辣椒细菌性叶斑病病原鉴定及室内药剂筛选

2012-2015年,在海南省辣椒主产区发生了一种严重危害辣椒的叶斑病,从发病叶片中分离到一种细菌,通过致病性测定、柯赫氏法则验证及分子生物学鉴定,将引起该病害的病原鉴定为野油菜黄单胞菌辣椒斑点病致病变种(Xanthomonas campestris pv.vesicatoria)。选用8种杀菌剂通过平板对峙法对该病原进行了室内毒力测定,结果表明:其中6种药剂对该病原有抑菌效果,效果最好的为72%农用硫酸链霉素可湿性粉剂,47%春雷·王铜可湿性粉剂次之,2%春雷霉素水剂和20%噻菌铜悬浮剂没有抑菌效果,不同药剂之间或同一种药剂不同剂型之间抑菌效果差异较大。病原鉴定和药剂筛选可为该病害的诊断和防治提供科学依据。     

番茄灰叶斑病原菌的鉴定及抗性种质资源的筛选

<正>番茄灰叶斑病是一种世界性病害,近年来,该病在北京、台湾、海南、山东和浙江等地均有发生,极大地限制了我国番茄生产的竞争力~([1])。该病害主要由Stemphylium spp.不同的4个种引起~([2])。本研究采用形态观察法、ITS和gpd序列分析法对从番茄病株分离得到的病原菌进行鉴定,并对番茄灰叶斑病抗性鉴定方法进行研究,以期为番茄灰叶斑病的抗病育种研究及其利用提供理论依据。     

新疆加工型辣椒细菌性斑点病的发生和病原鉴定

 新疆加工型辣椒主要产区巴音郭楞蒙古自治州发生了一种严重危害辣椒的细菌性病害。从发病辣椒叶片中分离细菌,通过烟草过敏性反应、马铃薯软腐试验和接种辣椒等致病性测定,确定了13个致病菌株,各菌株之间致病力无明显差异。通过菌体形态、培养性状观察、生理生化反应、寄主范围测定,结合16S rDNA和rpoD基因扩增、序列测定和系统发育分析,将病原菌鉴定为丁香假单胞菌丁香致病变种(Pseudomonas syringae pv. syringae)。病原菌人工接种还能侵染番茄、茄子、马铃薯及黄瓜、四季豆、白菜、萝卜、芹菜等植物。P. syringae pv. syringae引起加工型辣椒细菌性斑点病在国内属首次报道。     

辣椒细菌性疮痂病病原菌分类、检测及综合防治研究进展

辣椒细菌性疮痂病是一种世界性分布的细菌性病害,该病能引起辣椒严重的产量损失和品质下降。国外特别是美国对该病害研究较早且较深入,国内相关研究几乎是空白。本文主要围绕病原菌的分类、检测和病害综合防治等研究进展做一概述。     

番茄棒孢叶斑病病原鉴定及生物学特性研究

本研究采用PSA培养基对上海地区的番茄棒孢叶斑病的病原菌进行分离、培养及致病性测定,通过形态学和分子生物学方法鉴定病原菌,并对该病菌的生物学特性进行研究。结果表明:番茄棒孢叶斑病病原菌菌落呈灰褐色,菌丝质地致密、绒毛状;分生孢子单生或串生,圆柱形或倒棍棒形、顶端钝圆,基部平截,呈半透明至浅褐色,假隔膜4～10个,直或稍弯曲,基脐加厚,深褐色。结合病原菌rDNA ITS序列测定、比对后,确定该病原为多主棒孢Corynespora cassiicola。该病菌的菌丝生长最适温度为25～30℃,最适pH范围为4～8,产孢的最适温度约为25℃,最适pH范围为5～9,麦芽糖和乳糖为碳源的培养基适宜菌丝生长,可溶性淀粉和乳糖溶液中孢子萌发率最高,光暗交替条件适宜菌丝生长,孢子在水滴中极易萌发。这是该病在上海地区的首次报道。     

Selective detection of Pseudomonas syringae pv. tomato using dot blot hybridization and real-time PCR

Two rapid detection methods based on dot blot hybridization with a nonradioactive DNA probe and molecular beacon-PCR were developed for the specific detection of Pseudomonas syringae pv . tomato , the causal agent of bacterial speck of tomato. A 1378 bp DNA fragment (Acc. No. AM039892), obtained from the extension of a 255 bp fragment generated by a RAPD protocol, was used to find a suitable combination of primers specific for the tomato pathovar. A 138 bp fragment from the genome of P. syringae pv. tomato DC 3000 was used as DNA probe. In dot blots of DNA extracted from either pure cultures or artificially contaminated seeds washes, the probe recognized specifically the tomato pathovar. A molecular beacon was designed from the same region for the specific detection and quantification of P. syringae pv . tomato by real-time PCR. A highly significant correlation was observed between the amount of target DNA and the cycle threshold (Ct). Using a fast protocol for DNA extraction, from pure cultures and from washes of artificially contaminated seeds, the limit of detection was about 1 × 10² CFU. The diagnostic tools developed proved highly specific for P. syringae pv. tomato and simple to use. They can therefore be applied to large-scale testing of tomato seeds and seedlings for the assessment of their phytosanitary condition in nurseries.     

Monitoring the occurrence of tomato bacterial spot and range of the causal agent Xanthomonas perforans in Iran

A 2‐year comprehensive field survey was conducted across major tomato‐growing areas of Iran. Two hundred and thirty‐four tomato fields and six tomato‐producing greenhouses were surveyed for the potential presence of bacterial spot disease. Five hundred and ninety‐six tomato samples with and without symptoms were analysed. While Xanthomonas spp. were found in association with tomato plants both with and without symptoms from five surveyed counties, the bacterial spot disease was observed only in plants from three of them. Only strains isolated from plants with symptoms induced disease symptoms on tomato, while those isolated from symptomless plants caused symptoms only on cabbage and common bean. None of the isolates caused disease symptoms on pepper and eggplant. Phylogenetic analysis showed that X. perforans is the causal agent of tomato bacterial spot in Iran, although X. campestris and X. axonopodis were also associated with symptomless tomato plants. All X. perforans isolates in this study were sensitive to streptomycin, copper sulphate and copper oxychloride at concentrations of 50 mg L^?1, 200 mg L^?1 and 0.8 g L^?1, respectively. Unlike the type strain of X. perforans, isolates in this study did not produce bacteriocin against other Xanthomonas spp., nor were they detected using the usual species‐specific primer pair Bs‐XpF/Bs‐XpR. This suggests an atypical nature of X. perforans strains in Iran, which leads to the hypothesis that X. perforans strains in Iran may have a separate origin to those causing disease epidemics elsewhere. The aggregated dispersal pattern of the diseased tomato fields signifies the seedborne introduction of the pathogen into the country.     

Comparison of extraction procedures and determination of the detection threshold for Clavibacter michiganensis ssp. michiganensis in tomato seeds

Several seed extraction procedures, used for detection of Clavibacter michiganensis ssp. michiganensis ( Cmm ) in naturally infected and artificially infested tomato seed lots were evaluated. Extraction methods that included grinding the seeds were significantly better at detecting the pathogen in three different seed lots than methods that used only soaking. The detection threshold of Cmm in relation to seed sample size was determined by adding naturally infected seeds into samples of three different sizes. Cmm was detected by agar plating assay, on three media (CNS, mSCM, D₂ANX), and by direct PCR from seeds and Bio-PCR (bacteria cultured on agar media prior to PCR). In samples of 10 000 seeds containing one infected seed, Cmm could be detected only by Bio-PCR and in only one replicate out of five. In samples containing five or 10 infected seeds per 10 000 seeds, three of five and five of five replicates, respectively, were detected by the three detection methods. In samples of 5000 seeds, one infected seed could be detected in all five replicates only after adding a concentration step. A high correlation ( R ² = 0·9448) between artificially infested seeds and the disease incidence was found. Seed lots infested with less than 58 colony-forming units (CFU) per g did not cause disease under glasshouse conditions, whereas lots with about 1000 CFU g⁻¹ caused disease in 78 plants out of 2000.     

海南槟榔细菌性叶斑病病原鉴定

In 2008, a severe bacterial disease was observed in Wenchang, Hainan Province. Symptoms consisted of small circular to elongated brown lesions surrounded by yellow halos. All isolates, which formed white colonies, were gram-negative and each had a single, polar, sheathed flagellum. Isolates were oxidase and catalase positive, negative for arginine dihydrolase, gelatin hydrolysis and starch hydrolysis, and acid production from levan. Sequences of the 16S rDNA gene of the isolates shared 98% sequence identity with that of Burkholderia andropogonis strain LMG2129 (GenBank accession No. NR118985). Pathogenicity tests were conducted and fulfilled Koch's postulates, indicating that these isolates are causative agent of the disease. The results led to the conclusion that the causal pathogen of betel palm bacterial leaf spot was B. andropogonis. It is the first report of B. andropogonis causing bacterial leaf spot of betel palm in mainland China.     

黑龙江省水稻品种资源对水稻细菌性褐斑病的抗性鉴定

选育和应用抗病品种是防治水稻细菌性病害最经济有效的措施。本研究采用苗期喷雾法和针刺法对黑龙江省42个水稻主栽品种进行抗性鉴定。结果表明,利用针刺法鉴定的各水稻品种中表现中抗以上的品种有14个,其中高抗品种2个,对接种水稻品种病斑长度进行差异显著性分析,42个水稻主栽品种间抗性存在显著差异;利用喷雾法鉴定各品种,病级表现1级的品种有7个,占16.7%。对数据进行比较分析,针刺法和喷雾法对抗病品种鉴定的结果基本一致。本研究鉴定的2个高抗稻种资源对于抗细菌性褐斑病生产实践提供重要价值。因此,加大对水稻品种资源的深入研究,对实现水稻细菌性褐斑病的可持续控制有着重要意义。     

玉米灰斑病菌的可溶性蛋白质及同工酶多态性

利用聚丙烯酰胺凝胶电泳技术,对采自北方玉米主产区的23个玉米灰斑病菌菌株进行可溶性蛋白质和同工酶电泳图谱分析及聚类分析,从蛋白质和酶学的多态性水平上分析玉米灰斑病菌的生理分化特征.研究表明,玉米灰斑病菌在可溶性蛋白质和SOD、MDH、PPO、POD、EST、CAT等的同工酶谱存在差异,不同菌株之间某些同工酶谱带数和同一迁移率谱带的亮度和色泽差异非常显著,说明菌株间的多态性可在同工酶水平上得到反映.研究还发现,来自不同地区的菌株同工酶谱带无明显的变化规律,反映出病菌同工酶的变异与地理位置关系不密切,也表明该病菌可能具有较广泛的地域适应性.     

玉米麦根腐平脐蠕孢叶斑病的发生与病原鉴定

在河北省春播玉米WN2000和山东省夏播普通玉米上发现一种叶斑病,其病斑圆形或椭圆形,中心灰白色,边缘褐色,有黄褐色褪绿圈,糯玉米WN2000病斑较大,(3~10)mm×(3~5)mm;普通玉米病斑较小,(1~2)mm×(2~3)mm。通过病原菌分离培养、致病性测定、形态特征观察及r DNA-ITS和1,3,8-三羟基萘还原酶基因(3HNR)序列分析,证明两种大小病斑是由同一种病原菌—麦根腐平脐蠕孢(Bipolaris sorokiniana)引起。田间调查结果表明,在山东省夏播玉米各品种苗期叶片上均有不同程度的发病,发病率最高的为冠玉6号,病株率52%,最低的为五岳88,病株率2%;鲜食玉米仅在河北春播玉米WN2000上发生,发病率为32%。麦根腐平脐蠕孢侵染不同玉米品种产生两种大小不同的病斑在国内外尚属首次报道。     

辣椒褐斑病菌生物学特性研究

本文对辣椒褐斑病病原菌Cercospora capsici Heald et Wolf的生物学特性进行了研究,结果表明,不同的营养、温度、pH等条件对菌丝生长和孢子萌发有显著影响。病菌菌丝的生长以PDA培养基为最适;适宜温度为20~25℃,最适温度为25℃;最适pH为8.0~9.0,光照对菌丝生长没有明显的促进作用,菌丝致死温度及时间为55℃ 10 min。分生孢子萌发适宜碳源为1%的蔗糖溶液,适宜氮源为1%的甘氨酸溶液;孢子萌发适宜温度为20~30℃,最适温度25℃;最适pH为5~6,光照对孢子萌发没有明显的促进作用,分生孢子致死温度及时间为52℃10 min。     

番茄叶片漆腐病病原菌鉴定

 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.     

茭白胡麻叶斑病病原菌分离与鉴定

茭白胡麻叶斑病发生普遍且严重,已成为茭白的常见病害。为了明确病原菌,先后从茭白主产区采集胡麻叶斑病病叶,各产区分离纯化5～10株病原菌。通过对菌株DNA的内转录间隔区ITS基因、3-磷酸甘油醛脱氢酶GPDH基因、延伸因子EF-1α基因的序列进行分析。结合形态学观察、致病力测定等方法对茭白胡麻叶斑病病菌进行鉴定。结果表明,茭白胡麻叶斑病病原菌为稻平脐蠕孢Bipolaris oryzae,与水稻胡麻叶斑病病原菌一致。本文的研究成果可为茭白胡麻叶斑病的防治提供一种新的思路。     

甘草叶斑病的发生与病原菌鉴定

近年来,在甘肃省兰州市及渭源地区甘草产区发现一种甘草新病害—甘草叶斑病。2006－2008年对其发病情况进行了调查,甘草叶斑病在6月下旬开始发病,9月中旬为发病高峰期,发病率达80%。高燥地发病率低于低洼地,采用轮作可减轻甘草叶斑病的发生。经过对甘草叶斑病病原菌形态、培养性状和致病性等方面的测定表明,病原菌为豆链格孢（Alternaria azukiae）。通过寄主范围测定表明,该病原菌亦可侵染独活、羌活等药用植物。对大黄、苍耳等不具侵染性。     

中华常春藤细菌性叶斑病病原菌的鉴定

 2015年春季在昆明市区绿化带种植的中华常春藤上发现一种由细菌侵染而引起的病害,称为中华常春藤细菌性叶斑病。通过发病症状、菌落形态观察、致病性测定、Biolog分析,16S rDNA序列和核糖体DNA内转录间隔区(internal trans-cribed spacer, ITS )序列分析比较,对昆明地区的常春藤叶斑病病原菌及其系统进化关系进行研究。分离病原菌接种中华常春藤叶片完成科赫法则验证,发病初期在叶片表面形成带有黄色叶晕的不规则褐色斑点,后期叶片边缘形成倒V字型坏死并起皱。将菌株CCT1和CCT6测序结果与现有的黄单胞菌菌株的16S rDNA序列和核糖体DNA的ITS序列构建进化树,结果均显示病原菌与野油菜黄单胞菌的序列相似度最大,属于同一支。研究确定该病原菌为野油菜黄单胞菌(Xanthomonas campestris)。这是中国首次报道由X. campestris 引起的中华常春藤叶斑病。     

高粱链格孢叶斑病病原鉴定

正高粱是世界上重要的禾谷类作物之一,仅次于小麦、水稻、玉米和大麦,居第五位~([1])。高粱也是我国最早栽培的作物之一。在生产上,高粱病害是制约产量提高的重要因素。目前己报道的高粱病害有60余种~([2]),较普遍发生的有15种~([3])。2014年,在内蒙古民族大学农场发现一种新病害,在农场种植的     

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。     

西藏八角莲叶斑病鉴定及其生物学特性研究

从西藏农牧学院药材基地的西藏八角莲(Dysosma sayuensis Ying)发病叶片上分离得到病原菌,对病原菌进行形态特征观察、致病性测定及其生物学特性研究。结果表明,八角莲叶斑病为八角莲上一种新病害,其病原菌为壳针孢属(Septoriasp.)真菌。该菌能利用多种碳源和氮源,其中以山梨醇为最适碳源,蛋白胨为最适氮源,在无碳源和氮源条件下,菌丝生长不良,长势较差;菌丝生长最适温度为20～30℃;最适pH为7～9;不同光照条件对菌丝生长无显著影响。本试验明确了引起西藏八角莲叶斑病的病原,为该病的深入研究和防治提供了依据。