广东省烟草青枯茵的菌系和遗传多样性

从广东省韶关和梅州2个烟草产区采集了来自8个县市的共38个烟草青枯菌菌株，对病菌的生物型、致病型和遗传多样性进行了测定，结果表明广东烟草青枯菌全部为生物型Ⅲ。通过在红花大金元，K326，岩烟97和系3等4个烟草鉴别品种上的致病性反应，将广东烟草青枯菌分为强致病力、中等致病力和弱致病力3种致病型，其中以中等致病力菌株为主，占78．9％；强致病力菌株占13．2％；弱致病力菌株占7．9％。从116个RAPD通用引物中筛选出10条引物，用于对38个烟草青枯菌株DNA的RAPD分析，扩增条带表现出明显的多态性，条带主要聚集在0．3～4．0kb之间。聚类分析这38个菌株可分为2个组群，即组群A和组群B。组群A中又可以分为7个亚组（A1、A2、A3、A4、A5、A6、A7）；组群B也含有4个亚组（B1、B2、B3、B4）。研究结果还表明，广东烟草青枯菌菌株RAPD组群的划分与菌株的寄主来源有一定的相关性，但与地理区域、生物型和致病型没有明显的相关性；生物型不能反映烟草青枯菌菌系的差异。 04—0463—06     

桑细菌性青枯病病原及其生化型鉴定

为有效治理浙江省杭州地区桑树上严重发生的新病害,开展了病原及其特性研究。从临安市和桐庐县的青枯桑树上分离出15株致病细菌,经25项主要细菌学特性、菌落形态、致病性、Bi-olog、脂肪酸分析、RAPD-PCR鉴定及与4株标准菌株比较,证实了这种桑青枯病是由Ralstoniaso-lanacearum引起,属于生化型Ⅴ和Ⅳ,它们在15株分离菌中分别占73.3%和26.7%。同时,在该病原细菌的鉴定中,建立了简便易行的桑青枯病病原细菌致病性测定新方法。     

Isolation and characterisation of phages against Pseudomonas syringae pv. actinidiae

Purpose: Pseudomonas syringae pv. actinidiae causes bacterial canker of kiwifruit and is responsible for severe economic losses and emergence of drug-resistant bacteria. Bacteriophages are viruses that infect target bacterial hosts and may be the best strategy to prevent and control kiwifruit canker disease. The objective of this experiment was to monitor the prevalence of Pseudomonas syringae pv. actinidiae and provide insight for the use of phages in biological control.

Materials and methods: In this study, 52 strains of Pseudomonas syringae pv. actinidiae were isolated from 68 stem samples of kiwi plant (cv. Hongyang & Jinkui). Following polymerase chain reaction (PCR) analysis, 15 isolates belonging to biovar 3 were identified, one of which was named XWY0007 and used as the target strain to isolate the phages. Thirty-six phages were isolated and purified from a total of 51 surface water samples collected in Shanghai. All phages were identified by transmission electron microscopy (TEM) and their host ranges were evaluated. Three phages, designated φXWY0013, φXWY0014 and φXWY0026 were selected and further characterised using one-step growth curve and stability at different temperatures and pH.

Results and conclusions: The isolated phages are promising for use as antimicrobials against bacterial canker in kiwi. This report is regarding Pseudomonas syringae pv. actinidiae and its phages from major areas of kiwifruit cultivation.     

华鲮烂尾病病原的分离鉴定及药敏分析

从患有烂尾病的华鲮(Sinilabeo rendahli)体内分离到两个优势菌株(编号:BB090516C-1和BB090516C-2),对分离菌株形态特征、主要理化特性等生物学性状鉴定后测定其16S rRNA序列并构建系统发育树,同时采用K-B琼脂扩散法进行药敏实验。结果显示:经人工感染试验,证实分离菌对华鲮有较强的致病性,该菌为革兰氏阴性、兼性厌氧发酵型短杆菌。经形态特征、培养特性和生理生化反应等指标分析鉴定结果显示该菌为维隆气单胞菌温和生物型(Aerom onas veronii biovar sobria)。用16S rRNA基因序列比对结果显示分离菌株与维隆气单胞菌的同源性高达99%,系统进化树中与维隆气单胞菌自然聚为一支,表明该菌为维隆气单胞菌温和生物型。用22种抗菌类药物进行药敏实验发现,该菌对头孢曲松、硫酸庆大霉素等9种抗菌类药物高度敏感,对新霉素等6种药物中敏,对氨苄青霉素等7种药物耐药。     

桉树青枯病病原菌的分子鉴定及其致病性测定

对华南4个地区的桉树青枯病病原菌进行了分子鉴定和致病性测试研究,以期为有效预防其发生提供依据.分别对来源于不同区域的11株桉树青枯病病原菌进行分子鉴定和致病力强弱测试比较,11株病原菌与Ralstonia solanacearum序列形成支持率为86％分枝,表明其病原菌16S rRNA基因序列与Ralstonia solanacearum具有高度同源性.以桉树无性系DH32-29组培苗为材料,采用伤根法对11个菌株进行致病力测试,不同菌株间在致病性上差异显著(P＜0.01或P＜0.05),以发病高峰期、高峰期发病率和平均发病率进行K-均值聚类分析(K=3),结果表明来源于广东、广西、海南的各菌株间致病性存在差异,HY01、HY02、DA01、HP04菌株与HP05分为1类,DA02和DA03为第2类,而YJ01、HP01、HP02和HP03为第3类.对4个不同地区(采样点)的青枯病菌株平均发病率进行方差分析,结果发现来源广东阳江菌株致病性显著地低于其他来源菌株.不同地理来源菌株之间在致病性上无明显相关性,同一地区内同时存在着致病力强弱菌株,HP03的生物型4-1菌株与其他生物型3菌株在致病性亦无明显的差别.     

Phylogenetic relationships and population structure of Ralstonia solanacearum isolated from diverse origins in Taiwan

Bacterial wilt caused by Ralstonia solanacearum is a destructive disease for many crops. The aim of this study was to investigate the phylogenetic relationships and genetic structure of an R. solanacearum population from diverse origins in Taiwan. All 58 tested isolates belonged to phylotype I, except the two biovar 2 isolates from potato. These belonged to phylotype IIB sequevar 1 and were identical to known potato brown rot strains, which were probably introduced. Phylotype I isolates were grouped into 10 sequevars. Sequevar 15 was predominant (34 out of 56 isolates). Its distribution covered the whole island and it was largely associated with solanaceous crops such as tomato, and with tomato field soil. A total of 14 haplotypes were identified based on a partial endoglucanase gene sequence. Parsimony network analysis revealed that haplotype A was the oldest haplotype in the local population. It encompassed all but one of the sequevar 15 isolates. Large variation in virulence on tomato was observed among the 58 isolates, and seven pathotypes were identified. Significant genetic differentiation was detected among pathotypes. Moreover, genetic differentiation was detected between biovar 3 and biovar 4 subgroups and between the strains associated with solanaceous and non‐solanaceous species, but none was detected between strains from different geographic origins. The results suggest that the phylotype I population in Taiwan is homogeneous, while mutation and local adaptation to specific ecological niches keep shaping the population.     

罗汉果青枯病病原菌株的鉴定

 The pathogen of Siraitia grosvenorii wilt was identified.Six strains were isolated from diseased plants of S.grosvenorii collected from Yongfu County in Guangxi.By using bacteriological identification,pathogenicity tests assay,16S rDNA sequence analysis,the S.grosvenorii wilt was caused by Ralstonia solanacearum.The results of pathogenicity and carbohydrate utilized test demonstrated that the pathogen belonged to race 1 and biovarⅢ of R.solanacearum.     

台州猕猴桃溃疡病病原菌分子鉴定及药剂筛选

为了针对性地防治猕猴桃细菌性溃疡病,采用平板涂布分离法从台州主要猕猴桃种植果园分离到1株丁香假单胞猕猴桃致病变种(Pseudomonas syringae pv. actinidae,Psa)。用生物型(biovar)特异性引物检测发现,试验分离到的Psa菌株是biovar 3型。通过抑菌圈大小比较,对23种农用杀菌剂进行了筛选。结果表明,不同农用杀菌剂对Psa菌株的抑菌效果相差很大,且以四环素、乙蒜素和丁子香酚的抑菌效果较好(抑菌圈直径≥15 mm)。此外,用最小抑菌浓度(MIC)和时间-杀菌试验研究了纳米氧化锌对Psa菌株的杀菌效果。结果显示,纳米氧化锌的MIC值为50 μg·mL^-1,且杀菌效果随纳米氧化锌的浓度和作用时间的增加而增强。     

Cultivar × rhizobial strain interactions in peas with respect to early symbiosis, nodule initiation and N uptake

Sixteen pea cultivars differing in their phenotypic characteristics were studied in order to characterize the cultivars' variability in symbiotic characters using three Rhizobium leguminosarum biovar viceae strains. The peas did not show any cultivar × strain interactions with the rhizobial strains used with respect to root hair infection frequency or day of nodule initiation. Three cultivars had high root hair infection frequencies, and five had only a few infected root hairs. Four of five leafless/semi-leafless varieties investigated and one cultivar with normal leaves formed nodules quickly, whereas three varieties with normal leaves formed nodules slowly. Ten of the cultivars showed cultivar × strain interactions with respect to N uptake, indicating that the performance of the N2-fixing symbioses depends on the rhizobial partner. Six cultivars showed cultivar × strain interactions with respect to dry matter production and the nodule proportion of the dry matter. Of the cultivars investigated, we conclude that some of the leafless/semi-leafless varieties (‘Capella’, ‘Filby’ and ‘Solara’) are suitable for inclusion in a future breeding programme, since these cultivars combined several important symbiotic characters.     

Inhibition of crown gall formation by Agrobacterium radiobacter biovar 3 strains isolated from grapevine

Graft unions of nursery stock of grapevine (Vitis vinifera L.) collected in Japan yielded pathogenic and nonpathogenic strains of Agrobacterium. On the basis of classical diagnostic tests, a sequence analysis, and a multiplex polymerase chain reaction method previously reported, the pathogenic strain was identified as Agrobacterium tumefaciens biovar 3, whereas the nonpathogenic strains were assigned to Agrobacterium radiobacter biovar 3. Stems of tomato (Lycopersicon esculentum Mill.) seedlings were inoculated with both A. tumefaciens biovar 3 strain G-Ag-27 as a pathogen and one of the control strains isolated from grapevine or A. radiobacter biovar 2 strain K84 as competitors to assay the suppression of gall formation caused by the pathogen. In a test with a 1 : 1 pathogen/nonpathogen cell ratio, all A. radiobacter biovar 3 strains reduced gall incidence and size compared to that of the positive control inoculated only with the pathogen. Strain VAR03-1 was especially effective in reducing the incidence of gall formation on grapevine and reduced gall size by 84%–100% of those on the positive control. Many tested nonpathogenic biovar 3 strains were bacteriocinogenic, causing an inhibition zone against A. tumefaciens biovar 3 strains on YMA medium. Strain VAR03-1 was the most effective against indicator strains and appears to be a promising agent for controlling crown gall of grapevine.