枣果病原真菌的分离鉴定及特性分析

对云南省蒙自市6个主要枣树栽培品种储藏期的腐烂果实表面的病原真菌进行分离和鉴定,得到25株优势病原真菌,分别扩增得到核糖体DNA转录间隔区(Internal transcribed spacer,ITS)序列,通过对ITS的生物信息学和系统发育分析,并结合菌落形态特征,鉴定病原真菌分别为链格孢属(Alternaria sp.)、曲霉属(Aspergillus sp.)、葡萄座腔菌属(Neofusicoccum sp.)和脉纹孢菌属(Neurospora sp.)。     

2013年粳稻4t实仓储藏一年后优势菌种的分离与鉴定

稻谷极易受到微生物的侵染,粮食微生物的活动不仅会影响到粮食的安全储藏,导致粮食腐坏变质,还会严重影响食用者的身体健康.采用GB 4789.15-2010对稻谷进行传统培养,并从中分离得到两株优势菌株.对分离出的优势菌株进行形态学观察,观察菌株在不同生长时期的形态及菌丝、孢子形态,初步判定两菌株均为曲霉.同时运用聚合酶链式反应对两菌株的ITS及28S片段进行扩增,扩增得517bp的ITS片度及560 bp的28S片段,将基因序列在Genbank中进行相似性比对,最终确定两菌株为溜曲霉(Aspergillus tamarii)和亮白曲霉(Aspergillus candidus).     

From the micro-scale to the habitat: Assessment of soil bacterial community structure as shown by soil structure directed sampling

Natural structural units of a luvisol under maize crop were studied to assess if soil structure directed sampling could improve the understanding of arrangements of bacteria in spatially constraint location. Three habitats were defined: (i) soil around fine lateral roots (rhizo-aggregates), (ii) soil close to basal roots (core clods) and (iii) unplanted soil between rows (bare soil clods). These habitats were also investigated with maize plants resulting from Azospirillum lipoferum CRT1 inoculated seeds as a model of enhanced fine root system. Rhizo-aggregates were clearly separated from each other (disconnected habitat) in contrast to micro-samples (fragments) from clods, which belong to cohesive macro-structures. Genetic fingerprints on metagenomic extracts were used to characterize the structure of bacterial communities on 95 micro-samples from the three habitats. For eubacteria, automated RISA (Ribosomal Intergenic Spacer Analysis) of ITS (Internal Transcribed Spacer) profiles were performed. PCR-RFLP on nifH gene were used to describe the N-fixer guilds. Exploratory multivariate analyses (PCA and MDS) revealed bacterial community patterns in the sampled habitats. On the basis of ITS profiles, rhizo-aggregates harboured closely related communities, distant from those of the unplanted soil, and each sampled rhizo-aggregate could therefore be considered as a sub-unit of the whole macro-habitat, comprising all the fine roots. The observed low dissimilarity of disconnected rhizo-aggregates is likely to result from the direct influence of maize root tips on the recruitment of rhizosphere bacteria. Molecular fingerprints of nifH from basal root clods (core) were more similar to bare soil than to rhizo-aggregates, indicating similar ecological conditions without, or with, at least, poor maize exudating root influence. Although our study was performed on a limited number of situations, the distribution of bacteria was revealed to be patterned by soil structure units, which is a first step to improve the modelling of microbial ecology in soils.     

A combination of Lactobacillus buchneri and Pediococcus pentosaceus extended the aerobic stability of conventional and brown midrib mutants–corn hybrids ensiled at low dry matter concentrations by causing a major shift in their bacterial and fungal community

We evaluated the effects of applying a combination inoculant to four corn hybrids harvested at high moisture on their nutritive value and microbial populations. The treatment design was the factorial combination of corn hybrids ensiled with (INO) and without (CON) inoculant. The hybrids were TMF2R737 (MCN), F2F817 (MBR), P2089YHR (PCN), and PI144XR (PBR), ensiled at dry matter (DM) concentrations of 30.5%, 26.3%, 31.1%, and 31.5%, respectively; MBR and PBR were brown midrib mutants (BMR). The inoculant contained Lactobacillus buchneri and Pediococcus pentosaceus (4 × 10⁵ and 1 × 10⁵ cfu/g of fresh corn). The experiment had a complete randomized design with treatments replicated six times. Corn was treated or not with inoculant, packed into 7.6 L bucket silos, and stored for 100 d. At d 0, the relative abundance (RA, %) of Enterobacteriaceae was lower in PBR vs. the other hybrids [51.3 vs. x¯ = (average of) 58.4] and in the case of fungi, incertae sedis (i.s.) Tremellales and Mucoraceae were more and less abundant, respectively, in conventional hybrids vs. BMRs (x¯= 25.8 vs. x¯ = 13.9 and x¯ = 3.64 vs. x¯ = 7.52; P < 0.04). After ensiling, INO had higher LAB (9.3 vs. 7.1 log cfu/g of fresh corn) and acetic acid (3.44% vs. 1.32% of DM) and lower yeast (3.1 vs. 4.6) and molds (1.5 vs. 3.0), and also extended the aerobic stability (582 vs. 111 h) but decreased DM recovery (95.6% vs. 97.4%) vs. CON (P < 0.02). Inoculation reduced bacterial phylogenetic diversity (6.75 vs. 14.4) but increased fungal observed taxonomical units (46 vs. 20) vs. CON (P < 0.01). Also, a higher relative abundance (RA) for Lactobacillaceae (99.2% vs. 75.7%) and lower for Enterobacteriaceae (0.28 vs. 9.93) was observed due to inoculation (P < 0.001). For fungi, INO had a lower RA compared to CON for Monascaceae (12.6 vs. 44.7) and increased i.s. Tremellales (8.0 vs. 1.2) and i.s. Saccharomycetales (6.4% vs. 0.3%; P < 0.006). Inoculation changed the diverse bacterial community found in the phyllosphere across hybrids to a taxonomically uneven one dominated by Lactobacillaceae. In the case of fungi, INO application increased the fungal diversity at d 100 mainly by reducing the dominance of Monascaceae vs. CON. In conclusion, the INO treatment overwhelmed the disparate microbial populations found across BMR and conventional hybrids ensiled at low DM concentrations and ensured a significant concentration of acetic acid that modified fungal populations and in turn extended the aerobic stability of all hybrids.     

两株水牛伊氏锥虫内转录间隔区基因序列的测定与系统进化关系

为研究从水牛分离到的伊氏锥虫广西株和湖北株的分类学地位,对2株伊氏锥虫的核糖体基因内转录间隔区(ITS1-5.8S-ITS2)基因进行了克隆和测序分析.用CLUSTALXl.83软件多重比对分析了序列差异,应用MEGA4.0软件绘制系统进化树.结果表明,这2株水牛伊氏锥虫的ITS1-5.8S-ITS2 rRNA基因扩增产物分别为1 102 bp和1 095 bp,序列存在多态性,其中广西株的ITS-1序列长为340 bp,ITS-2序列长为582 bp,湖北株的ITS-1序列长为339bp,ITS-2序列长为587 bp.系统进化分析显示,这2株伊氏锥虫分布在同一大枝的不同亚群中.     

Data Access Technique and Implementation in ITS Virtual Common Information Platform

The framework of "ITS Virtual Common Information Platform"is introduced,which integrates and shares of data between ITS subsystems by integrating distributed storage and centralized control of data.The distributed data(access) engine of the platform is designed and implemented.Based on the distributed data access engine,the flow of data providing and accessing is addressed in detail.In the end a typical application of the distributed data access engine is presented.     

两种新疆羊肚菌的分类学鉴定

[目的]对新疆霍城县大西沟采集的两种形异羊肚菌(XJURML4和XJURML5)进行分类鉴定.[方法]用真菌专一引物ITS1f和ITS4扩增两菌ITS区并进行序列分析.[结果]XJURML4 ITS片段1 130 bp( EU086775),XJURML5 ITS片段1 127 bp(EU086776).两菌的ITS2区100;相似,而ITS1区仅有4 bp差异.BLAST比对测序结果发现XJURML4和XJURML5与近缘种宽圆羊肚菌(MorcheLLa esculenta (L.:Fr.)Pers.var.rotunda Pers.:Fr.)的同源性为99.28;和98.65;;NJ法构建进化树分析ITS序列,两菌也跟宽圆羊肚菌类聚在一起.随机引物OPA3进行RAPD扩增,共产生出六条带,二条相同,剩余四条为多态性条带,表明两菌基因组上也有差异.[结论]ITS区序列和进化树分析、OPA3 - RAPD标记技术均表明两菌是宽圆羊肚菌种内的新变种,而与传统形态学鉴定结果不一致.     

<Emphasis Type="Italic">Platanus</Emphasis> × <Emphasis Type="Italic">acerifolia</Emphasis> genotypes surviving to inoculation with <Emphasis Type="Italic">Ceratocystis platani</Emphasis> (the agent of canker stain): first screening and molecular characterization

Canker stain, caused by the fungus Ceratocystis platani, is a destructive disease in Platanus spp. It has been recently proved that resistant accessions can be produced and grown in Europe. However, additional resistant genotypes are still needed in order to avoid the onset of virulent pathogen strains favoured by the selection pressures exerted by genetically homogeneous resistant plane tree plantings. In this study we present the results of two parallel experiments performed on 975 accessions of P. × acerifolia seedlings and P. × acerifolia clones derived by cutting propagation from mature trees grown in the urban environments. The selection process was based on inoculation with C. platani and yielded 13 accessions that showed different types of resistant reactions and survived in a stable manner thoughout the period of observation. Selected accessions were characterised by sequencing the rDNA-ITS region and by developing PCR procedures capable of detecting P. orientalis and P. occidentalis LEAFY homoeologues. These molecular analyses enabled us to confirm the identification of the species, its hybrid origin and to assess an evident genetic variability among the accessions, which therefore have to be considered as different genotypes. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

月季ITS反应体系的优化

以古老月季品种‘玉玲珑'为试材,利用CTAB法对月季进行总DNA提取,对其PCR扩增条件中的主要因素进行了多梯度的优化,总反应体系为25μL,其中引物浓度为0.4μmol/L、Mg~(2+)浓度为2.0 mmol/L、dNTP为0.2 mmol/L、Taq酶用量为1.0 U、DNA模板用量为70 ng.利用该优化体系对部分月季品种和野生种进行PCR扩增和电泳检测,扩增条带清晰,结果稳定,测序结果理想,表明该体系适用于月季的ITS序列分析.     

我国梨和部分国外梨果实上链格孢菌的鉴定研究

本文从链格孢的产孢表型、分生孢子特征和分子生物学方面研究了从河北鸭梨、新疆库尔勒香梨、美国和日本以及智利的梨上分离的链格孢菌的种类.经对115支Alternaria菌株的形态学鉴定,共确定了3个种,即Alternaria alternata、A.tenuissima和A.infectoria.ITS系统发育分析研究表明,Alternaria大孢子种彼此间及大孢子种与小孢子种间已发生明显分化,可以根据ITS序列差异明确区分.而供试的多数小孢子种在ITS序列上差异很小,因而不能根据ITS序列差异来区分.