Incidence,distribution and limited genetic variability among Turkish isolates of Grapevine Pinot gris virus from different grapevine cultivars

Journal of Plant Diseases and Protection - Grapevine Pinot gris virus (GPGV) was firstly identified in northern Italy by deep sequencing from grapevine cv. Pinot gris, exhibiting mottling and...     

Identification of sugarcane grassy shoot-associated phytoplasma and one of its putative vectors in India

During a survey of sugarcane fields at the Sugarcane Research Institute, Shahjahanpur, Uttar Pradesh, India, in August–September 2012, 6% to 28% incidence of sugarcane grassy shoot disease was observed in different fields of sugarcane variety CoS 7250. The association of phytoplasma with symptomatic sugarcane was confirmed by direct and nested PCR amplification of phytoplasma ribosomal gene. Four different delphacid leafhopper species, viz. Cofana unimaculata Signoret, Exitianus indicus (Distant), Sogatella kolophon Kirkaldy and Hishimonus phycitis (Dist.) were the prevalent feeding species of the Auchenorrhyncha fauna in the symptomatic sugarcane fields. Out of these four leafhopper species, only E. indicus tested positive for phytoplasma presence. Phylogenetic analysis suggested that the phytoplasmas from sugarcane and E. indicus in the present study were members of 16Sr XI. The confirmation of association of sugarcane grassy shoot phytoplasma in E. indicus population is important to understand the secondary spread of this phytoplasma in sugarcane plants.     

人工培养液在植原体昆虫介体筛选中的适用性

研究了一种人工培养液对各种常见的昆虫(主要是叶蝉)的亲和性和适用性.结果表明,该人工培养液适于本试验中大多数昆虫的人工饲养.用此方法,悬钩子广头叶蝉Macropsis.ftscula Zetterstedt和桤树广头叶蝉Oncopsis alniSchrank分别被再次确认为悬钩子矮化植原体和桤树黄化植原体的传播介体;田旋花麦蜡蝉Hyalesthes obsoletus Signoret再次被确认为葡萄黄化(stolbur)植原体的传播介体.此前,上述三种叶蝉已被传统的人工接种方法鉴定为相应植原体的传播介体.危害桤树的河谷树叶蝉Allygus modestus Scott尽管虫体DNA检测结果经常为阳性,但迄今其人工培养液的检测结果都是阴性,因此,我们认为河谷树叶蝉不是桤树黄化植原体的传播介体.Eppendorf管人工培养液饲养法不仅适用于潜在的植原体介体昆虫的筛选鉴定,而且可用于介体昆虫的生物防治研究.此外,本研究首次发现自然感染了葡萄上的一种被德国人称为"Vergi-lungskrankheit"植原体(AY组)的草地脊冠叶蝉Aprodes makarovi Zachvatkin.     

中国不同地区亚洲韧皮杆菌遗传多样性分析

 柑桔黄龙病是最严重的柑桔病害之一,现已威胁世界柑桔产业。已被证实在中国南部该病害通过嫁接和木虱传播。因此黄龙病的流行病学研究就显得尤为重要。本研究中,基于外膜蛋白(omp)基因的PCR-RFLP的方法被用来研究中国南部各柑桔主产区的亚洲韧皮部杆菌的遗传多样性。研究采用不同的限制性内切酶消化各地区的亚洲韧皮部杆菌的omp基因产生了不同的RFLP指纹图谱,并对各地亚洲韧皮部杆菌的omp基因进行克隆测序然后构建系统发育树。结果显示:中国不同地理区域的亚洲韧皮部杆菌具有一定的遗传多样性,即便在某一特定的地区也有一定的差异。序列分析显示中国地区的亚洲韧皮部杆菌有很高的同源性。这些结果对揭示中国柑桔黄龙病的流行病学及制定科学有效的防治策略具有一定的指导意义。     

中国不同地理来源稻曲病菌rDNA-IGS的初步分析

 The rDNA-IGS of 11 Ustilaginoidea virens isolates from different geographical regions in China were amplified by PCR and sequenced. The results showed that the rDNA-IGS fragments consisted of one cen-tral variable region and two lateral conservative regions. The two conservative sequences among the11 isolates shared the sequence similarity of 99.44%. The variable region consisted of 2, 4, or 6 of 77 bp-length-repeats in the isolates and the sequences of the repeat units were high conservative. Primers UIGS Ⅲ and UIGS IV were designed for amplification of the variable region in the rDNA-IGS.     

宁夏中部干旱带药用植物区系特点及开发利用

通过对宁夏具有很高药用价值的中药材生境和分布特点的分析,提出了加强资源保护、开发利用的措施与途径。     

Effects of stolbur phytoplasma infection on DNA methylation processes in tomato plants

DNA methylation was investigated as a possible mechanism for regulation of floral gene expression in stolbur phytoplasma‐infected tomato buds. Expression of methylase and demethylase genes was found to be globally down‐regulated in tomato plants infected with stolbur isolate PO, but not in those infected with isolate C. These results are consistent with the finding that SlDEF, a gene orthologous to arabidopsis APETALA3 which is involved in petal formation, was down‐regulated in stolbur PO‐infected buds and remained unaffected in stolbur C‐infected buds, and with the fact that the two stolbur phytoplasma isolates C and PO induce distinct symptoms. Because of variations between the different cell‐types of the flower buds, the DNA methylation status of SlDEF could not be clearly established. However, the finding that treatment of stolbur PO‐infected plants with 5‐azacytidine partially restored SlDEF gene expression strongly suggests that DNA methylation is involved in down‐regulation of floral development genes in stolbur PO‐infected tomatoes.     

Biological diversity of citrus ringspot isolates in Spain

Eight isolates of citrus ringspot were selected by symptoms induced in field trees and compared with a citrus psorosis isolate for symptom expression on several citrus species under temperature-controlled glasshouse conditions. Symptom expression in each host-isolate combination was quantified by a pathogenicity index (PI) that considered symptom intensity and the number of plants showing each symptom. A general pathogenicity index (GPI) was defined for each isolate as a weighted mean of the different PI. A wide range of symptoms could be observed depending on host-isolate combination and incubation temperature. On the basis of symptoms induced in the glasshouse, cross protecting reaction against psorosis B challenge inoculation, mechanical transmissibility to Chenopodium quinoa , and presence of a c. 48-kDa protein associated to fractions of a sucrose gradient infective on C. quinoa (Navas-Castillo et al, 1993), six of the ringspot isolates (RS-ALC, RS-SOR, RS-GR, RS-INV, RS-CV and RS-SR) could not be distinguished from the psorosis isolate used as control, whereas the other two isolates (RS-ALM and RS-BUR) were clearly different. Field symptoms induced by these two isolates also differed from those induced by psorosis or by the other ringspot isolates.     

Aggressive and mild Potato virus Y isolates trigger different specific responses in susceptible potato plants

Differences in the early responses of two potato cultivars, Igor and Nadine, to two isolates of Potato virus Y (PVY), the aggressive PVY^NTN and the mild PVY^N, were monitored. Microarray and quantitative real‐time PCR analyses were carried out to identify differentially expressed genes after inoculation with each virus isolate. Additionally, symptom severity and development was observed and the amount of virus isolate accumulated in systemically infected leaves was evaluated, where a significantly higher amount of PVY^NTN was detected. Microarray analysis revealed 572, 1288 and 1706 differentially expressed genes at 0·5, 12 and 48 h post‐inoculation, respectively in cv. Igor, with a similar pattern observed in cv. Nadine. Microarray and quantitative real‐time PCR results implied an earlier accumulation of sugars and lower photosynthesis in leaves inoculated with the aggressive isolate than in leaves inoculated with the mild isolate. The PVY^NTN isolate did not activate early differential expression of the Fe‐superoxide dismutase and pectin methylesterase inhibitor (PMEI) genes, indicating a delay in plant response relative to that following PVY^N inoculation. Differences in the expression of the β‐glucanase‐I gene were also observed in early plant responses to inoculation with each virus isolate.     

Geographic distribution and disease severity of different Rhizoctonia species pathogenic towards maize in different regions in Germany and France

Journal of Plant Diseases and Protection - The geographic distribution and DNA-disease severity of the anastomosis groups (AG) 1 IA, AG 1 IB and AG 2-2 IIIB within the teleomorph Thanatephorus...     

Spatiotemporal distribution of flavescence dorée phytoplasma in grapevine

In this study quantitative real‐time PCR was used to follow the seasonal changes of flavescence dorée phytoplasma (FDp) titre in grapevines of cv. Modra frankinja (syn. Blaufränkisch) and cv. Refo?k (syn. Refosco'd'Istria) from two vineyards located in climatically different vine‐growing regions of Slovenia. Besides its known presence in the leaf veins, FDp was also detected in flowers, berry tissues and tendrils. In plants with high concentrations of FDp in tissues with symptoms, phytoplasma was also detected in symptomless tissues. A trend of decreasing FDp titre in all examined symptomless tissues from June to July and an increasing one throughout the growing season in tissues with symptoms was recorded. Accordingly, FDp was present in detectable amounts in flowers, petioles and veins of almost all infected plants in the late spring, and was detected in all examined tissue types in summer, with the highest titre in berries in August. The study showed that in the absence of plant health measurements an FDp infection may spread exponentially by a factor of 40 per year.     

西北旱区枣疯病植原体核糖体蛋白基因的生物信息学分析

枣疯病是枣树上的一种具有毁灭性的植原体病害,几乎分布于国内所有的枣树栽培区,造成了巨大的经济损失.对我国陕西、宁夏、甘肃3省枣疯病样品植原体核糖体蛋白基因进行克隆和测序,获得枣疯病植原体的核糖体基因片段为1 196bp,包含部分rps19,rpl22和rps3三个基因,其中rpl22和rps3大小分别354bp和753bp,分别编码118和251个氨基酸,且这两个基因为非重叠基因.序列同源性比较结果表明:我国陕西、宁夏、甘肃的枣疯病植原体的核糖体蛋白rp基因大小一致,归属于植原体16S rⅤ-B组;该植原体核糖体蛋白基因特性与樱桃致死黄化(CLY5)和桃树黄化印度分离株系(PY-In)植原体相似.首次报道了我国枣疯病核糖体蛋白基因rp基因的序列,把枣疯病植原体归到16S rⅤ-B组,为枣疯病植原体提供了新的分类依据.     

Comparative host reactions and Frankliniella occidentalis transmission of different isolates of tomato spotted wilt tospovirus from Spain

The abilities of different isolates of tomato spotted wilt tospovirus (TSWV) collected from northeastern and eastern Spain to infect 10 host species, and to be acquired and transmitted by the western flower thrips (WFT), Frankliniella occidentalis were compared. Two isolates of TSWV from a single source plant could be separated according to the different type of local lesions they induced in Nicotiana glutinosa. Host ranges of the studied TSWV isolates were very similar, but differences were found in the symptoms induced and in their capacity to infect specific hosts systemically. Lycopersicon esculentumDatura stramonium were evaluated for their potential as virus acquisition host species. The proportion of transmitter adult thrips obtained from WFT larvae fed on L. esculentum was greater than from D. stramonium. No differences were detected between TSWV isolates in their ability to be acquired and transmitted by WFT. No evidence was obtained of alterations in TSWV particles which could affect WFT transmissibility due to the repeated mechanical transfers used to clone the isolates. Our findings do not support the existence of pathological effects of TSWV on WFT.     

Possible correlations between the characteristics of Potato leafroll virus isolates occurring in different geographical regions in Tunisia

Biological and molecular characterization supported by transmission efficiency, symptom expression and Open Reading Frame 0 (ORF0) nucleotide sequence analysis were carried out to assess nine isolates of Potato leafroll virus (PLRV) collected from three Tunisian geographic and bioclimatic zones. Plant-to-plant transmission by Tunisian Myzus persicae aphid clones showed high transmission efficiency for all isolates tested. Symptom expression analysis on a Physalis floridana plant test distinguished viral isolates as very severe, severe and mild. The ORF0 sequences of the Tunisian PLRV isolates showed an assignment to two aggregates when compared with GenBank PLRV sequences. A significant correlation between symptom severity and ORF0 nucleotide sequence or between symptom severity and geographic origins of the PLRV isolates was established. However, the transmission efficiency and the ORF0 sequence were not affected by the bioclimatic origin. No significant correlation between transmission and symptom or between transmission and the ORF0 sequence was detected.     

四川省不同地域的核盘菌遗传多样性分析

 核盘菌(Sclerotinia sclerotiorum)属于世界性分布的植物病原真菌,可以危害油菜等多种经济作物。研究不同地域核盘菌的遗传多样性对了解核盘菌的遗传演化过程和指导病害防控具有重要意义。实验采用序列相关扩增多态性(sequence-related amplified polymorphism,SRAP)标记对四川省17个不同地理来源的66株核盘菌菌株的遗传多样性进行了分析。10对检测引物共获得129个位点,其中123个为多态位点,占95.35%。UPGMA聚类结果显示,在相似性系数为0.7时,66个核盘菌菌株分为5类(Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ),分别包含60、2、2、1和1个菌株。在相似性系数为0.74时,第Ⅰ类又可分为3个亚类(Ⅰ-1、Ⅰ-2、Ⅰ-3),分别包含21、37和2个菌株。聚类及组成分分析结果显示,四川省各地区的核盘菌菌株具有较高的遗传多样性,但其遗传变异与菌株地理来源无明显相关性。     

Expression of engineered antibodies in plants: a possible tool for spiroplasma and phytoplasma disease control

ABSTRACT In an attempt to develop maize plants with resistance to corn stunt spi-roplasma (CSS), a single-chain Fv fragment (scFv) gene that was constructed from antibodies with strong inhibitory activity against CSS, was expressed in a cell line of maize. However, plants regenerated from this transgenic cell line showed no distinct resistance to CSS infection under the greenhouse conditions. The affinity and functionality of scFv in vivo and the locations of CSS and expressed scFv in maize could be a part of the contributing factors affecting this result. Progress of expressing antibodies in plants for plant pathogen resistance is also discussed.     

Diaspore characteristics and ecological adaptation of Bromus tectorum L. from different distribution regions

The invasion of nonnative plants is considered one of the main threats to the structure and function of North American ecosystems.Moreover,they can alter ecosystem processes and reduce biodiversity.In arid and semi-arid region of North America,the species of European annual grass Bromus tectorum L.is an outstanding example of these problems,which not only increase the fire density and change the fire regime,but replace native communities.Therefore,there are amount of researches on B.tectorum,including resource acquisition,water use efficiency and growth.Whereas the relevant research on the morphology of diaspore is scare.Diaspores have a fundamental role in seed germination and seedling establishment.Besides,as an important link between different generations,diaspores have a vital significance on individual reproduction and population extension.Hence,diaspores under selection for studying have an important implication.This study compares differences in seed morphology for Bromus tectorum collected from the United States,Kazakhstan,and Xinjiang of China.The following indices of B.tectorum diaspores were analyzed:size,thickness of covering layers,and micromorphological characteristics of the base,middle and transition area of diaspores as well as of the awn.Micromorphology of the lemma and the cross-section of the diaspore were observed by scanning electron microscopy.Results showed that thickness of the lemma and the palea of diaspores from B.tectorum-infested grasslands in the United States were reduced(P<0.05),likely because of environmental influences.This reduction facilitated the germination of diaspores and lowered the resistance of B.tectorum to adverse environmental conditions.The length of the awn also increased significantly(P<0.05),which helped in dispersal and anchoring of diaspores.Therefore,B.tectorum adapted ecologically to its new environment in the United States by strengthening its establishment ability.However,the defense capability of B.tectorum decreased.These results fit the evolution of increased competitive ability hypothesis(EICA)of invasive species.Analysis of various cells on the lemma revealed that prickle densities and collapsed,long epidermal cells were easily influenced by environmental factors such as temperature and moisture because of the physiologic function of these structures on silicon accumulation.However,the form and the position of silica cells,which were not greatly influenced by environmental factors,might be genetically controlled.Studying these structures at the microscopic level helps define the relationship between the diaspore and its environment.This study has a reference value for future studies on B.tectorum.     

Seasonal detection of pear decline phytoplasma by nested-PCR in different pear cultivars

Seasonal detection of pear decline phytoplasma was studied in three pear cultivars: Bartlett, Limonera and Blanquilla. Samples from 43 infected trees were collected monthly over 2 years and analysed by nested PCR. The three cultivars each showed a different pattern of phytoplasma detection. The maximum detection rate of pear decline phytoplasma occurred in December in the three orchards, and it remained high throughout the winter months. In spring, when new buds appeared and sap was produced, the detection rate decreased. Leaf midribs, buds and stems were compared to determine which sample was more reliable for phytoplasma detection. The best indicators were stems. The presence of phytoplasma in sieve tubes during the dormant season was determined by grafting. The results suggest that phytoplasmas could overwinter in shoots, with the implication that vegetative propagation during this period could also disseminate the disease.     

Detection and identification of the maize bushy stunt phytoplasma in corn plants in Brazil using PCR and RFLP

Plants of corn (Zea mays L.) exhibiting symptoms of stunting and leaf reddening were assayed for the presence of phytoplasma gene sequences through the use of phytoplasma rRNA and ribosomal protein gene and maize bushy stunt (MBS) phytoplasma-specific oligonucleotide primers in polymerase chain reactions (PCR). Polymorphisms in 16S rDNA amplified from diseased plants were those characteristic of phytoplasmas classified in the16S rRNA gene group 16SrI, subgroup IB, of which MBS phytoplasma is a member. Amplification of ribosomal protein (rp) gene sequences in PCR primed by phytoplasma-specific primers confirmed presence of a phytoplasma in the diseased plants. Restriction fragment length polymorphism (RFLP) patterns of the amplified phytoplasma rp gene sequences were similar or identical to those observed for a known strain of MBS phytoplasma. In separate PCR, an MBS-specific oligonucleotide pair primed amplification of a MBS-characteristic DNA from templates derived from the diseased corn. Our data provide the first firm evidence for the presence of maize bushy stunt phytoplasma in corn in Brazil.     

Incidence and natural spread of apple mosaic ilarvirus in hazel in north-east Spain

Apple mosaic ilarvirus (ApMV) was detected in all (160) hazel ( Corylus avellana ) cv. Negret orchards assayed in Catalonia, north-east Spain, during 1997 and 1998. In a preliminary survey, using enzyme-linked immunosorbent assay (ELISA), only 9 of the 320 samples collected appeared free from ApMV and three of these individuals indexed ELISA positives when re-tested. The spread of ApMV was studied in a plantation of Negret hazel in which virus-free trees were randomly interspersed with graft-inoculated individuals. When tested by ELISA 10 years after planting, 15% of the ungrafted trees contained ApMV. At that time, 8% of seeds on ungrafted and virus-free trees and 54% of seeds on ApMV-inoculated trees indexed ApMV-positive. These results are discussed in terms of pollen-mediated inoculation.