小麦赤霉病菌多菌灵抗性群体的扩散路径分析——基于致病菌种类及所产毒素化学型鉴定和抗药性检出的时序性

通过对江苏、安徽、山东、河南、湖北、河北和四川7省小麦赤霉病菌对多菌灵抗性及敏感菌株Fusarium asiaticum和F.graminearum的鉴定、所产生毒素的化学型及多菌灵抗性菌株检出时序性的分析,初步推测了小麦赤霉病菌对多菌灵抗性群体在中国麦区的扩散路径。结果表明:江淮流域的江苏、安徽、湖北3省和四川省小麦赤霉病菌对多菌灵的抗性或敏感菌株优势群体均是F.asiaticum,而黄淮流域的山东、河南2省及河北省小麦赤霉病菌对多菌灵的敏感菌株优势群体为F.graminearum,抗性菌株优势群体则为F.asiaticum。江苏、安徽、山东和河南抗多菌灵菌株F.asiaticum产生毒素的化学型为3-AcDON和NIV,并以3-AcDON为主。江苏省连续使用多菌灵防治小麦赤霉病长达20多年后才检测到田间抗性菌株,而近年来检测到田间抗性菌株的山东、河南2省用多菌灵防治赤霉病的历史较短,且为偶尔使用,药剂的选择压力相对较小,因此推测山东和河南麦区出现的小麦赤霉病菌抗多菌灵菌株可能是通过种子调运及联合收割机跨区作业等方式从抗药性发生较早的江淮麦区流入的。     

山东省小麦赤霉病菌种群组成及其致病力分化

由禾谷镰孢菌群Fusarium graminearum clade引起的赤霉病是小麦的重要病害。为明确山东省小麦赤霉病菌的种群组成及其致病力,于2011年和2012年从山东省15地市分离了95株小麦赤霉病菌,在形态和分子生物学鉴定种的基础上,采用鉴定B型毒素化学型的特异性引物进行毒素化学型分析。在95个菌株中,93株分离物为禾谷镰孢菌F.graminearum,2株为燕麦镰孢菌F.avenaceum。94株分离物为脱氧雪腐镰孢菌烯醇(deoxynivalenol,DON)化学型,1株为雪腐镰孢菌烯醇(nivalenol,NIV)化学型。在94株DON毒素化学型菌株中,90株为15-乙酰脱氧雪腐镰孢菌烯醇(15-acetyldeoxynivalenol,15-AcDON)化学型,4株为3-乙酰脱氧雪腐镰孢菌烯醇(3-acetyldeoxynivalenol,3-AcDON)化学型。在小麦扬花期,采用单花滴注接种法对29个菌株进行了致病力测定,供试菌株的致病力分化明显。表明在山东省冬小麦产区,产15-AcDON毒素的F.gra-minearum是小麦赤霉病菌的优势种群。     

我国小麦茎基腐病菌致病力比较分析

为明确我国不同种、地理来源和毒素化学型小麦茎基腐病菌的致病力分化情况,采用纸塔法对来自全国9个省市80个采样点分离的224株小麦茎基腐病菌进行致病力分析。结果表明,不同种镰刀菌的致病力不同,黄色镰刀菌Fusarium culmorum,禾谷镰刀菌F.graminearum,假禾谷镰刀菌F.pseudograminearum及亚洲镰刀菌F.asiaticum致病力强于其他种。F.culmorum致病力显著高于F.pseudograminearum和F.asiaticum,而F.pseudograminearum,F.graminearum及F.asiaticum三者之间无显著性差异。中华镰刀菌F.sinensis,木贼镰刀菌F.equiseti,锐顶镰刀菌F.acuminatum致病力较弱,三者间苗期致病力无显著性差异;多数省份F.pseudograminearum群体间致病力无显著差异,仅山东F.pseudograminearum群体的致病性显著低于河南群体;此外,产毒类型为3ADON的F.pseudograminearum群体致病力显著高于15ADON群体。     

北京市及河北省小麦赤霉病菌群体遗传结构及生物学特性

本文分析了北京市与河北省小麦赤霉病菌群体遗传结构以及基础生物学特性。结果表明所有菌株均为禾谷镰刀菌(Fusarium graminearum),属于一个大的单一群体,群体内具有较高的遗传多样性。毒素化学型测定表明,北京与河北地区小麦真菌毒素污染的主要风险为DON与15ADON毒素。表型测定显示,与F.asiaticum群体相比,F.graminearum具有较高的产孢能力,而生长速率和产毒能力较低。该群体对主要杀菌剂多菌灵、戊唑醇和氰烯菌酯均无抗药性。     

长江中下游地区亚洲镰刀菌NX-2毒素群体的检测

由禾谷镰刀菌复合种(Fusarium graminearum species complex,FGSC)引起的麦类赤霉病,是农业生产上的重要病害。为明确中国长江中下游冬小麦主产区小麦赤霉病菌种的构成及其地理分布,对2008年从江苏、浙江和湖北3省采集的656株小麦赤霉病菌株进行了分类鉴定。结果显示,其中558个菌株为亚洲镰刀菌(Fusarium asiaticum),98个菌株为禾谷镰刀菌(Fusarium graminearum sensu stricto),表明中国长江中下游冬小麦主产区小麦赤霉病的主要致病菌是亚洲镰刀菌。选择亚洲镰刀菌(F.asiaticum)为研究对象,通过PCR-RFLP的方法对其进行产NX-2毒素菌株的检测。结果没有检测到产NX-2毒素菌株,表明中国长江中下游冬小麦主产区并未出现NX-2毒素群体。本研究旨在了解NX-2毒素群体在中国长江中下游地区的地理分布,为进一步研究麦类赤霉病菌群体遗传多样性和演化趋势奠定基础,为麦类赤霉病的防治和毒素污染的控制提供理论依据。     

Fusarium graminearum sensu stricto和F.asiaticum在玉米秸秆和水稻秸秆上子囊壳形成的比较

为探究麦田前茬作物玉米秸秆和水稻秸秆上中国小麦赤霉病菌的两个优势种Fusarium graminearum sensu stricto (F.graminearum)和F.asiaticum子囊壳形成和发育成熟过程的差异,分别选择3株F.graminearum菌株(SE81,3-ADON型;LcA-2,15-ADON型;HX5-1,NIV型)和2株F.asiaticum菌株(M4A,3-ADON型;M31-2,NIV型),用其孢子悬浮液接种玉米秸秆和水稻秸秆,放置于花泥和地表两种环境条件下,比较接种不同菌株秸秆上子囊壳的形成和发育成熟情况及5个菌株对‘郑麦366’(高感小麦赤霉病品种)的致病力。结果表明：花泥环境下产生子囊壳的速度要快于地表,以玉米秸秆为载体子囊壳产生速度比水稻秸秆快且多数密度大,3个F.graminearum菌株(SE81、LcA-2和HX5-1)在同种秸秆相同环境下比2个F.asiaticum菌株(M4A和M31-2)产生子囊壳的速度快且子囊壳密度大。5个菌株在不同环境的不同秸秆上产生的子囊壳的成熟度均无规律,接种‘郑麦366’后其病情指数间有极显著差异(P<...     

小麦赤霉病菌拮抗菌筛选及最适培养条件初步研究

<正>小麦赤霉病(Fusarium head blight)是我国小麦生产上最重要的真菌病害之一。已有的研究表明,在我国其致病菌的优势种主要为Fusarium graminearum和F. asiaticum。此病害具有爆发速度快、流行范围广等特点[1,2],而且其致病菌还可产生脱氧雪腐镰刀菌烯醇(deoxynivalenol, DON)和玉米赤霉烯酮(zearalenone, ZEN)等毒素,人畜吃过会产生不同程度的中毒反应[3]。目前小麦     

利用Tri8基因区分中国小麦赤霉病菌的毒素化学型

由镰孢菌(Fusarium spp.)引起的小麦赤霉病在全世界均有发生,我国小麦赤霉病主要由禾谷镰孢菌复合种(Fusarium graminearum species complex,FGSC)所引起,根据产生单族毒素种类分为三种化学型,分别为3-乙酰脱氧雪腐镰孢菌烯醇(3-AcDON)、15-乙酰脱氧雪腐镰孢菌烯醇(15-AcDON)和雪腐镰孢菌烯醇(NIV)化学型。为了更方便地对禾谷镰孢菌复合种进行化学型的区分,本研究利用三对引物分别扩增了三种化学型菌株的Tri8基因并进行了序列测定,根据不同化学型菌株的基因序列差异,进行了特异性引物设计和筛选。利用设计的引物同时扩增三种化学型的Tri8基因并用AvaI内切酶进行酶切,根据酶切片段的大小区分三种化学型。研究表明该方法可准确有效地区分我国小麦赤霉病菌的化学型。     

小麦四种镰刀菌毒素的ELISA检测及毒素污染分析

为明确镰刀菌毒素在小麦中的污染与分布状况,选取以四种镰刀菌毒素的特异性抗体制备的ELISA试剂盒,检测了来自全国范围内的183个冬小麦、46个春小麦品种(系)样品中的镰刀菌毒素含量,并对不同麦区小麦样品中脱氧雪腐镰刀菌烯醇(DON)、伏马(fumonisin)、单端孢霉烯醇(T-2)和玉米赤霉烯酮(ZON)四种毒素含量及污染率进行比较分析.结果表明,我国小麦品种(系)中镰刀菌毒素污染比较普遍,但污染水平较低,均符合国家规定标准.四种镰刀菌毒素中以DON毒素为主,其平均含量为73.87μg/kg.不同麦区间DON、fumonisin和T-2毒素的含量存在显著差异(P<0.01),而ZON毒素的含量在麦区间差异不显著(P>0.05),东北春麦晚熟组、长江中下游冬麦组和黄淮冬麦区南片冬水组镰刀菌毒素总体含量较高.DON与ZON毒素污染率在麦区间存在显著差异(P<0.01),而fumonisin与T-2毒素污染率差异不显著(P>0.05).在不同麦区间,东北春麦晚熟组的DON毒素的含量和污染率均为最高.     

河南省首次发现梨孢镰刀菌引起的小麦赤霉病

2015年从河南省田间小麦赤霉病病穗上分离得到一种生长速度较慢的镰刀菌, 通过形态学和分子鉴定明确其分类地位, 通过田间单小花滴注法和喷雾法接种测定其致病力, 并通过高效液相色谱串联质谱分析对麦穗中的毒素种类进行测定, 明确其产毒特征?结果表明:分离得到的8个菌株均为梨孢镰刀菌, 在马铃薯葡萄糖琼脂(PDA)培养基上为白色菌落, 菌落底部产生少量红色色素, 平均生长速度为13.3 mm/d; 小型分生孢子为椭球形葡萄状, 平均大小为7.1 μm×5.8 μm, 未见大型分生孢子和厚垣孢子; 致病力弱, 且不侵染穗轴, 单小花滴注法接种条件下平均病级为0.1, 喷雾法接种条件下平均病小穗率为6.5%; 供试的8个镰刀菌菌株均不产生T-2和HT-2毒素, 均产生雪腐镰刀菌烯醇(NIV)毒素, NIV毒素含量水平为371.74~5 282.80 μg/kg, 其中3个菌株产生少量脱氧雪腐镰刀菌烯醇(DON)毒素(86.13~227.22 μg/kg)?     

Molecular characterization of the Fusarium graminearum species complex in Japan

Members of the Fusarium graminearum species complex are important cereal pathogens worldwide and belong to one of at least nine phylogenetically distinct species. We examined 298 strains of the F. graminearum species complex collected from wheat or barley in Japan to determine the species and trichothecene chemotype. Phylogenetic analyses and species-diagnostic polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLPs) revealed the presence and differential distribution of F. graminearum sensu stricto (s. str.) and F. asiaticum in Japan. F. graminearum s. str. is predominant in the north, especially in the Hokkaido area, while F. asiaticum is predominant in southern regions. In the Tohoku area, these species co-occurred. Trichothecene chemotyping of all strains by multiplex PCR revealed significantly different chemotype compositions of these species. All 50 strains of F. graminearum s. str. were of a 15- or 3-acetyl deoxynivalenol type, while 173 (70%) out of 246 strains of F. asiaticum were of a nivalenol type. The possibility of gene flow between the two species was investigated by use of 15 PCR-RFLP markers developed in this study. However, no obvious hybrids were detected from 98 strains examined, including strains collected from regions where both species co-occur.     

Geographic distribution and genetic diversity of Fusarium graminearum and F. asiaticum on wheat spikes throughout China

A large number of Fusarium graminearum and F. asiaticum isolates were collected from wheat spikes from all regions in China with a history of fusarium head blight (FHB) epidemics. Isolates were analysed to investigate their genetic diversity and geographic distribution. Sequence characterized amplified region (SCAR) analyses of 437 isolates resolved both species, with 21% being F. graminearum (SCAR type 1) and 79% being F. asiaticum (SCAR type 5). AFLP profiles clearly resolved two groups, A and B, that were completely congruent with both species. However, more diversity was detected by AFLP, revealing several subgroups within each group. In many cases, even for isolates from the same district, AFLP haplotypes differed markedly. Phylogenetic analyses of multilocus DNA sequence data indicated that all isolates of SCAR type 1, AFLP group A were F. graminearum , whilst isolates of SCAR type 5, AFLP group B were F. asiaticum , demonstrating that it is an efficient method for differentiating these two species. Both species seem to have different geographic distributions within China. Fusarium graminearum was mainly obtained from wheat growing in the cooler regions where the annual average temperature was 15°C or lower. In contrast, the vast majority of F. asiaticum isolates were collected from wheat growing in the warmer regions where the annual average temperature is above 15°C and where FHB epidemics occur most frequently. This is the first report of the distribution of, and genetic diversity within, F. graminearum and F. asiaticum on wheat spikes throughout China.     

Fusarium populations on Chinese barley show a dramatic gradient in mycotoxin profiles

We report on a large gene bank of Fusarium isolates established by a broad survey conducted in 2005 in which infected barley ears were collected in 23 counties of seven provinces and two municipalities along the Yangtze River in China. In total, 1,894 single spore isolates were obtained. The isolates were characterized at the species level by a newly developed and robust set of diagnostic primers based on single nucleotide polymorphisms (SNPs) among members of the F. graminearum clade. In addition, we determined their chemotype using previously described polymerase chain reaction (PCR) primers. The results showed that in all regions F. asiaticum was the predominant species causing Fusarium head blight (FHB) on barley in China (N = 1,706), while in the upper valleys of the Yangtze River also F. graminearum sensu stricto, F. meridionale, and F. proliferatum were found. Major differences in the chemotypes were found in the F. asiaticum populations, from very high to exclusive nivalenol (NIV) chemotypes in the mountainous upper valleys of the Yangtze River to predominantly deoxynivalenol (DON) chemotypes in the middle and lower valleys. In contrast to the F. asiaticum isolates from three counties in Sichuan province, which were largely NIV producers (278 of 291), F. graminearum isolates from these sampling sites were for the vast majority (27 of 28) DON producers, indicating that despite sharing the same habitat, these sympatric species apparently have unique mycotoxin chemotypes.     

安徽省玉米穗腐病主要致病镰孢菌的分离与鉴定

为明确安徽省玉米穗腐病主要致病镰孢菌的种类,采用单孢分离、形态学鉴定和分子生物学鉴定的方法,对2017年采集自安徽省6个玉米主产区——合肥市、蚌埠市、淮北市、阜阳市、亳州市和宿州市的玉米穗腐病病样进行了分离鉴定。在获得的455株镰孢菌中,拟轮枝镰孢菌、禾谷镰孢复合种、层出镰孢菌、尖孢镰孢菌和黄色镰孢菌的分离频率分别为59.13%、21.28%、13.68%、5.12%和0.79%。其中拟轮枝镰孢菌在各地的分布最广,属于安徽省的优势致病镰孢菌。     

小麦赤霉病菌对多菌灵的抗药性研究

测定了1976 年从江苏南京采集的54 株小麦赤霉病菌菌株对多菌灵的敏感性,EC50值在0. 2617～ 0. 6544 μg. mL - 1 之间, 1983 年在同一地点采集的76 株, EC50 值相应为0. 2517～ 0. 7050 μg. mL- 1 , 而1998 年从浙江、湖北、上海、福建、安徽、江苏各地采集的104 株菌株EC50值为0. 2478～ 6. 4574 Lg. mL- 1 , 表明湖北、上海、浙江等地田间已检测到小麦赤霉病菌抗多菌灵菌株。紫外光诱导分生孢子也获得了该病菌抗多菌灵突变株, 其EC50 值为14. 1993 Lg. mL - 1 。抗药性突变体JPR 与敏感亲本菌株JPS 相比, 在菌丝生长、产孢量方面无明显差异, 但JPR 孢子萌发率为57. 1% , 而JPS 为100% , 而且50% 孢子萌发的时间较野生亲本菌株滞后12 h。JPR 产生脱氧雪腐镰刀菌烯醇(DON 毒素) 3. 90 μg mL - 1 , 而JPS 产生的DON 毒素为9. 28μg. mL - 1 。     

河南省玉米茎基部镰刀菌的形态和分子鉴定

 为明确河南省玉米茎基部镰刀菌(Fusarium spp.)的种群组成及分布, 2011和2012年, 我们采集了河南省14个地市42个县(区)的玉米茎基腐病病样, 分离得到163个镰刀菌单孢菌株。首先对菌株进行了形态学鉴定, 在此基础上使用镰刀菌种特异性引物进行了PCR检测;对于部分PCR检测未能确认的菌株进行了Translation elongation factor 1-α(TEF-1α)基因片段测序及BLAST分析, 最终将163个菌株鉴定到种。结果表明在163个菌株中, 禾谷镰刀菌F. graminearum为优势种, 占44.2%(72株), 其次为层出镰刀菌F. proliferatum和轮枝镰刀菌F. verticillioides, 分别占28.8%(47株)和27.0%(44株)。豫西北的焦作(47.6%)、洛阳(50.0%)及豫中的许昌(45.5%)均以F. verticillioides为主, 豫东的商丘(42.9%)、开封(57.1%)以F. proliferatum为主, 豫北的安阳(66.7%)、濮阳(71.4%)、新乡(62.5%)和豫南的南阳(57.1%)、驻马店(45.0%)以及中部的郑州(57.1%)、漯河(66.7%)则均以F. graminearum为优势种。