蝴蝶兰细菌性褐斑病病原菌初步鉴定

对蝴蝶兰褐斑病病原细菌进行了分离、致病性测定、形态观察、染色反应、培养性状观察和生理生化特性测定。结果表明,该菌与黄单胞菌属野油菜黄单胞菌种性状、特征完全相同。因此,可以初步断定,引起蝴蝶兰褐斑病的病原菌是黄单胞菌属野油菜黄单胞菌[Xamthomonas campestris(Pammel)Dowson.]。     

黄单胞菌毒性因子调控的研究进展

黄单胞属植物病原细菌寄主非常广泛,能引起许多种重要经济作物发生病害。黄单胞菌在寄主组织内的侵染和繁殖取决于分泌的多糖、脂多糖、吸附素和III型分泌系统等毒性因子。细菌毒性因子的协调表达是通过群体感应途径、双组分系统和Clp、Zur、FhrR、HrpX和HpaR等转录后调控因子精细调控的。此外,毒力基因表达还受RNA结合蛋白RsmA转录后控制。在本综述中,我们对黄单胞菌控制分泌毒性因子的调控网络的研究进展进行了概括。     

转Xa21基因水稻的根际细菌群落研究

在水稻不同生育期抽样,采用稀释平板分离法对转Xa21基因水稻、受体皖A和常规稻汕优63三个水稻品种的根际细菌种群动态变化及其多样性进行了初步研究。研究结果表明,在水稻不同的生育期,分离出的细菌有棒杆菌属、芽孢杆菌属、黄单胞菌属、根瘤菌属、Novosphingobium属、欧氏杆菌属、葡萄球菌属、假单胞菌属8个属。3个水稻品种之间的根际细菌种类基本相同,总数量和多样性指数随着时间的变化,同一水稻品种不同时间之间差异显著,而同一时间不同品种间差异较小。转Xa21基因水稻对根际细菌影响小。     

哈密瓜内生细菌菌群密度及分布动态

分离哈密瓜植株组织中的内生细菌,对不同品种、不同栽培环境、不同组织器官、不同生育期的哈密瓜内生细菌的菌群组成、菌群密度的动态分布特点进行研究.结果表明:哈密瓜不同品种、不同种植地区、不同组织器官、不同生育期内生细菌的菌群密度有较大差异.抗病品种中的内生细菌显著高于感病品种.哈密瓜各组织中,种子的内生细菌最多,其次为根,再次为茎、叶片、叶柄,果实中最少.在不同生长阶段,随着哈密瓜生育期的延长,内生细菌数量变化呈"高-低-高"的变化特点,即从苗期到花期,植株内生细菌总量下降,至成熟期数量显著增加的特点.对所分离的内生细菌进行菌群分析表明,哈密瓜内生细菌包括芽孢杆菌属、黄单胞菌属、假单胞菌属、欧文氏菌属、葡萄球菌属、鞘氨醇单胞菌属、黄色单胞菌属和类芽孢杆菌属(Paenibacillus)等,其中芽孢杆菌属出现的频率最高,在所分离的哈密瓜品种中都是优势菌群,约占总分离菌株的54.58%;但菌群多样性在不同哈密瓜品种和种植地区间差异不显著;种子中类群最少,随着植株生长,类群增加.     

沿黄灌区饲用油菜与苏丹草间作对土壤酶活性及细菌群落结构的影响

为探究沿黄灌区饲用油菜与苏丹草间作对土壤酶活性及细菌群落结构的影响,于2022年在内蒙古鄂尔多斯市达拉特旗,采用高通量测序技术结合土壤酶活性,比较分析饲用油菜单作、苏丹草单作、饲用油菜苏丹草间作3个种植模式下的土壤细菌群落结构及酶活性的差异。结果表明：饲用油菜苏丹草间作显著提高土壤脲酶、蔗糖酶活性,较饲用油菜单作和苏丹草单作分别提高了8.09%~13.93%和13.01%~58.02%。饲用油菜苏丹草间作使土壤细菌OTUs数目和细菌群落多样性指数提高。不同种植模式下土壤细菌群落组成和相对丰度表现不同,其中变形菌门、拟杆菌门、放线菌门、芽单胞菌门为优势菌门,Salinimicrobium、海杆菌属、革兰菌属为优势菌属;间作种植模式提高了变形菌门、放线菌门、芽单胞菌门、革兰菌属的相对丰度。相关性分析表明：土壤脲酶、蔗糖酶与变形菌门相对丰度呈显著正相关关系,与酸杆菌门相对丰度呈显著负相关关系;土壤碱性磷酸酶与蛭弧菌门相对丰度呈显著正相关关系,与放线菌门相对丰度呈显著负相关关系;土壤过氧化氢酶与蛭弧菌门相对丰度呈显著正相关关系,与厚壁菌门相对丰度呈极显著负相关关系。综上可知,饲用油菜苏丹草间作改变了土壤细菌群落结构,提高土壤酶活性和有益菌物种丰度,可作为改善沿黄灌区盐碱土壤微生态环境的有效措施。     

海南一品红细菌性叶斑病病原菌的分离与鉴定研究

 为明确海南省近几年发生的一品红细菌性叶部病害的病原菌特别是该病原菌的分子特征,为该病害的治理提供可行的依据,本文描述了2005年海南省部分花圃一品红植株上发生的细菌性叶斑病症状,并通过致病性测定、BIOLOG分析和16SrDNA序列比较将分离的病原菌鉴定为黄单胞菌属(Xanthomonas);部分碳源利用测定进一步显示2005年海南分离的菌株与2004年海南报道的细菌性疫病病原菌存在差异,但与杭州地区报道的一品红细菌性叶斑病菌基本一致。利用黄单胞菌模式种典型菌株的核糖体DNA内转录间隔区(internal transcribed spacer,ITS)序列构建系统发育树,结果显示3个海南菌株与巴豆黄单胞菌(Xanthomonas codiaei)和地毯草黄单胞菌(X.axonopodis)单独聚合成群,其中与巴豆黄单胞菌亲缘关系最近。     

红掌细菌性疫病的病原菌初步鉴定

 在云南西双版纳的红掌上发现一种由细菌侵染引起的病害,从叶片的病组织上分离到具有致病性的杆状细菌,将分离的病原菌接种于健康的红掌上,表现出与田间一致的症状。感病初期在叶缘或叶脉间出现水渍状小点,后期病斑多呈棕褐色至黑色坏死,病健交界处多呈黄色。从接种发病的病斑与田间标样上分离的病原菌菌落形态完全相同。通过菌株的培养性状、常规生理生化特性及透射电镜下菌体形态观察结果,将病原菌初步鉴定为黄单胞菌属(Xanthomonas)。BIOLOG鉴定和致病性测定结果进一步鉴定病原菌为地毯草黄单胞菌花叶万年青致病变种Xanthomonas axonopodis pv. dieffenbachiae(Xad)(McCulloch & Pirone) Vauterin et al.     

多肽抗生素apidaecin对一些植物致病细菌的抗菌活性及对烟草原生质体毒性的研究

 以多肽抗生素Shiva-I为对照,利用平板抑菌圈法测定了多肽抗生素apidaecin对我国一些重要植物致病细菌的抗菌活性。发现apidaecin对供试棒状杆菌属、黄单胞菌属的菌株没有活性,但是对供试假单胞菌属、欧文氏菌属和土壤杆菌属菌株的抗菌活性比Shiva-I强2~5倍,其最小抑菌浓度一般在0.4~5.9μmol/L之间。apidaecin C-末端酰胺化后,抗菌活性略有升高。Shiva-I对烟草原生质体的最小抑制浓度为86.2μmol/L,2 0 mmol/L浓度的apidaecin对烟草原生质体依然没有毒性。     

不同种植年限对特殊药材土壤化学性质和微生物多样性的影响

以不同种植年限特殊药材根际土壤为材料,采用IlluminaNovaSeq高通量测序法和常规分析方法,分析了种植年限(0 a, 1 a, 4 a, 24 a)对根际土壤化学性质及微生物群落多样性的影响。结果表明：连续种植24 a后,土壤电导率、硝态氮含量显著高于其他种植年限。特殊药材连作对土壤细菌α-多样性指数均无显著影响,但土壤细菌丰富度和多样性减少;连续种植24 a真菌α-多样性指数中ACE指数、Chao1指数显著高于种植1 a,真菌丰富度和多样性增加。细菌群落Proteobacteria(变形菌门)、Acidobacteria(酸杆菌门)、Actinobacteria(放线菌门)为优势菌门,Sphingomonas(鞘氨醇单胞菌属)、Xanthomonadaceae(黄单胞菌科)相对丰度随着种植年限增加呈明显上升趋势,未知菌属RB41、Lysobacter(溶杆菌属)相对丰度呈下降趋势。土壤真菌群落Mortierellomycota(被孢霉门)、Ascomycota(子囊菌门)、Basidiomycota(担子菌门)为优势菌门,Mortierella(被孢霉属)相对丰度随种植年限增...     

利用PCR-DHPLC技术检测柑橘溃疡病菌

柑桔溃疡病是由柑桔溃疡病菌Xanthomonas axonopodis pv.citri(Xac)引起的重要的国际国内检疫性病害.柑桔溃疡病菌属黄单胞菌属,由于该属的种类及变种繁多,对Xac的生化鉴定十分繁杂和困难.     

Synopsis on the taxonomy of the genus xanthomonas

ABSTRACT The genus Xanthomonas exhibits a high phytopathogenic diversity in contrast to a phenotypic uniformity, which has hampered the genesis of a stable classification for a long time. In past decades, a large number of Xanthomonas strains have been characterized by a variety of phenotypic and genotypic methods in a multitude of studies. Extensive DNA hybridization studies and repetitive sequence-based polymerase chain reaction and amplified fragment length polymorphism genomic fingerprinting have clearly revealed the genomic diversity and relationships within the genus. A review of the current classification of the genus Xanthomonas based on the synopsis of these studies is given here.     

Polyphasic characterization of xanthomonas strains from onion

ABSTRACT Xanthomonas leaf blight has become an increasingly important disease of onion, but the diversity among Xanthomonas strains isolated from onion is unknown, as is their relationship to other species and pathovars of Xanthomonas. Forty-nine Xanthomonas strains isolated from onion over 27 years from 10 diverse geographic regions were characterized by pathogenicity to onion and dry bean, fatty acid profiles, substrate utilization patterns (Biolog), bactericide resistance, repetitive sequence-based polymerase chain reaction fingerprinting, rDNA internally transcribed spacer (ITS) region, and hrp b6 gene sequencing. Multiplication of onion Xanthomonas strain R-O177 was not different from X. axonopodis pv. phaseoli in dry bean, but typical common bacterial blight disease symptoms were absent in dry bean. Populations from each geographical region were uniformly sensitive to 100 mug of CuSO(4), 100 mug of ZnSO(4), and 100 mug of streptomycin sulfate per ml. Biolog substrate utilization and fatty acid profiles revealed close phenoltypic relatedness between onion strains of Xanthomonas and X. axonopodis pv. dieffenbachiae (57% of strains) and X. arboricola pv. poinsettiicola (37% of strains), respectively. A logistic regression model based on fatty acid composition and substrate utilization classified 69% of strains into their geographical region of origin. Sequencing of a portion of the hrp B6 gene from 24 strains and ITS region from 25 strains revealed greater than 97% sequence similarity among strains. DNA fingerprinting revealed five genotype groups within onion strains of Xanthomonas and a high degree of genetic diversity among geographical regions of origin. Based on pathogenicity to onion, carbon substrate utilization, fatty acid profiles, rDNA genetic diversity, and genomic fingerprints, we conclude that the strains examined in this study are pathovar X. axonopodis pv. allii. Implications of genetic and phenotypic diversity within X. axonopodis pv. allii are discussed in relation to an integrated pest management program.     

A comprehensive species to strain taxonomic framework for xanthomonas

ABSTRACT A comprehensive classification framework was developed that refines the current Xanthomonas classification scheme and provides a detailed assessment of Xanthomonas diversity at the species, subspecies, pathovar, and subpathovar levels. Polymerase chain reaction (PCR) using primers targeting the conserved repetitive sequences BOX, enterobacterial repetitive intergenic consensus (ERIC), and repetitive extragenic palindromic (REP) (rep-PCR) was used to generate genomic fingerprints of 339 Xanthomonas strains comprising 80 pathovars, 20 DNA homology groups, and a Stenotrophomonas maltophilia reference strain. Computer-assisted pattern analysis of the rep-PCR profiles permitted the clustering of strains into distinct groups, which correspond directly to the 20 DNA-DNA homology groups(genospecies) previously identified. Group 9 strains (X. axonopodis) were an exception and did not cluster together into a coherent group but comprised six subgroups. Over 160 strains not previously characterized by DNA-DNA hybridization analysis, or not previously classified, were assigned to specific genospecies based on the classification framework developed. The rep-PCR delineated subspecific groups within X. hortorum, X. arboricola, X. axonopodis, X. oryzae, X. campestris, and X. translucens. Numerous taxonomic issues with regard to the diversity, similarity, redundancy, or misnaming were resolved. This classification framework will enable the rapid identification and classification of new, novel, or unknown Xanthomonas strains that are pathogenic or are otherwise associated with plants.     

水稻白叶枯病菌和细菌性条斑病菌的分子标记筛选及检测

 水稻白叶枯病菌（Xanthomonas oryzae pv. oryzae）和细菌性条斑病菌(Xanthomonas oryzae pv. oryzicola)是水稻种子产地检疫中最重要的两种检疫对象,且同属于水稻黄单胞杆菌。本研究基于生物信息学技术构建比较基因组学算法对两种病原的全基因组序列比对分析,得到一系列能够区分两种病原的特异性PCR引物。结合简单的PCR技术及全自动DNA分析系统,我们选取了12对引物分别对23株水稻白叶枯病菌和5株水稻细菌性条斑病菌及其它相关菌株进行验证。结果获得了2对显性标记(Xoo-Hpa1和Xoc-ORF2)以及3对共显性分子标记(M568、M897和M1575)可以达到理想的区分检测两种病原的效果。分子标记的检测灵敏度从5×10⁴到5 × 10⁷cfu·mL^-1不等,且从水稻种子浸提液中也能成功地检测水稻白叶枯病菌和细菌性条斑病菌。本研究丰富了检测标记的靶位点,并有效的结合了高通量检测的手段对多位点联合分析,增强了检测的可靠性,有望在今后的植物检疫及病原鉴定中发挥着重要的作用。     

Rep-PCR技术对中国水稻条斑病菌的遗传多样性初析

 采用Rep-PCR技术,对30个水稻细菌性条斑病菌株(Xanthomonas oryzae pv.oryzicola)进行遗传多样性分析,同时对李氏禾条斑病菌等其它10个参试菌株也进行了比较。Rep-PCR是利用一些基于细菌的短的重复序列引物(ERIC和BOX)的DNA扩增特性,2种引物组合的电泳图谱结合并分析,以水稻细菌性条斑病菌各自的指纹谱型在相似率80%时可分为6簇,初步表明我国水稻细菌性条斑病菌群体的遗传分化明显;发现自然界存在的弱或无毒性菌株与毒性菌株的Rep-PCR指纹图谱差异很大;毒性菌株的遗传分簇与其致病性具有一定的相关性。用ERIC扩增水稻条斑病菌基因组DNA的指纹比BOX更为多样,两者对菌株的分辨率不同。因此,Rep-PCR技术可有效地用于监测水稻细菌性条斑病菌的遗传变异,还可应用于菌株的鉴定和分类学研究。     

Identification and Characterisation of Xanthomonas campestris pv. campestris Strains from Tanzania by Pathogenicity Tests, Biolog, rep-PCR and Fatty Acid Methyl Ester Analysis

Black rot, caused by Xanthomonas campestris pv. campestris (Xcc), is a major disease constraint to cabbage production by smallholder farmers in Africa. Variability exists within the pathogen, and yet differentiation of Xcc strains from other closely-related xanthomonads attacking crucifers is often difficult. The Biolog system, fatty acid methyl ester analysis using microbial identification system (MIS), rep-PCR and pathogenicity tests were used to identify and characterise Xcc strains from Tanzania. Great diversity was observed among Xcc strains in their Biolog and rep-PCR profiles. Specific rep-PCR genomic fingerprints were linked to some geographical areas in the country. Most of the Xcc strains were clustered in two groups based on their fatty acid profiles and symptom expression in cabbage although some deviant strains were found. Each of the methods allowed a degree of identification from species, pathovar to the strain level. Biolog and MIS identified all Xcc strains at least to the genus level. Additionally, Biolog identified 47% of Xcc strains to the pathovar and 43% to strain level, whereas MIS identified 43% of the strains to pathovar level. In the absence of a database, the utility of rep-PCR for routine diagnosis of strains was limited, although the procedure was good for delineation of Xcc to the strain level. These findings indicate the existence of Xcc strains in Tanzania that are distinct from those included in Biolog and MIS databases. The limitations noticed warrant continued improvement of databases and inclusion of pathogenicity testing, using universally susceptible cultivars, as an integral part of strain identification.     

High Genetic Stability of the Begomovirus Tomato yellow leaf curl Sardinia virus in Southern Spain Over an 8-Year Period

ABSTRACT The evolution of the plant single-stranded DNA virus Tomato yellow leaf curl Sardinia virus (TYLCSV) (genus Begomovirus, family Geminiviridae) has been monitored for 8 years after its appearance in southern Spain. Variation within three genomic regions of 166 TYLCSV isolates collected from three locations was assessed by single-strand conformation polymorphism (SSCP) analysis. According to SSCP, the intergenic region (IR) was the most variable. Low genetic diversity was found within the population and geographical or temporal differences were not evident. Nucleotide sequences of specific genomic regions of haplotypes identified by SSCP indicated close relationships among them. Therefore, the Spanish TYLCSV population appears to represent a single, undifferentiated population. The analysis of IR sequences for a subsample of 76 randomly chosen isolates confirmed the limited genetic diversity revealed by the SSCP analysis. A tendency to a lineal increase in diversity over time was observed in Málaga and Almería subpopulations; however, no accumulation of mutations in single isolates was evident. Negative selection to variation seems to operate to conserve certain regions of the genome. Thus, the low genetic diversity found in the studied TYLCSV population might be the result of a founder effect with subsequent selection against less fit variants arising by mutation.     

Chromosomal Gene Transfer by Conjugation in the Plant Pathogen Xanthomonas axonopodis pv. vesicatoria

ABSTRACT Genes for copper resistance, located on the chromosome of strain XvP26 of Xanthomonas axonopodis pv. vesicatoria, were transferred by conjugation to a recipient strain of the bacterium. The chromosomal gene transfer was verified by analyses of the genomes of donor, recipient, and putative transconjugants for plasmid profiles, by polymorphism of DNA bands obtained by digesting total genomic DNA by a rare-cutting endonuclease and pulsed-field gel electrophoresis, and by Southern hybridization with a probe containing the copper genes. Transfer of kanamycin resistance to a recipient strain, associated with Tn5 insertion into the chromosome of another strain of the bacterial spot pathogen, was also verified. The frequency of kanamycin resistance transfer to recipient was more than 75 times greater in pepper leaves than in vitro. The transfer of chromosomal sequences containing the hypersensitive reaction and pathogenicity (hrp) genes and pigmentation (pig) genes was linked with transfer of kanamycin resistance (Tn5). Horizontal transfer in planta of the chromosomal genes (i.e., cop, pig, hrp, and Tn5 sequences) among strains of X. axonopodis pv. vesicatoria means that horizontal chromosomal gene transfer is possible in nature. This type of gene transfer may explain the presence of great diversity among strains of the bacterial spot pathogen in terms of DNA polymorphism and may also explain the apparent horizontal transfer of hrp sequences among pathovars of Xanthomonas.     

Variation in Albicidin Biosynthesis Genes and in Pathogenicity of Xanthomonas albilineans, the Sugarcane Leaf Scald Pathogen

ABSTRACT Total genomic DNA from 137 strains of Xanthomonas albilineans from worldwide locations was hybridized with two DNA probes that together harbor the entire 49-kb albicidin biosynthesis gene cluster and two additional 3-kb genomic regions required for albicidin production. Fourteen haplotypes and two major genetic groups (albicidin [ALB]-restriction fragment length polymorphism [RFLP] A and ALB-RFLP B) were identified, and strains that were isolated after recent outbreaks of leaf scald disease belonged to group ALB-RFLP B. Albicidin genetic diversity was very similar to the previously described genetic diversity of the pathogen based on the whole genome. No relationship was found between variability of albicidin biosynthesis genes and the amount of albicidin produced in vitro by X. albilineans. Leaf scald-susceptible sugarcane cv. H70-144 was inoculated with 20 strains of the pathogen belonging to different ALB-RFLP haplotypes. Among them, 10 strains from Guadeloupe belonged to the same ALB-RFLP group but differed in the amount of albicidin produced in vitro. Strains were distributed in at least three different pathogenicity groups based on symptom severity and pathogen population density in the stalk. These two pathogenicity factors varied concurrently; however, no relationship between variation in albicidin biosynthesis genes, variation in the amount of albicidin produced in vitro, and variation in pathogenicity of X. albilineans was found. Further investigation is necessary to identify other genes involved in pathogenicity of X. albilineans.