贡柑黑腐病发生规律研究

2011—2012年采用盆栽苗人工接种和田间调查相结合的方法,研究了贡柑产区近年为害严重的黑腐病(病原菌为Alternaria citri)的发生规律。结果表明:在人工接种条件下,其病原菌可以从没有伤口的叶面侵入,但更易从伤口入侵;贡柑和默科特桔最感病,其次是椪柑和南丰蜜桔,在有伤口和连续下雨的条件下马水桔、沙糖桔和温州蜜柑的嫩芽也偶被感染,橙类、柚类、柠檬和金柑抗病。在田间,贡柑、默科特桔最感病,其次是南丰蜜桔、椪柑,马水桔、沙糖桔、温州蜜柑未见发病。刚抽发的嫩梢和幼果易感病,组织越嫩越易感病,老叶、成熟果实感病少,幼树和刚结果树以及嫩梢多的树发病重。该病原菌孢子萌发需要高湿,下雨、高温、高湿有利于该病的发生和流行。夏梢发病最重,其次是春梢和幼果,秋梢发病轻。在人工接种条件下,只要下雨和温度达到15～30℃,1～3天即可显症,病斑沿着叶脉快速扩展。发病快、扩展快是其难防治的最大原因。     

生防菌Alternaria amaranthi-3对反枝苋的防治效果

反枝苋Amaranthus retroflexus是一种世界性恶性杂草。为了确定微生物除草剂候选菌Alternaria amaranthi-3防除反枝苋的潜力,通过盆栽试验研究了接种浓度、露期和水乳剂型对A.ama-ranthi-3菌株致病力的影响。结果显示,接种浓度显著影响菌株的致病力,在48h露期条件下,接种孢子浓度为105个/mL时,菌株水剂对反枝苋幼苗生长抑制率为35.55%;浓度为107个/mL时,生长抑制率达到75.25%。露期对该菌株的致病力也有较大影响,在不保湿条件下,菌株水剂对反枝苋的生长抑制率为26.43%,而保湿48h处理的生长抑制率达到77.96%。Span80∶Tween80=1∶3的复配乳化剂和大豆油制备的水乳剂可显著降低露期对菌株防效的影响和提高菌株的致病力,无露期时,菌株水乳剂对反枝苋的生长抑制率达到88.35%,显著高于水剂;48h露期条件下,菌株水乳剂处理的生长抑制率为90.59%,而菌株水剂处理为77.96%。表明通过剂型的改进菌株Alternaria amaranthi-3能有效防除反枝苋。     

Fine mapping of the gene for susceptibility to black spot disease in Japanese pear (Pyrus pyrifolia Nakai)

Black spot disease, which is caused by the Japanese pear pathotype of the filamentous fungus Alternaria alternata (Fries) Keissler, is one of the most harmful diseases in Japanese pear cultivation. We mapped a gene for susceptibility to black spot disease in the Japanese pear (Pyrus pyrifolia Nakai) cultivar ‘Kinchaku’ (Aki gene) at the top of linkage group 11, similar to the positions of the susceptibility genes Ani in ‘Osa Nijisseiki’ and Ana in ‘Nansui’. Using synteny-based marker enrichment, we developed novel apple SSR markers in the target region. We constructed a fine map of linkage group 11 of ‘Kinchaku’ and localized the Aki locus within a 1.5-cM genome region between SSR markers Mdo.chr11.28 and Mdo.chr11.34. Marker Mdo.chr11.30 co-segregated with Aki in all 621 F₁ plantlets of a ‘Housui’ × ‘Kinchaku’ cross. The physical size of the Aki region, which includes three markers (Mdo.chr11.28, Mdo.chr11.30, and Mdo.chr11.34), was estimated to be 250 Kb in the ‘Golden Delicious’ apple genome and 107 Kb in the ‘Dangshansuli’ Chinese pear genome. Our results will help to identify the candidate gene for susceptibility to black spot disease in Japanese pear.     

文山三七根腐病病原真菌的鉴定与药剂防治

从云南文山三七药材基地感染根腐病的三七组织获得分离物,并对其进行培养和纯化,并运用科赫法则（Koch’s Postulate）进行病原菌回接感染,得3株致病明显的菌株,后对此菌株进行生物学和18S-rDNA全序列分析鉴定及室内药效防治。结果表明,此3株菌分别是尖孢镰刀菌（Fusarium oxysporum）,茄病镰刀菌（Fusarium solani）和链格孢菌（Alternaria alternata）。甲基硫菌灵、根康、多菌灵对茄孢镰孢菌、尖镰孢菌的抑菌效果最好,腐霉利、甲基硫菌灵对链格孢菌的抑菌效果最好。该研究可为尖孢镰刀菌、茄病镰刀菌和链格孢菌引起的三七根腐病的防治提供参考。     

Occurrence of the F129L mutation in Alternaria solani populations in Germany in response to QoI application,and its effect on sensitivity

Early blight caused by Alternaria solani is a highly destructive disease of potatoes. Control of early blight mainly relies on the use of preventive fungicide treatments. Because of their high efficacy, azoxystrobin and other quinone outside inhibitors (QoIs) are commonly used to manage early blight. However, loss of sensitivity to QoIs has previously been reported for A. solani in the United States. Two hundred and three A. solani field isolates collected from 81 locations in Germany between 2005 and 2011 were screened for the presence of the F129L mutation in the cytochrome b gene; of these, 74 contained the F129L mutation. Sequence analysis revealed the occurrence of two structurally different cytb genes, which differed in the presence (genotype I) or absence (genotype II) of an intron, with genotype I being the most prevalent (63% of isolates). The F129L mutation was detected only in genotype II isolates, where it occurred in 97%. Sensitivity to azoxystrobin and pyraclostrobin was determined in conidial germination assays. All isolates possessing the F129L mutation had reduced sensitivity to azoxystrobin and, to a lesser extent, to pyraclostrobin. Early blight disease severity on plants treated with azoxystrobin was significantly higher for A. solani isolates with reduced fungicide sensitivity in the conidial germination assay compared with sensitive isolates. Data suggest an accumulation of F129L isolates in the German A. solani population over the years 2009–2011. It is assumed that the application of QoIs has selected for the occurrence of F129L mutations, which may contribute to loss of fungicide efficacy.     

三七黑斑病抗性人工鉴定方法的建立*

 三七黑斑病(Alternaria panax Whetz)是影响三七生产尤其是GAP规范化生产的重要病害,建立三七黑斑病抗性人工鉴定方法是开展三七抗病育种必备环节。分别以室内离体叶片和田间在体叶片进行不同接种方法的试验,明确了通过针刺叶片接种浓度为1.0×10⁷个/mL的三七黑斑病孢子悬浮液是诱导发病有效途径。室内离体接种和田间接种对比,同一植株室内离体叶片接种与田间接种的相关系数0.9548^**,达到极显著水平。对个体基因型还有变异的三七栽培群体进行人工抗性鉴定的初步研究发现,不同个体三七对黑斑病接种的发病程度有明显的差异,说明黑斑病抗病性与三七个体遗传特性有关。测定田间接种发病程度不同的三七叶片的苯丙氨酸解氨酶活性,发现三七对黑斑病的抗性与三七植株叶片苯丙氨酸解氨酶活性有明显的关系。     

烟草赤星病菌遗传多样性ISSR和RAPD标记比较分析

对分离出的链格孢菌株用ISSR和RAPD分子标记技术分析研究其遗传多样性,比较2种分子生物学方法在链格孢遗传分析中的优劣,为研究烟草赤星病菌遗传多样性及烟草抗病品种的培育奠定基础。采用ISSR和RAPD分子标记方法对来自不同地区的28份烟草赤星病菌进行遗传多样性分析,筛选出10个ISSR引物和10个RAPD引物;ISSR扩增出多态性条带112条,多态性条带比率为86.82%,菌株间相似性系数为0.53~0.97;RAPD引物扩增出多态性条带70条,多态性条带比率为81.39%,菌株间相似性系数为0.57~0.94。用SPSS 17.0 软件对2种标记遗传距离进行相关性分析,发现2种分子标记结果呈显著正相关,表明2种分子标记方法都适合于烟草赤星病菌遗传多样性研究,ISSR是一种多态性优于RAPD的标记技术。根据2种标记的结果,利用NTSYS软件按UPGMA方法进行聚类分析,发现烟草赤星病菌遗传多样性与地理差异没有显著相关性。     

刺五加黑斑病菌孢子萌发的影响因子

为了探讨刺五加黑斑病菌孢子萌发的影响因素,在刺五加不同器官汁液等基质培养条件下及在不同温度处理条件下,研究了刺五加黑斑病菌在不同时间的孢子萌发率.刺五加黑斑病菌在刺五加不同器官汁液基质培养条件下及在不同温度处理条件下,孢子的萌发率都随着时间的延长而升高;其中在不同刺五加器官汁液基质处理条件的孢子萌发率比清水对照高,表明其各器官汁液基质更有利于病原菌孢子的萌发,其中茎基质处理的孢子萌发率最高;而在不同温度处理条件下,各调查时间段中,病原菌孢子萌发率都是在25℃时最高.即刺五加黑斑病菌孢子萌发的最适温度为25℃.     

一株铰链孢属真菌培养条件及激活蛋白的提取

本研究以PD培养液为测定培养基,对一株铰链孢属(Alternaria spp.)真菌的培养条件进行了初步研究,通过正交试验设计,确定其最佳培养条件为26℃,160 r/min,装液量120 mL/250 mL,培养72 h时菌丝干重量最高。使用超声波破碎法,从所培养的铰链孢属真菌中提取到分子量为42～43kD的激活蛋白。     

温度对刺五加黑斑病菌的影响

[目的]探讨温度对刺五加黑斑病菌的影响。[方法]在不同温度条件处理下,对刺五加黑斑病菌的生长速率和病原菌孢子的萌发状况进行系统的研究和分析。[结果]刺五加黑斑病病原菌对温度条件的适应性比较强,在5—35℃温度范围内均可以生长,超过40℃不能生长．而且在60℃以下的温度条件下均可以存活,最适宜的生长温度范围为25—30℃。其孢子萌发率在25℃时随着时间的延长而升高。刺五加黑斑病病原菌的生长速率和病原菌孢子的萌发率都是在25℃温度条件处理下为最高,表明25℃是刺五加黑斑病菌生长发育和侵染的最佳温度。[结论]该研究证明温度对刺五加黑斑病菌的生长速率和孢子萌发率均具有较大影响,该研究为刺五加黑斑病的进一步研究奠定了理论基础。