青海地区萝卜黑心病病原

2018-2021年自青海西宁及其周边地区采集萝卜黑心病样品66份,采用组织分离法对样品进行分离并纯化,获得80株轮枝菌属菌株,编号为VL1-VL80。供试菌株在PDA上生长十分缓慢,培养后产生微菌核和轮枝状分生孢子梗。微菌核不规则形,53.91-164.42×8.91-29.81 μm,平均长宽比大于2。分生孢子透明且较长,5.35-9.31×2.12-4.78 μm,不产生厚垣孢子和休眠菌丝。ITS和ACT序列测定分析表明,供试菌株均属于长孢轮枝菌A1/D1株系。测定4株代表性菌株的ITS、ACT、EF、GPD、OX和TS的片段序列,分别构建ITS片段和多基因系统发育树,ITS发育树表明供试菌株与长孢轮枝菌位于同一个分支,多基因系统发育分析表明供试菌株与长孢轮枝菌A1和D1株系位于同一个分支。致病性测定结果表明,可侵染萝卜引起黑心病症状。以上研究结果表明,青海地区萝卜黑心病菌病原为长孢轮枝菌A1/D1株系,这是我国首次报道长孢轮枝菌A1/D1株系。     

西宁市萝卜黑心病的病原

萝卜黑心病是近年来在青海省西宁地区发生的一种新病害,重病田发病率高达100%,严重影响了萝卜的产量和品质。罹病萝卜肉质根的根尖及毛根颜色变暗,纵向剖开病根,可见肉质根的中柱组织自根尖向上呈不同程度的黑色;横向剖开病根,可见肉质根的中柱组织呈现均匀或不均匀放射状分布的黑色变色点。经病原菌分离培养、科赫氏法则证病,及病原菌形态学和分子生物学鉴定,明确该病的致病菌包括长孢轮枝菌Verticillium longisporum、三体轮枝菌V. tricorpus和瓜小织球壳菌Plectosphaerella cucumerina,其中长孢轮枝菌的组织侵染率达100%,为优势病菌。这3种病菌的适宜生长温度分别为20℃、20-25℃和25℃。本文是这3种病菌为害萝卜的首次报道,萝卜是这3种病菌的新寄主植物。     

Biosurfactants are involved in the biological control of Verticillium microsclerotia by Pseudomonas spp

AIMS: To examine the effect of previously described bacterial antagonists on the viability of Verticillium microsclerotia in vitro and to elucidate the possible modes of action of bacterial strains in the suppression of Verticillium microsclerotia viability. METHODS AND RESULTS: A microplate assay was developed to test the suppressive effect of well-defined Pseudomonas spp. on the viability of Verticillium microsclerotia in vitro. Experiments using phenazine- and biosurfactant-deficient mutants indicated that biosurfactants and phenazine-1-carboxylic acid play a role in the suppression of microsclerotia viability by Pseudomonas spp. In addition, microsclerotia colonization tests revealed that Pseudomonas spp. are able to colonize the surface of the microsclerotia, but not the inner matrix. Growth response curves showed that the population levels of Pseudomonas spp. increased when they were in the vicinity of Verticillium microsclerotia, indicating that Pseudomonas spp. may utilize nutrients from the microsclerotia for their growth. CONCLUSIONS: Pseudomonas spp. seem to be good candidates for Verticilllium microsclerotia biocontrol. Biosurfactant production is one of the main mechanisms involved in their mode of action. SIGNIFICANCE AND IMPACT OF THE STUDY: This line of work may contribute to a better understanding of biological control agents and their working mechanisms.     

一株鸡传染性贫血病毒野毒株致病性及其全基因组序列比较

从山东某商品代肉鸡场表现生长迟缓的14日龄病鸡群分离到一株鸡传染性贫血病毒(CAV)C14株。C14株感染1日龄SPF鸡能抑制对禽流感病毒(AIV)的抗体反应,还能与禽网状内皮增生病病毒(REV)在免疫抑制上起协同作用。用PCR方法分段扩增出C14基因组的三条部分重叠片段,分别克隆于T载体并进行测序,拼接后得到其全基因组序列。测序结果表明,CAV-C14株基因组全长2298bp,含有3个互相重叠的开放阅读框和1个调控区。将C14与国内外已发表的CAV参考株基因组比较,同源性为97.2%~99.2%。序列比较表明CAV非编码区中含有的多个与复制及转录调控相关已知基序的序列都非常保守。CAV的3个编码基因VP1、VP2和VP3均有一定程度变异,以VP1变异性最大,且不同毒株间的3个蛋白质氨基酸序列的变异是互不相关的。     

青枯菌致病性与基因组之间的关系

青枯菌是引起植物毁灭性青枯病的病原菌。青枯菌基因组约5.8Mb,具有高(G C)含量和约5120个可能的编码基因。该菌基因组由3.7Mb的染色体和2.1Mb的大质粒所组成,其独特的基因组构成与Ⅲ型分泌系统等主要的致病因子密切相关。综述了青枯菌的致病性与其基因组之间关系的新近研究进展。     

从功能基因到生物学性状:大丽轮枝菌致病性形成的分子基础

大丽轮枝菌为典型的土传维管束病原真菌,针对其致病相关基因的挖掘与功能解析一直是植物病理学研究的热点.大丽轮枝菌具有定殖维管束、"毒素"致萎、形成微菌核、种群分化多元化等特征,这些性状最终支撑或决定了病原对寄主的致病性基础.从进化角度来说,功能基因决定生物学性状.截至目前,在大丽轮枝菌中已鉴定出上百个功能基因;但针对其与...     

Evolution of paralogous genes: Reconstruction of genome rearrangements through comparison of multiple genomes within Staphylococcus aureus

Analysis of evolution of paralogous genes in a genome is central to our understanding of genome evolution. Comparison of closely related bacterial genomes, which has provided clues as to how genome sequences evolve under natural conditions, would help in such an analysis. With species Staphylococcus aureus, whole-genome sequences have been decoded for seven strains. We compared their DNA sequences to detect large genome polymorphisms and to deduce mechanisms of genome rearrangements that have formed each of them. We first compared strains N315 and Mu50, which make one of the most closely related strain pairs, at the single-nucleotide resolution to catalogue all the middle-sized (more than 10 bp) to large genome polymorphisms such as indels and substitutions. These polymorphisms include two paralogous gene sets, one in a tandem paralogue gene cluster for toxins in a genomic island and the other in a ribosomal RNA operon. We also focused on two other tandem paralogue gene clusters and type I restriction-modification (RM) genes on the genomic islands. Then we reconstructed rearrangement events responsible for these polymorphisms, in the paralogous genes and the others, with reference to the other five genomes. For the tandem paralogue gene clusters, we were able to infer sequences for homologous recombination generating the change in the repeat number. These sequences were conserved among the repeated paralogous units likely because of their functional importance. The sequence specificity (S) subunit of type I RM systems showed recombination, likely at the homology of a conserved region, between the two variable regions for sequence specificity. We also noticed novel alleles in the ribosomal RNA operons and suggested a role for illegitimate recombination in their formation. These results revealed importance of recombination involving long conserved sequence in the evolution of paralogous genes in the genome.     

亚比棉基因组原位杂交及核型分析

亚比棉异源四倍体是山西农业大学棉花育种组于上个世纪80年代用A染色体组亚洲棉(Gossypium.arboreum)(迁西小黑籽)与G染色体组野生棉比克氏棉(G.bickii)杂交成异源二倍体后,又经过加倍而获得的.亚比棉异源四倍体不仅育性得到恢复、结铃正常,而且成功地将比克氏棉的优异性状--种子腺体延缓形成转育到亚比棉中.这为实现棉花综合利用和提高抗虫性创育了新的育种材料.在随后的多年中,山西农业大学棉花育种组对亚比棉异源四倍体进行了广泛的细胞形态学研究,对其核型做了分析.然而,仅依据形态学和普通的核型图像,还不能确定该异源四倍体棉种中比克氏棉G染色体(亚)组在核型中的表现.该文以比克氏棉gDNA为探针,亚比棉异源四倍体根尖体细胞染色体为靶细胞染色体,封阻材料为亚洲棉(迁西小黑籽),进行亚比棉基因组原位杂交(Genome in situ hybridization,GISH)及核型分析.从获得的图像中可以清晰地发现有52条染色体,其中有/无杂交信号的各一半,这直观地证实了人工复合亚比棉杂交种确为异源四倍体,而且是双二倍体.A亚组与G亚组染色体长度存在交替排列.亚比棉异源四倍体基于GISH图像的核型公式为2n=4x=52=46m(4sat)+6sm(4sat).A亚组和G亚组染色体上各有2对随体.G亚组染色体中至少有5对双重显色明显的染色体,意味着可能有A亚组染色体的交换,而A亚组染色体中只观察到或多或少的探针红色荧光信号,由于分辨率不够而难于定量分析.进一步以45SrDNA为探针,以鲑鱼精DNA作为封阻DNA,对亚比棉异源四倍体进行45SrDNA-FISH,实验表明,亚比棉异源四倍体有14个NOR(核仁组织区)信号,说明亚比棉异源四倍体有14个随体,即7对随体.比克氏棉对亚洲棉的GISH结果显示,在有亚洲棉DNA封阻的条件下,亚洲棉靶细胞染色体无任何杂交信号,说明比克氏棉与亚洲棉染色体之间不存在较大的同源或相似序列.     

Genome comparison of Bifidobacterium longum strains NCC2705 and CRC-002 using suppression subtractive hybridization

Because probiotic effects are strain dependent, genomic explanations of these differences will contribute to understanding their mechanisms of action. The genomic sequence of the Bifidobacterium longum probiotic strain NCC2705 was determined, but little is known about the genetic diversity between strains of this species. Suppression subtractive hybridization (SSH) is a powerful method for generating a set of DNA fragments differing between two closely related bacterial strains. The purpose of this study was to identify genetic differences between genomes of B. longum strains NCC2705 and CRC-002 using PCR-based SSH. Strain CRC-002 produces exopolysaccharides whereas NCC2705 is not known for reliable exopolysaccharide production. Thirty-five and 30 different sequences were obtained from the SSH libraries of strains CRC-002 and NCC2705, respectively. Specific CRC-002 genes found were predicted to be involved in the biosynthesis of exopolysaccharides and metabolism of other carbohydrates, and these genes were not present in the genome of strain NCC2705. The identification of an endo-1,4-beta-xylanase gene in the CRC-002 SSH library is an important difference because xylanase genes have previously been proposed as a defining characteristic of the NCC2705 strain. The results demonstrate that the SSH technique was useful to highlight potential genes involved in complex sugar metabolism that differ between the two probiotic strains.     

斑点杂交生物素法检测流行性出血热病毒RNA

为寻找一种用于检测流行性出血热病毒的分子杂交方法,以生物素-7-dATP标记流行性出血热病毒(EHFV)R_(22)株M片段的cDNAR_3克隆作探针,与人源性的EHFVH-114、H-435株RNA基因组进行斑点杂交,得到阳性结果,可检出5pg的cDNA或RNA。此探针与疱疹病毒DNA不出现杂交信号。以上结果说明这种标记探针具有EHFV特异性,可以扩大应用范围,结果还表明动物源性和人源性EHFV均具有共同的保守核苷酸序列。