吉林省烟草野火病菌群体分子多态性分析

为研究烟草野火病菌的群体遗传多样性,采用正交试验方法,对5个多态性引物(REP,ERIC,BOX,J3,IS1112)的Rep-PCR反应体系分别进行了优化,同时优化了体系中的Mg2+、dNTP、rTaq酶等几个因素,并在此基础上对来源于吉林省不同地区的70株烟草野火病菌进行了分子多态性分析。结果表明:优化的PCR反应体系电泳图谱清晰,多态性丰富,重复性好,适合烟草野火病菌群体分子标记研究。通过聚类分析发现,吉林省烟草野火病菌群体存在分子多态性,在遗传相似系数为0.9时,70个菌株被聚为4个类群(G1~G4),其中G1类群最大(39个菌株,占55.71%),而G3类群最小,仅包括2个菌株。G1和G2类群分别包含了5个和3个亚群。不同类群和亚群的菌株地理来源存在多样性。     

Effects of cropping history and origin of seed potatoes on population structure of Phytophthora infestans

The effects of origin of seed potatoes and the cropping history on the phenotypic structure of Phytophthora infestans populations was studied in northern Hessia, central Germany, from 2000 to 2002. Populations originating from fields with a history of potato cropping with only short or no rotation (old fields) were compared with populations from new fields, i.e., where no potatoes had been grown for at least 30 years and seed potatoes were either imported from breeders or produced on-farm (certified). The main goal was to determine the importance of seed potato infection in the establishment of new P. infestans populations. Isolates were characterized for mating type, virulences and rep- (repetitive extragenic palindromic) PCR fingerprints. Among a total of 639 isolates sampled from 31 sites, mating types A1 and A2 co-existed in all three years in 60–92% of the sites. Over all three years, 53 pathotypes were detected in a subsample of 272 isolates. Isolates originating from the new fields had significantly higher frequencies of the virulences v1, v2, v3, v6 and v7, indicating general effects of seed introduction into a new region. Thirty-six fingerprints were detected in a subsample of 281 isolates of which 22 were unique while four occurred in all three years and in many sites. Pathogen populations from potato fields that were grown from seed tubers of geographically different origin differed significantly based on χ ² tests. While the Nei genetic distances were less than .1 among the local populations, distances to the US lineages US-1, US-6, US-7 and US-8 ranged from .22 to .47; however, the bootstrap values were not significant. Populations from old fields were more diverse and 14 of the 22 rep-PCR types occurred there among 132 isolates tested in comparison to six in the new fields (n = 140 isolates) and two among six isolates from volunteers. The results also suggest that both sexual and asexual reproduction play a role.     

西南地区水稻细菌性条斑病菌的遗传多样性分析

选取云南、贵州、四川不同省市地区的水稻条斑病菌菌株141株,利用Rep-PCR技术筛选出多态性较好的引物对其DNA进行多态性分析.引物J3、ERIC、JELIR的扩增多态性较好,不同毒性菌株间的Rep-PCR指纹图谱差异很大.通过聚类分析显示,我国西南地区的水稻细菌性条斑病菌群体遗传分化明显,菌株的遗传族群与其致病性具有一定的相关性,强、中、弱毒性菌株大致归为不同的类群.因此,Rep-PCR技术可用于检测水稻细菌性条斑病菌的遗传变异与鉴定研究.     

鸭疫里默氏菌基因分型

在鸭疫里默氏菌 (RA)血清分型的基础上 ,以提取的 RA基因组 DNA为模板 ,进行 Rep- PCR扩增。根据扩增产物的琼脂糖凝胶电泳图谱 ,将分属于 8个血清型的 4 0株 RA区分为 12个不同的基因型 ,不同血清型 RA的图谱有明显差异 ,相同血清型 RA的图谱也存在差异。这些结果显示 ,RA基因组中重复的基因外的回文结构可用于区别相同血清型的不同分离株 ;Rep- PCR作为一种实用的分子生物学方法 ,在分子流行病学研究中可对 RA分离株进行基因定型     

Virulence and Molecular Diversity within Colletotrichum lindemuthianum Isolates from Andean and Mesoamerican bean Varieties and Regions

Virulence on a standard set of 12 common bean differential varieties, DNA sequence of repetitive-elements (Rep-PCR) and random amplified microsatellites (RAMS) were used to assess the genetic variability of 200 Colletotrichum lindemuthianum isolates collected from Andean and Mesoamerican bean varieties and regions. High levels of pathotypic (90 pathotypes) and genetic diversity (0.97) were identified among 200 isolates, revealing that C. lindemuthianum is a highly diverse pathogen. Although a significant number of pathotypes were common to Andean and Mesoamerican regions, many more were only found in the Mesoamerican region. Cluster analysis of virulence and molecular data did not separate isolates into groups that were structured with common bean gene pools. No genetic differentiation (G _ST=0.03) was apparent between Andean and Mesoamerican isolates of C. lindemuthianum. The diversity exhibited by C. lindemuthianum does not appear to cluster according to common bean gene pools, and the high diversity found in the Mesoamerican region seems to indicate that C. lindemuthianum originated and was disseminated from this region. Due to the high genetic variation exhibited by C. lindemuthianum, stacking major resistance genes appears to be the best option for developing cultivars with durable anthracnose resistance.     

Genetic diversity and symbiotic efficiency of population of rhizobia of <Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> L. in Brazil

The genetic diversity and symbiotic efficiency among indigenous rhizobia isolates obtained from native field with or without organic fertilization and superficial mineral fertilization were investigated. Eighty-six indigenous rhizobia were isolated from these fields using four common bean varieties as trap-host. The common bean varieties Mexico 309 and Rio Tibagi selected the most efficient rhizobia strains because they showed the best yields and N contents results. The genetic characterization of 36 rhizobia isolates was evaluated by using electrophoretic profiles of amplification products using primers ERIC1-R and ERIC-2. Our results demonstrated that besides the large diversity in the indigenous rhizobial community, the genotype of the trap-host probably influences the selection of the most efficient strains.     

苹果树根际高效解钾菌的筛选及鉴定

为探明苹果树根际解钾菌的种群特征和解钾性能,从苹果树根际土壤中分离获得解钾能力较强的高效解钾菌。本研究以钾长石为唯一钾源,分离获得118株具有解钾活性的菌株,重复片段PCR基因指纹分析(Repetitive-element PCR,rep-PCR)聚为29个类群。利用火焰分光光度法测定菌株解钾能力,获得5株高效解钾菌,平均解钾活性达到41.47mg·L-1,其中K105的解钾能力最强,有效态钾增长23.09%。采用H2O2消煮后测得的K+浓度升高,有效态钾增长最高达31.22%。采用形态特征观察、生理生化特性检测和基于16S r RNA基因序列的系统发育分析对高效解钾菌株进行鉴定。结果表明:K1为不动杆菌属(Acinetobacter sp.)、K98、K105和K115为假单胞菌属(Pseudomonas sp.)、K168为芽孢杆菌属(Bacillus sp.)。研究结果可为开辟新型高效解钾菌,为连作土壤的改良和苹果专用微生物菌肥的开发提供依据。     

福建省马铃薯黑胫病病原菌的鉴定及其遗传多样性分析

为明确福建省马铃薯黑胫病病原菌的种类、分布情况及群体遗传特性,通过16S rDNA序列分析法对分离自福建省各马铃薯产区的90株黑胫病病原菌进行鉴定,并利用细菌基因组重复序列PCR(repetitive sequence-PCR,Rep-PCR)方法分析福建省不同地区的马铃薯黑胫病病原菌群体的遗传变异情况。结果表明,经16S rDNA序列分析鉴定,引起福建省马铃薯黑胫病的病原菌包括黑腐果胶杆菌Pectobacterium atrosepticum、胡萝卜果胶杆菌巴西亚种P. carotovorum subsp. brasiliense、胡萝卜果胶杆菌胡萝卜亚种P. carotovorum subsp. carotovorum、帕曼蒂氏果胶杆菌P. parmentieri、P. polaris和达旦提狄基氏菌Dickeya dadantii。对90株菌株的Rep-PCR检测得到47多态性条带条,多态性比率为100.00%。按采集地区将菌株划分为8个群体,其等位基因数平均为1.48,有效等位基数平均为1.30,Nei’s基因多样性指数平均为0.17,Shannon信息指数平均为0.26,多态...     

红掌细菌性疫病病原菌遗传多样性的 RAPD 和 Rep-PCR分析

由地毯草黄单胞菌花叶万年青致病变种引起的细菌性疫病是红掌生产中最具破坏性的病害。本研究对来自全国 8 个省市主产区的 93 个红掌细菌性疫病菌株，采用 RAPD 和 Rep-PCR（ERIC、BOX、REP）分子标记进行了遗传多样性分析。筛选出的 13 个 RAPD 引物共扩增出 131 个清晰、重复性好的条带，而 Rep-PCR 共扩增出65 个条带，所有位点多态性比率为 100%。基于 RAPD 和 Rep-PCR 结果计算的群体遗传距离显著相关，综合 2 种标记类型的分型结果进行 UPGMA 聚类分析，表明我国红掌细菌性疫病具有高度的遗传多样性，在遗传相似性系数范围 0.519 1~0.984 7，在 0.63 水平时可以将所有菌株划分为 3 个组群，分别包括 58、28、7 个个体，病菌群体的遗传多样性与其寄主品种、地理来源没有明显相关性。本研究为红掌细菌性疫病监测和抗病育种提供了科学依据。     

DNA fingerprinting of Pyricularia grisea by rep-PCR using a single primer based on the terminal inverted repeat from either of the transposable elements Pot2 and MGR586

We investigated the use of single primers complementary to sequences in the terminal inverted repeat (TIR) of either Pot2 or MGR586, transposable elements found in Pyricularia grisea, for DNA fingerprinting by repetitive-element-based polymerase chain reaction (rep-PCR). Under standard amplification conditions, rep-PCR with each single primer generated distinct fingerprint patterns among rice-infecting P. grisea isolates collected in Japan. With the Pot2-TIR primer, bands ranging in size from 0.2 to 8 kb and in number from 8 to 13 per isolate were amplified. Although fewer bands were amplified with the MGR586-TIR primer, this molecular technique should be more reliable to identify and classify P. grisea isolates by combining the data of fingerprint patterns from each TIR primer. In a cluster analysis based on DNA fingerprints from this rep-PCR with the Pot2-TIR primer, 10 reference isolates and 12 field isolates from Saga Prefecture in 2002 were separated into six clonal lineages. We also demonstrated that the 12 field isolates belonged to one clonal lineage. Thus, this rep-PCR method using the single primer Pot2-TIR will be useful for the analysis of the population structure of rice blast pathogens.