应用RAPD标记技术鉴定香蕉褐缘灰斑病菌

采用Mycosphaerella fijiiensis和M.musicola的RAPD标记技术鉴定海南香蕉褐缘灰斑病菌,结果表明,分离自海南儋州、乐东、文昌、东方、澄迈、临高、琼海、昌江、琼山、三亚香蕉上的10个菌株均为M.fijiensis,引起香蕉黑叶条斑病。     

Molecular diagnostics for the sigatoka disease complex of banana

ABSTRACT The Sigatoka disease complex of banana involves three related ascomycetous fungi, Mycosphaerella fijiensis, M. musicola, and M. eumusae. The exact distribution of these three species and their disease epidemiology remain unclear, because their symptoms and life cycles are rather similar. Disease diagnosis in the Mycosphaerella complex of banana is based on the presence of host symptoms and fungal fruiting structures, which hamper preventive management strategies. In the present study, we have developed rapid and robust species-specific molecular-based diagnostic tools for detection and quantification of M. fijiensis, M. musicola, and M. eumusae. Conventional species-specific polymerase chain reaction (PCR) primers were developed based on the actin gene that detected DNA at as little as 100, 1, and 10 pg/mul from M. fijiensis, M. musicola, and M. eumusae, respectively. Furthermore, TaqMan real-time quantitative PCR assays were developed based on the beta-tubulin gene and detected quantities of DNA as low as 1 pg/mul for each Mycosphaerella sp. from pure cultures and DNA at 1.6 pg/mul per milligram of dry leaf tissue for M. fijiensis that was validated using naturally infected banana leaves.     

Reactions of plantain and banana cultivars to black sigatoka leaf spot disease in three farming systems in the Nigerian guinea savanna

Journal of Plant Diseases and Protection - An assessment of the severity of black sigatoka (BS), Mycosphaerella fijiensis, leaf spot disease on plantain and banana was undertaken in three farming...     

Black leaf streak (Mycosphaerella fijiensis) in Uganda

The occurrence of black leaf streak caused by Mycosphaerella fijiensis is reported on a number of banana cultivars in Uganda, demonstrating the spread of the disease from West and Central to East Africa since the early 1980s.     

利用香蕉水培系统测定枯萎病病原菌致病力

 为建立一种评价香蕉枯萎病病原菌(Fusarium oxysporum f.sp. cubense)菌株间致病力差异的体系,在改进的香蕉水培系统基础上,对影响香蕉枯萎病菌致病力的接种菌液浓度、类型及处理方式等因素开展分析,同时与不同菌株盆栽测定结果进行比较以验证该方法的可靠性。结果表明:在香蕉水培系统下,将摇培5 d的菌液稀释10倍以上,使孢子初始浓度为1×10⁶ cfu·mL^-1,直接加入该菌液50 mL即可用于致病力评价,且不同菌株用该测定方法与盆栽测定的致病力结果基本一致。该方法的建立为香蕉枯萎病菌致病力的评价提供了快速简便的方法,也为下一步解析尖孢镰刀菌致病相关基因功能和致病机理及抗病品种的选育奠定了基础。     

The use of species-specific DNA probes for the identification of Mycosphaerella fijiensis and M. musicola, the causal agents of Sigatoka disease of banana

The polymerase chain reaction (PCR)-based technique of random amplification of polymorphic DNA (RAPD) was used to differentiate DNA from species of the genus Mycosphaerella. DNA from two pathogens which cause Sigatoka leafspot diseases of banana, M. fijiensis and M. musicola , and two other Mycosphaerella species which are commonly found on banana, M. musae and M. minima , gave distinct RAPD banding patterns with all PCR primers tested. PCR, using primer RC07, amplified a 1250bp RAPD fragment from all isolates of M. fijiensis obtained from 11 geographical origins. This fragment was absent from the other species of Mycosphaerella. In Southern blots of genomic DNA, this band hybridized exclusively to DNA from M. fijiensis , and the pattern of hybridization suggested that it was binding to repeated DNA. A RAPD band amplified with primer PM06 obtained from M. musicola was also found to be species-specific. Southern analysis suggested that the fragment hybridized to a single-copy sequence in the M. musicola genome. Total genomic DNA from M. musicola was found to be a species-specific hybridization probe. Dot-blots confirmed the specificity of these probes, and could be used to identify isolates of Mycosphaerella which cause Sigatoka disease of banana in south-east Asia.     

Effect of Juglone on Active Oxygen Species and Antioxidant Enzymes in Susceptible and Partially Resistant Banana Cultivars to Black Leaf Streak Disease

The black leaf streak disease (BLSD), caused by Mycosphaerella fijiensis, is the most destructive disease of bananas and plantains around the world. Breeding for resistance is the most promising strategy to fight this disease especially in small farmer plantations. Mycosphaerella fijiensis produces many phytotoxins such as juglone, which can be used, jointly with field and inoculations under controlled conditions, for screening banana cultivars for BLSD-resistance. This non-host specific phytotoxin has been shown to act on chloroplasts and disturbs the proton electrochemical gradient across the plasmalemma membrane. Moreover, an involvement of the oxidative burst during the interaction has been suggested. The present study was carried out using two cultivars that differed for either their juglone-responses or their resistance to BLSD (cv. Grande Naine susceptible to BLSD and juglone and cv. Fougamou partially resistant to BLSD and highly tolerant to juglone). The production of active oxygen species (AOS) and the enhancement of the enzymatic and/or non-enzymatic AOS-scavenging systems were investigated after treatment of the two cultivars with juglone. The time-course of AOS-production and AOS-scavenging was shown to be the key difference between these two tested cultivars after treatment with juglone. Thus, an early release of AOS (O₂⁻ radical and H₂O₂) and a quick stimulation of a preferment anti-oxidant system (superoxide dismutases, catalases, and peroxidases) was observed for cv. Fougamou as compared to cv. Grande Naine for which a late and weak generation of AOS accompanied by a late stimulation of the anti-oxidant systems were detected.     

云南省水稻细菌性条斑病菌的致病型划分和水稻抗性资源的鉴定

 为明确云南省水稻细菌性条斑病菌(Xanthomonas oryzae pv. oryzicola, Xoc)的致病力分化以及不同类型水稻品种对Xoc的抗感特性,通过针刺接种法将云南省8个稻区采集的86株Xoc菌株,接种于6个携带不同抗性基因的水稻鉴别品种(IRBB4、IRBB5、IRBB14、IRBB18、IRBB21和IR24)。根据这些菌株在鉴别品种上的毒力差异进行了UPGMA聚类分析,将其划分为9个致病型(Ⅰ型 ~ Ⅸ 型)。其中,Ⅰ型为优势菌群,分布频率为50.5%。对不同稻区的优势菌群进行分析,发现云南省各稻区Xoc的致病型呈多样性分布,以强毒力的Ⅰ型为高频率致病型。选用 Ⅰ 型、Ⅱ 型和 Ⅵ 型代表菌株对云南省的80个主栽和区试水稻品种进行抗性评价,对3个致病型表现抗性的材料比例分别为30.0%、35.0%和57.5%。筛选出9个对3种致病型都表现为抗性的品种,其中“Deyou16”和“Changgui2”表现为高抗。研究结果可为云南省防治水稻细菌性条斑病的水稻区域性布局和抗性品种的利用提供理论依据。     

Genetic structure of Mycosphaerella fijiensis populations from Australia, Papua New Guinea and the Pacific Islands

Single-copy restriction fragment length polymorphism (RFLP) markers were used to determine the genetic structure of Mycosphaerella fijiensis , the cause of black leaf streak (black Sigatoka) disease of banana and plantain, in the Torres Strait, Papua New Guinea (PNG), and the Pacific Islands. A moderate level of genetic variation was observed in all populations with genotypic diversity values of 60–78% of the theoretical maximum, and gene diversity ( H ) values between 0·269 and 0·336. All populations were at gametic equilibrium, and with the high level of genotypic diversity observed this indicated that sexual reproduction has a major role in the genetic structure of the M. fijiensis populations examined. Population differentiation was tested on several hierarchical scales. No evidence of population differentiation was observed between sites on Mer Island. A moderate level of population differentiation was observed within the Torres Strait, between Badu and Mer Islands ( F _ST = 0·097). On a regional scale, the greatest differentiation was found between the populations of the Torres Strait and the Pacific. Populations from these regions were more closely related to the PNG population than to each other, suggesting they were founded in separate events from the same population.     

Tackling black leaf streak disease and soil fertility constraints to enable the expansion of plantain production to grassland in the humid tropics

In Central Africa, plantain is traditionally grown after a forest fallow. Given increasing urban demand and a lack of forest fallows near urban centres, as well as poor roads and environmental concerns to reduce pressure on forests, research is needed to identify suitable shade, fertility management and cultivars to shift production of plantain to grasslands and to reduce losses to diseases such as black leaf streak disease (BLSD). Effects of light level (full, 67%, 33% light), and nitrogen (N)-amendment on BLSD-tolerant (FHIA-21) and BLSD-susceptible (Batard) cultivars planted on soil from paired grassland and forest sites were determined. BLSD and growth were monitored until 5 months after planting. Three months after planting, leaf area attacked on cultivar FHIA-21 was less than half that on Batard. Plants grown under 33% and 67% light had less leaf area attacked (2.9% and 4.6%, respectively) than those grown in full light (7.3%). Leaf area and dry matter (DM) were higher under shade and when grown on forest soils. Compared to growing BLSD-susceptible plantain on forested land under shade, a shift onto grasslands and a reduction in shade use is predicted to reduce yields. Using cultivar FHIA-21 may limit, but not eliminate, yield loss.     

香蕉花叶病的酶联免疫检测技术研究

 用分离自香蕉病株的黄瓜花叶病毒香蕉毒株(CMV-B)和烟草花叶病毒香蕉毒株(TMV-B)制备抗血清,其效价As-CMV-B为1:5000,As-TMV-B为1:8000。用这两种抗血清检测香蕉花叶病,间接ELISA和PAS-ELISA法可检测香蕉病叶汁液的最大稀释度均为1:1280,健叶汁液无非特异性反应,Dot-ELISA可检测病叶汁液的最大稀释度为1:640,健叶汁液在稀释度为1:20时有轻度非特异性反应。用间接ELISA法检测了田间香蕉病株和香蕉试管苗标样共158份,其中CMV的带毒率为58%,TMV的带毒率为19%,二者复合感染率为8.5%。     

Molecular Typing and Presence of Genetic Markers Among Strains of Banana Finger-Tip Rot Pathogen, Burkholderia cenocepacia, in Taiwan

ABSTRACT Burkholderia cenocepacia (genomovar III of B. cepacia complex), the causal agent of banana finger-tip rot, is a common plant-associated bacterium but also an important opportunistic pathogen of humans. To better understand the nature of B. cenocepacia from banana, the genetic variation among B. cenocepacia isolates from various banana-growing regions in southern Taiwan was examined. Forty-four serial isolates recovered from diseased banana stigmata from three banana-growing regions during the periods ranging from 2002 to 2004 were investigated. All B. cenocepacia isolates picked from quinate-yeast extract tetracycline-polymyxin semiselective medium could cause onion maceration and were polymerase chain reaction (PCR) positive for bcscV, which is a type III secretion gene present in all members of the B. cepacia complex except B. cepacia (formerly genomovar I). Genetic diversity was assessed using recA PCR restriction fragment length polymorphism, recA nucleotide sequence analysis, and pulsed-field gel electrophoresis assays. The assays revealed the genetic variability among the isolates and also allowed us to trace the relationship among isolates. The isolates all were assigned to genomovar III and consisted of two groups, A and B, which corresponded to recA lineage IIIA and IIIB. The group B strains were separated into B1 and B2 subgroups and the B1 strains were further divided into distinct lineages. The B1 strains were the most frequently detected and occurred in all regions tested. There was no significant difference between strains from each subgroup in the virulence on banana fingers of cv. Cavendish. PCR assays were further used to determine whether B. cenocepacia from banana contained the cable pilus subunit gene (cblA), IS1356, and B. cepacia epidemic strain marker (BCESM), which are DNA markers associated with epidemic B. cepacia clinic strains. The results indicated that cblA and IS1356 were absent but the BCESM was found in all isolates. The present study revealed that banana is a natural reservoir of genetically diversified B. cenocepacia strains.     

应用单克隆抗体检测香蕉束顶病

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     

Effects of inoculum concentration, wetness duration and plant age on development of early blight (Alternaria solani) and on shedding of leaves in tomato plants

Effects of inoculum concentration, wetness duration and plant age on the development of tomato early blight were evaluated in relation to host susceptibility under controlled environmental conditions. The main effect of early blight was premature defoliation, which was linearly related to the percentage of leaf area showing symptoms. As ln(inoculum concentration, conidia mL⁻¹) increased from 6·2 to 11·5, the percentages of leaf area affected and of defoliation increased linearly. Four h of leaf wetness after inoculation were sufficient to initiate the disease on plants of hybrid Skala RZ but not on those of cv. Rio Rojo, for which at least 6 h leaf wetness were needed. As wetness duration increased up to 24 h, there was an increase in the percentage leaf area showing symptoms and in the percentage of defoliation, but thereafter there was no significant increase in either parameter. Tomato plants were susceptible to Alternaria solani at all growth stages, but susceptibility increased as plants matured. There were no significant differences in susceptibility between tomato cultivars and hybrids.     

黄淮区大豆花叶病毒株系组成与分布

 用8个大豆品种作为一组株系鉴别寄主,将黄淮豆区8省市202个毒株划分为7个株系(y₁~y₇)。y₁及y₂为弱毒株系,仅侵染感病品种(1138-2、文丰5号);y₃、y₄、y₅为中毒株系,分别侵染感病和中抗品种(齐黄10号、徐豆1号、诱变30、鲁豆4号);y₆及y₇为强毒株系,除侵染以上品种外,还侵染高抗品种(齐黄22、密荚1号)。全区弱毒株系占55.9%,中毒株系占28.7%,强毒株系占15.3%。江苏、山东、河南、河北、安徽和北京市以弱毒株系为主(43.5%~88.9%),山西省以中毒株系为主(50.0%),陕西省弱毒株系、中毒株系各占50.0%,山东、河南、河北3省强毒株系所占比例较高(22.4%~33.3%)。株系分布具有一定的稳定性。     

Virulence factors analysis of native isolates of Xanthomonas albilineans and Xanthomonas sacchari from Tucumán,Argentina, reveals differences in pathogenic strategies

Xanthomonas albilineans (Xa) and X. sacchari (Xs) are both sugarcane pathogens. Xa is the causal agent of leaf scald disease, but there is limited information about the pathogenicity of Xs. The aim of this work was to study virulence factors of native strains of Xa (Xa32, Xa33, and XaM6) and Xs (Xs14 and Xs15) previously isolated from sugarcane with leaf scald symptoms, to gain insight into the biology of each microorganism. We analysed epiphytic survival, sensitivity to oxidative stress, extracellular degradative enzymes, cell motilities, exopolysaccharide (EPS) characteristics, cell adhesion, biofilm development, and control of stomatal regulation of the five strains. We observed that each species presented similar phenotypes for every factor analysed. Xa strains appeared to be more sensitive to oxidative stress and presented lower epiphytic survival than Xs. All strains presented endoglucanase activity; however, we could only detect protease and amylase activities in Xs strains. Swimming and sliding were higher in Xs, but twitching was variable among species. We also observed that only Xs strains produced a xanthan-like EPS, presented a strong cell adhesion, and structured biofilm. We detected some intraspecific variations showing that higher amounts of EPS produced by Xs14 correlated with its higher sliding motility and its homogenous and more adhesive biofilm. In addition, EPSs of Xs14 and Xs15 presented differences in strand height and acetyl percentage. Finally, we found that strains of both species were able to interfere with stomatal aperture mechanism. All these differences could influence the colonization strategies and/or disease progression in each species.     

广西香蕉真菌性叶斑病病原菌种群结构分析

2009年2-3月对广西香蕉主产区真菌性叶斑病病原进行抽样鉴定。结果显示,广西香蕉真菌性叶斑病病原至少有10种,主要病原为Cordana musae,分布广,检出率为56.77%~92.19%;Corynespora cassiicola和Deightoniella torulosa为次,两种病原菌均主要集中在南宁地区,检出率分别为32.34%和22.61%;叶斑病类型以单一病原侵染为主,检出率为78.13%~92.19%;复合侵染的叶斑病类型以2种病原共同侵染居多。     

江苏省水稻品种对水稻细菌性条斑病抗性鉴定及评价

为明确江苏省水稻条斑病菌致病力分化状况和不同类型水稻品种对条斑病抗感特性,在孕穗期采用针刺接种法对徐淮稻区2007—2009年采集分离获得的45株条斑病菌进行致病力测定,根据病原菌在6个水稻鉴别品种IRBB4、IRBB5、IRBB14、IRBB21、IR24和金刚30上的抗感反应划分致病型,从中选择具有代表性的3种不同致病型条斑病菌,并鉴定了240个不同类型的水稻品种对条斑病的抗感性。根据菌株在鉴别品种上的抗感反应将供试菌株划分为8个致病型,其中优势致病型为C3致病型,所占比例为35.5%;大多数菌株与鉴别品种间表现出弱互作关系,少数菌株表现出强互作关系。粳稻品种对条斑病的抗性明显高于籼稻,但常规粳稻和杂交粳稻对强致病力菌抗性比例仅为59.8%和37.5%。表明水稻细菌性条斑病流行仍具有潜在的威胁。     

Patterns of development of Septoria nodorum and S. tritici in some winter wheat crops in Western Europe, 1981-83

In data collected at 19 sites in Western Europe during 1981-83. two patterns of development of Septoria nodorum and S. tritici on foliage of winter wheal were distinguished. In sudden outbreaks, lesions appeared simultaneously on the upper leaf layers of crops, usually after the end of stem extension; these outbreaks were ascribed to short, heavy rain storms in which pycnidiospore inoculum in basal leaves was elevated up to 60 cm through the crop canopy. Gradual epidemics were characterized by disease arising on successive leaf layers as they appeared during sustained periods of weather suitable for inoculum transport and infection.
The data indicate incubation periods of 2-4 weeks for S. nodorum and 3-5 weeks for S. tritici. it is suggested that a leaf layer cannot normally sustain more than one pathogen generation and that its infection arises from inoculum borne on leaves older than in the layer situated immediately below it. The potential level of disease in a crop may relate to the amount of inoculum present in spring. The proportions of disease caused by the two Septoria species varied greatly between sites and years, but the data provided no explanation.
It is concluded that a septoria forecast scheme needs to recognise the importance of sudden disease outbreaks and to include not only weather but also host growth and inoculum factors.     

Pathogenic and genetic variability of Ralstonia solanacearum strains from the Philippines

In the Philippines, bacterial wilt caused by Ralstonia solanacearum is one of the most important diseases affecting vegetables and banana. In this study, 89 strains of R. solanacearum isolated from various hosts were screened for their biovar, phylotype, pathogenicity, and genetic diversity. Foreign strains were included for comparison with these Philippine strains. Results of the biochemical and multiplex-PCR tests divided the Philippine strains into five biovars (1, 2, 3, 4, and N2) and three phylotypes (I, II, and IV). Three potato strains belonged to biovar N2/phylotype IV. Pathogenicity tests divided the strains into five pathogenicity types based on their virulence in tomato, potato, eggplant, sweet pepper, and tobacco. Strains classified as biovar N2 were weakly pathogenic to potato (pathogenicity type III) and almost all strains isolated from banana were not pathogenic to the test plants except potato (pathogenicity type V). The results of AFLP analysis divided the strains into four clusters. Cluster 1 was composed of strains isolated from solanaceous crops, ginger (Zingiber officinale), and Morus sp. from the Philippines and other Asian countries. Cluster 2 grouped the potato strains (biovar N2) from the Philippines and Japan and blood disease bacterium strains from Indonesia. Cluster 3 contained the local and foreign strains isolated from potato (biovar 2) and banana (biovar 1). Cluster 4 consisted only of the tomato strain from the USA.