可可毛色二孢菌侵染后的烟草酵母双杂交cDNA文库构建

正葡萄是世界上最重要的经济果树之一,据联合国粮食及农业组织统计,截止到2013年,中国葡萄栽培面积已达715.5万hm2,葡萄总产量超2 300万kg。长期以来,葡萄病害是影响葡萄产业健康发展的重要因素。近年来研究发现由葡萄座腔菌科真菌(Botryosphaeriaceae)引起的葡萄溃疡病(Botryosphaeria dieback)是葡萄上的主要枝干病     

肉桂枝枯原因研究

肉桂泡盾盲蝽吸食肉桂枝条的汁液，造成微小伤口，并携带可可毛色二孢菌，使肉桂枝条容易被该菌侵染，造成皮层坏死，引发极严重的枝枯病。在某些林分，其它枝梢害虫如肉桂双瓣卷蛾、肉桂小峰、肉桂瘿蚊、肉桂木蛾、肉桂天牛，它们的为害也能不同程度地引起肉桂的枝梢枯死。     

芒果可可球二孢蒂腐病菌生物学培养特性

对芒果可可球二孢蒂腐病菌(Botryodiplodia theobromae Pat.)的生物学培养特性进行了研究.结果表明:菌丝生长最适温度30℃;孢子萌发最适温度30℃;菌丝生长最适pH值5～6,孢子萌发最适pH值7～10;菌丝生长的最佳碳源是棉子糖、葡萄糖,最佳氮源是牛肉浸膏、蛋白胨:光照及振荡培养对菌丝生长无显著影响;菌丝的致死温度为60℃,30 min或65℃,10 min,孢子的致死温度50℃,15min.     

可可毛色二孢全基因组分泌蛋白的预测及分析

【目的】可可毛色二孢(Lasiodiplodia theobromae)是一种世界性分布的重要植物病原真菌,可引起严重的葡萄溃疡病(Botryosphaeria dieback),影响果木品质并造成巨大的经济损失。本研究预测并分析可可毛色二孢基因组范围内的分泌蛋白,并明确其基本特征,为该病菌分泌蛋白致病机理的研究打下基础。【方法】依据已公布的可可毛色二孢全基因组序列,利用信号肽预测软件SignalP v5.0、跨膜结构分析软件TMHMM v2.0、细胞器定位分析软件ProtComp v9.0、GPI锚定预测软件big-PI Fungal Predictor和亚细胞器定位分析软件TargetP v2.0生物信息学软件对该菌中的典型分泌蛋白进行筛选。对分泌蛋白N端信号肽的长度、氨基酸使用频率及其切割位点进行统计分析。依据蛋白序列的同源性,应用BLASTP程序对分泌组蛋白进行功能注释分析,预测其生物学功能。采用蔗糖酶缺陷的酵母分泌系统,对所选分泌蛋白的信号肽进行活性检测。利用qRT-PCR方法检测所选分泌蛋白基因在可可毛色二孢侵染葡萄中的表达情况。【结果】在可可毛色二孢全基因组编码蛋白中共筛...     

桉树枯枝病病害研究

按树枯枝病 Botryodiplodia theobromae Pat主要为害 2～ 3龄雷林 1号桉 3 3 ,造成枯枝落叶和死亡。病原菌周年为害 ,以当年 1 2月份至翌年 3月份表现最严重。     

Relationship between incidence and severity of cashew gummosis in semiarid north-eastern Brazil

The incidence–severity relationship for cashew gummosis, caused by Lasiodiplodia theobromae , was studied to determine the feasibility of using disease incidence to estimate indirectly disease severity in order to establish the potential damage caused by this disease in semiarid north-eastern Brazil. Epidemics were monitored in two cashew orchards, from 1995 to 1998 in an experimental field composed of 28 dwarf clones, and from 2000 to 2002 in a commercial orchard of a single clone. The two sites were located 10 km from each other. Logarithmic transformation achieved the best linear adjustment of incidence and severity data as determined by coefficients of determination for place, age and pooled data. A very high correlation between incidence and severity was found in both fields, with different disease pressures, different cashew genotypes, different ages and at several epidemic stages. Thus, the easily assessed gummosis incidence could be used to estimate gummosis severity levels.     

Control of postharvest diseases of rambutan using controlled atmosphere storage and potassium metabisulphite or<Emphasis Type="Italic">Trichoderma harzianum</Emphasis>

Botryodiplodia theobromae, Colletotrichum gloeosporioides andGliocephalotrichum microchlamydosporum are the causal fungi of three rambutan postharvest diseases, stemend rot, anthracnose and brown spot, respectively. Two different treatments of rambutan fruits to control the diseases were investigated: application of potassium metabisulphite (250 ppm) or culture filtrate ofTrichoderma harzianum (TrH 40) followed by controlled atmosphere storage (CA) at 13.5°C and 95% r.h. Potassium metabisulphite at 250 ppm under CA effectively controlled the incidence and severity of the three postharvest diseases and maintained the eating quality and color of the fruit for 21 days. The greatest effect of this treatment was on brown spot disease, caused byG. microchlamydosporum. Application of TrH 40 was less effective than potassium metabisuphite. http://www.phytoparasitica.org posting July 8, 2002.     

茶树病害病原菌Lasiodiplodia theobromae生物学特性研究

采用皿内测定方法对贵州省惠水县茶树叶斑病的病原菌Lasiodiplodia theobromae进行了病原生物特性研究。结果表明,其病原菌可在SM、PDA、OA和MEA培养基上生长,菌株在MEA培养基上的生长速率显著快于SM培养基、PDA培养基和OA培养基。病原菌适宜在OA培养基上产孢。碳素和氮素营养可影响菌株的生长及菌丝色素的形成。在PDA培养基上,菌株在33℃条件下的生长速度最快,可适应酸性和碱性条件。但在弱碱条件下生长最为迅速。菌株可在PDB培养基中生长,接种菌碟后生长88 h进入稳定期。     

可可毛色二孢β-1,4-葡聚糖内切酶LtEg1信号肽的鉴定及其酶活性分析

为明确可可毛色二孢β-1,4-葡聚糖内切酶LtEg1是否能够分泌至胞外及其活性,采用酵母互补试验分析了内切酶LtEg1的信号肽活性,利用3,5-二硝基水杨酸法检测了内切酶LtEg1活性,通过实时荧光定量PCR测定了LtEg1基因在不同侵染阶段的表达量。内切酶LtEg1的信号肽和酵母蔗糖酶的融合表达能够引起酵母蔗糖酶的外泌,使得酵母转化子能够在以棉子糖为唯一碳源的培养基上生长。内切酶LtEg1能够以羧甲基纤维素钠为底物,发挥其酶活功能。与营养菌丝阶段相比,LtEg1基因在病原菌侵染阶段的表达水平显著升高;且在接种后48 h,LtEg1基因的表达量达到高峰,约为营养菌丝阶段的12倍,表明内切酶LtEg1作为重要的外泌蛋白酶,在可可毛色二孢侵染致病过程发挥了重要作用。     

九里香内生细菌HBS-1的鉴定及其对芒果采后病害的防效

通过组织表面消毒法对九里香内生菌进行分离,以芒果炭疽病菌(Colletotrichum gloeosporioides Penz)和芒果蒂腐病菌(Botryodiplodia theobromae Pat)为靶标菌对获得菌株进行皿内拮抗能力筛选,利用浸果法测定菌株发酵滤液对芒果采后病害的防治效果,结合菌株形态、生理生化特征及16S rDNA序列鉴定菌株。结果从九里香中分离到28株内生菌,其中有8株真菌、9株细菌对2种病原菌均有拮抗活性,以HBS-1抑菌活性最好,对芒果炭疽病菌及芒果蒂腐病菌抑制率分别为42.0%和50.0%;菌株HBS-1发酵滤液能明显降低芒果采后病害的发病率,10倍稀释液的防效优于化学药剂特克多800倍液,50倍稀释液的防效与化学药剂特克多800倍液相当;经鉴定,内生菌HBS-1为芽孢杆菌属枯草芽孢杆菌(Bacillus subtilis)。