Characterization,phylogenetic analyses and pathogenicity of Bipolaris setariae associated with brown stripe disease on sugarcane in Yunnan,China

Brown stripe disease is a severe foliar fungal disease of sugarcane worldwide and is widespread in all sugarcane planting areas in China. Brown stripe is a major disease that seriously affects the output and quality of the sugarcane industry in Yunnan Province, China's second-largest sugar base, while the pathogen of this disease remains not yet fully understood. To address this, we isolated and identified the fungi associated with 68 leaf samples showing typical symptoms of brown stripe from 22 sugarcane varieties in different areas of Yunnan Province. A total of 113 isolates were obtained, which were morphologically similar. Of these, 64 representative isolates were sequenced for the internal transcribed spacer region (ITS), GAPDH and EF-1α loci. All representative isolates grouped with the type strain of Bipolaris setariae in the phylogenetic trees inferred with individual and concatenated sequences of ITS, GAPDH and EF1-α. Pathogenicity test results showed that B. setariae strains were able to induce typical symptoms of brown stripe. The results obtained in this study clarify that only B. setariae is associated with sugarcane brown stripe in Yunnan, China. It is recorded here for the first time as a pathogen causing sugarcane brown stripe in Yunnan, and it is able to infect many major cultivars and new varieties, posing a new threat to the sugar industry in Yunnan Province. In addition, these results provide the scientific basis for the future breeding of disease-resistant varieties and effective prevention and control of sugarcane brown stripe disease.     

Early detection of Neophysopella tropicalis in grapevine leaves and on spore traps by qPCR

Neophysopella tropicalis, one of the causal agents of Asian grapevine leaf rust (AGLR), can cause severe epidemics in Brazil that lead to yield losses in commercial vineyards. An early detection of the pathogen by air sampling of urediniospores on spore traps or in symptomless leaves would be valuable to multiple studies, such as epidemics modelling, risk forecasting, monitoring of pathogen introductions in rust-free areas, and predicting the beginning of epidemics. This study developed a quantitative PCR (qPCR) protocol to quantify N. tropicalis urediniospores attached to adhesive tapes and in grapevine leaves before symptom appearance. A specific primer pair was designed based on the internal transcribed spacer (ITS) sequence region of the AGLR pathogen. Standard amplification curves using genomic DNA from urediniospores of N. tropicalis and from urediniospores attached to adhesive tapes were established. Grapevine leaves inoculated with N. tropicalis were collected at 2, 5, and 7 days postinoculation (dpi). One primer pair (580F/720R) amplified a 140 bp product in all AGLR isolates but did not amplify products of other rust genera, such as Phakopsora, Puccinia, Hemileia, Tranzschelia, Cerotelium, and Coleosporium. As little as 0.1 pg DNA and 10 urediniospores of N. tropicalis attached to adhesive tapes could be detected. qPCR enabled the detection of the pathogen as early as 2 dpi, before symptom appearance. This method can be used to monitor N. tropicalis inoculum in grapevine-growing areas and to quantify symptomless infections of the AGLR pathogen.     

Evaluation of the combination of 1,3-dichloropropene and dazomet as an efficient alternative to methyl bromide for cucumber production in China

BACKGROUND: The combination of 1,3‐dichloropropene (1,3‐D) and dazomet (DZ) offers a potential alternative to methyl bromide (MB) for soil disinfection. MB is scheduled to be withdrawn from routine use by 2015 in developing countries. Combination treatments of 1,3‐D + DZ were evaluated in a laboratory study and in two commercial cucumber fields. RESULTS: Laboratory studies found that nearly all of the tested combinations of 1,3‐D and DZ displayed positive synergistic activity on root‐knot nematodes (Meloidogyne spp.), two major soilborne fungi (Fusarium spp. and Phytophthora spp.) and the seeds of two major weed species (Digitaria sanguinalis and Abutilon theophrasti). Field trials revealed that the combination of 1,3‐D and DZ (at 10 + 25 g m^?2) successfully suppressed Meloidogyne spp. root galling, sharply reduced Fusarium spp. and Phytophthora spp. and maintained high cucumber yields. The combination treatment of 1,3‐D + DZ was more effective than 1,3‐D or DZ alone and provided results similar to methyl bromide with respect to pest control, plant mortality, plant height, yield and income. All of the treatments were significantly better than the non‐treated control. CONCLUSION: The results indicate that the tested combination of 1,3‐D and DZ offers an efficient alternative to methyl bromide for cucumber production. Copyright © 2012 Society of Chemical Industry     

东省不同地区玉米内生真菌的群落组成和多样性分析

为探究山东省玉米内生真菌的资源分布、群落组成及多样性,采用叶片组织分离法对山东省52个市县的玉米内生真菌进行分离纯化,通过内转录间隔区（internal transcribed spacer,ITS）序列分析技术进行内生真菌的种属鉴定,并分析内生真菌的群落组成和多样性。结果表明,共分离到857株内生真菌,隶属25个属,其中链格孢属Alternaria的菌株数量最多,为583株,占总菌株数的68.03%,其次为镰刀菌属Fusarium有71株,占总菌株数的8.28%;大部分地区的优势属为链格孢属,少数地区的优势属为镰刀菌属、炭疽菌属Colletotrichum和弯孢属Curvularia。不同地区的内生真菌定殖率变化很大,在12.50%~100.00%之间,平均定殖率为76.15%;陵城的多样性指数和均匀度指数最高,分别为1.74和0.97;莱芜的丰富度指数最高为2.22。山东省玉米内生真菌可以分为5个类群,其中黄河流域地区内生真菌定殖率相对较高。表明山东省玉米内生真菌种类丰富,不同地区间多样性水平存在差异。     

八种杀虫剂对黑粪蚊的防治效果及残留分析

选择8种杀虫剂,分别在实验室和菇房条件下研究其对黑粪蚊的防治效果和对平菇菌丝生长的影响,并分析施药后不同时间平菇中的农药残留.结果表明,40%辛硫磷EC、40%氧乐果EC、48%毒死蜱EC对黑粪蚊具有很好的防治效果,但对平菇菌丝的抑制率明显高于其它药剂;10%吡虫啉WP、4.5%高效氯氰菊酯EC不仅对黑粪蚊成、幼虫具有良好的防治效果,而且使用后对平菇菌丝生长的影响较小;2.5%高效氯氟氰菊酯EW、4.5%高效氯氰菊酯EC、1.8%阿维菌素EC等使用5天后均未检测到农药残留,5%氟虫腈SC和10%吡虫啉WP药后15天仍能检测到残留.     

一种改进的尖孢镰刀菌的实时荧光定量PCR检测方法

为实现荧光定量PCR对土壤病原真菌数量更为高效、灵敏的检测,本研究将液体培养的孢子悬浮液和长年耕作的水稻土制作成孢子浓度为4×10~1~4×10~6 spore/g的带菌土,采用MoBio PowerSoil@DNA Isolation Kit提取模拟带菌土壤总DNA,引入常规PCR预扩增包含qPCR目标序列的1 446bp片段,以预PCR产物为模板进行qPCR,构建荧光定量PCR标准曲线。研究结果表明:以试剂盒提取的带菌土壤总DNA为模板绘制的qPCR标准曲线相关系数r为0.985,检测下限为4×10~3 spore/g土;引入预PCR后,qPCR标准曲线相关系数r为0.974,检测下限为4×10~2 spore/g土,较未引入时提高了10倍,结合不同土壤病原真菌的特异性引物,该检测方法可为土壤病原真菌的有效定量检测提供技术支撑。     

不同季节土庄绣线菊根围丛枝菌根真菌群落差异性研究

土庄绣线菊(Spiraea pubescens)是内蒙古大青山干旱生态系统重要的护土灌木,研究季节变化对土庄绣线菊根围丛枝菌根真菌(arbuscular mycorrhizal fungi,AMF)群落的影响,有助于进一步探讨AMF在土庄绣线菊生长发育过程中发挥的作用。采用传统染色检测与Illumina Mi Seq测序相结合的方法,研究季节变化对内蒙古大青山土庄绣线菊AMF侵染率、孢子密度、丰富度、多样性和群落组成的影响。结果显示:在春季、夏季和秋季,AMF侵染率和丛枝丰度不存在显著差异,而孢子密度却存在显著差异。AMF丰富度和多样性在夏季显著低于春季和秋季,但在春季和秋季间差异并不显著;主成分分析(principal coordinate analysis,PCo A)和PERMANOVA分析表明,春季和秋季AMF群落组成与夏季存在显著差异,而在春季和秋季间差异并不显著。     

海洋细菌B-9987菌株产生的抑菌物质及对几种植物病原真菌的作用

 从海洋生境中分离得到317株菌株,筛选获得1株芽孢杆菌B-9987菌株,其胞外分泌的代谢产物对植物病原真菌有显著的抑制作用,其中对茄交链孢菌、大丽轮枝菌、黄枝孢菌、立枯丝核菌主要是造成真菌孢子或菌丝末端膨大成球状,继而胞壁崩解,原生质外泄;对尖孢镰刀菌和莴苣霜霉菌的抑制作用,则表现为孢子萌发率降低和芽管长度明显变短。该抑菌物质经乙醚萃取,硅胶G薄层层析纯化,官能团显色反应初步推断为酚类化合物。     

应用均匀设计法筛选稗草病原菌产孢最佳配方培养基

 筛选生防菌最佳产孢培养基是高效、大量生产生防菌孢子的基础。应用均匀设计法结合二次多项式回归分析,从众多因素中筛选出稗草病原菌弯孢菌(Curvularia lunata)和互格交链孢菌(Alternaria alternata)最佳产孢培养基组分,建立多元二次回归数学模型。弯孢菌最佳产孢组分为胡萝卜、芹菜、欧芹、水芹、莴苣、蔗糖和Vc;互格交链孢菌最佳产孢组分为胡萝卜、芹菜、水芹、莴苣和蔗糖。根据回归模型计算理论(拟和)产孢值。比较实测产孢量和理论产孢值,结果表明,稗草病原菌的实测产孢量和回归模型拟合产孢值非常接近,拟合误差小。说明采用均匀设计法从众多因素中筛选培养基最佳成分是可行的。