Spatial patterns of leprosis and its mite vector in commercial citrus groves in Brazil

In order to describe the dissemination of Citrus leprosis virus (CiLV) and Brevipalpus phoenicis , the Tenuipalpidae mite which transmits it, as well as to generate data for future development of better sampling and disease control procedures, analyses were conducted at three levels of spatial hierarchy. Over 100 distribution maps of mite-infested plants and plants with CiLV symptoms were constructed after evaluation of 174 sweet orange commercial grove blocks from four citrus regions of São Paulo State, Brazil. Spatial correlation between maps of mite-infested plants and those with CiLV symptoms was very low and mostly not significant. Spatial dependency between adjacent plants was incipient as ordinary runs analysis showed that percentages of aggregated sequences within or across rows were very low for both mite-infested and diseased plants. Index of dispersion ( D ) values for all quadrat sizes suggested aggregation of plants with symptoms within quadrats, but much less aggregation for mite-infested plants. Values of log( A ) and b were higher than 0 and 1, respectively, indicating a general and significant aggregation of infested/diseased plants inside quadrats. However, the degree of aggregation of plants with symptoms was higher than that of infested plants. Aggregation in each grove was positively correlated to the incidence of infested/diseased plants. Spatial autocorrelation also showed a higher strength of aggregation for plants with symptoms than for infested ones. This is the first time that spatial patterns of leprosis and B. phoenicis have been described.     

The influence of citrus rootstocks on the relationship between the mite Brevipalpus phoenicis and citrus leprosis disease

BACKGROUND: Leprosis is one of the most serious citrus plant diseases. Leprosis‐affected plants, especially sweet orange [Citrus sinensis (L.) Osbeck], which is the most widely cultivated citrus fruit worldwide, show reduced photosynthetic capacity and severe defoliation. The aim was to evaluate the relationship between the Brevipalpus phoenicis (Geijskes) vector mite and citrus leprosis disease in Pera sweet orange plants grafted on different rootstocks. Data were analysed using numerical classification and conventional statistical analysis (ANOVA). RESULTS: Both viruliferous and non‐viruliferous B. phoenicis populations increased in number on plants maintained at low soil water content. Among the evaluated rootstocks, Sunki mandarin proved least favourable for mite population increase. Furthermore, the viruliferous mite population increased more rapidly than the non‐viruliferous mite population. CONCLUSION: The Cleopatra rootstock showed low variability in leaf nitrogen content, low mite number and low leprosis severity, thereby producing the most favourable results. Under appropriate moisture conditions, the Sunki rootstock produced the best results, showing the least severe leprosis. Rangpur lime rootstock grown in soil with varying water content showed the greatest variation in foliar nitrogen content, mite number and severity of leprosis and is considered to be the most difficult rootstock to manage. Copyright © 2012 Society of Chemical Industry     

运用一步法RT-PCR检测柑橘鳞皮病毒及其在橘橙不同部位中的全年分布

2005年5月到2006年4月逐月采样,运用一步法RT-PCR检测Citrus psorosis virus(CPV)在Dweet橘橙苗木叶片和枝皮中的分布。保存在控温温室中的Dweet橘橙病株中老叶、老皮、嫩叶和嫩皮全年都可以检测出CPV;保存在网室中的Dweet橘橙病株中老叶、老皮全年均能检测到CPV,而夏梢的嫩叶、嫩皮不能稳定地检测出CPV,春、秋梢的嫩叶、嫩皮均可检测到CPV,表明一步法RT-PCR检测CPV最佳取样部位为老叶和老皮。     

来源于猕猴桃的柑橘叶斑驳病毒的RT-PCR检测及外壳蛋白基因序列分析

柑橘叶斑驳病毒(Citrus leaf blotch virus,CLBV)为柑橘病毒属(Citrivirus)代表种。本研究采用RT-PCR技术从我国栽培猕猴桃上首次检测到CLBV,总检出率为13.3%。测定了5个CLBV分离物的外壳蛋白基因(coat protein gene,cp)序列,全长均为1 092核苷酸(nt),分离物间cp基因核苷酸和编码氨基酸序列相似性分别为83.2%~99.7%和91.9%~98.9%,这些分离物与新西兰报道的CLBV猕猴桃分离物M3-A核苷酸序列相似性仅为83%左右,与来源于柑橘及近缘种的CLBV分离物cp基因核苷酸序列相似性为84%~86%。在基于该病毒cp基因核苷酸序列的系统进化树中,本研究所测定的CLBV猕猴桃分离物与新西兰报道的除M3-A外的猕猴桃分离物聚在同一组群。研究结果为进一步明确CLBV在我国猕猴桃上的侵染和危害特点及建立该病毒的分子检测技术提供了重要信息。     

高通量测序鉴定柑橘黄化脉明病毒诱导柠檬差异表达microRNA

 柑橘黄化脉明病毒(Citrus yellow vein clearing virus,CYVCV)是我国新发生的危害柠檬的重要病毒。本研究对接种和未接种CYVCV的柠檬叶片样品进行了高通量小RNA测序,以甜橙基因组作为参考,采用生物信息学软件进行miRNA鉴定和靶基因预测。结果显示CYVCV侵染对13个保守miRNA和12个新鉴定的非保守miRNA的表达具有调控作用,对这些差异表达miRNA的靶基因进行预测和功能分析的结果显示,保守miRNA的靶基因功能在植物逆境反应及光合作用和能量代谢相关通路中具有明显的富集现象,推测与CYVCV侵染柠檬植株及诱导病害症状表现有关,而多数新鉴定的非保守miRNA的靶基因功能未能注释到KEGG通路。采用实时荧光定量PCR对部分miRNA的表达水平进行了分析,结果表明miRNA的表达水平在接种CYVCV后30~90 d呈时间动态变化。研究结果为进一步揭示CYVCV与柠檬互作机制提供了重要信息。     

猕猴桃植株中柑橘叶斑驳病毒实时荧光定量PCR检测技术的建立及应用

 柑橘叶斑驳病毒(Citrus leaf blotch virus, CLBV)在陕西省栽培猕猴桃中发生普遍。为监测CLBV发生情况,本研究建立了CLBV的实时荧光定量PCR(Real-time fluorescent quantitative polymerase chain reaction,RT-qPCR)检测方法。该方法特异性强,可准确检测目的病毒,标准曲线斜率为-3.378,决定系数R²=0.997 9,扩增效率为97.7%,比普通RT-PCR灵敏度高100倍,可用于猕猴桃植株CLBV的批量检测或低丰度病毒样本(如猕猴桃休眠枝条)的检测。为苗木携带CLBV病毒的早期诊断、果园病毒病预测预报和防控奠定了基础。     

应用常规RT-PCR和荧光定量RT-PCR检测柑桔衰退病毒

 根据柑桔衰退病毒(CTV)P20基因序列设计cquctv9/cquctv10特异引物对,以柑桔RNApolymeraseⅡ基因作为内参照,建立了柑桔衰退病的常规RT-PCR快速检测体系;依据柑桔衰退病毒P20基因序列设计cquctv1/cquctv2特异引物对和TaqMan探针cquctvp1,建立了柑桔衰退病荧光定量RT-PCR快速检测体系。常规RT-PCR检测下限是含有CTV的50pg总RNA,荧光定量RT-PCR法的检测下限是2fg纯CTV片段。荧光定量RT-PCR的灵敏度相对比常规RT-PCR高100倍。利用常规RT-PCR和荧光定量RT-PCR体系对从2005年3月到2006年7月采自田间的样品进行检测,结果表明,2种检测体系都有很好的特异性和准确性;对183个田间柑桔苗木样品带毒率检测结果表明,荧光定量RT-PCR检出率为(82.5%),比常规RT-PCR检出率(73.2%)高。     

Selective transmission of the seedling yellows-type genotypic variants of Citrus tristeza virus by Aphis gossypii in northern Iran

Citrus tristeza virus (CTV) has existed in northern Iran for more than five decades. The long-time interaction of different virus genotypes with Aphis gossypii, as the only aphid vector of CTV in northern Iran, has led to the emergence of highly virulent subpopulations, among others, in the established foci. Here, we studied the population structure of the originally established CTV isolates present in Satsuma mandarin (Citrus unshiu) trees imported from Japan, and subisolates thereof, formed following experimental transmission by A. gossypii, as well as those evolved through natural transmission by this aphid species in the groves. Symptoms of the naturally spread and the experimentally aphid-transmitted isolates were similar to those of the Satsuma CTV source isolates for all indicator plants except for sour orange (Citrus aurantium) and grapefruit (Citrus paradisi), with the aphid-transmitted isolates additionally inducing severe seedling yellows and stunting in these two indicators. Studies on the population heterogeneity of these isolates through comparison of their single-strand conformational polymorphism profiles and nucleotide sequences of the 25 kDa capsid protein gene from the predominant haplotypes, and dot-blot hybridization signals, revealed the presence of two major T36- and SY568- (or NUagA-) like genotypes along with a minor poorly characterized one in the originally infected Satsuma trees; in contrast, only a certain genomic variant having the highest similarity to the isolate SY568 (and NUagA) was predominant both in the naturally infected trees and in those infected experimentally by A. gossypii. It seems that transmission by A. gossypii to sweet orange (Citrus sinensis) has led to the preponderance of the CTV genomic variants inducing severe seedling yellows in northern Iran.     

湖南柑橘衰退病调查分析及弱毒系筛选

 The investigation showed that stem-pitting Citrus tristeza virus (CTV)occurred commonly in citrus production areas in several varieties of Hunan Province. Accurate detection of CTV strains was performed by p23/PCR method, PCR and the results indicated that the most samples were infected with several CTV isolates. Three mild strains were isolated and their pathogenicity was identified by biological identification, it indicated that p23/PCR groups had uniformity with the pathogenicity of CTV isolates. Furthermore, three mild isolates were tested in the cross protection by analysis of biological symptoms and composition of p23 gene. Different protecting effects were observed among these strains and W17 mild isolate was effective.     

茶园天敌对花蓟马和茶短须螨的跟随效应研究

为了合理保护和利用自然天敌进行害虫的综合防治,对安徽茶园主要害虫与天敌进行系统调查,进行了天敌对花蓟马和茶短须螨跟随效应的研究。运用地学统计学方法求得天敌与害虫半变异函数的变程,用灰色关联度分析方法分析变程之间的关联度,与害虫关联度值越大的天敌对害虫的跟随关系越密切。本文分析了花蓟马种群数量在全年最多时段(4月13日—7月8日)、茶短须螨种群数量在全年最多时段(5月13日—8月5日)内,茶园中5种主要天敌对两种害虫跟随关系的密切程度。结果表明,花蓟马与斜纹猫蛛、八点球腹蛛、粽管巢蛛、龟纹瓢虫和异色瓢虫半变异函数变程的关联度分别为0.658 3、0.614 6、0.607 1、0.753 8和0.647 1,茶短须螨与斜纹猫蛛、八点球腹蛛、粽管巢蛛、龟纹瓢虫和异色瓢虫半变异函数变程的关联度分别为0.718 9、0.615 6、0.701 0、0.676 5和0.697 6。表明与花蓟马跟随关系密切的天敌依次是龟纹瓢虫、斜纹猫蛛和异色瓢虫,与茶短须螨跟随关系密切的天敌依次是斜纹猫蛛、粽管巢蛛和异色瓢虫。     

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.     

柑桔病毒类有害生物及其检疫重要性

本文针对柑桔上发生的数十种病毒类有害生物，列举了具有明确分类地位的６种病毒、５种类病毒、１种类菌原体及１种类细菌，初步分析了其所引起的柑桔病害的危险性及其检疫重要性     

小麦黄花叶病毒外壳蛋白基因RNAi表达载体的构建及遗传转化

小麦黄花叶病毒(Wheat yellow mosaic virus,WYMV)是马铃薯Y病毒科(Potyviridae)大麦黄花叶病毒属(Bymovirus)的成员,是危害我国小麦生产的一种重要病毒病害.RNA干扰是由dsRNA介导的,通过核酸序列特异性的相互作用来抑制基因表达的一种基因沉默现象,这种调控机制在植物抗病毒基因工程育种中已得到广泛应用.以RNA干扰为原理、以病毒的外壳蛋白基因为靶标,构建了抗WYMV的植物表达载体.采用基因枪方法共转化‘扬麦12’的幼胚愈伤组织,得到了再生植株.对To代进行PCR检测,得到15株阳性植株.     

Engineering of a Single Chain Variable Fragment Antibody Specific for the Citrus tristeza virus and its Expression in Escherichia coli and Nicotiana tabacum

Citrus tristeza virus (CTV) is one of the most destructive citrus virus diseases in the world. The construction of an engineered antibody, EMBL accession number AJ278109, able to specifically recognize its antigen, i.e. the coat protein of CTV, directly on infected plant material without any purification or manipulation of the entire woody plant. The potential uses of this engineered antibody are discussed.     

Quantifying inherent predictability and spatial synchrony in the aphid vector Myzus persicae: field-scale patterns of abundance and regional forecasting error in the UK

Background

Sugar beet is threatened by virus yellows, a disease complex vectored by aphids that reduces sugar content. We present an analysis of Myzus persicae population dynamics with and without neonicotinoid seed treatment. We use 6 years' yellow water trap and field-collected aphid data and two decades of 12.2 m suction-trap aphid migration data. We investigate both spatial synchrony and forecasting error to understand the structure and spatial scale of field counts and why forecasting aphid migrants lacks accuracy. Our aim is to derive statistical parameters to inform regionwide pest management strategies.

Results

Spatial synchrony, indicating the coincident change in counts across the region over time, is rarely present and is best described as stochastic. Uniquely, early season field populations in 2019 did show spatial synchrony to 90 km compared to the overall average weekly correlation length of 23 km. However, 70% of the time series were spatially heterogenous, indicating patchy between-field dynamics. Field counts lacked the same seasonal trend and did not peak in the same week. Forecasts tended to under-predict mid-season log₁₀ counts. A strongly negative correlation between forecasting error and the proportion of zeros was shown.

Conclusion

Field populations are unpredictable and stochastic, regardless of neonicotinoid seed treatment. This outcome presents a problem for decision-support that cannot usefully provide a single regionwide solution. Weighted permutation entropy inferred that M. persicae 12.2 m suction-trap time series had moderate to low intrinsic predictability. Early warning using a migration model tended to predict counts at lower levels than observed. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Spatiotemporal dynamics and modelling support the case for area-wide management of citrus greasy spot in a Brazilian smallholder farming region

Citrus greasy spot (CGS), caused by Zasmidium citri, induces premature defoliation and yield loss in Citrus spp. The epidemiology of CGS is well understood in high humidity areas, but remains unaddressed in Brazil, despite differing climatic conditions and disease management practices. The spatiotemporal dynamics of CGS were characterized in the Recôncavo of Bahia (Brazil) at four hierarchical levels (quadrant, plant, grove, and region). A survey conducted in 19 municipalities found the disease to be present throughout the region with an incidence of 100% in groves and plants, and higher than 70% on leaves. Index of dispersion (D) values suggest the spatial pattern of units with symptoms lies between random and regular. This was confirmed by the parameters of the binary power law for plants and their quadrants (log[A] < 0 and b < 1). No consistent differences were observed in the disease incidence at different plant heights. We introduce a compartmental model synthesizing CGS epidemiology. The collected data allow such a model to be parameterized, albeit with some ambiguity over the proportion of new infections that result from inoculum produced within the grove versus external sources of infection. By extending the model to include two populations of growers—those who control and those who do not—coupled by airborne inoculum, we investigate likely performance of cultural controls accessible to citrus growers in northeastern Brazil. The results show that control via removal of fallen leaves can be very effective. However, successful control is likely to require area-wide strategies in which a large proportion of growers actively manage disease.     

Biological and genetic characterization of carrot red leaf virus and its associated virus/RNA isolated from carrots in Hokkaido,Japan

Carrot motley dwarf (CMD) is caused by mixed infection of carrot red leaf virus (CtRLV) with either carrot mottle virus (CMoV) or carrot mottle mimic virus, and additional infection with CtRLV-associated RNA (CtRLVaRNA). Here, the author investigated the viruses or virus-like RNA isolated from carrots with reddening symptoms in Hokkaido, the northern island of Japan. Three types of infections were mainly detected: single infection with CtRLV, which was most prevalent; double infection with CtRLV and CMoV; and triple infection with CtRLV, CMoV, and CtRLVaRNA. Fields with the three agents were severely affected, with diseased plants showing mottling, whereas in fields where disease incidence was low and sporadic, CtRLV was often found alone in plants with mild symptoms. Inoculation tests using carrot plants showed that CMoV enhanced disease severity, and the RNA accumulation of CtRLV. However, in the presence of CtRLVaRNA (+ CMoV), distinct symptoms such as systemic mottling and stunting developed, while the enhancement of CtRLV accumulation was abolished. These results imply that CtRLVaRNA (+ CMoV) antagonizes CtRLV despite its dependence on CtRLV for aphid transmission, and that mixed infection with CtRLVaRNA is involved in the development of the conspicuous mottling. All agents detected in Hokkaido were very similar to European and American isolates in terms of their genomic sequences and host range. This represents the first report of CMD in Japan, and provides further information on the genetic and biological properties of CMD-associated agents, as well as the aetiology of the disease.