Interactions of fungicide-herbicide combinations against plant pathogens and weeds

The fungicide cyproconazole and four herbicides, DNOC, dicamba, ioxynil and bromoxynil, were tested singly and in mixtures against Rhizoctonia cerealis (in vitro and on wheat) and against Pseudocercosporella herpotrichoides (in vitro, as well as against two weeds, Avena fatua and Sinapis alba. Cyproconazole showed very strong fungicidal acitivity against both fungi. DNOC was inhibitory to the fungi in vitro but had little effect on the disease; dicamba had low activity against the fungi in vitro but prevented the disease on wheat. Ioxynol and bromoxynil were moderately fungitoxic both in vitro and against the disease. Cyproconazole-herbicide mixtures were synergistic in their action against both fungi in vitro and against the disease. The level of synergism depended on the relative proportion of fungicide and herbicide components in the mixtures. Mixtures of herbicides with cyproconazole against weeds were at best additive in their activity.     

小麦纹枯病菌拮抗细菌的稳定性及对小麦纹枯病菌致病作用的影响

从江苏省51个市(县)采集的550余份小麦纹枯病病叶鞘上共分离获得441个细菌分离物,其中35个菌株对小麦纹枯病菌产生了拮抗作用,占总菌株数的7.94%。经300 d转代保存后仍有28株菌保持了对小麦纹枯病菌的拮抗或抑制作用。拮抗细菌与纹枯菌混合接种小麦,对纹枯菌的侵染有抑制作用,其中GC4菌株对纹枯病的抑制率达63%。     

小麦纹枯病生防菌株的筛选及小区试验

从小麦根系获得1株对小麦纹枯病菌和另外6种病原真菌和3种病原细菌有显著抑制活性的菌株B_3(Bacillus sp.)。在NA平板上,不同方法测得B_3对小麦纹枯病菌的生长抑制率为73.3～99.5％。PDA培养基中加入12.5％B_3无细胞培养滤液时,纹枯病菌的菌丝生长减少80％以上。种子处理后B_3能在小麦根系及地面以下茎杆和叶鞘组织中定殖。B_3培养液在1:10～1:10000浓度下可提高小麦发芽率5.7～21.7％,并增加根和芽的生长,提高根/芽比。在盆钵及小区试验中,B_3粉剂种子处理后小麦出苗快而齐,幼苗生长茁壮。在人工接种病菌的灭菌土和自然土盆钵中,防病效果分别为30.1％和49.4％。小区试验以B_3粉剂拌种拔节期平均防效62％。处理比对照增加有效穗6.6％,千粒重增加0.92克,增加小麦产量13.7％。     

小麦纹枯病菌菌核形成条件研究

研究了小麦纹枯病菌在不同光照、温度、培养基条件下菌核数量形成的差异.结果表明,小麦纹枯病菌在不同的光照条件下都可以形成菌核,但是形成菌核数量有差异.小麦纹枯病菌在24 h光照(即G24)条件下产生的菌核数量最多,其次是光照12h,而在全黑暗(即G0)的条件下产生的菌核数量较少.小麦纹枯病菌在15～35℃条件下都可以形成菌核,且在15～30℃的条件下,随着温度的升高,菌核数量也在增加,而在高于30C的条件下形成的菌核数量在减少;小麦纹枯病菌在25～30℃条件下最有利于形成菌核且数量较多.最适合小麦纹枯病菌形成菌核的培养基是PDA培养基.     

小麦纹枯病生防菌株S 024的诱变育种研究

采用5种诱变方法(紫外线、微波、LiCl,LiCl 紫外线、LiCl 紫外线 微波)对出发菌株S 024进行了诱变处理.结果表明,几种诱变方法对S 024的诱变效果差异较大,紫外线(UV)照射和LiCl处理均未能筛选到正突变菌株;微波(MV)处理(50,100 s)可选育出高效菌株,LiCl和UV复合诱变处理的效果最好,共筛选到3个高效菌株(LiU-024-1,LiU-024-4,LiU-024-9),其中LiU-024-4菌株抑菌率比出发菌株S 024提高了34.44%;连续3代转代培养,其产生抗生素的能力基本稳定.用MV对3株高效菌株再进行诱变处理,其抑菌活性基本没有提高.     

选择性培养基在小麦纹枯病菌分离及土壤菌核检测中的应用

由禾谷丝核菌(Rhizoctonia cerealis,Rc)引起的纹枯病是黄海麦区的重要病害[1]。对病菌分离主要采用组织分离法[2]。该法费时、费力、操作要求严格,且无法对土壤中的病菌数量进行检测。选择性培养基(Selective medium,SM)是根据不同真菌对化学药剂敏感性的不同,通过向基础培养基中添加特定的抑菌物质,达到专一性分离目标真菌而有选择地抑制杂菌生长的目的。     

利用荧光定量PCR检测禾谷丝核菌的相对生物量

小麦纹枯病是以禾谷丝核菌(Rhizoctonia cerealis)侵染为主的小麦土传病害。为建立检测禾谷丝核菌在寄主小麦(Triticum aestivum)中的相对生物量的可靠方法,促进小麦抗纹枯病机制的研究,本研究克隆了禾谷丝核菌肌动蛋白基因RcActin的部分(3′端)cDNA,并设计了RcActin的特异引物。该引物不仅能区分禾谷丝核菌与寄主小麦,还能区分全蚀病菌(Gaeumannomyces graminis)、根腐病菌(Bipolaris sorokiniana)和立枯丝核菌(Rhizoctonia solani)等常见小麦土传病害的病原菌,表明该引物能用于小麦纹枯病的分子检测,也能用于相对表达量的测定。利用相对定量法,以RcActin相对于寄主管家基因的相对表达量作为禾谷丝核菌相对生物量的指标,结果表明,此方法能准确反映禾谷丝核菌在寄主中的相对生物量和对小麦纹枯病抗性程度进行快速鉴定。     

冬青抑菌活性的初步研究

采用生长速率法,测定冬青(Ilex purpurea Hassk)不同器官的提取液对小麦纹枯病菌(Rhizoctonia cerealis)、番茄褐斑病菌(Helminthosporiurn carposaprum)、番茄早疫病菌(Alternaria solani)、黄瓜褐斑病菌(Corynespora cassiicolai)的抑菌作用.结果表明,当浓度为0.1 g/mL时,冬青不同器官的提取液对4种病原菌菌丝生长都有一定的抑制作用,但不同部位提取液的抑菌活性不同,其中冬青果实和枝条提取液的抑菌活性较弱,树皮提取液的抑菌活性次之,叶片提取液的抑菌活性较强.     

Evaluation of diagnostic and quantitative PCR for the identification and severity assessment of eyespot and sharp eyespot in winter wheat

Diagnostic and quantitative polymerase chain reaction (PCR) provided clarification of the causes of symptoms and the extent of infection by eyespot ( Tapesia spp.) and sharp eyespot ( Rhizoctonia cerealis ) on winter wheat at early growth stages. Disease assessments made before stem extension, when decisions to apply fungicides are usually made, often did not agree with the pathogen diagnoses using PCR, suggesting that such early visual diagnoses may be unreliable. Visual and PCR diagnoses made on stems in summer generally supported each other, but there were often discrepancies in relating disease severity to amounts of pathogen present when determined by regression analyses of incidence or severity of symptoms on amount of pathogen DNA. Mixed symptoms caused by different pathogens may sometimes have been confounded. Relationships between symptoms and DNA of eyespot pathogens were less clear on some cultivars, often those with least disease. Sharp eyespot symptoms had a stronger relationship to DNA of its pathogen. Significant regressions often accounted for a small percentage of the variance, suggesting either that pathogens not assayed were contributing to symptoms or that lesions were in some cases persisting longer into the season than pathogen DNA. The frequency of pathogen detection before stem extension was a poor predictor of the amounts of pathogen DNA measured later in the season.     

应用real-time PCR定量检测土壤中小麦纹枯病菌方法的建立

根据小麦纹枯病菌Rhizoctonia cerealis AG-D融合群ITS区的DNA序列设计特异性引物对WKF-8/WKR-8,对引物的特异性和灵敏性进行检测,建立并优化基于SYBR GreenⅠ荧光染料法的real-time PCR反应体系,绘制标准曲线。检测范围在1.0×10-3~10 ng/μl之间有良好的线性关系,相关系数R2为0.995,扩增效率为99.7%,灵敏度比常规PCR方法高100倍。结果表明,该方法具有快速、特异性强、敏感度高等特点。