodis pv. citri to new groves.     

Survival of Xanthomonas axonopodis pv. citri in Leaf Lesions Under Tropical Environmental Conditions and Simulated Splash Dispersal of Inoculum

ABSTRACT Asiatic citrus canker (ACC) is a severe disease of several citrus species and hybrids in many tropical and subtropical areas. Populations of Xanthomonas axonopodis pv. citri in leaf and twig lesions are the most important inoculum source for secondary infections. In areas with a marked winter season (e.g., Argentina and Japan), low temperatures induce a decrease of 10(2) to 10(4) in population sizes in lesions, thus creating a discontinuity in the X. axonopodis pv. citri life cycle. The purpose of this study was to evaluate the dynamics of X. axonopodis pv. citri populations in leaf lesions exposed to the mild winter temperatures prevailing in a tropical environment. Internal X. axonopodis pv. citri population levels in Mexican lime leaf lesions reached 10(6) to 10(7) CFU lesion(-1) whatever the lesion size. These densities, however, were not strongly negatively affected by winter temperatures prevailing under experimental conditions. The estimated decrease in internal X. axonopodis pv. citri population sizes was approximately 10-fold. When exposed to 35 mm h(-1) of simulated rainfall, internal population sizes decreased over time by approximately 1 log unit for lesions 1 and 2 months old, but did not for older lesions. A microscopic examination indicated that lignin-like compounds are present in lesions more than 6 months old. The slow decrease over time of X. axonopodis pv. citri population sizes in leaf lesions may be the balanced result of defense reactions by the host at late stages of disease development, and the concomitant multiplication of the pathogen at the margin of old lesions. We conclude that the epidemiological significance of overwintered leaf lesions in the tropics is higher than that reported in other areas.