Phylogenetic analysis based on ITS sequences and conditions affecting the type of conidial germination of Bipolaris oryzae

One taxonomic characteristic of Bipolaris species is the bipolar germination of conidia, but conidia of Bipolaris oryzae , the causal pathogen of brown spot in rice, are regularly observed to show intercalary germination, a characteristic of Drechslera species. The effect of selection, culture media and culture age on type of conidial germination was determined for three brown spot isolates from Cavinti, San Pablo and Palawan in the Philippines, obtained from infected leaves showing typical disease symptoms. Based on the analyses of their ITS1, ITS2 and 5·8S rDNA nucleotide sequences, the local isolates were clearly identified as B. oryzae . Selection for colonies of the three isolates derived from single conidia with either bipolar or intercalary germination had no effect on the number of spores showing bipolar germination in subsequent cultures. Germination on seven different culture media was tested; of these, rabbit food agar and water agar increased the percentage of bipolar germination of conidia, although this varied between isolates. Incubation of the cultures of all three isolates for longer periods prior to harvesting conidia increased the percentage of bipolar-germinating conidia from c . 40 to c . 90% with 5-day-old and 30-day-old cultures, respectively.     

Population structure,genetic diversity,and sexual state of the rice brown spot pathogen Bipolaris oryzae from three Asian countries

Bipolaris oryzae causes brown spot in rice (Oryza sativa) inflicting substantial grain yield losses worldwide. Knowledge of the population structure, genetic diversity and sexual recombination of the fungal pathogen can help to implement effective disease management strategies. In this study, B. oryzae isolates sampled from Iran, the Philippines and Japan were analysed with 12 simple‐sequence repeat (SSR) markers, newly developed from the genome sequence of the fungus. Among the 288 B. oryzae isolates genotyped, 278 unique haplotypes were identified. High genotype numbers (richness) with even distribution (evenness) were found within the collection sites. Both mating types, MAT1‐1 and MAT1‐2, were present in each collection area, and the sexual state was induced under controlled conditions with production of viable ascospores. However, the tests of linkage disequilibrium rejected of the hypothesis of random mating. Discriminant analysis of principal components (DAPC) revealed that the B. oryzae collection formed three clusters, each consisting of isolates from different collection sites. Analysis of molecular variance (amova ) showed that genetic variation among clusters was 18.7%, with the rest of the variation distributed within clusters (R_ST = 0.187, P < 0.001). Statistically significant pairwise genetic differentiation was found between the clusters. These results show that Asian B. oryzae isolates are genetically diverse, and, overall, distributed in three groups. These findings will be helpful in managing the disease and guide the use of representative isolates needed for selection of resistant rice varieties.     

Population genetic structure of four regional populations of the barley pathogen Pyrenophora teres f. maculata in Iran is characterized by high genetic diversity and sexual recombination

The leaf spot form of the barley disease net blotch, caused by the fungus Pyrenophora teres f. maculata (PTM), is an increasingly important foliar disease of barley. Studies of population genetic structure and reproductive mode are necessary to make predictions of the evolutionary potential of the pathogen. Sources of resistance to PTM have been found in Iranian landraces, which may have the potential to improve plant breeding efforts. However, little is known about the population genetic structure of this fungus in Iran. In this study, we analysed the frequency of the mating type genes to assess the potential for sexual mating of PTM collected from four provinces—Khuzestan, Hamadan, Golestan, and East Azerbaijan—and we investigated the population genetic structure using seven simple sequence repeat markers. High genotype diversity, linkage equilibrium, and equal ratios of mating types frequencies in the PTM populations at Khuzestan and Hamadan support the occurrence of sexual reproduction in these populations, while in Golestan and East Azerbaijan populations, significant gametic disequilibrium and relatively low genotype diversity suggest a higher incidence of clonality or different demographic histories. Unequal mating type frequencies in Golestan confirm a predominance of asexual reproduction. Finally, we found significant evidence for strong population structure with most of the genetic variation represented within regional populations (89%). Overall, our study provides evidence for high genetic variation in Iranian PTM populations, which may be the basis for rapid adaptive evolution in this pathosystem. This highlights the need for integrated efforts to control the disease.     

Assessing new SSR markers for utility and informativeness in genetic studies of brown rust fungi on wheat,triticale, and rye

The aim of the present study was to validate new simple-sequence repeat (SSR) markers and use them to assess genetic variability among 24 isolates of Puccinia triticina collected from wheat (Pt-wheat) and triticale (Pt-triticale), and 15 isolates of P. recondita f. sp. secalis (Prs) collected from rye. The Pt and Prs isolates were tested for virulence on a set of 35 Thatcher wheat near-isogenic lines, eight rye lines with known resistance genes, and 53 triticale cultivars with uncharacterized leaf rust resistance. Molecular genotypes were determined using a newly developed set of 34 SSR microsatellite primer pairs. All SSR markers tested on Pt isolates successfully amplified fragments of appropriate size. When tested on the Prs isolates, 21 out of the 34 Pt SSRs amplified expected fragments. Sixteen of these 21 SSRs were polymorphic, providing for the first time microsatellite markers to study genetic variation in Prs. Based on virulence data, variation among Prs isolates was low, probably due to the small number of rye differential lines available. Much higher variation for virulence was observed within the collection of Pt isolates from wheat and triticale, and two separate groups were established with mixed host origin. Substantial genetic variation was detected among the isolates studied with the SSR markers, assuming two different models of SSR evolution (infinite alleles model and stepwise mutation model). The newly developed set of SSR markers proved their effectiveness in detecting genetic variation and should be useful in further population genetics investigations of the two pathogens.     

湖北省随州地区稻瘟病菌群体结构分析及生态模式菌群的建立

采用人工接种方法,用7个中国鉴别品种和95个自选资源材料对2014年至2015年间采自湖北省随州地区的76个稻瘟病菌单孢菌株进行致病型鉴定。分别利用传统生理小种种群划分方法和近邻传播聚类法(affinity propagation clustering,AP聚类法)进行菌群结构分析,比较两种方法的差异。结果表明,传统生理小种种群划分方法将76个菌株分为6个菌群,14个中国小种, ZB菌群为优势种群,占比65.79%,ZB15为优势生理小种,占比达到60.53%;近邻传播聚类法将76个菌株分为14个菌群,Cluster 2、Cluster 6、Cluster 7、Cluster 8为随州地区优势菌群,占比分别为18.42%、19.74%、22.37%、17.11%,Cluster 6、Cluster 7和Cluster 8具有相对较高的致病力;两种方法划分结果虽不完全一一对应,但划分趋势相同,近邻传播聚类法在稻瘟病菌种群结构研究中具有一定应用前景;根据试验结果筛选出1个代表随州地区稻瘟病总致病谱的菌群,菌群由12个菌株组成,将其命名为“随州生态模式菌群”。     

湖北省小麦品种(系)对赤霉病、条锈病、白粉病和纹枯病的抗性评价

为了解湖北省小麦品种(系)的抗病性水平,对2001年以来湖北省审定的46个小麦品种和在湖北省进行区域试验的447份小麦品系进行赤霉病、条锈病、白粉病和纹枯病等4种主要病害的抗病性评价。结果表明:1)参试品系中有1份材料对赤霉病表现高抗,未发现对纹枯病表现高抗或免疫的材料;对赤霉病和纹枯病的平均抗性评价分值分别为14.30和3.49,介于中感和高感之间;对条锈病和白粉病的平均抗性评价分值分别为13.43和8.34,介于中抗和中感之间。2)审定品种的综合抗性水平不高,对条锈病的抗性评价分值为12.47,介于中抗和中感之间,对其他3种病害的抗性介于中感和高感之间;审定品种的单产呈增加趋势,株高在不断降低,生育期相应延长,综合抗病性也在增强。3)审定品种的综合抗病性与对条锈病抗性的相关系数最高,为0.738;对纹枯病的抗性与株高呈显著正相关,与单产呈显著负相关;对赤霉病的抗性与株高无显著相关。鉴于此,湖北省需加强抗病育种力度,同时应充分做好小麦病害的防控。     

Population genetic diversity and structure of the pecan scab pathogen,Venturia effusa,on cv. Desirable and native seedlings,and the impact of marker number

Scab (Venturia effusa) is the major cause of economic loss in pecan in the south-eastern USA. We explored population genetic diversity and structure among orchards of cv. Desirable and native seedlings, and within-orchard variability among trees of all cultivars sampled. We compared the ability of 30, 15 and 7 previously developed microsatellites to characterize the population genetic diversity and structure of V. effusa. Analyses of molecular variance (AMOVA) provided little evidence of structure dependent on cultivar, but there was some evidence of structure between orchards of a cultivar based on distance. Individual orchard AMOVA showed that three of 11 orchards had between-tree population structure. Among six populations from cv. Desirable, a Mantel test showed that geographic distance was related to the pairwise genetic divergence (R² = 0.84). Among 11 orchards of various cultivars there was little difference in diversity using 30, 15 or 7 markers, or population structure based on AMOVA. Some minor differences in population structure were seen based on discriminant analysis of principal components, or dendrograms. Thus, depending on the objectives, future studies may use as few as 15 or 7 markers without losing ability to discern population genetic diversity or structure. More populations exhibited linkage disequilibrium when using 15 or 30 markers compared to when using seven markers. Knowledge of population genetics of V. effusa in relation to host genotype is needed to understand pathogen population interactions and gene flow, knowledge that will help underpin future breeding efforts to develop durable resistance in this long-lived orchard tree.     

Effects of temperature on germination and hyphal growth from conidia of Ramularia rhei and Ascochyta rhei, causing spot diseases of rhubarb (Rheum rhaponticum)

Rhubarb leaf and petiole spot disease, caused by Ramularia rhei and Ascochyta rhei , has gradually become more noticeable in the UK field crop. Conidial germination and subsequent colony growth of R. rhei and A. rhei were investigated under in vitro conditions on potato dextrose agar and in vivo on leaf discs. Results indicated that the two fungi responded differently to temperature. Ramularia rhei was better adapted to temperatures ≤ 25°C, with an optimum around 20°C, whereas A. rhei was more adapted to temperatures ≥ 15°C, with an optimum > 25°C. Overall, conidia of R. rhei germinated and subsequent colonies grew at greater rates than those of A. rhei on leaf discs at temperatures ≤ 25°C. These results indicated that it is important to identify the causal agent of leaf and petiole spot diseases in rhubarb field crops in order to estimate disease risks accurately.     

Epidemiology and management of Leptosphaeria maculans (phoma stem canker) on oilseed rape in Australia, Canada and Europe

Phoma stem canker (blackleg), caused by Leptosphaeria maculans , is an important disease on oilseed rape (canola, rapeseed, Brassica napus , Brassica juncea , Brassica rapa ) causing seedling death, lodging or early senescence in Australia, Canada and Europe, but not in China. The two forms of L. maculans (A group and B group) that occur on oilseed rape are now considered to be separate species. The epidemiology and severity of phoma stem canker differs between continents due to differences in the pathogen population structure, oilseed rape species and cultivars grown, climate and agricultural practices. Epidemics are most severe in Australia, where only the A group occurs, and can be damaging in Canada and western Europe, where both A and B groups occur, although their proportions vary within regions and throughout the year. Epidemics are slight in China, where the A group has not been found. Dry climates (Australia, western Canada) lengthen the persistence of infected debris and may synchronize the release of airborne ascospores (after rain) with seedling emergence. L. maculans spreads from cotyledon and leaf infections down petioles to reach the stem, with infections on cotyledons and leaves early in the season producing the most damaging stem cankers at the stem base (crown). Development of both crown cankers and phoma stem lesions higher up stems is most rapid in regions with high temperatures from flowering to harvest, such as Australia and Canada. Breeding for resistance (genetic, disease escape or tolerance), stubble management, crop rotation and fungicide seed treatments are important strategies for control of phoma stem canker in all areas. Fungicide spray treatments are justified only in regions such as western Europe where high yields are obtained, and accurate forecasts of epidemic severity are needed to optimize their use.     

基于两性生命表的以朱砂叶螨为猎物的加州新小绥螨四川种群生长发育、繁殖及捕食率

为全面掌握加州新小绥螨四川种群的种群特征,本试验运用两性生命表开展了以朱砂叶螨为猎物的加州新小绥螨四川种群的生长发育、繁殖与捕食率研究。结果表明,加州新小绥螨四川种群不同发育阶段重叠,在雌成螨产卵期结束前雌成螨特定年龄-阶段存活率S_xj明显高于雄成螨。单位时间（12 h）特定年龄-阶段繁殖力f_x5、特定年龄繁殖力m_x最大值分别为1.74和1.12粒/雌,单雌总产卵量为49.28粒。种群内禀增长率为0.27/d、周限增长率为1.31/d、净增殖率为26.61粒/个体、世代平均周期为12.34 d。在未成熟阶段,加州新小绥螨各螨态特定年龄-阶段捕食率C_xj均呈双峰型,雌成螨和雄成螨捕食率均随时间明显波动。加州新小绥螨取食朱砂叶螨的转化率为4.55、内禀捕食率为0.54、周限捕食率为1.72头。以朱砂叶螨为猎物时,加州新小绥螨四川种群表现出较好的个体发育、种群增长和捕食能力,可作为控制朱砂叶螨的生防资源,值得进一步研究和开发利用;同时朱砂叶螨可作为加州新小绥螨四川种群人工繁殖选择的猎物之一。     

Effects of cultural conditions on fungal biomass,blastospore yields and toxicity of fungal secreted proteins in batch cultures of Metarhizium anisopliae (Ascomycota: Hypocreales)

BACKGROUND: Recently, two fungal proteins with apparent molecular masses of 11 and 15 kDa and insecticidal activity against Ceratitis capitata (Wied.) have been purified from the crude soluble protein extract (CSPE) secreted by the entomopathogenic fungus Metarhizium anisopliae (Metsch.) Sorokin (strain EAMa 01/58‐Su) in Adamek's liquid medium. The feasibility of culturing this strain in fermentation facilities in order to harvest and formulate the insecticidal proteins for C. capitata control is mainly dependent on the ability to produce high concentrations of the active proteins at a reasonable cost. RESULTS: These studies report that, in batch cultures of EAMa 01/58‐Su strain, the carbon (C) and nitrogen (N) ratios and sources are important considerations with respect to fungal biomass production, blastospore yield and secretion of insecticidal proteins against C. capitata adults. The data indicate that the type and concentration of N source in the medium influence the production of insecticidal protein and thus the toxicity of the CSPEs. The electrophoretic analysis suggests that the monomer of 11 kDa plays an important role in the insecticidal effect described. Concerning biomass production, no clear differences were found between media with different C and N sources and C:N ratios in total biomass production at day 7. Conversely, important differences were found among the media in terms of blastospore yields. CONCLUSIONS: By optimising the culture media, the insecticidal effect of the CSPE against C. capitata can be improved. In the CSPE from G₄₀:P₂₀ (40 g L^?1 glucose and 20 g L^?1 peptone in dH₂O), the LC₅₀ and the LT₅₀ were 7 and 4.5 times lower than in the CSPE obtained from Adamek. Copyright © 2010 Society of Chemical Industry     

Effect of compost and casing treatments of insecticides against the sciarid Bradysia ocellaris (Diptera: Sciaridae) and on the total yield of cultivated mushrooms,Agaricus bisporus

BACKGROUND: Bradysia ocellaris (Comstock) is one of the major pests of cultivated mushrooms, Agaricus bisporus (Lange) Imbach. The feeding of sciarid fly larvae causes damage to the cultivation substrates, compost and casing. A comparative study of insecticidal treatments incorporated into compost and casing against B. ocellaris was conducted during 2003–2004. Treatments with an organophosphate, diazinon, a pyrazole, fipronil, and an insect growth regulator (IGR), triflumuron, were incorporated into casing. Another IGR, cyromazine, was incorporated into compost. Insecticidal incorporation into compost and casing has been associated with reductions in mushroom yield. Therefore, the effect of these insecticidal treatments on the overall yield was evaluated. RESULTS: A combined treatment, incorporation of cyromazine at 10 mg kg^?1 into compost and of triflumuron at 20 mg kg^?1 into casing, provided 100% control against B. ocellaris. There was no apparent reduction in yield as a result of incorporation of the tested chemicals into either compost or casing. CONCLUSION: The results obtained from this study suggest that incorporation of insecticides into both compost and casing is required to control a moderate to high pressure of insect pests. There were no significant yield reductions as a result of the incorporation of pesticides in the Australian mushroom‐growing system. Copyright © 2009 Society of Chemical Industry     

光波长、刺激时长和性别结构对龟纹瓢虫趋光性的影响

为揭示天敌昆虫趋光性中光源因子与虫源因子的作用,采用室内多向行为选择方法研究光波长、刺激时长和性别结构对龟纹瓢虫Propylea japonica趋光性的影响。结果显示:在365～630 nm波长范围内,光波长对龟纹瓢虫雌雄性比为1∶1的两性种群的趋光性有显著影响,其对365 nm处紫外光的趋光率最高,为27.90%;对625～630 nm处红光的趋光率最低,为6.50%。在120 min范围内,光刺激时长对龟纹瓢虫雌雄性比为1∶1的两性种群的趋光性有显著影响,且该影响与光波长有关。利用紫外光刺激龟纹瓢虫雌雄性比为1∶1的两性种群5 min时的趋光率(41.07%)显著高于其它刺激时长处理,且其它处理之间均无显著差异。随着光刺激时长的增加,龟纹瓢虫雌雄性比为1∶1的两性种群对紫外光、蓝光短波长的趋光率降低,对红光和黑暗处理的趋光率增加。在紫外光、蓝光、绿光及橙光处理下性别结构对龟纹瓢虫有显著影响,雌性单性种群偏嗜紫外光和绿光;雄性单性种群偏嗜绿光和橙光;雌雄性比为1∶1的两性种群偏嗜紫外光、蓝光和橙光。表明光波长是龟纹瓢虫趋光性的核心诱因,性别结构及光刺激时长具有重要的协同调控作用。