Use of real-time PCR to examine the relationship between disease severity in pea and <Emphasis Type="Italic">Aphanomyces euteiches</Emphasis> DNA content in roots

Aphanomyces euteiches causes severe root rot of peas. Resistance is limited in commercial pea cultivars. Real-time fluorescent PCR assay specific for A. euteiches was used to study the relationship between disease severity and pathogen DNA content in infected peas. Five pea genotypes ranging in levels of resistance were inoculated with five isolates of A. euteiches. Plants were visually rated for disease development and the amount of pathogen DNA in roots was determined using the PCR assay. The susceptible genotypes Genie, DSP and Bolero tended to have significantly more disease and more pathogen DNA than the resistant genotypes 90-2079 and PI 180693. PI 180693 consistently had less disease, while 90-2079 had the lowest amount of pathogen DNA. The Spearman correlation between pathogen DNA quantity and disease development was positive and significant (P < 0.05) for three isolates, but was not significant for two other isolates. This suggests that the real-time PCR assay may have limited application as a selection tool for resistance in pea to A. euteiches. Its utility as a selection tool would be dependent on the correlation between disease development and pathogen DNA content for a given pathogen isolate. The accuracy and specificity of the real-time PCR assay suggests considerable application for the assay in the study of mechanisms of disease resistance and the study of microbial population dynamics in plants.     

Stem-base disease and fungal colonisation of winter wheat grown in compost inoculated with Fusarium culmorum, F. graminearum and Microdochium nivale

Fungal colonisation of winter wheat cv. Cadenza by Fusarium culmorum, F. graminearum and Microdochium nivale was studied under conditions designed to avoid the splash dispersal of conidia from infested compost, to evaluate the possibility that systemic growth may transfer infection from the stem-base to the head. At decimal growth stages 33, 59, 77–87 and 95 the extent of fungal growth was assessed using a sample of 72 plants, by the recovery of fungal species from the stem-base, from each node and from the ear. Each of the fungi was recovered from stem tissues above soil level in some, apparently symptomless, plants. Symptoms of Fusarium foot rot were seen in an increasing proportion of plants during grain-fill and desiccation. There was an inverse relationship between recovery and the height above stem-base from which the stem tissue was excised. F. culmorum was the most frequently isolated fungus and it was also recovered from the highest position in plants. Only 3% of plants were colonised above the second node and none of the fungal species were recovered from either the fifth node or the ear. This suggests that colonisation and systemic growth from Fusarium infested compost is unlikely to contribute to the development of ear blight symptoms in winter wheat.     

8个WRKY转录因子在水稻叶片生长和抗病过程中的表达分析

 WRKY是植物体内最大的转录因子家族之一, 其成员在植物生长发育和抗逆过程中发挥重要作用。为了解水稻WRKY的功能, 本研究利用蛋白质免疫印迹(western blotting, WB)技术, 检测了8个WRKY转录因子在水稻叶片生长和Xa21介导的抗白叶枯病过程中的表达丰度变化。结果表明:OsWRKY12、43、55和86在苗期表达量较低, 随叶片的生长表达逐步增加;OsWRKY50和84在苗期叶片中的表达量相对较高, 成熟期下降;未检测到OsWRKY40/64和52在叶片中的表达。在Xa21介导的白叶枯病抗性反应中, OsWRKY40/64、50和52受白叶枯病菌诱导表达, OsWRKY84受侵染后表达量上调, OsWRKY12、43、55和86在抗病过程中没有检测到表达丰度的变化。进一步比较这些转录因子在抗、感和对照(Mock)反应中的表达, 发现抗、感反应间蛋白质的表达变化是相似的。研究结果提示:OsWRKY12、43、55和86可能在叶片正常生长中发挥作用;OsWRKY40/64和52可能在水稻-白叶枯病菌互作过程中发挥作用;OsWRKY50和84则在叶片生长和抗白叶枯病过程中都有一定的功能。     

油菜菌核病生防芽孢杆菌的分离鉴定及其脂肽化合物分析

采用平板拮抗筛选,分别从西藏日喀则地区和拉萨地区杂草根围土壤中筛选到2个对油菜菌核病菌有显著拮抗活性的芽孢杆菌菌株RJGP16和YBWC43。通过生理生化鉴定、16S rDNA序列分析和BOX-PCR指纹图谱分析,鉴定菌株RJGP16为萎缩芽孢杆菌Bacillus atrophaeus, 菌株YBWC43为解淀粉芽孢杆菌Bacillus amyloliquefaciens。离体叶片试验结果显示,菌株RJGP16和YBWC43对油菜菌核病菌防治效果分别为50.24%和100.00%。脂肽化合物种类分析显示,菌株RJGP16产生脂肽化合物表面活性素和芬枯草菌素,菌株YBWC43产生杆菌霉素D和芬枯草菌素。表明菌株RJGP16和YBWC43对油菜菌核病的防治效果与其产生的脂肽化合物有关。     

Concepts and Direction of Induced Systemic Resistance in Plants and its Application

Resistance to plant disease is often specific and metabolites and receptors contributing to this specificity may have specific structures. However, simple, structurally-unrelated compounds induce systemic resistance in unrelated plants to diverse pathogens including fungi, bacteria and viruses. Both resistance and induced systemic resistance (ISR) are associated with the rapid accumulation of the same structurally unrelated putative defense compounds that have diverse functions. It has been suggested that cultivar (race)-specific resistance is initiated by the specific interaction of a pathogen product (or pathogen induced product) and a plant receptor. However, restricted infection by pathogens can result in ISR and many different compounds can cause ISR. It is thus evident that there are both specific and non-specific routes to the master switch for ISR and there may be more than one master switch. Are reactive oxygen species and free radicals regulating the master switch(es) via both routes? It is also evident there are many switches, other than the master switch. Adding to the complexity of resistance and ISR are the observations that different compounds and pathways may mediate different biochemical resistances. Activation of one of the pathways may antagonize or enhance the activation or effectiveness of another. The review will address these complexities and questions and propose directions of research which require high priority. Factors which encourage and suppress the application of ISR in agriculture will also be addressed.     

广西玉米纹枯病菌培养性状及其致病力研究

从广西玉米主产区的8个县市秋玉米的纹枯病病株中分离出8个玉米纹枯病菌株,经生物性状的观察和菌丝融合群测定表明,它们都属于立枯丝核菌AGk-1-IA融合群;腐生定殖能力和致病力测定表明,菌株GX4、GX1的腐生定殖能力和致病力最强。     

Repression of deoxynivalenol accumulation and expression of Tri genes in Fusarium culmorum by fungicides in vitro

A defined medium was developed in which to monitor deoxynivalenol (DON) accumulation, fungal growth and expression of genes involved in trichothecene biosynthesis (designated Tri genes). In liquid culture, DON accumulated shortly after maximum expression of Tri6 and coincident with expression of Tri5. This was generally 96 h after inoculation. The effects of sublethal concentrations of the fungicides azoxystrobin, trifloxystrobin, kresoxim-methyl and tebuconazole on biosynthesis of the trichothecene DON by Fusarium culmorum were studied using this medium. The strobilurin fungicides trifloxystrobin and azoxystrobin significantly reduced the accumulation of DON in culture medium at a range of concentrations. Kresoxim-methyl, also a strobilurin, and tebuconazole, a triazole, did not significantly reduce the accumulation of DON, although levels were lower than those in nonamended cultures. Trifloxystrobin significantly reduced the accumulation of DON when added to cultures before initiation of trichothecene biosynthesis. RT-PCR assays of the expression of Tri6 and Tri5 genes indicated that trifloxystrobin acted by inhibiting the initiation of trichothecene biosynthesis.     

两个玉米自交系对纹枯病的抗病反应机制初探

玉米纹枯病是由立枯丝核菌(Rhizoctonia solani Kühn)引起的一种重要病害。具有优良性状的自交系是开展玉米抗病品种选育的重要种质资源。本试验以西南地区两个优良玉米自交系R08与18-599R为材料,采用人工接种R.solani菌丝融合群AG1-IA,对成株期玉米进行了抗性鉴定,并对病原菌诱导后苗期玉米叶片上的活性氧(ROS)、抗病相关基因(PR1、ZmDREB2A、ZmERF3和WIP1)表达及细胞坏死情况进行了动态检测。结果显示,R08对纹枯病表现为中抗,而18-599R为高感材料;受R.solani侵染后,玉米叶片ROS的积累在R08中较18-599R多;两个自交系中基因表达量及表达时间存在显著差异;R08叶片出现显著的细胞坏死反应,而18-599R相对较轻。本研究表明两个自交系对R.solani的抗性差异主要体现在相关抗病反应的时间和强度上,这为进一步研究玉米纹枯病抗病机理提供了依据。     

中国小麦农家品种‘老白麦’抗条锈性遗传分析

小麦条锈病是小麦生产上重要的气传叶部病害。不断发掘和利用新抗源是持续控制条锈病流行危害的重要基础研究工作。‘老白麦’是我国小麦农家品种,对我国当前主要流行小种和致病类型均表现为高抗水平。本研究采用常规杂交分析方法,对‘老白麦’及其与感病品种‘Taichung 29’的杂交后代在成株期和苗期分别接种CYR32号小种和CYR33号小种,进行抗条锈性鉴定和统计分析。结果表明,‘老白麦’对CYR33号小种在苗期和成株期均表现近免疫,其全生育期抗条锈性由1对显性基因控制;对CYR32号小种在成株期表现近免疫,苗期表现高感,成株抗条锈性由1对显性基因控制,属细胞核遗传。研究结果表明‘老白麦’至少含有2对显性抗病基因,分别控制‘老白麦’对CYR33号小种的全生育期抗性和对CYR32号小种的成株抗性。基因推导分析认为‘老白麦’对CYR33的全生育期抗性基因可能为未知新基因。建议在抗病育种中加以有效合理利用,促进小麦品种中抗病基因多样化布局。     

多因子联合诱导系统的构建及对黄瓜白粉病防治试验

由多个不同的抗病诱导因子组成植物抗病诱导系统的1个子系统。每个因子都是这个系统中的1个要素。系统的性质不但取决于系统中的要素,还取决于要素的结构。合理的配伍可以使要素与要素之间、要素与系统之间的关系得以优化,使系统的功能得以强化。把筛选得到的多个不同的抗病诱导因子进行配伍,组合成新的"诱导组合体",其诱导效应比单一因子显著提高;用不同的"诱导组合体"进行"相继诱导"和"循环诱导",则进一步提高诱导效应,对黄瓜白粉病的田间防治效果达到78.16%,高于常规杀菌剂的防治效果。