马铃薯疮痂病菌致病相关基因的克隆及表达

 A pathogenic-related gene nec1 was cloned in Streptomyces scabies CPS-1, potato scab strain. Analysis results showed that the length of open reading frame(ORF) for nec1 gene was 666 bp, and the GC content was 54.2%. Sequence alignment indicated that a 650 bp up-stream sequence shared 91% similarity with IS 256 family transposase nucleotide sequences by BLASTn searches against GenBank. The segments obtained by PCR amplification were digested by enzymes SphⅠand SacⅠ, and linked to the expression vector pIJ702. The recombinants were transformed into nonpathogenicity strain Streptomyces lividans 66 TK24. Bioas-say results suggested that the transformants possessed the same symptoms as pathogenic strain on potato tuber slices and radish seedlings, which implied that nec1 gene was associated with the pathogenicity of S. scabies CPS-1.     

马铃薯疮痂病菌毒素及其致病性的研究

 本文对分离自我国不同地区的马铃薯疮痂病菌产生的毒素进行了研究。通过对不同菌株进行毒素提取、纯化和产毒分析发现各致病菌株均能产生同一类毒素,而非致病菌株均未见毒素产生。毒素的活性检测结果表明,毒素能使马铃薯薯片和萝卜幼苗产生与病菌作用相同的症状,说明毒素是疮痂病菌侵染过程中的主要致病因子。     

黑龙江省马铃薯干腐病菌种类鉴定及致病性

本研究将采自黑龙江省不同地区的马铃薯干腐病病样进行分离和病原菌纯化,得到27个镰刀菌菌株,通过致病性鉴定,其中的18个菌株具有致病性。运用培养性状和形态特征综合分析的方法,对上述18个菌株进行鉴定,结果显示为6种镰刀菌,分别为拟枝孢镰孢(Fusarium sporotrioides)、茄镰孢(F.solani)、接骨木镰孢(F.sam-bucinum)、拟丝孢镰孢(F.trichothecioides)、燕麦镰孢(F.avenaceum)和茄病镰孢蓝色变种(F.solanivar.coerule-um)。同时对上述6种镰刀菌进行致病性测定,结果表明不同种类镰刀菌致病性不同,以接骨木镰孢、燕麦镰孢和拟丝孢镰孢致病力最强,拟枝孢镰孢致病力最弱。     

我国马铃薯疮痂病及其防治研究进展

马铃薯疮痂病(potato common scab)是由放线菌目链霉菌属的链霉菌Streptomyces spp.引起的土传兼种传病害,广泛分布于世界各马铃薯种植区,不仅影响马铃薯的外观品质和销售价格,严重时还会导致马铃薯出苗延迟甚至引起幼苗死亡,造成产量下降,给马铃薯产业带来巨大的经济损失,已经成为全球危害马铃薯生产的第四大病害。2015年我国确立马铃薯主粮化战略,推动了马铃薯产业的发展。近年来,随着种植区域和规模不断扩大,马铃薯疮痂病在我国很多省(自治区)有不同程度的发生,并有逐年扩大和加重的趋势,严重影响商品薯、加工原料薯和种薯的生产,成为制约我国马铃薯生产的主要病害。本文对马铃薯疮痂病症状、发病因素、传播规律、致病机理、分类方法以及我国马铃薯疮痂病发生情况、种类及分布进行归纳,并对马铃薯疮痂病防治措施进行总结,以期为我国马铃薯疮痂病的研究和防治奠定理论基础。     

云南省马铃薯疮痂病致病链霉菌种类组成研究

 为明确云南省马铃薯疮痂病病原菌(Streptomyces spp.)的种类及其生物学特征,自2013年从云南省13个马铃薯主产区采集疮痂病病样,共分离到200株链霉菌,通过温室盆栽致病性试验筛选出67株致病菌。通过形态学、生理生化指标、致病性测定及16S rDNA序列分析对获得的菌株进行鉴定。结果显示,引起云南地区马铃薯疮痂病的病原为10种链霉菌,分别为S. caviscabies、S. anulatus、S. scabies、S. turgidiscabies、S. acidiscabies、S. europaeiscabiei、S. luridiscabiei、S. enissocaesilis、S. griseus和S. aureofaciens。其中S. enissocaesilis和 S. anulatus为优势种群,S. caviscabies、S. anulatus和S. luridiscabiei为国内首次报道的病原菌。因此,认为云南省马铃薯疮痂病菌种类复杂多样。     

甘肃马铃薯疮痂病病原初步鉴定

对甘肃不同地区马铃薯疮痂病病原进行了分离鉴定.采用盆栽方法进行致病性测定、形态特征、生理生化特性测定和16S rDNA序列分析.结果表明:共有6株菌能使马铃薯块茎出现疮痂症,分别为GP-1、GP-2、GH-1、GH-2、JB-1和JB-2.经鉴定菌株GP-1、JB-2的形态特征、生理生化特性及16S rDNA序列与Streptomyces scabies 87 22菌株均一致;GP-2、JB-1的16S rDNA序列与S.scabies菌株相似性为99％.菌株GH-2和GH-1与S.griseus的16S rDNA序列相似性分别为100％和99％,形态特征一致,但不能以棉籽糖和肌醇为单一碳源,暂定为S griseus.     

西北地区马铃薯疮痂病病原菌鉴定及其生物学特性

为明确西北地区马铃薯疮痂病病原菌的种类和生物学特性,分别采用常规组织分离法和土壤混悬液分离法从宁夏、陕西和甘肃3个省区采集的29份疮痂病发病薯块和8份发病地块土壤中进行病原菌分离,并利用形态特征、生理生化特性和16S rDNA序列分析对病原菌进行鉴定。结果表明,从发病薯块和发病土壤中共分离到50株链霉菌Streptomyces spp.,通过回接法验证获得6株马铃薯疮痂病致病菌株。6株致病菌株的培养特性和形态特征差别较大;其中菌株G4-1、G9和SYN13不能以果糖和木糖为单一碳源,菌株SYNT3不能以棉子糖为单一碳源;除菌株NLG4-1外,其余5株菌株均能在络氨酸琼脂培养基上产生黑色素。经16S rDNA序列分析,菌株G4-1、G9与疮痂病链霉菌S. scabiei的相似率分别达99.47%和99.34%,菌株NLG4-1、SYNT3与S. enissocaesilis的相似率分别达97.90%和98.18%,菌株GBH2与加利利链霉菌S. galilaeus的相似率达99.93%,菌株SYN13与S. turgidiscabies的相似率达97.56%,表明西北地区马铃薯疮痂病病原菌至少存在4个种。     

义乌市马铃薯疮痂病发生原因及防控措施

马铃薯疮痂病已成为威胁义乌市马铃薯生产的最主要病害。当田间有大量病原菌存在时,该病的发生程度与土壤酸碱度、土壤湿度、品种抗性等因素密切相关。选用抗病品种、水旱轮作、施用酸性肥料、调节土壤酸碱度、利用太阳能杀灭土壤病菌,块茎生长期间保持土壤湿度、选用无病种薯并进行消毒、防治好地下害虫等防控措施,可有效地抑制疮痂病的发生。     

马铃薯疮痂病菌在植株和田间的分布与动态分析

为探明马铃薯疮痂病菌在植株和土壤中的分布情况及种群动态变化特点,利用常规PCR和定量PCR(qPCR)技术对不同环境的马铃薯疮痂病株和田间植株不同生育期的土壤样品进行病原菌的定性定量检测.结果 表明,病田、温室盆栽和微型薯苗床中马铃薯疮痂病重度发病植株的根、匍匐茎、块茎、地上茎、叶片等组织样品均可检测到184 bp的疮...     

甘肃省定西市马铃薯疮痂病新病原Streptomyces galilaeus的分离、鉴定及生物学特性研究

对甘肃省定西市安定区的马铃薯疮痂病病原进行了分离、鉴定和生物学特性研究。结果表明,菌株5T-1具有较强致病性,菌落表面呈灰色,有金属光泽,平均直径为4.68mm,可产生黄褐色素,孢子圆形或圆柱形,孢子丝松散,革兰氏染色呈阳性。5T-1的16SrDNA序列与加利利链霉菌Streptomyces galilaeus菌株的相似度为99%,结合形态特征将菌株5T-1鉴定为Streptomyces galilaeus,为甘肃省新报道病原菌。菌株5T-1生长最适温度为30℃,最适光照条件为全黑暗,最适pH为8.5,最佳碳源和氮源分别为肌醇和天冬氨酸。该研究结果为甘肃省马铃薯疮痂病诊断和综合防治提供了依据。     

Detection and characterization of Streptomyces causing potato common scab in Western Europe

A PCR-based diagnostic method was developed for direct detection from tuber lesions of pathogenic Streptomyces causing common scab of potato. Primers were designed to amplify a fragment of the txtAB ( txtA and txtB ) genes, which are pathogenicity determinants in the main pathogenic Streptomyces species. The method was evaluated on 84 naturally infected potato samples, comprising 19 potato cultivars, harvested in the years from 2000 to 2004 in the Netherlands, the UK, France, Germany and Spain. Pathogenic Streptomyces in tuber lesions were detected by PCR in 70 samples and were also successfully isolated from these 70 samples. All pathogenic isolates showed the basic general phenotypic traits of the S. scabiei phenetic cluster. RFLP analysis of amplified rRNA sequences, together with carbon source utilization and repetitive BOX profiles, allowed most isolates to be assigned to S. europaeiscabiei , which emerged as the main cause of potato common scab in Western Europe.     

Phytotoxin Production and Aerial Mycelium Formation by Streptomyces scabies and S. acidiscabies in Vitro

Culture conditions (pH, Ca²⁺, phosphate, and temperature) on phytotoxin production (thaxtomin A and concanamycins A and B) and aerial mycelium formation were examined for two strains each of Streptomyces scabies and S. acidiscabies. Thaxtomin production decreased at pH 7.5 and increased at 15°C in S. scabies and increased at 30 mM phosphate in S. acidiscabies. Concanamycin production was stimulated by the addition of CaCl₂ and was correlated with formation of aerial mycelium. Aerial mycelium formation in S. scabies was greater the higher the pH, Ca²⁺ concentration and temperature, whereas that in S. acidiscabies was unaffected. Received 2 November 2000/ Accepted in revised form 27 April 2001     

拮抗链霉菌防治马铃薯疮痂病的大田试验研究

 本文报道了利用拮抗链霉菌防治马铃薯疮痂病的大田试验研究。拮抗链霉菌接种在0.8公顷的病田中。不同接种量和接种方法均显著地影响防治效果。3年的试验表明:拮抗菌系以蛭石接种形式防效优于种薯浸泡。一般情况下,连续2年接种或高接种量的防效优于1年或低接种量的防效。不同菌系的防效差异明显,单个拮抗菌的防效优于2个拮抗菌混合使用的效果。甜玉米和大豆轮作结合拮抗菌显著降低了马铃薯疮痂病的危害。     

Occurrence and survival of potato scab pathogens (Streptomyces species) on tuber lesions: quick diagnosis based on a PCR-based assay

A time-saving and cost-effective polymerase chain reaction (PCR)-based method was developed for species-specific detection of the scab pathogens ( Streptomyces scabies and S. turgidiscabies ) prevalent in potato ( Solanum tuberosum ) in northern Scandinavia. Species specificity of primers was verified using a collection of previously characterized Streptomyces strains isolated from potato scab lesions in Finland and Sweden. A total of 1245 scab lesions was tested from potato cvs Matilda and Sabina grown in the field in two geographic regions of Finland in 2000 and 2001. Freshly harvested or stored potato tubers were incubated at room temperature (18–21°C) under humid conditions for a few days. Bacterial growth was collected from scab lesions for DNA isolation and PCR. The two scab pathogens were detected in the same potato fields, tubers and scab lesions. The relative incidence of S. scabies was high in freshly harvested tubers but was much lower than that of S. turgidiscabies following storage. Both pathogens were seed-transmitted in Matilda and Sabina after 24 weeks of storage at 4°C.     

Cumulative and residual effects of different potato cropping system management strategies on soilborne diseases and soil microbial communities over time

Soilborne potato diseases and soil microbial community characteristics were evaluated over 8 years in different potato cropping systems designed to address specific management goals of soil conservation, soil improvement and disease suppression. Results were compared to a standard rotation and non‐rotation control in field trials in Maine. Standard rotation consisted of barley underseeded with red clover, followed by potato (2‐year). Soil‐conserving system (SC) featured an additional year of forage grass and reduced tillage (3‐year, barley/timothy–timothy–potato). Soil‐improving system (SI) added yearly compost amendments to SC, and the disease‐suppressive system (DS) featured crops with known disease‐suppressive capability (3‐year, mustard/rapeseed–sudangrass/rye–potato). Systems were established in 2004, evaluated with and without irrigation, and actively managed until 2010, with potato also planted in 2011 and 2012 to examine residual effects. All rotations reduced soilborne diseases black scurf and common scab, and increased yield after one rotation cycle (3 years), but diseases increased overall after two rotation cycles. DS maintained lower soilborne disease levels than all other rotations, as well as high yields, throughout the study. Cropping system effects became more pronounced after multiple cycles. SI system and irrigation both resulted in higher yields, but also higher levels of soilborne disease. Cropping system and irrigation effects were significant even after systems were no longer maintained. Soil microbial community data showed significant changes associated with cropping system, and differences increased over time. Cropping system strategy had significant and lasting effects on soil microbiology and soilborne diseases, and can be used to effectively enhance potato production.     

Elimination of common scab sensitive progeny from a potato breeding population using thaxtomin A as a selective agent

Common scab is one of the most important soil‐borne diseases of potato and is difficult to control. Selection of potato breeding lines for resistance to common scab is also cumbersome due to environmental factors influencing symptom development and an erratic spatial distribution of the scab pathogens (Streptomyces spp.) in the field. The bacterial phytotoxin thaxtomin A, which causes scab symptoms, can be used to screen large numbers of potato seedlings for tolerance in vitro, but few studies have investigated whether the results correspond to resistance to common scab observed in the field. In this study, 120 F1 potato progeny from a single cross were screened in vitro by exposing the seedlings to thaxtomin A added to the culture medium. Eighteen genotypes were selected based on high sensitivity or tolerance using shoot growth as the criterion, multiplied in vitro, and tested for resistance to common scab caused by S. turgidiscabies and S. scabies in a glasshouse and in three different fields. Evaluation of ca. 6500 tubers showed that the 18 potato genotypes differed in scab indices and disease severity (P < 0·0001). The relative shoot height in vitro (thaxtomin A used at 0·5 μg mL^?1) and the scab index in the field showed significant correlation (r_s = ?0·463, P = 0·0528, n = 18), also consistent with the results obtained under controlled conditions in the glasshouse. Hence, the in vitro bioassay may be used to discard scab‐susceptible genotypes and elevate the overall levels of common scab resistance in the potato breeding populations.