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。     

25%乙嘧酚SC对香蕉褐缘灰斑病菌的室内毒力测定

采用生长速率法测定结果表明，25％乙嘧酚(ethirmol)对香蕉褐缘灰斑病菌具有较好的抑制作用，其毒力回归方程为：y=1．3084x 1．1806，EC50为822．26mg／L，EC90为7918．72mg／L。     

大豆灰斑病菌生理小种鉴定

大豆灰斑病菌生理小种鉴定胡国华，于凤瑶，程显伟，勇健康，栾怀海（黑龙江友谊县农垦科学院红兴隆所１５５８１１）大豆灰斑病（ＣｅｒｃｏｓｐｏｒａｓｏｊｉｎａＨａｒａ）广泛发生于世界大豆产区。１９１５年日本首次发现，我国以黑龙江东部地区发生严重，近年向西移...     

玉米灰斑病菌生物学特性研究

 玉米灰斑病菌(Cercospora zeae-maydis Tehon and Doniels)适合分离的培养基有花生叶斑病尾孢菌培养基、PDA、V8汁、V8汁+蔗糖20 g和Richard培养基。适宜菌丝生长的pH 4~12,最适宜的pH 6~8,适宜温度为20~25℃。病菌对葡萄糖、麦芽糖、乳糖和对酵母膏、硝酸钾、牛肉膏的利用好于其它碳源或氮源。分生孢子萌发的温度为20~30℃,pH范围广,RH ≥ 81%以上分生孢子萌发,随湿度增加萌发率增高。营养对孢子萌发无影响。     

大豆灰斑病菌毒素生物活性分析

 大豆灰斑病菌[Cercosporium sojina(Hara) Liu&Guo]可以产生有毒代谢物。滤液透析及热稳定性测试结果表明:该毒素可以穿过半透膜,为较小分子化合物并具有很强的热稳定性。该菌培养物浸提液经浓缩,硅胶柱层析可得粗毒素。生物测定结果表明:该毒素对大豆幼苗及叶片具有致萎作用。针刺叶片可产生类似真菌感染的病斑。浸渍处理后,可使叶组织失绿、萎蔫最后坏死,同时毒素对抗性不同的品种有鉴别作用。     

云南省玉米灰斑病菌的分子鉴定

本研究利用ITS区、tef1基因、cmdA基因和HIS基因4个基因部分序列对2009-2011年间采自云南省主要灰斑病发生区域和吉林省部分区域的玉米灰斑病菌进行比较分析。结果表明,无论是4个基因序列单独聚类还是拼接序列聚类均把云南省灰斑病发生区域采集的菌株与Cercospora zeina聚为一群,而吉林省采集的灰斑病菌则与Cercospora zeae maydis聚为一群。聚类分析还表明云南省不同玉米产区的灰斑病菌遗传背景单一,在所有聚类树中均聚为一群,且自举支持值均>99%。     

草莓角斑病菌检测鉴定技术研究进展

本文综述了近年来草莓角斑病菌(Xanthomonas fragariae)检测鉴定技术的研究进展.传统的鉴定方法主要有依赖田间症状识别、寄主接种鉴定、培养性状、生理生化鉴定、脂肪酸分析法(FAP)等;应用血清学鉴定检测方法主要有酶联免疫吸附法(ELISA)、菌落免疫荧光染色法(IFAS)等,分子生物学检测方法主要有限制性片段长度多态性(RFLP)、聚合酶链反应(PCR)、巢式PCR、实时荧光PCR等.对于XF的检测鉴定不能单单依靠一种方法,应将多种方法结合起来进行最终判定,因此建立一套完整的检测鉴定技术体系十分必要.     

玉米灰斑病抗性鉴定技术

对玉米灰斑病菌孢子产生、病菌接种和寄主抗病性测定技术的研究结果表明，应用玉米叶粉碳酸钙琼脂和玉米叶粉琼脂两种培养基，温度24～25℃，培养5天，病菌可大量产生分生孢子；于植株喇叭口期，用注射器将病菌孢子悬浮液注射于植株喇叭口中，获得了理想的发病效果；对20个玉米自交系注射接种鉴定结果表明，玉米自交系间抗病性差异明显，但未发现免疫自交系。     

毛豆灰斑病的发生与防治

近几年来 ,用于速冻加工出口创汇的毛豆在我县发展迅速 ,但灰斑病发生较为普遍 ,尤其是１９９８年 ,一般田块因病而无法用于加工的豆荚占２０%～３０% ,重病田达到８０%以上 ,经济损失严重。为此 ,笔者对其发生进行了调查 ,并对防治技术进行了探索 ,现介绍如下 :１症状与病原１．１症状幼苗及成株均可染病。幼苗期发病 ,子叶上出现圆形或半圆形稍凹陷的红褐色病斑 ,病情严重时 ,可导致死苗。成株期叶片、茎秆、豆荚、籽粒也可发病。叶片上病斑初期为红褐色小点 ,以后扩展成圆形 ,边缘深褐色 ,中央灰白色 ,潮湿天气 ,背生灰色霉层 ,后期病斑…     

香蕉黑腐病菌(Botryodiplodia theobromae)的PCR检测

 根据香蕉黑腐病菌可可球二孢菌(Botryodiplodia theobromae)与其它香蕉病原真菌核糖体基因转录间隔区(rDNA-ITS)ITS1和ITS2间序列差异,设计了特异引物Bth-S(5'-TCTCCCACCCTTTGTGAAC-3')和Bth-A(5'-AAAAGT-TCAGAAGGTTCGTC-3'),利用此引物对包括可可球二孢菌在内的21个菌株基因组DNA进行PCR扩增,结果只有4个可可球二孢菌菌株扩增到422bp特异带,其它17个菌株无扩增产物。灵敏度测试结果表明此特异引物能对1pg的可可球二孢菌基因组DNA进行扩增。对自然感染黑腐病的香蕉果实组织和接种可可球二孢菌或多种香蕉病原真菌混合接种的果实组织进行检测,Bth-S和Bth-A引物对不仅能够在自然感染黑腐病果实组织中特异检测到可可球二孢菌,而且能在未显症和发病的接菌香蕉果实组织中特异检测得到可可球二孢菌。这为香蕉可可球二孢菌潜伏侵染检测提供了技术支持。     

Population differentiation in the banana leaf spot pathogen Mycosphaerella musicola, examined at a global scale

Single-copy restriction fragment length polymorphism (RFLP) markers were used to determine the genetic structure of the global population of Mycosphaerella musicola , the cause of Sigatoka (yellow Sigatoka) disease of banana. The isolates of M. musicola examined were grouped into four geographic populations representing Africa, Latin America and the Caribbean, Australia and Indonesia. Moderate levels of genetic diversity were observed for most of the populations ( H  = 0·22–0·44). The greatest genetic diversity was found in the Indonesian population ( H =  0·44). Genotypic diversity was close to 50% in all populations. Population differentiation tests showed that the geographic populations of Africa, Latin America and the Caribbean, Australia and Indonesia were genetically different populations. Using F _ST tests, very high levels of genetic differentiation were detected between all the population pairs ( F _ST > 0·40), with the exception of the Africa and Latin America-Caribbean population pair. These two populations differed by only 3% ( F _ST = 0·03), and were significantly different ( P  < 0·05) from all other population pairs. The high level of genetic diversity detected in Indonesia in comparison to the other populations provides some support for the theory that M. musicola originated in South-east Asia and that M. musicola populations in other regions were founded by isolates from the South-east Asian region. The results also suggest the migration of M. musicola between Africa and the Latin America-Caribbean region.     

Genetic variability of Pseudocercospora fijiensis,the black Sigatoka pathogen of banana (Musa spp.) in Mexico

Pseudocercospora (previously known as Mycosphaerella) fijiensis causes black Sigatoka disease in banana (Musa spp.) and is considered to be the most devastating pathogen of this crop worldwide. To improve knowledge of its evolutionary patterns, this study determined the genetic variability of populations from two regions of Mexico: Central Pacific (Colima and Michoacan) and Southern (Chiapas, Tabasco and Oaxaca), using 10 simple sequence repeat (SSR) loci and the MAT-specific PCR assay. Both mating types were present in all regions under study, with frequencies of 63% MAT1-1 and 37% MAT1-2. The SSR markers showed an average of three alleles per locus, resulting in 34 alleles in total. The genetic diversity (H_T) was 0.3308, but at the local level (H_S) ranged from 0.0976 (Colima) to 0.2228 (Oaxaca). However, the genotypic diversity was usually high (H′ > 2.4, S > 0.89). Cluster analysis grouped the isolates into five clusters with high statistical support (au > 80%), suggesting a geographic organization of the genetic variability of P. fijiensis; AMOVA, the minimum spanning tree and the population structure analysis supported this result, and all data indicated that the major genetic differences were between the different populations under analysis. Thus, the high level of genetic variability in P. fijiensis is attributed partly to a high rate of sexual reproduction, and also to a strong evolutionary capacity coupled with isolation due to limited genetic flow between distant populations. Both possibilities could be playing a relevant role in population differentiation of the pathogen.     

Distribution of Pseudocercospora species causing Sigatoka leaf diseases of banana in Uganda and Tanzania

Sigatoka leaf diseases are a major constraint to banana production. A survey was conducted in Tanzania and Uganda to assess the distribution of Pseudocercospora species and severity of Sigatoka leaf diseases. Pseudocercospora species were identified using species-specific primers. Sigatoka-like leaf diseases were observed in all farms and on all cultivars, but disease severity varied significantly (P < 0.001) between countries, districts/regions within countries, altitudinal ranges and banana cultivars. In all regions except Kilimanjaro, P. fijiensis, the causal agent of black Sigatoka, was the only pathogen associated with Sigatoka disease. Mycosphaerella musae was associated with Sigatoka-like symptoms in Kilimanjaro region. Black Sigatoka disease was more severe in Uganda, with a mean disease severity index (DSI) of 37.5%, than in Tanzania (DSI = 19.9%). In Uganda, black Sigatoka disease was equally severe in Luwero district (mean DSI = 40.4%) and Mbarara district (mean DSI = 37.9%). In Tanzania, black Sigatoka was most severe in Kagera region (mean DSI = 29.2%) and least in Mbeya region (mean DSI = 11.5%). Pseudocercospora fijiensis, the most devastating sigatoka pathogen, was detected at altitudes of up to 1877 m a.s.l. This range expansion of P. fijiensis, previously confined to altitudes lower than 1350 m a.s.l. in East Africa, is of concern, especially for smallholder banana farmers growing the susceptible East African Highland bananas (EAHB). Among the banana varieties sampled, the EAHB, FHIA hybrids and Mchare were the most susceptible. Here, the loss of resistance in Yangambi KM5, a banana variety previously resistant to P. fijiensis, is reported for the first time.     

Relationship between the Spray Droplet Density of Two Protectant Fungicides and the Germination of Mycosphaerella fijiensis Ascospores on Banana Leaf Surfaces

The effect of fungicide spray droplet density (droplet cm^-2), droplet size, and proximity of the spray droplet deposit to fungal spores was investigated with Mycosphaerella fijiensis ascospores on the banana (Musa AAA) leaf surface for two contact fungicides: chlorothalonil and mancozeb. When droplet size was maintained at a volume median diameter (VMD) of 250 μm while total spray volume per hectare changed, M. fijiensis ascospore germination on the leaf surface fell below 1% for both fungicides at a droplet deposit density of 30 droplet cm^-2. At a droplet deposit density of 50 droplet cm^-2, no ascospores germinated in either fungicide treatment. When both droplet size and droplet cm^-2 varied while spray volume was fixed at 20 litre ha^-1, ascospore germination reached 0% at 10 droplet cm^-2 (VMD=602 μm) for both fungicides. At lower droplet densities (2–5 droplet cm^-2 VMD=989 μm and 804 μm respectively), ascospore germination on the mancozeb-treated leaves was significantly lower than on the chlorothalonil-treated leaves. The zone of inhibition surrounding a fungicide droplet deposit (VMD=250 μm) on the leaf surface was estimated to extend 1·02 mm beyond the visible edge of the spray droplet deposit for chlorothalonil and 1·29 mm for mancozeb. The efficacy of fungicide spray droplet deposit densities which are lower than currently recommended for low-volume, aerial applications of protectant fungicides was confirmed in an analysis of leaf samples recovered after commercial applications in a banana plantation. Calibrating agricultural spray aircraft to deliver fungicide spray droplets with a mean droplet deposit density of 30 droplet cm^-2 and a VMD between 300 and 400 μm will probably reduce spray drift, increase deposition efficiency on crop foliage, and enhance disease control compared to aircraft calibrated to spray finer droplets. © 1997 SCI.     

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.     

Sources of resistance to Pseudocercospora fijiensis,the cause of black Sigatoka in banana

Black Sigatoka, caused by Pseudocercospora fijiensis, is one of the most devastating diseases of banana. In commercial banana-growing systems, black Sigatoka is primarily managed by fungicides. This mode of disease management is not feasible for resource-limited smallholder farmers. Therefore, bananas resistant to P. fijiensis provide a practical solution for managing the disease, especially under smallholder farming systems. Most banana and plantain hybrids with resistance to P. fijiensis were developed using few sources of resistance, which include Calcutta 4 and Pisang Lilin. To broaden the pool of resistance sources to P. fijiensis, 95 banana accessions were evaluated under field conditions in Sendusu, Uganda. Eleven accessions were resistant to P. fijiensis. Black Sigatoka symptoms did not progress past Stage 2 (narrow brown streaks) in the diploid accessions Pahang (AA), Pisang KRA (AA), Malaccensis 0074 (AA), Long Tavoy (AA), M.A. Truncata (AA), Tani (BB), and Balbisiana (BB), a response similar to the resistant control Calcutta 4. These accessions are potential sources of P. fijiensis resistance and banana breeding programmes can use them to broaden the genetic base for resistance to P. fijiensis.     

香蕉叶鞘腐败病病原鉴定

 香蕉叶鞘腐败病为近年来发生在广东、广西、海南等地蕉园的一种新病害。经病原菌分离及柯赫氏法则检测,证明引起该病的病原菌为一种细菌,根据致病性、寄主范围测定、菌体形态、培养特性、生理生化反应及16S rDNA序列分析将该病原细菌初步鉴定为成团泛菌(Pantoea agglomerans)。成团泛菌危害香蕉,引起叶鞘腐败病在国内尚属首次报道。     

柑桔黄龙病病原和检测方法研究进展

介绍了柑桔黄龙病病原认识过程、病害田间诊断、指示植物鉴定、病原显微镜观察、血清学、DNA-DNA杂交和PCR检测等方法。