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1.
应用单克隆抗体检测香蕉束顶病   总被引:5,自引:0,他引:5  
应用香蕉束顶病毒(BBTV)的单克隆抗体(McAb)建立了检测香蕉束顶病的间接ELISA方法,其检测香蕉病组织汁液的最大稀释度为1:1280,健康香蕉组织汁液无非特异性反应。用间接ELISA方法检测了采自广东、广西9个市(县)的香蕉产区的80个大田香蕉束顶病样本和25个荫棚香蕉试管苗样本,其中大田样本BBTV的带毒率为76.5%,试管苗样本的带毒率为8.0%。香蕉束顶病株不同部位BBTV含量检测表  相似文献   

2.
During regular surveys of banana plantations in the Sindh province of Pakistan for banana bunchy top nanavirus (BBTV), sporadic plants showing mosaic, chlorotic spots intermingled with a dark green area were observed at a few places in the districts of Thatta, Hyderabad and Nawabshah. The infected young plants were uprooted and grown in pots for 2 years to ascertain the cause of the disease. Apart from studying the development of symptoms, the infected plants were tested for the presence of BBTV, cucumber mosaic cucumovirus (CMV), chilli veinal mottle potyvirus (CVMV), tomato mosaic tobamovirus (TMV), potato X potexvirus (PVX) and potato Y potyvirus (PVY) by DAS-ELISA. Virions were also partially purified. The infected plants developed typical symptoms of CMV. All tested plants were ELISA-positive only for CMV, and a partially purified preparation revealed virus particles measuring 25–28 nm in diameter. Based on the characteristic field symptoms, serology and particle morphology, the presence of CMV in banana in Pakistan was established.  相似文献   

3.
香蕉束顶病毒海口分离物基因组的克隆及序列分析   总被引:1,自引:0,他引:1  
 以海口香蕉束顶病株的幼嫩假茎和叶片总DNA为模板,通过反向PCR法克隆了香蕉束顶病毒(Banana bunchy top virus,BBTV)海口分离物(命名为BBTV-HaiKou)的6个DNA组分的全长序列。结果表明,DNA1~6序列全长分别为1106、1040、1058、1040、1013和1082nt。各组分非编码区各包含一个茎环共同区和一个主要共同区,同源性分别为91.55%和88.45%。各组分编码区均编码一个开放可读框(ORF),其中DNA1 ORF内部还有一个小的ORF。序列分析表明,BBTV-HaiKou分离物与其它分离物间的DNA1最为保守,DNA2变异最大。DNA1组分核苷酸序列与亚洲组、南太平洋组各分离物及ABTV (Abacá bunchy top virus)分离物同源性分别为93.1%~99.1%,89.6%~90.7%和76.2%~77.4%,相应的编码蛋白氨基酸序列同源性在BBTV和ABTV中分别为93.4%~100%和85.7%。根据Karan等的分类方法,确定BBTV-HaiKou分离物属于亚洲组成员。  相似文献   

4.
香蕉束顶病毒株系生物学特性的研究   总被引:7,自引:1,他引:6  
 在广东香蕉束顶病株症状基本相同的情况下,在8个产区分别随机采集11~15个标样共111个分离物,接种在香蕉苗上,其后又从8个产区的分离物中各选取1个代表分离物用于进行各项研究。寄主范围试验结果,这8个代表分离物可分为能侵染粉蕉的NSP株系(以广州天河分离物为代表的7个分离物)和不能侵染粉蕉的NS株系(高州代表分离物)。用Eco RI对8个毒源地的111个分离物DNA组分1进行酶切分析,结果表明:所有高州分离物共15个和信宜分离物14个中的9个都可以被酶切,应属NS株系;而其余6个毒源地的所有分离物及信宜分离物中的另外5个都不能被酶切,应属NSP株系。在病害潜育期方面,2个株系在大蕉上的差异达到显著水平;在香蕉4个品种上,2株系也存在较明显的差异,但其中有些差异未达到显著水平。在病毒增殖、运转速度及病毒达到高浓度所需的时间上,2个株系也存在显著的差异。  相似文献   

5.
The study of the transmission biology of insect-borne plant viruses is important to develop disease control practices. We characterized the transmission of a nanovirus, Banana bunchy top virus (BBTV), by its aphid vector Pentalonia nigronervosa Coquerel (Hemiptera, Aphididae) with respect to temperature, vector life stage, and plant access time. Adult aphids transmitted BBTV more efficiently than third instar nymphs at all temperatures tested. Adult aphids transmitted the virus more efficiently at 25 and 30 degrees C than at 20 degrees C, but temperature had no impact on transmission efficiency by nymphs. By decoupling the relationship between temperature and aphid BBTV acquisition or inoculation, we determined that temperature affected inoculation events more strongly than acquisition. Longer plant access periods increased viral acquisition and inoculation efficiencies in a range of 60 min to 24 h. Both BBTV acquisition and inoculation efficiencies peaked after 18 h of plant access period. We also show that BBTV transmission by P. nigronervosa requires a latent period. Our results demonstrate that vector transmission of BBTV is affected by temperature, vector life stage, and plant access period.  相似文献   

6.
香蕉束顶病毒(BBTV)侵染对寄主内源激素的影响   总被引:11,自引:0,他引:11  
 本试验用ELISA方法测定了香蕉感染束顶病毒(BBTV)后植株体内的3种内源激素,赤霉素(GA3)、玉米素类(iPAs)和脱落酸(ABA)的变化。结果表明,感株的GA3水平在侵染过程中虽有微弱增加,但含量和增长速度显著低于健株对照;iPAs在接种BBTV第14天后明显下降,并维持较低水平;ABA在BBTV侵染后被大量诱导增加并不断积累,在接种后第35天测定含量最高,为对照的3.34倍。试验还同时检测了BBTV在侵染过程中的运转。用间接ELISA测定的接种叶和顶叶的BBTV含量显示:接种21天后BBTV在接种叶和顶叶中还大量增殖,呈系统性分布。但寄主的症状在接种35天后才逐渐在顶叶表现。以上结果表明:香蕉束顶病的症状表现似乎主要与病株中的内源激素的失调有关,而与BBTV在体内的运转并不直接相关  相似文献   

7.
[目的] 对香蕉束顶病毒(Banana bunchy top virus,BBTV)海口分离物起始蛋白(replication initiation proteins, Rep)基因进行克隆、原核表达、抗血清制备,为BBTV有效的检测及分子流行病学等研究奠定基础。[方法] 以海口地区染病香蕉幼嫩假茎和叶片的总DNA为模板,通过PCR技术克隆BBTV海口分离物的起始蛋白基因,连接到表达载体并进行大肠杆菌原核表达,制备高效价特异性抗血清。[结果] 应用PCR方法从染毒香蕉幼嫩假茎和叶片总DNA中扩增复制rep基因,回收目的片段后经酶切,获得了含BBTV Rep基因的重组质粒pET32b Rep。将重组质粒转化E.coli BL21(DE3),经不同的时间、温度及IPTG浓度优化,12% SDS PAGE电泳分析,在20 ℃、0.1 mmol/L IPTG条件诱导4h,最终获得大量的可溶性融合蛋白。将可溶性蛋白上清液经Ni2+ NTA亲和层析柱纯化,得到高纯度的融合蛋白。用纯化后的融合蛋白免疫家兔,获得了BBTV Rep蛋白抗血清。以融合蛋白做抗原,间接ELISA法测定抗血清效价大于125 000。以田间样品做抗原时,结果表明抗血清最佳工作浓度为1∶1 000。Western blot鉴定结果表明抗血清能与融合蛋白特异性结合。[结论] 利用Rep基因制备的特异性抗血清在BBTV病毒粒体的组装机制研究及病毒病诊断上具有重要的应用价值。  相似文献   

8.
根据香蕉束顶病毒(Banana bunchy top virus,BBTV)、黄瓜花叶病毒(Cucumber mosaic virus,CMV)和香蕉线条病毒(Banana streak virus,BSV)3种病毒核酸保守序列设计相应特异性引物,通过体系优化,建立了可以同时检测香蕉束顶病毒、黄瓜花叶病毒、香蕉线条病毒的多重PCR体系。  相似文献   

9.
Pseudomonas fluorescens strains CHA0 and Pf1 were investigated for their biocontrol efficacy against Banana bunchy top virus (BBTV) in banana (Musa spp.) alone and in combination with chitin under glasshouse and field conditions. Bioformulation of P. fluorescens strain CHA0 with chitin was effective in reducing the banana bunchy top disease (BBTD) incidence in banana under glasshouse and field conditions. In addition to disease control, the bioformulation increased the economic yield significantly compared to the untreated control. Increased accumulation of oxidative enzymes, peroxidase (PO), polyphenol oxidase (PPO), phenylalanine ammonia lyase (PAL), pathogenesis-related (PR) proteins, chitinase, β-1,3-glucanase and phenolics were observed in CHA0 bioformulation amended with chitin-treated plants challenged with BBTV under glasshouse conditions. Indirect ELISA indicated the reduction in viral antigen concentration in P. fluorescens strain CHA0 with chitin-treated banana plants corresponding to reduced disease ratings. The present study revealed that induction of defence enzymes by P. fluorescens with chitin amendment reduced the BBTD incidence and increased bunch yield in banana.  相似文献   

10.
Journal of Plant Diseases and Protection - The banana bunchy top disease is a serious threat to banana production. Caused by a DNA virus, its detection using specific primers is the only sensitive...  相似文献   

11.
Isolates of Banana bunchy top virus (BBTV), which causes bunchy top disease in bananas, were collected in field surveys on seven islands in Okinawa Prefecture, Japan. From 44 banana samples, one isolate from each island was selected, and the DNA-1 and DNA-3 components were sequenced. Analysis of the major common region of DNA-1 showed that BBTV in Okinawa belongs to the Asian group of BBTV. DNA-1 and DNA-3 analysis revealed that Okinawan BBTV had a closer relationship with isolates from Taiwan and the Philippines than with some isolates from China and Vietnam. All the Okinawan BBTV isolates had high homology in the nucleotide sequences of DNA-1 and DNA-3 (%) because of a single, recent BBTV invasion of this area.The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession numbers AB108449 – AB108458  相似文献   

12.
香蕉束顶病毒的寄主及其在病害流行中的作用   总被引:3,自引:0,他引:3  
 接种测定结果表明:香蕉、粉芭蕉和大蕉等是香蕉束顶病毒的寄主,但美人蕉、芋头、艳山姜、芭蕉芋、姜黄、长春花、甜瓜等其它24种供试植物不是该病毒的寄主。研究结果还认为:在福建省香蕉产区,香蕉束顶病流行的主要侵染源为香蕉病株,而粉芭蕉和大蕉等病株则可在病区中病害的长期定殖和流行起重要的作用。本文还讨论了这些研究结果对该病害的流行学和防治的重要性。  相似文献   

13.
This study on the epidemiology of banana bunchy top disease (BBTD) was carried out in the context of small‐scale farms in Burundi for an integrated management approach. Banana trials were established in farmers’ fields comparing different plot locations, while spatial and seasonal occurrence of aphid vectors was evaluated at three different altitudes. In addition, serological tests were performed on banana leaf samples to confirm the presence and titre of the virus. The results showed that BBTD incidence varied among banana cultivars and locations. Nine months after plot establishment, BBTD incidence ranged from 21·8% to 56·4% in plots within affected fields, while a range of 0–12·3% was reported in plots located between 5 and 30 m away from affected banana fields. Aphid numbers were highest in the dry season. These aphids were able to acquire and transmit the virus irrespective of altitude. A mean incubation period of 21 and 84 days was observed at low (780 m a.s.l.) and high (2090 m a.s.l.) altitude, respectively. Thus, a holistic approach, taking into account banana cultivar, plot location, disease‐free planting material and regular field sanitation, should be promoted for long‐term BBTD management.  相似文献   

14.
由香蕉线条病毒(Banana streak virus,BSV)引起的香蕉线条病,严重影响了香蕉产量及种质资源交流。本研究根据BSV ORF Ⅲ基因保守序列设计引物和探针,建立了BSV的实时荧光定量PCR检测方法。该法检测BSV时,与黄瓜花叶病毒(Cucumber mosaic virus,CMV)和香蕉束顶病毒(Banana bunchy top virus,BBTV)无交叉反应;不论是检测质粒DNA还是香蕉病株总DNA,其灵敏度都比普通PCR高,检测质粒DNA的灵敏度比普通PCR高100倍,检测香蕉病株总DNA的灵敏度比普通PCR高10倍,且具有良好的重复性。利用建立的实时荧光定量PCR方法检测我国主栽的不同香蕉品种‘大蕉’、‘粉蕉’和‘粤优抗一号’香蕉组培苗中BSV的浓度,发现不同品种BSV传递规律不同。‘大蕉’第3代、第9代组培分化芽中BSV含量较原代显著减少;‘粉蕉’中BSV含量随继代数增加表现为先增加后减少;‘粤优抗一号’中BSV含量随继代数增加呈增加趋势。BSV在‘粉蕉’、‘大蕉’和‘粤优抗一号’第10代到第12代组培苗中以顶部第1片或(及)第2片叶中含量最高。直接结合PCR分析初步表明不同品种在原代和组培至第12代,其植株中的BSV均为游离状态。本文通过对带毒香蕉组培苗中BSV的含量监测,明确了BSV在不同品种带毒香蕉组培苗中的传递和分布规律,为研究BSV与寄主互作及BSV检测提供了理论依据。  相似文献   

15.
The cause of a banana disease in the province of Sindh, Pakistan, was attributed to banana bunchy top virus. Identification was based on symptomatology, size/morphology of the virus particles (20-22 nm) detected in diseased tissue, and serology (DAS-ELISA).  相似文献   

16.
This study combined the micro‐cross‐section cultural system with in vitro mutagenesis induced by ethyl methanesulphonate (EMS) to screen for fusarium wilt‐resistant lines of Brazil banana (Musa spp., AAA). The results indicated that the optimum EMS concentration and duration for the treatment of micro‐cross‐sections cut from the pseudostem of tissue‐cultured plantlet were 300 mm and 60 min, respectively. Under the optimal treatment, an average of 2·2 regenerated shoots were produced from each explant. One hundred regenerated plantlets were used for screening for fusarium wilt‐resistant lines by the early screening technique. The initial disease symptom – yellowing in lower leaves of susceptible plantlets – was observed 2 weeks after inoculation. After 2 months, only six plants survived – the putative fusarium wilt‐resistant lines. The fusarium wilt pathogen Fusarium oxysporum f. sp. cubense race 4, was identified in the preliminary test field by a SCAR marker technique. Of the six putative resistant lines, five survived the preliminary field test. The regenerated plantlets from these five fusarium wilt‐resistant lines were subjected to early screening again, where they showed markedly reduced disease incidences compared with regenerated plantlets of Brazil banana (control). It was concluded that EMS‐induced mutation of banana through the micro‐cross‐section cultural system is potentially useful for banana improvement.  相似文献   

17.
Vegetatively propagated crops suffer from yield loss and reduced stand density and longevity caused by the build-up of certain pests and pathogens between successive plantings via infected planting material. Here, six seedborne phytosanitary problems of banana are reviewed to evaluate whether a seed degeneration framework is a useful tool to identify approaches to achieve healthier planting materials. Phytoparasitic nematodes and weevils generate gradual declines in yields and in sucker health. Fusarium wilt and banana bunchy top virus cause progressive mat collapse across the field. Symptomless suckers from any mat in infested fields represent a risk of transmitting the disease to a new field. Xanthomonas and ralstonia wilts, due to incomplete systemicity, are intermediate in their threat to yield loss and frequency of transmission in suckers. Losses to banana streak virus are triggered by abiotic stress, although sucker transmission of episomal banana streak virus also contributes. A qualitative equation described here for seed degeneration covers a cycle beginning with the quality and risk factors of the planting material used to plant a new field and ends with the quality and risk factors of the suckers extracted from the field to plant a new field. This review of five planting material multiplication methods commonly used in banana contrasts their differing usefulness to address seed degeneration in the small farm context. It is proposed that initiatives to offset banana seed degeneration should integrate the role of off-farm actors into decentralized initiatives rather than attempt to duplicate national seed certification frameworks from other true seed or vegetatively propagated crops.  相似文献   

18.
The removal of infected individuals is a common practice in the management of plant disease outbreaks. It minimizes the contact between healthy individuals and inoculum sources by reducing the infectious window of contaminated individuals. This requires early detection and consistent removal at landscape scale. Roguing of mats with symptoms of banana bunchy top disease (BBTD) in Cavendish banana production systems has been tested in Australia, using trained personnel, but has never been tested in smallholder systems. We studied the effectiveness of long-term consistent roguing in prolonging the productivity of banana orchards under smallholder farming systems in highland banana and plantain dominated production systems in Africa. We assessed the possibility of low-risk seed sourcing from the managed plots. Roguing reduced BBTD incidence to 2% in managed farmer fields and to 10% in experimental field plots, while a nonmanaged field eventually collapsed in the same period. With roguing, new infections decreased monthly compared to an exponential increase in a nonmanaged field. The emergence of new infections in both managed and nonmanaged farms followed a seasonal cycle. BBTD managed plots were a source of low-risk seed for replacing the rogued mats in the same fields, but perhaps not safe for use in nonendemic areas. We conclude that it is possible for smallholder farmers to recover and maintain banana productivity with rigorous roguing, which would entail early identification of symptoms and early removal of diseased mats. Studies are needed on the intensity of roguing under different disease and production conditions.  相似文献   

19.
Meristem domes excised from plantlets of Ornithogalum thyrsoides infected with ornithogalum mosaic virus (OrMV) in Kenya were cultured on basic Murashige and Skoog medium without added growth hormones. Of the resulting plantlets 30% were virus-free as determined by symptom appearance, mechanical inoculation to virus-free O. thyrsoides , electron microscopy and serological tests. When healthy plants were placed within a field of diseased O. thyrsoides , 35% became re-infected within 4 months and 100% infection was reached within 8 months. However, healthy plants placed in isolation, about 2 km downwind of infected commercial plantings, remained healthy over the 12-month test period.  相似文献   

20.
Latent infection of winter oilseed rape by Leptosphaeria maculans   总被引:2,自引:2,他引:0  
Plants of oilseed rape, cultivars Primer and Jet Neuf, were grown in a glasshouse and inoculated at G.S. 2.4–2.7 with pycnidiospores or ascospores of Leptosphaeria maculans. The plants were kept for a further 2–4 weeks at 14°C and then transferred, together with uninoculated plants, to a polythene tunnel in winter. The majority of stems of inoculated plants did not have macroscopic symptoms of L. maculans infection 6 weeks after inoculation. Examination of whole mounts of peripheral tissue and transverse sections of fixed and embedded portions of these stems revealed intercellular septate fungal hyphae, often deep in non-necrotic cortical tissue, in symptomless inoculated plants but not in uninoculated plants. L. maculans was recovered following surface sterilization of adjacent portions of the same stems. When symptomless inoculated plants were transferred to a glasshouse at 18–20°C, cankers soon developed. The significance of these latent mycelial infections to canker development in the field is discussed.  相似文献   

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