柑桔内生拮抗细菌的发酵液对油菜菌核病的田间防治效果

从柑桔健康组织中分离得到两株内生拮抗细菌"重桔1号"和"元石45",制成细菌悬浮液, 25℃恒温水浴振荡培养72h, 喷雾于接有油菜菌核病菌的油菜成株叶片上,以喷灭菌水为对照. 结果表明,喷有"重桔1号"和"元石45"两株细菌的发酵液的叶片菌核病菌病斑比对照叶片病斑扩展明显减小,与对照相比其病斑面积平均少扩大值分别为9.75cm2和10.62cm2 , 两株细菌的发酵液对油菜菌核病的绝对防治效果分别为19.5%和47.5%.     

向日葵品系对菌核病的抗性鉴定

在2002～2003年连续两年对经初步抗性筛选所获得的儿个向日葵品系进行了菌核病的抗性鉴定．结果表明：在所有的参鉴材料中没有发现免疫类型，但各品系间抗性有一定的差异．经两年的抗性筛选后发现AS02、AS03、AS04、AS07和AS08等5个品系对根腐型和盘腐型菌核病的抗性显著高于生产上中抗杂交种龙葵杂1号等对照品种．     

核盘菌交配型基因mat1-1敲除载体的构建及转化

利用根癌农杆菌介导真菌遗传转化方法对核盘菌的交配型基因mat1-1进行基因同源重组的敲除对比实验,获得了缺失mat1-1基因的转化菌株.先利用PCR方法获得mat1-1基因的左右两侧片段,将测序正确的两个侧翼片段分别重组到农杆菌转化载体PBI-G3C中,并将新霉素抗性标记引入农杆菌转化载体PBI-G3C中,构建成农杆菌介导转化核盘菌的打靶载体ΔPBI-G3CN-mat1-1,再将ΔPBI-G3CN-mat1-1质粒转化至根癌农杆菌EHA105中.利用核盘菌菌丝进行转化,将得到的转化菌株,利用PCR方法进行验证,证实有敲除菌株存在.通过对敲除菌株进行生理表型的测定发现,缺失mat1-1基因的敲除菌株其生长速度与野生核盘菌菌株相比无明显差异,但敲除菌株不能产生菌核与子囊盘.     

周口地区油菜核盘菌分离鉴定及侵染分析

从河南省周口市采集感染菌核病的油菜,对菌核进行分离和鉴定.采用单孢分离法分离得到1个单菌系,对该单菌系进行了菌落和显微形态分析,初步鉴定该菌系为Sclerotinia sclerotiorum.利用真菌通用引物ITS1和ITS4对所分离的单菌系内转录间隔区(inter transcribed spacer,ITS)进行PCR扩增,得到540bp大小的片段,经过克隆测序和在GenBank中进行BLASTn分析,发现与部分S.sclerotiorum的ITS序列同源性达到100%,进一步鉴定该分离菌株为S.sclerotiorum.进化树分析显示,该菌系与来自意大利、日本及我国武汉地区的菌系聚为一支,说明周口菌系来源广泛.对该菌系侵染油菜叶片的细胞学和组织化学分析表明,该菌系对耐病和感病油菜品种的细胞反应相似,都没有H2O2积累,差别主要表现在耐病油菜上菌丝长度较短.     

Transmissible hypovirulent element in isolate Ep-1PN ofSclerotinia sclerotiorum

The cause of virulence decline of hypovirulent isolate Ep-1PN ofSclerotinia sclerotiorum (Lib.) de Bary has been studied through transmitting test and analysing its offsprings derived from single ascospore. When the colony of a normal strain Ep lPNA 183 intermingled with the colony of Ep-lPN, its margin became abnormal, just like EplPN. Cultures isolated from toptip of the colony of Ep-1PNA 183 showed the hypovirulence and its associated properties of Ep-1PN. Moreover, a series of cultures which have characteristics between EplPN and normal strain could be isolated from stipes of Ep-1PN. More than 200 offsprings derived from single ascospore of Ep-1PN were normal strains. According to those results, there exists a transmissible cytoplasmic hypovirulent element in hypovirulent isolate EplPN of S.sclerotiorum.     

转基因油菜雄性不育系15A及其恢复系菌核病抗性的鉴定与筛选

采用苗期接种法对湖南油料所构建及选育的转基因油菜雄性不育系15A、胞质三系不育系742A,常规油菜恢复系695、780的抗耐油菜菌核病进行鉴定,结果表明,4个品种均为抗性品种,其中15A为中抗,742A、695、780为低抗。四个品系的病情指数均比对照中油821低。各品系单株之间抗性存在明显差异。     

菌核病接种方法及新疆核盘菌致病力差异的初步研究

以采自新疆6种不同作物上的核盘菌作为供试菌株,对植株苗期抗病性鉴定方法进行筛选,同时测定了不同寄主来源的菌株间在生物学特性、草酸的产生能力及致病力上的差异。结果显示:菌丝块贴接法和草酸浇灌法都能用于抗性植株的筛选,且草酸浇灌法可作为苗期鉴定抗病性的一种快速简便的方法。不同寄主来源的菌株的致病力、生物学特性及草酸的产生能力都存在明显的差异,来自马铃薯的菌株致病力最弱,但所有菌株对供试的各向日葵品种致病力均较强。     

一株产菌核青霉的生物学特性研究

从土壤样品中分离到73株青霉,从中筛选出能够产生菌核的Q1菌株.通过菌落形态和个体形态特征的观察,初步将Q1菌株鉴定到青霉属汤姆青霉系(Penicillium thomil series).实验结果表明Q1菌株形成菌核的最适温度为25℃,最适pH为6.5,麦芽汁培养基为最适培养基.供试的4种无机盐中,K_2HPO_4的单因子效应最好,说明K_2HPO_4对于Q1菌株的茼核形成有重要作用.培养基的碳氮源对Q1菌株的菌核生物量有较大影响,麦芽糖是最好的碳源,酵母膏是最好的氮源.培养基的含碳量保持在20 g/L,含氮量保持在0.24 g/L～0.48 g/L时,对Q1菌株菌核形成有利.     

Isolation and identification of Sclerotinia stem rot causal pathogen in Arabidopsis thaliana

A new stem rot disease is found to occur naturally on Arabidopsis plants in greenhouses of Fuzhou, China. In order to identify its pathogen, we conducted a series of fungal isolation and purification, plant reinoculation, and ascus and ascospore induction from the sclerotia. The isolate caused typical water-soaked lesions after reinoculation and produced sclerotia both on Arabidopsis plants and culture medium plates, and the sclerotia could be induced to produce discal apothecia and 8 binucleate ascospores per ascus. These disease symptom and fungal morphology data revealed that the fungus Sclerotinia sclerotiorum （Lib.） de Bary was the pathogen for Arabidopsis stem rot. To confirm this, we further amplified its large subunit ribosomal DNA （LSU rDNA） by polymerase chain reaction （PCR）, and compared the sequence with the known LSU rDNA sequences in GenBank. The results show that the sequence shares the highest identities with the LSU rDNAs of different S. sclerotiorum strains. Taking all these data together, we concluded that the fungus that caused the Arabidopsis stem rot is S. sclerotiorum （Lib.） de Bary. This is the first report that Arabidopsis is naturally infected by S. sclerotiorum.     

向日葵菌核病研究进展

对向日葵菌核病的症状、病原菌及侵染特性、抗病性及防治措施做了系统概述.