Molecular evidence supports hypervariability in Phytophthora colocasiae associated with leaf blight of taro

The oomycete Phytophthora colocasiae that causes taro leaf blight is the most devastating disease of taro and is widely distributed worldwide. Molecular and phenotypic techniques were employed for assessing and exploiting the genetic variability among four populations of P. colocasiae obtained from a fine spatial scale (multiple leaf blight lesions on single taro leaf). Phenotypic characters such as virulence, morphology and mating type showed no variation. ITS characterization revealed detectable polymorphism among isolates of P. colocasiae. The mean number of haplotypes (H), haplotype diversity (HD), nucleotide diversity (π), and nucleotide substitution rate (θ) among analyzed sequences were 6.75, 1.00, 0.069, and 0.088 respectively. High levels of inter and intra specific variation were detected by random amplified polymorphic DNA (RAPD) assays. Moderate genetic diversity (H?=?0.2651) was observed among populations of P. colocasiae. Analysis of molecular variance (AMOVA) confirmed that most of the genetic variability was confined to within a population (63.54 %). The coefficient of genetic differentiation among populations (G _ST ) was 0.2007 and estimates of gene flow (Nm) among populations was 1.991 migrants per generation. Cluster analysis using UPGMA revealed that individuals from the same population failed to cluster in one distinct group. The results of the study reveal considerable genetic diversity among and within populations of P. colocasiae obtained from fine spatial scale. The possible mechanisms and implications of this genetic variation are discussed.     

大豆疫霉菌单孢分离物生物学性状的遗传变异研究

 本文研究了大豆疫霉菌单游动孢子无性分离物和自交单卵孢子分离物的菌丝生长速率、菌落形态、同宗配合性状、产孢量以及对甲霜灵敏感性的遗传变异。结果表明:菌落形态、生长速率和同宗配合性状在单游动孢子后代和自交后代可稳定遗传,控制上述性状的遗传因子是纯合的;大豆疫霉菌的游动孢子产生能力和对甲霜灵的敏感性在单游动孢子后代和自交后代中均发生连续性变异,表明这两种性状可能是数量遗传性状,也可能控制这两种性状的基因为杂合基因或细胞质遗传因子。     

基于DNA条形码技术的湖南省柑橘主产区实蝇幼虫分子鉴定

为明确湖南省柑橘主产区的实蝇入侵为害现状,从该省7个市(自治州)22个地点收集柑橘蛆果中的幼虫,利用DNA条形码技术对其进行分子鉴定,并以DNA条形码序列作为分子标记探究橘大实蝇Bactrocera minax的20个中国地理种群(湖南省6个组群共16个种群、其它3省市4个种群)以及1个印度地理种群间的亲缘地理关系,分析橘大实蝇在我国的遗传进化关系。结果表明,仅采集自邵阳市城步苗族自治县柑橘蛆果中的5头幼虫被鉴定为蜜柑大实蝇B. tsuneonis,其余21个地点采集的595头幼虫均被鉴定为橘大实蝇。21个橘大实蝇地理种群的平均单倍型多样性为0.75,核苷酸多样性为0.0032,核苷酸差异数为2.13,中国所有地理种群均具有较高的遗传多样性;单倍型网络进化图显示湖南、重庆、贵州种群共享的单倍型H3为原始单倍型,表明其为比较原始的种群;AMOVA分析结果显示种群内个体间遗传变异占总体变异的59.04%,是遗传变异的主要来源;遗传分化结果表明湖南省6个组群间均出现了中度至高度的遗传分化,FST在0.0521～0.7795之间。表明DNA条形码技术可用于蜜柑大实蝇和橘大实蝇幼虫的分子鉴定及其种群遗传进化分析。     

Toxicological and biochemical response to azinphos-methyl in Cydia pomonella L. (Lepidoptera: Tortricidae) among orchards from the Argentinian Patagonia

BACKGROUND: Azinphos‐methyl is the main insecticide used to control codling moth on apple and pears in Northern Patagonia. The aim of this study was to evaluate the toxicological and biochemical response of diapausing larvae of codling moth in orchards subjected to different insecticide selection pressure. RESULTS: Dose–mortality assays with azinphos‐methyl in diapausing larvae of Cydia pomonella L. showed significant differences between the LD₉₅ from a population collected in one untreated orchard (2.52 µg moth^?1) compared with that in a laboratory‐susceptible population (0.33 µg moth^?1). Toxicity to azinphos‐methyl in field populations of diapausing larvae collected during 2003–2005 was evaluated by topical application of a discriminating dose (2.5 µg moth^?1) that was obtained from larvae collected in the untreated orchard (field reference strain). Significantly lower mortality (37.71–84.21%) was observed in three out of eight field populations compared with that in the field reference strain. Most of the field populations showed higher esterase activity than that determined in both the laboratory susceptible and the field reference strains. Moreover, there was a high association between esterase activity and mortality (R² = 0.64) among the field populations. On the other hand, a poor correlation was observed between glutathione S‐transferase activity and mortality (R² = 0.33) among larvae collected from different orchards. CONCLUSIONS: All the field populations evaluated exhibited some degree of azinphos‐methyl tolerance in relation to the laboratory susceptible strain. Biochemical results demonstrated that esterases are at least one of the principal mechanisms involved in tolerance to this insecticide. Copyright © 2008 Society of Chemical Industry     

Random amplified polymorphic DNA (RAPD) analysis of Phytophthora infestans isolates collected in Canada during 1994 to 1996

Mating type, glucose-6-phosphate isomerase ( Gpi ) allozyme banding patterns, response to the fungicide metalaxyl and random amplified polymorphic DNA (RAPD) markers were used to characterize genetic variability among 141 Canadian isolates of Phytophthora infestans collected between 1994 and 1996. Multiple correspondence analysis of RAPD profiles separated isolates into 21 groups that were not correlated to groups defined by mating type, Gpi allozyme banding patterns or response to metalaxyl. Population subdivision analysis showed that 97% of the total genetic variation was found among individuals within populations, compared with 3% among populations. The average similarity coefficient among isolates was 80%. No significant differences in haplotypic diversity were observed among the years under study, but levels of genetic diversity among local populations of P. infestans were high (0.76). All classes of response to the fungicide metalaxyl were observed, with 55% of isolates displaying moderate levels of insensitivity. The high level of genetic diversity detected within populations indicates that migration and sexual recombination probably play important roles in the population biology of P. infestans in Canada.     

小菜蛾种群抗性分子遗传与进化分析

小菜蛾 Plutella xylostella( L.)抗药性问题日益严重 ,对其抗性机理和治理策略的研究也较为广泛 ,但用淀粉电泳的试验方法从群体遗传学的角度研究小菜蛾的抗性发生、发展及进化规律的报道 ,国内尚不多见。为此作者用淀粉电泳结合 Genepop ( V 3.1 6 )软件同时分析了小菜蛾群体中酯酶 ( EST)、6 -磷酸葡萄糖脱氢酶 ( GPT)、苹果酸脱氢酶 ( MDH)和苹果酸酶 ( ME)的特征 ,旨在为抗性治理提供理论依据。1　材料与方法1 .1　供试小菜蛾及药品北京种群由中国农科院植保所吴奎华教授惠赠 ,采自北京郊区菜地 ;武汉、河北和云南种群由中国农业…     

Structure of the Cochliobolus sativus population variability

The genetic diversity of several local populations of the fungal pathogen Cochliobolus sativus, collected over 3 years from different regions of the Czech Republic, was examined using amplified fragment length polymorphism (AFLP). A high level of variability was found even among isolates from one lesion. Measures of multilocus linkage disequilibrium suggested that recombination has a minor impact on the genetic structure of populations. Cochliobolus sativus forms genetically divergent populations (F_ST = 0·33), indicating a low level of geneflow between populations. This was supported by a significant correlation between genetic diversity and geographical distances up to 80–100 km. The most likely explanation for the genetic variability is that the fungus forms conidia with highly variable chromosomal rearrangements. The differentiation observed among local populations implies that genetic drift, including a founder effect, combined with restricted migration generates the structure of C. sativus populations.     

Hemocyte diversity of the American bollwormHelicoverpa armigera

The hemocytes of the American bollwormHelicoverpa armigera (Hübner) were studied in the last instar larvae by phase contrast microscopy to investigate the differences among 14 populations, collected from ten locations, spread over a distance of approximately 3000 km in six different states of India. The cluster analysis of differential hemocytes showed as many as 11 clusters of populations at 5% and six clusters at 10% homogeneity level. The cluster analysis of hemocytes of seven populations from cotton crops showed five and three clusters at 5% and 10% homogeneity levels, respectively. Diversity of hemocytes of four populations from chickpea was wider than that from cotton. The differences in hemocytes of various populations may be due to space, time, host plant, insecticide use, other agroecological conditions, and insect genetic variabilityper se. The field populations ofH. armigera heavily treated with insecticides appeared to contain more spherulocytes at the expense of plasmatocytes and granulocytes. The treatment of 6-day-old larvae ofH. armigera with cypermethrin decreased plasmatocytes and granulocytes, and increased prohemocytes and spherulocytes in the hemolymph of last instar larvae. Thus, differential hemocyte counts may serve as an indicator of insecticide exposure and the diversity of insect populations. http://www.phytoparasitica.org posting Dec. 20, 2004.     

Variability and divergence in horsegram (Dolichos uniflorus)

Horsegram is an important and unexploited tropic and sub-tropic legume crop grown mostly in dry land agriculture.The study involving 23 cultivars of horsegram (Dolichos uniflorus) was conducted at the Seed Research & Technology Centre in Rajendranagar,Hyderabad,India during the early spring seasons of 2008 and 2009 to assess the nature and magnitude of variability in the existing species and also to identify diverse parents for use in further breeding programmes.The results indicated significant differences among the 23 cultivars for all characters studied,indicating the presence of sufficient genetic variation.High variability and heritability coupled with high genetic advance were reported for seed yield per plant and pod hulm per plant,indicating additive gene action and a possible scope for the improvement of these characters.Mahalanobis D 2 statistics grouped all the 23 cultivars of horsegram into six clusters.The maximum inter cluster distance (62.39) was noticed between cluster IV (HG 50) and cluster V (HG 11).The maximum intra cluster distance ranged from 0 (clusters III,IV,V and VI) to 15.17 (cluster),indicating that the genotypes in these clusters were relatively more diverse than the genotypes within other clusters.Seed yield per plant contributed the maximum (33.20%) to the genetic divergence.     

Molecular Variability Within Diaporthe/Phomopsis helianthi from France

ABSTRACT Diaporthe/Phomopsis helianthi causes brown stem canker of sunflower (Helianthus annuus) and is responsible for considerable yield loss. This species shows considerable variation for morphological characters, growth, and pathogenicity. Molecular variability of two sample groups was assessed with amplified fragment length polymorphism (AFLP) markers. Isolates of the first sample were collected from infected sunflower tissues from the main regions in France where the crop is grown, whereas isolates from the second sample came from stems within a single field of sunflower. A soybean strain was taken as an outgroup for AFLP analyses. Within sample one, the greatest genetic distance among isolates was 0.97, whereas it was 0.44 within sample two isolates. For the whole of France, the average genetic distance was 0.68, whereas in the one field it was 0.12. Nei's genetic diversity indices were 0.20 and 0.06 for France and for one field, respectively. The greatest genetic distance was found between isolates from the most northern crops. The greatest genetic distance between D. helianthi isolates and the strain isolated from soybean was similar to that observed for D. helianthi isolates from different geographical areas. The problems in defining the genus Phomopsis are discussed. It is shown that internal transcribed spacer sequencing could be a useful criteria for Diaporthe/Phomopsis species determination. The considerable genetic variability of the pathogen could lead to the occurrence of new strains that could be more aggressive or more resistant to chemical control.     

Assessing genetic diversity in the web blight pathogen <Emphasis Type="Italic">Thanatephorus cucumeris</Emphasis> (anamorph = <Emphasis Type="Italic">Rhizoctonia solani</Emphasis>) subgroups AG-1-IE and AG-1-IF with molecular markers

Web blight, an important foliar disease of dry beans in the Americas, is a challenge to manage. We studied genetic variation of 92 isolates of Rhizoctonia solani subgroups AG-1-IE and AG-1-IF using DNA fingerprinting methods and mycelial compatibility grouping. The isolates were collected over 13 years from bean fields in the Dominican Republic, Honduras and Puerto Rico. Cluster and AMOVA analysis of combined data from two universal rice primers and two internal sequence repeats revealed significant genetic variation among and within populations of both subgroups. Variation was influenced by geographic origin and sampling year for AG-1-IE isolates and geographic origin for AG-1-IF isolates. Mycelial compatibility of paired isolates was mostly scored as incompatible in both subgroups and supported many unique phenotypes. Only two isolates of AG-1-IE displayed mycelial compatibility and DNA fingerprints, suggesting clonal origin. Genetic variation in these AG-1-IE and AG-1-IF isolate populations may explain the lack of durable resistance to web blight reported in dry beans.     

Variation and Heritability of Phenology in the Fungus Monilinia vaccinii-corymbosi on Blueberry

ABSTRACT The germination of field-collected pseudosclerotia and the development of apothecia from eight New Jersey populations of the mummy berry fungus Monilinia vaccinii-corymbosi were evaluated under controlled conditions in the greenhouse. Development data for apothecia were used to describe the timing of apothecium formation and to estimate broad- and narrow-sense heritabilities of fungal phenology. Mean development times for the formation of apothecia ranged from 35.4 to 54.7 days. The mean development times for populations collected from early-season cv. Weymouth ranged from 35.4 to 39.6 days and were significantly shorter than the development times for three of the four populations collected from late-season cv. Jersey (46.9 to 54.7 days) or for the population collected from mixed stands of cultivated blueberries (42.7 days). The development of populations from late cultivars planted in very close proximity to early cv. Weymouth was early (36.5 to 39.0 days) and not significantly different from the development of populations collected from cv. Weymouth. Phenotypic and genetic variances of apothecium development for individual populations ranged from 18.9 to 44.8 and 7.2 to 30.9, respectively. Broad-sense heritabilities of apothecia development for each fungal population, calculated by partitioning phenotypic variation into genetic and environmental components, ranged from 0.31 to 0.78. Narrow-sense heritabilities of apothecia development, based on parent-offspring regression, ranged from 0.58 to 0.78. These results indicate that populations of M. vaccinii-corymbosi differ in phenology and that a significant portion of the phenological variation within populations is genetic. Thus, it is plausible to propose that the phenology of apothecium development is a component of fungal fitness and that host phenology can influence the timing of pathogen development.     

Genetic variation and structure in native and invasive Solidago canadensis populations

Solidago canadensis is native to North America, but has become a noxious invasive plant in China. We know only a little about its invasion history and the effects of introductions on its genetic composition. Here, we investigated genetic variation and structure between 15 North American and 13 Chinese populations of S. canadensis using AFLP makers. Four AFLP loci suggested relatively high genetic diversity of this weed and similar genetic variation between the invasive range and the native range. Most genetic variation was within populations across two ranges, but the Chinese range had a higher degree of among‐population variation than the North American range. Multiple tests, including Bayesian assignment, UPGMA analysis, PCoA and analysis of ‘isolation by distance’, showed that the Chinese populations originated from at least two distinct native sources and that secondary introduction or dispersal should be common in China. Also, North American populations were possibly a single genetic group. Overall, S. canadensis in China was probably founded from multiple introductions and then spread through long‐distance dispersal associated with human activities. Genetic variability in the species in the invaded range appears to have favoured establishment and spread and may well provide a challenge to successful control.     

ISSR markers detect high genetic variation among <Emphasis Type="Italic">Fusarium poae</Emphasis> isolates from Argentina and England

Fusarium poae is one of the Fusarium species isolated from cereal grains infected by Fusarium head blight (FHB), and in recent years it has been identified as a major FHB component. In this study, 97 F. poae isolates from Argentina (n = 62) and England (n = 35) were analysed by inter-simple sequence repeats (ISSR) to examine the genetic diversity and to determine whether intraspecific variation could be correlated with geographic and/or host origin. The molecular analysis showed high intraspecific variability within F. poae isolates, but did not reveal a clear relationship between variability and the host/geographic origin. Fusarium poae isolates from the same geographic region or host appeared in different subclusters. Conversely, isolates with the same haplotype were also collected from different geographic regions. However, we did observe subclusters consisting of isolates from Argentina only or from England only. Furthermore, a single seed sample was found to host different haplotypes. Analysis of molecular variance (AMOVA) indicated a high genetic variability in F. poae, with most of the genetic variability explained by differences within, rather than between Argentinean and English populations. This is the first report on genetic diversity of F. poae using ISSR markers. Moreover, ISSR fingerprinting generates highly polymorphic markers for F. poae and proved to be a useful and reliable assay for genetic variability studies.     

Genetic variation within and between southern Ontario populations of Sclerotinia homoeocarpa

Restriction fragment length polymorphisms (RFLP) of the intergenic spacer region (IGS) of rDNA and random amplified polymorphic DNA (RAPD) markers were used to survey genetic variability among 181 isolates of Sclerotinia homoeocarpa from Ontario and 10 isolates from Japan. RAPD and IGS-RFLP analyses revealed polymorphisms within and between populations of S . homoeocarpa , distinguishing 151 genotypes. Both types of markers gave similar results in phenetic analysis of genetic distances between populations. Cluster analysis showed that Japanese isolates of S. homoeocarpa were genetically distinct from Ontario isolates, demonstrating significant intraspecific differentiation. An average genetic similarity of 0.66 was found between Japanese isolates. Among Ontario isolates, average genetic similarity was 0.86, and genotypic diversity analysis showed that 49.3% of the total genetic variation observed within Ontario populations occurred among individuals within populations compared to 50.7% between populations. Gametic linkage disequilibrium analysis within Ontario populations revealed an average 15.6% significant nonrandom associations between putative RAPD loci, and that half of the populations showed signs of significant linkage disequilibrium. These results suggest that both clonal propagation and recombination events occurred in local populations of S. homoeocarpa . The high level of genetic similarity between populations and the low levels of intraspecific genetic variation may reflect a small founding population for southern Ontario isolates of S. homoeocarpa .     

甘肃省二斑叶螨地理种群的遗传分析

为探讨甘肃省二斑叶螨Tetranychus urticae种群遗传多样性及遗传分化,通过mtDNA-COI基因DNA条形码技术对采自甘肃省8个不同生境35个二斑叶螨地理种群的582个样品进行序列分析及地理种群的遗传分化分析。结果表明,获得二斑叶螨mtDNA-COI基因片段大小为424 bp,其中保守序列336个,变异位点66个,简约信息位点45个,单突变位点21个;碱基(A+T)含量明显高于(C+G)含量,有明显的A/T碱性偏倚性;在35个地理种群中共检出16个单倍型,单倍型指数为0.906;Mantel检测结果表明种群间的遗传距离与地理距离无显著相关性;35个地理种群间的遗传分化指数Fst为0.012,种群间变异为1.200。表明二斑叶螨的遗传变异主要来自于种群内部,种群间还未发生明显的遗传分化。     

Genetic and morphologic diversity of <Emphasis Type="Italic">Echinochloa crus-galli</Emphasis> populations from different origins

Echinochloa crus-galli (L.) P. Beauv. (barnyardgrass) is an annual weed that is native to Asia and found throughout the world. The broad ecological tolerance and competitive ability of E. crus-galli makes it the most important weed species in rice. Genetic studies of plants are becoming increasingly common because reliable information is necessary to better understand population dynamics, occurrence of herbicide resistance, and demographic data. Echinochloa crus-galli populations from 34 different locations in Turkey were compared with respect to morphological differences and genetic variation. For morphological variation, five seeds of each population were sown in pots and grown in a screenhouse using a randomized block design. Morphological parameters such as germination speed, flowering time, leaf area, plant height, spikelet length, above-ground biomass, root dry weight and number of seeds were measured. Distinct differences among populations with respect to hierarchical cluster analysis were observed. Genetic variations among populations were performed using random amplified polymorphic DNA (RAPD) markers. The seven RAPD primers amplified 55 bands whose molecular weight varied between 200 and 4000 bp. The percentage of polymorphic bands was 74.54%. Results showed high morphological and genetic variability among individual genotypes within geographic locations. Phenotypic and genetic variability among E. crus-galli populations would be influenced by agricultural practices, crop characteristics, geographic location and herbicide pressure. Differences between weed populations may affect response to chemical or biological control.     

甘肃中部及周边地区小麦条锈菌种群的遗传结构分析

为明确甘肃中部与周边地区小麦条锈菌种群的遗传结构及关系,利用SSR分子标记技术对采自甘肃、陕南、青海及新疆等7个地区共369份小麦条锈病菌标样的群体遗传结构进行了分析。结果表明,小麦条锈菌群体Nei’s基因多样性指数为0.39、Shannon信息指数为0.57,各地区条锈菌群体遗传多样性较为丰富,且在不同地区之间存在明显差异,7个条锈菌群体中以天水种群的遗传多样性相对较高,其Nei’s基因多样性指数为0.42、Shannon信息指数为0.61。该地区小麦条锈菌群体间和群体内都存在着一定的遗传分化,群体间遗传变异占总变异的2.24%,群体内遗传变异占总变异的97.76%。表明甘肃中部及周边地区小麦条锈菌群体存在一定的遗传分化,但遗传变异主要发生在群体内部;甘肃中部、陇南及陕南3地的小麦条锈菌种群遗传相似度较高,菌源交流密切。     

Genetic diversity and population structure analysis of isolates of the rice false smut pathogen Ustilaginoidea virens in India

Genetic diversity assessment and population structure analysis are essential for characterization of pathogens and their isolates. Markers are essential tools for exploring genetic variation among the isolates. False smut of rice caused by Ustilaginoidea virens, formerly Villosiclava virens, is a major emerging disease of rice in India. A high level of variability is observed at the field level, but no information is available from India on genetic diversity and population structure. This is the first report of genetic diversity and population structure of U. virens from India that included 63 isolates distributed across the vast geographical area of eastern and north-eastern India (18.9 to 26.7°N and 82.6 to 94.2°E). Seventeen RAPDs and 14 SSRs were identified as polymorphic and a total of 140 alleles were detected across the populations. The average number of alleles per locus for each primer was 4.5. All the isolates were grouped into two major clusters, with partial geographical segregation that was supported by principal coordinate analysis. Mantel test suggested genetic distance within the isolates increased with increasing geographical distance. Analysis of molecular variation showed more genetic variation within populations and less among populations. This outcome will help in understanding genetic diversity of U. virens from eastern and north-eastern India and in planning effective management strategies.     

Genetic Characterization of Cercospora sorghi from Cultivated and Wild Sorghum and Its Relationship to Other Cercospora Fungi

ABSTRACT Genetic variability and population structure of Cercospora sorghi from wild and cultivated sorghum were investigated to gain insight into their potential impact on epidemics of gray leaf spot of sorghum in Africa. Population structure was examined using data derived from amplified fragment length polymorphism (AFLP) of C. sorghi by Nei's test for population differentiation, G(ST), and analysis of molecular variation (AMOVA). Two ecological populations of C. sorghi in Uganda were devoid of population structure (G(ST) = 0.03, small ef, CyrillicF(ST) = 0.01, P = 0.291). AMOVA revealed that genetic variability was due mainly to variations within (99%) rather than between (0.35%) populations, and Nei's genetic distance between the two populations was 0.014. Phenetic analysis based on AFLP data and polymerase chain reaction-restriction fragment length polymorphism analyses of the internal transcribed spacer regions of rDNA and mitochondrial small subunit rDNA separated Cercospora cereal pathogens from dicot pathogens but did not differentiate among C. sorghi isolates from wild and cultivated sorghum. Our results indicate that Ugandan populations of C. sorghi compose one epidemiological unit and suggest that wild sorghum, while not affecting genetic variability of the pathogen population, provides an alternative host for generating additional inoculum.