薄壳山核桃疮痂病菌TaqMan荧光定量PCR检测方法的建立及应用

疮痂病是薄壳山核桃上最具毁灭性的病害,带菌植物材料是传播疮痂病的重要来源。准确、灵敏、快速的检测方法可为该病害流行规律调查和防控提供有力的依据。本文通过比较薄壳山核桃疮痂病菌Venturia effusa及其近似种之间的ITS序列差异,设计了特异性引物和TaqMan探针,建立了薄壳山核桃疮痂病菌的荧光定量PCR检测方法。特异性检测结果表明,该方法可以检测不同地区的薄壳山核桃疮痂病菌菌株,而对其近似种以及薄壳山核桃上的其他真菌均没有信号。本研究建立的检测方法对薄壳山核桃疮痂病菌DNA的最低检测限可达0.5 pg/μL。该方法用于田间样品检测时,检测时间仅需1 h,远快于常规的分离培养法。本研究建立的基于TaqMan探针的荧光定量PCR检测方法为薄壳山核桃疮痂病菌的快速检测和监测提供了有力工具。     

内蒙古马铃薯黑胫病病原菌的分离和鉴定

 马铃薯是我国的第四大主粮。内蒙古是我国最大的马铃薯生产基地,马铃薯产业是当地农民收入的主要来源。2017年8月,内蒙古锡林郭勒盟部分马铃薯田块发生了一种细菌性黑胫病,发病面积约为200亩,发病率约为20%。发病初期,植株的茎基部变色、发黑,伴有臭味,严重时茎秆腐烂,植株死亡。为明确该地区马铃薯细菌性黑胫病的病原菌种类,本研究对该病原菌进行了分离,并利用生理生化测定,Biolog分析和分子生物学手段将该病原菌鉴定为胡萝卜软腐果胶杆菌巴西亚种(Pectobacterium carotovorum subsp. brasiliense)。据以往报道,我国马铃薯黑胫病主要由黑腐果胶杆菌(P. atrosepticum)和胡萝卜软腐果胶杆菌胡萝卜亚种(P. carotovorum subsp. carotovorum)两种病原菌引起,本研究是P. carotovorum subsp. brasiliense引起马铃薯黑胫病在国内的首次报道。     

葡萄根瘤蚜与常见农业蚜虫种类识别

介绍葡萄根瘤蚜及其他农业蚜虫共2科10属15种,编制葡萄根瘤蚜与15种常见农业蚜虫的分种检索表。     

免疫捕捉PCR和经典PCR方法检测水稻细菌性谷枯病菌灵敏性比较

将免疫吸附富集(ISE)和经典PCR结合以提高水稻细菌性谷枯病的检测效率,并且利用多克隆抗体技术成功地获得了4种抗血清,即抗Burkholderia glumae全菌体血清ASBg14,ASBg32,ASBg26和ASBg04,ODD法测定该4种抗血清的效价均达到1∶32以上,利用凝聚法测定的所有抗血清的效价也均达到了1∶5 120 以上,通过两种方法结合测定效价均显示为合格抗体。通过对4种抗血清的专化性进行研究,结果显示ASBg14和ASBg32专化性较高,抗血清ASBg26 和ASBg04专化性不是很理想。用直接PCR技术和免疫捕捉PCR技术检测谷枯病菌,结果表明,所有谷枯病菌都能产生500 bp左右的特异性片段,非谷枯病菌的8个菌株均无特异性片段产生。两种检测方法的灵敏度比较发现,直接PCR技术能检测到1×105 cfu·mL-1左右的悬浮液,免疫捕捉PCR技术能检测到103 cfu·mL-1 左右悬浮液,免疫捕捉PCR比直接PCR灵敏性提高102倍。检测人工接种的病稻种实验中,直接PCR技术能检测到2粒及以上带菌种子制得浸悬液,而免疫捕捉PCR技术仅1粒带菌种子即可。     

甜瓜细菌性叶枯病菌胶体金ICA检测试纸条的研发与应用

甜瓜(Cucumis melo)作物上会存在甜瓜细菌性叶枯病菌(Pseudomonas syringae pv.lachrymas)和瓜类细菌性果斑病菌(Acidovorax citrulli)两种细菌病害,二者混合发生,症状极其相似,不易区分.为了快速、准确地在田间检测区分出两种病害,本研究利用甜瓜细菌性叶枯病菌单克隆抗体制备了免疫层析试纸条,并建立一种快速、简便地现场检测甜瓜细菌性叶枯病菌的胶体金免疫层析分析方法(immunochromatography assay, ICA).在定性检测中,本方法对甜瓜细菌性叶枯病菌的定性检测限为2.5×103cfu/mL,与细菌性果斑病菌不存在交叉反应,具有很强的特异性.在半定量检测中,甜瓜细菌性叶枯病菌在2.5× 103～1×105 cfu/mL浓度范围内,T线和C线读数的比值与甜瓜细菌性叶枯病菌浓度的对数值呈显著的线性关系,线性方程=0.392 2x-1.263 8,相关系数R2达到0.988 1.因此,本研究制备的快速、简便、经济的高特异性甜瓜细菌性叶枯病菌ICA是一种非常实用的检测方法,不仅可以在田间实时原地快速检测甜瓜细菌性叶枯病菌,同时还能与瓜类细菌性果斑病菌进行有效区分,为甜瓜等瓜类种植业的发展带来很大帮助.     

平茹菌砖与草皮垡相间筑墙高产栽培技术

通过深入调查，总结种菇户经验，整理出一套适于高寒地区的平菇菌砖夹草皮垡筑墙高产栽培技术。文中较详细地介绍了建棚、制种，制菌砖、挖草垡、筑墙及相应的出菇管理技术。     

pyrG基因对西瓜噬酸菌致病性和组氨酸利用的影响

为研究pyr G基因与西瓜噬酸菌(Acidovorax citrulli)致病相关性,利用同源重组的方法构建pyr G基因突变体(Δpyr G),对pyr G基因的功能进行分析。结果显示,pyr G基因影响病原菌的致病性、游动性、生物膜形成和定殖能力以及激发烟草过敏反应能力;pyr G基因影响his D基因的表达;此外,pyr G基因突变影响西瓜噬酸菌对组氨酸的利用。     

红枣气体射流冲击干燥收缩特性研究

Chinese jujube originated in China and has a long historical standing. China is the only country which can produce a great amount of Chinese jujube in the world. Modern research showed that the nutrition of Chinese jujube is abundant. Chinese jujube has great medicinal and food value, with high economic value and good prospects for development. Shrinkage characteristics of dates generally refer to volume shrinkage characteristics of whole dates. Large fruit and plump grain appearance of dates is the first choice of consumers to buy elements. The volume of dried jujube points to the consumer’s heart, and is an important indicator of consumers whether to buy. Jujube processing enterprises grade the date in accordance with its size, different grades dates have different value and price. Therefore, drying is the most important steps during the dates processing as it affects directly the quality of the product, and the shrinkage characteristics is the most important quality characteristic of Chinese jujube after drying. However, the traditional natural desiccation and desiccation in the drying room of the red dates has many problems. The conventional drying method for Chinese jujube has some disadvantages of severe shrinkage, long drying time and large energy consumption etc. Therefore, in order to improve the quality of dried Chinese jujube and reduce the shrinkage, it is imperative to replace the traditional drying method with the modern technologies. Air impingement drying technology was applied to dry Chinese jujube. The drying characteristics of shrinkage, moisture effective diffusivity considering shrinkage characteristics and the activation energy of Chinese jujube shrinkage were investigated under different air temperatures (55, 60, 65 and 70℃) and air velocities (6, 9, 12 and 15m/s). The dynamic changes of the regular pattern for the shrinkage characteristic of Chinese jujube in the air impingement drying process were studied, the results showed that the volume relative shrinkage ratio of Chinese jujube appeared to increase and then decrease with the increase of air temperature and air velocity. Considering shrinkage characteristics, the moisture effective diffusivity of Chinese jujube in the air impingement drying process through the Fick’s second law was obtained. The diffusivity appeared to increase and then decrease with the decrease in moisture content in dry basis of Chinese jujube. The Arrhenius equation calculated the shrinkage activation energy of the Chinese jujube was 1281kJ/mol. This research provided technical basis for shrinkage characteristics of Chinese jujube using air impingement drying technology.     

鲜食玉米剥皮机的设计与试验

为了解决中国在鲜食玉米加工中的剥皮难题,在借鉴国外先进鲜食玉米剥皮技术的基础上研制出一种鲜食玉米剥皮机。该机采用一对转辊相向转动,转辊与玉米穗苞叶接触产生摩擦将玉米苞叶先搓起一角,之后一对转辊夹住苞叶旋转将其剥掉。该设备可适应玉米籽粒含水率介于58%～73%间的原料,对脆嫩易剥的鲜食玉米籽粒品种的破损率≤5%、剥净率≥90%、剥皮质量好。解决了中国鲜食玉米剥皮技术存在的剥净率偏低、破损率等主要性能指标不能满足使用要求的难题。经过对该设备试制、试验结果表明,完全适合高水分鲜食玉米穗剥皮。     

瓜类细菌性果斑病菌luxR基因的功能分析

luxR基因作为信号分子受体蛋白的编码基因,在革兰氏阴性菌群体感应(quorum sensing,QS)LuxI/LuxR环路中起重要作用.本研究采用PCR技术从瓜类细菌性果斑病菌(Acidovgorax avenae subsp.citrulli)xjL12中扩增出luxR同源基因,应用同源重组的方法构建了突变株(△luxR),并对其功能进行初步研究.实验结果表明,△luxR与野生型菌株相比产生的群体感应信号分子浓度降低,抗生素耐受水平有所提高,对哈密瓜(Cucuumis melo var.cantalupensis))幼苗的致病能力下降;RT-PCR实验表明,luxR基因的缺失对luxI基因的表达有抑制作用.本研究为今后对以luxR基因为靶标,利用信号干扰技术综合防治瓜类细菌性果斑病提供基础资料.