四川省玉米弯孢叶斑病菌的种类及生物学特性研究

为了明确引致四川省玉米弯孢菌叶斑病（Maize Curvularia leaf spot）的病原菌种类和优势种,以便为病害的防治和抗病育种提供依据,对所获得的78个弯孢菌（Curvularia）菌株采用形态学分类法进行种类鉴定,结果鉴定出6个种或变种,其中属于新月弯孢菌（Curvularia lunata）种类的菌株占种群的61．5％,新月弯孢菌是四川省玉米弯孢菌叶斑病的主要致病菌种类．对属于其中3个种的4个弯孢菌菌株进行生物学特性研究,结果表明,各供试菌株在不同条件下的生长速度都存在一定的差异．     

四川省玉米弯孢菌对玉米的致病性研究

为了明确四川省玉米弯孢菌叶斑病(Maize Curvularia leaf spot)病原菌的致病性分化情况,在四川省玉米弯孢菌种类鉴定和生物学特性研究的基础上,选用不同种类或不同来源的27个弯孢菌菌株,在感病品种川单13和抗病品种Mo17进行致病性测定.把病斑大小和产孢量作为弯孢菌致病性鉴定与评价的标准,结果发现不同种类的弯孢菌或同一种类不同来源的菌株对玉米均有致病性,而且各菌株之间存在致病性的分化,可初步划分为强、中等和弱致病力三种类型.     

根癌农杆菌介导的异角状拟盘多毛孢菌转化系统研究

以携带潮霉素B磷酸转移酶抗性基因(hph)的pBHt1作为转化载体,根癌农杆菌AGL-1作为转化介体,实施转化异角状拟盘多毛孢菌。研究发现,异角状拟盘多毛孢菌的最优转化体系为:异角状拟盘多毛孢菌分生孢子悬浮液浓度为1×106个/mL,根癌农杆菌OD600为0.3,共培养时间48 h,共培养温度为25℃,诱导培养基中乙酰丁香酮(AS)浓度为200μmol/L,选择培养基添加250μg/mL潮霉素B、250μg/mL头孢噻肟钠。1×106个异角状拟盘多毛孢菌分生孢子可以产生200～300个转化子,随机挑选10个转化子进行PCR检测,均能扩增出预期条带,且在不含潮霉素B的PDA培养基平板上转化子连续培养5代后,hph基因仍能稳定遗传,这表明T-DNA已经插入到异角状拟盘多毛孢菌的基因组中。此次建立的异角状拟盘多毛孢菌的转化体系可为该病菌的功能基因研究和寄主与病原菌的互作研究提供有效的工具。     

有机杀菌剂与苦皮藤素对玉米弯孢霉的联合毒力研究

采用生长速率法,测定了生产上常用防治玉米弯孢霉叶斑病的7种有机杀菌剂和1种植物源农药--苦皮藤素对玉米弯孢霉的毒力,得到8个毒力回归方程及相应EC50值.结果表明:供试有机杀菌剂中烯唑醇和扑海因对玉米弯孢霉属极强毒力杀菌剂,其抑制中浓度分别为1.4048E 1和2.2172E 1.苦皮藤素对玉米弯孢霉菌丝生长的抑中浓度为5.8235E 5.所筛选出的两种对玉米弯孢霉菌丝生长具极强抑制作用的有机杀菌剂分别与苦皮藤素的混配,试验结果表明:苦皮藤素与烯唑醇3∶ 7的混配比例对玉米弯孢霉菌丝生长的实际相对抑制率最大,达74.29%,且为增效作用组合,增效比为1.87.     

不同抗性玉米自交系对玉米弯孢菌侵染应答反应差异蛋白分析

采用Mg／NP-40及PEG预分离技术,在去除叶片高丰度表达蛋白主要是Rubisco的基础上,研究了鲁原92（高抗）,78599-1（抗）,E28（感）和黄早4（感）4个玉米自交系苗期叶片对玉米弯孢菌（Curvularia lunata）强致病株系CX-3侵染应答反应差异蛋白质图谱．接种病菌后抗、感自交系蛋白质点总数均明显增加;抗性自交系表达丰度上调二倍的蛋白点数多于感病自交系．对可能与抗、感病性明显相关的8个差异蛋白点进行了质谱鉴定,发现玉米萌发素类似蛋白GLP和翻译起始因子elF-5A与玉米对玉米弯孢菌叶斑病的抗性关系密切．     

提高根癌农杆菌介导黑曲霉转化效率的研究

为提高根癌农杆菌(Agrobacterium tumefaciens)介导黑曲霉(Aspergillus niger)转化(ATMT)效率,本文对影响转化效率的分生孢子与农杆菌的比例、转化媒介、诱导溶氧、诱导时间、诱导时乙酰丁香酮(AS)浓度这5个因素进行单因素实验,最终确定提高其转化效率的最优条件为:根癌农杆菌诱导过程在400,μmol/L乙酰丁香酮终浓度条件下,100,m L三角瓶中28,℃、100,r/min振荡培养5,h,分生孢子与根癌农杆菌的比例为1﹕100,在硝酸纤维膜媒介上24,℃共培养48,h.在此优化条件下,黑曲霉的转化效率达到35个/107分生孢子.相比于优化前,转化效率提升了78%,,并且整合至黑曲霉基因组的外源潮霉素B抗性基因在转接15代以后仍能稳定遗传.     

13种杀菌剂对玉米大斑病菌和弯孢霉叶斑病菌的毒力测定

为了筛选出高效、低毒的杀菌剂,采用生长速率法和孢子萌发法比较了13种杀菌剂对玉米大斑病菌和弯孢霉叶斑病菌的毒力作用。结果表明,不同杀菌剂之间对2种病菌菌丝生长和孢子萌发的抑制活性存在显著差异。其中氟啶胺、苯醚甲环唑、烯唑醇、丙环唑、腈苯唑、咪鲜胺锰盐等对病菌菌丝生长具有较强的抑制效果,其EC50值均低于1μg·mL~(-1),而代森锰锌的抑制作用较差,EC50值分别为5.3745和6.4650μg·mL~(-1);6种不同作用机理的杀菌剂中,氟啶胺和异菌脲对大斑病菌孢子萌发抑制效果较好,EC50值分别为0.3147和1.5156μg·mL~(-1),而丙环唑和吡唑醚菌酯则对弯孢霉叶斑病菌孢子萌发抑制作用较佳,EC50值分别为5.0490和18.8439μg·mL~(-1)。     

金叶女贞似尾孢叶斑病初步研究

金叶女贞在6－7月和9－10月间大量落叶，研究结果表明系叶片上发生叶斑病所致，其中一种为假尾孢菌。该菌的分生孢子宽为7．1－9．6μm，明显不同于已报道过的发生在木犀科中7种假尾孢菌，在发病初期连续3次喷施多菌灵、甲基托布津，可有效地控制叶斑病发生为害。     

金叶女贞假尾孢叶斑病初步研究

金叶女贞在 6 - 7月和 9- 10月间大量落叶 ,研究结果表明系叶片上发生叶斑病所致 ,其中一种为假尾孢菌 .该菌的分生孢子宽为 7.1- 9.6 μm ,明显不同于已报道过的发生在木犀科中 7种假尾孢菌 ,在发病初期连续 3次喷施多菌灵、甲基托布津 ,可有效地控制叶斑病发生为害 .     

菌酶共酵玉米秸秆生产微生物油脂的研究

以玉米秸秆为原料,利用产黄纤维单孢菌、皮状丝孢酵母与纤维素酶处理玉米秸秆,采用单因子试验与正交试验探索产黄纤维单孢菌、皮状丝孢酵母与纤维素酶共酵玉米秸秆生产微生物油脂最佳条件。研究结果表明:当维素酶添加量为1%,产黄纤维单孢菌与皮状丝孢酵母菌种配比为1∶1,混合菌接种量为9%,在35℃下发酵36h,每100ml玉米秸秆发酵液中可获微生物油脂0.116g。     

Analysis of the Protein Electrophoresis Patterns of Curvularia Lunata in Maize and Immunological Detection of the Pathogenic Culture

Twelve isolates of Curvularia lunata and six related species from maize or other host plants are examined for the analysis of polyacrylamide gel electrophoresis (SDS-PAGE) patterns of their soluble proteins,Differences exist significantly among different species and the same species.There is a special protein band for Curvularia at relative mobility value 0.177(Rf 0.177) and a special protein band for C.lunata at Rf 0.225,Two kinds of rpimary antibodies to C.lunata are used to indirect ELISA and Avidin-Biotion-peroxidase Complex(ABC) immunoassay to evaluate their detecting sensitivity and specificity,The sensitivity of ABC immuno- assay is higher than indirect ELISA.APAbs to C.lunata is special to C.lunata.     

Proteomic comparison of four maize inbred lines with different levels of resistance to Curvularia lunata(Wakker)Boed infection

Protein profiles of leaves in four maize inbred lines with different disease resistance to pathogen Curvularia lunata(Wakker)Boed were studied by two-dimensional electrophoresis(2-DE)and mass spectrometry.Proteins were extracted from the forth leaf of maize seedlings 24 h after fungal inoculation,and fractionated by polyethylene glycol to precipitate the most abundant leaf protein,Rubisco,before gel separation.Protein profiles from 2-DE showed that total numbers of protein spots were increased in all four inbred lines inoculated with C.lunata CX-3 strain compared with the control.The numbers of changed protein spots in abundance were higher in resistant inbred lines than in susceptible ones,which implied that resistant inbred lines were more sensitive than susceptible ones to pathogen infection.Among proteins identified by MALDI-TOF MS,germin-like protein GLP and translation initiation factor eIF-5A were supposed to play important roles in maize resistance against C.lunata infection.     

Proteomic comparison of four maize inbred lines with different levels of resistance to Curvularia lunata （Wakker） Boed infection

Protein profiles of leaves in four maize inbred lines with different disease resistance to pathogen Curvularia lunata(Wakker)Boed were studied by two-dimensional electrophoresis(2-DE)and mass spectrometry.Proteins were extracted from the forth leaf of maize seedlings 24 h after fungal inoculation,and fractionated by polyethylene glycol to precipitate the most abundant leaf protein,Rubisco,before gel separation.Protein profiles from 2-DE showed that total numbers of protein spots were increased in all four inbred lines inoculated with C.lunata CX-3 strain compared with the control.The numbers of changed protein spots in abundance were higher in resistant inbred lines than in susceptible ones,which implied that resistant inbred lines were more sensitive than susceptible ones to pathogen infection.Among proteins identified by MALDI-TOF MS,germin-like protein GLP and translation initiation factor eIF-5A were supposed to play important roles in maize resistance against C.lunata infection.     

Comparative proteomic analysis of the response in resistant and susceptible maize inbred lines to infection by Curvularia lunata

Proteins differentially expressed from maize leaves in response to the infection by Curvularia lunata strain CX-3 were identified through a high-resolution two-dimensional gel electrophoresis (2-DE) method. Two inbred lines, 78599-1 and E28, were used, respectively, as resistant and susceptible lines to CX-3 infection. Proteins were extracted from the fourth leaves of six- or seven-leaf stage plants sampled at 24, 36, 48, 60, and 72 h after inoculation with CX-3. Twenty-seven differentially expressed protein spots resolved on the 2-DE gels were identified by MALDI-TOF MS/MS. The results showed that these proteins are associated with photosynthesis, respiration,oxidative and drought stress tolerance as well as signal transduction in maize. Among stress-related proteins, the 22 kDa drought-inducible protein, putative glutathione peroxidase (GPX), and translation initiation factor (eIF-5A) were up-regulated in the resistant inbred line and were implicated in host defense response to C. lunata infection. It suggests that drought-inducible and oxidation stress-related proteins might directly contribute to maize resistance to C. lunata.     

化学农药对玉米中后期几种病虫害的田间防治效果

在玉米抽雄期喷施化学农药防治玉米生长中后期的几种病虫害,结果表明,40%氯虫·噻虫嗪WG 225 g/hm2剂量对玉米螟虫和蚜虫均有较高的防治效果,田间防效分别为81.08%和89.91%,小区产量比对照增加20.8%.10%吡虫啉WP 450 g/hm2剂量对蚜虫的防效为63.64%,对玉米螟虫效果较差.12.5%烯唑醇WP 450 g/hm2剂量对玉米锈病的防效为76.60%.30%苯甲·丙环唑对小斑病和弯孢霉叶斑病的防效分别为62.04%和51.72%.氯虫·噻虫嗪与烯唑醇混合喷施对中后期的玉米螟虫、蚜虫和锈病具有较好的控制作用.     

解淀粉芽孢杆菌Q426可湿性粉剂的研制

通过对载体、润湿剂、分散剂、紫外保护剂的筛选,确定了解淀粉芽孢杆菌Q426可湿性粉剂的最佳配方。最终所得制剂中载体D10％、润湿剂6％、分散剂6％、紫外保护剂0．5％。测得制剂的芽孢数量为2．82×10”efu·g-1,润湿时间为43s,悬浮率为76％,指标均达到国家标准及预期目标。     

荧光法检测根际土壤胞外酶活性的优化

为准确测定土壤胞外酶活性,对β-葡萄糖苷酶(βG)、纤维二糖水解酶(CBH)、β-N-乙酰氨基葡萄糖苷酶(NAG)和亮氨酸氨基肽酶(LAP)、碱性磷酸酶(AKP)5种有关C、N、P循环的关键酶的荧光法测定进行了优化.结果发现,使用Cytation5等高灵敏度的酶标仪测定时水土比值应高于300∶1(mL∶g).5种胞外酶活性的最佳测定参数为:AKP、βG用超纯水作缓冲溶液,XH-C漩涡混匀器(60 W)混匀土壤5 min,25 ℃培养4 h;CBH用乙酸钠(0.2 mol/L,pH 5.8)作缓冲溶液,XH-C漩涡混匀器(60 W)混匀土壤5 min,25 ℃培养2 h;NAG用乙酸钠(0.2 mol/L,pH 5.8)作缓冲溶液,摇床(150 r/min)混匀60 min,25 ℃培养3 h;LAP用超纯水作缓冲溶液,摇床(150 r/min)混匀60 min,25 ℃培养1 h.本研究结论可为提高荧光法准确测定土壤胞外酶活性提供依据.     

温州盘菜种子萌发特性研究

本文研究了温州特色蔬菜温州盘菜种子的萌发特性。结果表明 :温州盘菜种子属脂肪类种子 ,最适浸种时间为 6小时 ,种子萌发的适宜温度为 2 5℃～ 30℃之间的恒温 ,发芽床以 6 0 %～ 80 %含水量的砂床效果最好。     

悬铃木白粉病病原菌及其生物学特性

对近年来在南京、上海、杭州等地危害较为严重的悬铃木白粉病病原菌的无性型和有性型进行了形态学观察和分子生物学鉴定。结果表明,该病害病原的无性型为粉孢属一种(Oidium sp.),病原的有性型为孓蠹菌门的Erysiphe platani。病原菌生长的适宜温度为20~28 ℃,最适温度为26 ℃;湿度对孢子萌发的影响不大,一般环境湿度下都可萌发;分生孢子寿命约为13 d。     

高温环境下2种常用平菇生产菌种的耐热性和生存极限

采用PDA培养基,在体视显微镜下切取菌丝尖端2 mm接种,进行菌丝体平板培养,探讨平菇豫6,831菌丝体在持续高温环境下的生长能力和对极限高温环境的耐受能力.研究表明,40℃以上持续高温时,参试菌种的菌丝体均不能生长,在34℃下,豫6,豫831生长相对良好,回置25℃后的生长旺盛.供试菌种在高温处理之后其恢复生长所需要...