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南方水稻黑条矮缩病毒一步双重RT-PCR 检测技术及其应用 总被引：5，自引：0，他引：5
An one-step dual RT-PCR method was developed for Southern rice black-streaked dwarf virus (SRBSDV) detection from host plants and insect vector white-backed planthopper (Sogatella furcifera Horvath,Hemiptera: Delphacidae), from which two cDNA fragments of the viral genome S5 and S10 were amplified
simultaneously. Two primer pairs, S5-F1 / S5-R2 (5′-ttacaactggagaagcattaacacg-3′ / 5′-atgaggtattgcgtaactgagcc -3′) and S10-oF / S10-oR(5cgcgtcatctcaaactacag -3′ / 5′-tttgtcagcatctaaagcgc -3′), were selectedfrom 40 primer pairs based on SRBSDV genome sequences, and amplified viral S5 ORF1 fragment (819 bp) and cp gene fragment ( 682 bp), respectively. Using total RNA extracts from infected plant leaf tissue or individual planthopper adult as templates, one step RT-PCR reaction in the conditions of annealing temperature of 53℃ with the final concentrations of 240 nmol / L S5-F1 / S5-R2 and 120 nmol / L S10-oF / S10-oR generated a similar yield of two amplicons in argrose electrophoresis analysis. Sequence analysis confirmed the correct
result of the amplification. Additionally, several commercal RNA extraction kits was proven to be fit for the template preparation, and the protocol for total RNA extraction from plant leaf tissue and individual planthopper was simplified by sample grinding direct in eppendorf tube. This method can be used for SRBSDV detection of dried or 70% alcochol soaked planthopper samples stored over one month in room temprature. Key words: Southern rice black-streaked dwarf virus; Sogatella furcifera Horvath 相似文献
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Wheat blue dwarf(WBD) is a disease caused by phytoplasma and only reported from China. A fragment about 1.3 kb in protein translocation gene, secY was amplified by PCR from the total DNA of di-seased wheat sample with primer pair secYF/secYR, which was designed based on secY gene sequence of known 16SrI group members. Nucleotide acid sequence analysis of amplified fragment indicated that the length was 1 240 bp. A phylogenetic tree based on secY gene sequences was constructed and showed that wheat blue dwarf phytoplasma was clustered into the Candidatus Phytoplasma asteris, subgroup 16SrI-C. Wheat blue dwarf phytoplasma showed high homology with clover phyllody phytoplasma strains based on sequence comparison and phylogenetic analysis. 相似文献
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安徽桑黄花型萎缩病植原体16S rDNA序列分析及分子检测 总被引：1，自引：0，他引：1
Mulberry yellow dwarf(MYD)disease is an quarantine disease and the causal agent is a phytoplasma.Two pairs of published universal primer, P1/P7 and Rm16F2/Rm16R1, based on the 16S-23S rDNA sequence of phytoplasma and total DNA extracted from infected mulberry tissues were employed for PCR and nested-PCR detection.The results revealed that a phytoplasma-specific 1 830 bp fragment with a G+C content of 46.01% was sequenced(GenBank accession No.GQ249410).The sequence shared 99.7% and 99.8% identity with aster yellows, the representatiive phytoplasma in 16SrI group, and mulberry dwarf phytoplasma classified into subgroup B in 16SrI group and named as the MYD phytoplasma strain Anhui(MYD-Anh).A phylogenetic tree based on 16S rDNA sequences was constructed and showed that MYD-Anh was clustered into 16SrI group.Identity of 16S rDNA sequence between MYD-Anh and mulberry yellow dwarf phytoplasma strain Zhenjiang(MD-zj) was nearly 100%, and they might belong to the same strain.Nested-PCR was used to detect the pathogenic phytoplasma from the differential tissues of mulberry infected with MYD-Anh.The results showed that a phytoplasma-specific 1.4 kb fragment was amplified with total DNA extracted from bark and vein.Nested-PCR was more sensitive than PCR for detecting MYD phytoplasma. 相似文献
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Near the HMW-glutenin gene of wheat (Triticum aestivum), there is a locus (temporarily named TaXa) encoding LRR-receptor-like protein kinase, which is homologous to disease resistance protein Xa21 of rice (Oryza sativa). Through RT-PCR approach, a cDNA clone of ZS2002 was isolated from the orthologous locus of TaXa in Triticum turgidum. ZS2002 was 3 081 bp long and encoding a peptide composed of 1 026 amino acid. The protein included N-terminal conserved sequence, LRR domains, a transmembrane region and a serine/threonine protein kinase domain. ZS2002 was expressed in root, stem, leaf and spike. The transcribing in seedling leaves was significantly enhanced by Blumeria graminis f.sp. tritici. TaXa gene might play a role in powdery mildew resistance reaction in Triticum. 相似文献
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番茄叶片漆腐病病原菌鉴定 总被引：1，自引：1，他引：0
A new leave disease of tomato (Lycopersicon esculentum Miller) was observed in polyethylene film-covered greenhouse in Beijing, China. This disease spreaded rapidly in the greenhouse and caused serious loss of the production of tomatoes. In the study, a fungal strain isolated from the lesion was confirmed to be pathogen for this new disease, and identified as Myrothecium roridum Tode ex Ft. based on the morphology, cultural characters on PDA plate and sequence analysis of ribosomal DNA-ITS. This is the first report of myrothecium leaf spot on tomato occurring in commercial greenhouse in China. 相似文献
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江西甘蔗花叶病病原的分子鉴定 总被引：3，自引：0，他引：3
Sugarcane mosaic disease, caused by Sugarcane mosaic virus (SCMV), Sorghum mosaic virus (SrMV), Maize dwarf mosaic virus (MDMV) or Johnsongrass mosaic virus (JGMV) in Potyvirus, is one of the most important viral diseases of sugarcane. In the study, four primer pairs specific to SCMV, SrMV, MDMV and JGMV, respectively, were designed and used to detect 29 sugarcane leaf mosaic samples collected from 9 locations in Jiangxi province. The representative RT-PCR products were sequenced. The results showed that 22 samples were infected by SCMV, three by SrMV, and four were mix-infected by SCMV and SrMV. MDMV or JGMV were not identified in all samples. The result indicates that SCMV is the major pathogen of sugarcane mosaic disease in Jiangxi province, and SrMV is also a pathogen for the disease. 相似文献
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川芎茵核病的症状及病原菌鉴定 总被引：1，自引：0，他引：1
Symptoms of sclerotinia stem rot of Ligusticum chuanxiong were observed and the causal agent of the disease was identified. The results showed that the symptoms of this disease began with the formation of dark brown hygrophanous splotch on the basal leaves, followed by the appearance of dark brown circular spot on the basal stem. Finally, the whole plant stoped growing and died. White mycelium and/or sclerotia were formed under moisturized conditions. On PDA medium, the pathogen of this disease formed brown sclerotia with various shapes and 2-5 mm in diameter. The diameter of apothecia was 0.3-0.8 cm and the length of stipes was 2-8 cm. Asci were columniform in shape and (100-135)×9μm in size, each containing 8 ascospores sized by (6-12) μm×(3-7)μm. No asexual spore formation was observed. Based on the morphological characteristics and the sequence of ribosomal DNAITS, the pathogen was identified as Sclerotinia sclerotiorum (Lib.) de Bary. 相似文献
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Ants (Formicidae, Hymenoptera) play an important role in seed bank, seedling establishment andplant composition of arid ecosystems. Thus, knowing plant-ant interaction provides useful information formanagers to design restoration and conservation plans. In this study, the roles of desert harvester ants (Messorintermedius and Messor melancholicus) and scavenger ants (Cataglyphis nodus and Lepisiota semenovi) on plantcommunities were investigated in arid ecosystems of southeastern Iran. Two vegetation types weredistinguished in the study area and the nest density of ant species was determined. Furthermore, plantcomposition and soil seed bank were estimated at different distances from the ant nests. Results showed thatthe density of M. intermedius and M. melancholicus nests was higher in dwarf shrub-shrub vegetation type and thedensity of C. nodus and L. semenovi nests was higher in dwarf shrub vegetation type. The harvester and scavengerants had enhanced the seed bank to 55% and 70%, respectively. Therefore, the role of scavenger ants on theplant communities' seed bank was greater than that of harvester ants. Although the scavenger ants were moreinfluential on the annuals and the invasive plant species, the radius impact of the harvester ants on theperennials was greater, i.e., a positive interaction existed between the perennial plants and the harvester ants. C.nodus and L. semenovi played an important role in enhancing the ecosystem's potential for restoration throughestablishment of pioneer species in early stage of succession. The activity of M. intermedius is crucial for thedevelopment and maintenance of climax plant communities in arid ecosystems through assisting the plantspecies' establishment in late stage of succession. It is essential to preserve the diversity of these key ant speciesfor the maintenance and sustainability of shrubs in arid ecosystems. 相似文献
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The NS2 gene of Rice stripe virus (RSV) was amplified by RT-PCR, cloned into pGEM-T vector and sequenced. The NS2 gene was inserted into prokaryotic expression vector pET32a to produce recombinant plasmid pET32a-NS2. The recombinant plasmid was introduced into Escherichia coli strain BL21 (DE3) pLysS. SDS-PAGE and Western blot analysis confirmed that NS2 fusion protein was expressed after induction by IPTG. The recombinant NS2 protein was purified with Ni2+-NTA agarose affinity chromatography and the polyclonal antibody against NS2 protein was raised in rabbit. NS2 protein was successfully detected in small brown planthopper (Laodelphax striatellus) at 1:1 600 dilution of the total protein of single planthopper and in infected rice (Oryza sativa) at 1:800 dilution of 10 mg leave by dot immunobinding assay using the polyclonal antibody. 相似文献
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Fei Wang Guozheng Qin Zhenhua Sui Zhaohui Wang Zhaoyu Wang Jialin Yu Juren Zhang 《European journal of plant pathology / European Foundation for Plant Pathology》2006,116(4):289-300
The study of maize rough dwarf disease (MRDD) and breeding for resistance requires inoculation of maize plants by means of planthoppers. The plant age, insect density and inoculation duration are main factors in the success of maize rough dwarf disease inoculation. These parameters were tested using a susceptible maize inbred line Ye478. Using one or two-leaf plants, 15 planthoppers per plant and a five day inoculation duration, the line Ye478 was the most susceptible with 100% diseased plants; F112132 was moderately susceptible with 60% diseased plants and 90110 and F022411 were resistant without any disease. The results were consistent with those from six years of field studies. Using enzyme-linked immunosorbent assay (ELISA) and real-time quantitative RT-PCR, rice black-streaked dwarf virus was detected in severely diseased plants. The plants were rated from 0 to 3 according to their symptoms at the time of flowering. Plants scoring 0, 1 and 2 could not be distinguished by ELISA, only by real-time quantitative RT-PCR. All of the plants with a score of 3 were positive by ELISA and real-time quantitative RT-PCR. The significant differences in the average viral contents in plants with different symptom ratings could be distinguished by using real-time RT-PCR. 相似文献
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水稻黑条矮缩病暴发流行原因分析——以河南开封为例 总被引：1，自引：0，他引：1
水稻黑条矮缩病最早于1963年在浙江省余姚的早稻上发现,1964-1966年在浙江、江苏和上海等地流行或局部危害,1967年后华东地区发病迅速减轻,20世纪70年代在浙江病区难以找到病株,但1991-2002年浙江杂交稻区水稻黑条矮缩病又再次流行成灾,随后发病面积下降。2006年以来在江苏、浙江、山东等稻区大面积发生,并迅速上升为当地水稻主要病害之一,给水稻生产造成了巨大的经济损失。2013-2014年,水稻黑条矮缩病在河南沿黄部分稻区严重发生。本文从稻-麦连作的耕作制度,介体灰飞虱的越冬基数大、带毒率高,田间毒源丰富、易感水稻品种多,介体灰飞虱发生高峰与秧苗敏感期高度重合等方面,分析了水稻黑条矮缩病间歇性暴发流行的原因,以期为该病科学防控提供理论依据。 相似文献
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水稻新病害南方水稻黑条矮缩病发生特点及危害趋势分析 总被引：36，自引：1，他引：35
由斐济病毒属(Fijivirus)暂定新种南方水稻黑条矮缩病毒(Southern rice black-streaked dwarf virus,SRBSDV)引起的水稻矮缩病,自2001年在广东省阳西县被发现以来,已迅速扩散至我国南方广大稻区。2009年,该病害在我国南部及越南晚季稻上暴发成灾,造成严重的产量损失。本文简介了该病害的症状、危害特点、病原病毒及其传毒介体特征,对其发生趋势进行了讨论,提出了以秧苗期治虫为重点的病害防控应急措施。 相似文献
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为探明栽培管理措施与黄淮区水稻黑条矮缩病发生的关系,采用田间实地调查法研究了灌水、施氮、落谷密度和防虫网处理对水稻黑条矮缩病发生的影响,并对黄淮区主要水稻品种进行抗（耐）病筛选。结果表明,秧田期减小落谷量和施氮量,发病率最高可分别降低78.5%和77.7%;水稻抽穗前采用浅湿灌溉与深水处理相比,发病率最高可降低70.3%;秧田期防虫网处理与无防虫网相比,发病率最高可降低87.6%;分蘖期不施氮肥处理较过量施氮发病率最高可降低83.5%。黄淮区主要品种抗（耐）病性存在显著差异,抗病和产量性状都较好的品种为新稻20和苏秀10号,而金粳18可能对该病具有较强的耐害性。因此,在黄淮稻区,采用浅湿灌溉方式,适当减少施氮量和落谷量,秧田期采用防虫网处理,并选择抗（耐）病品种,可减轻黑条矮缩病对水稻生产的危害。 相似文献
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田间对比试验表明,作为一项应急补救措施,在病田进行水稻掰蘖补栽(丛),是当前黑条矮缩病发生流行以后最有价值的农业防治技术。掰蘖补栽处理所挽回的稻谷损失随病田发病率的上升而增加。丛病率为10%的杂交晚稻本田经掰蘖补栽后,可增产稻谷25.26kg/亩(约30元),而掰蘖补栽的工时费不足10元。因此,在病害流行年份,本田初期丛病率10%以上的杂交晚稻病田,均值得进行掰蘖补栽的防治工作。 相似文献