Molecular and morphological characterization of Pyricularia and allied genera

The phylogenetic relationships of Pyricularia species and species from related genera were established from sequences of the internal transcribed spacer ribosomal RNA gene. Phylogenetic analysis disclosed a consistent correlation with spore morphology. Most Pyricularia species studied, and two species of Dactylaria that have obpyriform conidia, fell within the Magnaporthaceae cluster with high bootstrap support. Pyricularia variabilis was more related to Dactylaria, Tumularia or Ochroconis species than to the Magnaporthaceae. Dactylaria and species of Nakataea, Ochroconis, Pyriculariopsis and Tumularia were distinct from the Magnaporthaceae, and the genus Dactylaria is polyphyletic. The combination of morphological and molecular characters, such as spore morphology and ITS ribosomal DNA sequences data, suggested that conidial shape could be a primary character to distinguish Pyricularia from related genera.     

Evolution of an avirulence gene, AVR1-CO39, concomitant with the evolution and differentiation of Magnaporthe oryzae

The significance of AVR1-CO39, an avirulence gene of the blast fungus corresponding to Pi-CO39(t) in rice cultivars, during the evolution and differentiation of the blast fungus was evaluated by studying its function and distribution in Pyricularia spp. When the presence or absence of AVR1-CO39 was plotted on a dendrogram constructed from ribosomal DNA sequences, a perfect parallelism was observed between its distribution and the phylogeny of Pyricularia isolates. AVR1-CO39 homologs were exclusively present in one species, Pyricularia oryzae, suggesting that AVR1-CO39 appeared during the early stage of evolution of P. oryzae. Transformation assays showed that all the cloned homologs tested are functional as an avirulence gene, indicating that selection has maintained their function. Nevertheless, Oryza isolates (isolates virulent on Oryza spp.) in P. oryzae were exceptionally noncarriers of AVR1-CO39. All Oryza isolates suffered from one of the two types of known rearrangements at the Avr1-CO39 locus (i.e., G type and J type). These types were congruous to the two major lineages of Oryza isolates from Japan determined by MGR586 and MAGGY. These results indicate that AVR1-CO39 was lost during the early stage of evolution of the Oryza-specific subgroup of P. oryzae. Interestingly, its corresponding resistance gene, Pi-CO39(t), is not widely distributed in Oryza spp.     

根际放线菌I06—03431产生的核苷类抗生素3431的研究

在抗稻瘟菌农用抗生素的筛选过程中,从60多种中药植物根际土壤获得的放线菌中发现,放线菌106—03431的发酵液具有强抗稻瘟菌的活性。经色谱分离纯化,获得活性成分3431,经光谱学数据分析,证明3431为已知核苷类抗生素,丰加霉素（Toyocamycin）：进一步生物活性研究表明,0．005μg／mL的3431能够抑制稻瘟菌孢子萌发和菌丝生长。另外,3431还具有与植物生长调节剂6-苄基腺嘌呤（6-BA）一样的抑制拟南芥根生长的活性。     

薇甘菊挥发油的化感潜力

外来植物薇甘菊 (MikaniamicranthaKunth .)已成为华南地区重要的杂草 ,其挥发油对植物、真菌和细菌均具有生物活性 ,对植物和水稻稻瘟病菌的抑制活性尤其显著 .随着薇甘菊挥发油浓度 (2 0 0、4 0 0、80 0、16 0 0mg·L-1)的增加 ,6种受试植物幼苗的生长随之明显减弱 .薇甘菊挥发油 (2 5 0 0 g·hm-2 )土壤处理 ,受试的 6种植物鲜重明显减少 ,出苗时间推迟 1～ 2d .薇甘菊挥发油在中等浓度 (40 0mg·L-1)时 ,对水稻稻瘟病菌的抑制作用最强 ,抑菌率为 5 3.38%;对香蕉枯萎病菌的抑制作用次之 ,抑菌率为2 8.6 6 %;对长春花疫病菌的抑制作用最弱 ,抑菌率为 18.6 9%.     

A novel enzyme immunoassay commonly applied for ten strains of Pyricularia oryzae

Antiserum against a strain of the rice blast fungus Pyricularia oryzae was elicited in rabbits immunized with its cell fragments emulsified with incomplete Freund's adjuvant. The fragments were also used as solid-phase antigens. A highly sensitive, competitive type enzyme-linked immunosorbent assay for P. oryzae was developed by using these two preparations as the immune reagents together with the use of beta-D-galactosidase-labeled anti-rabbit IgG as the tracer. Cross-reactivity of nine different strains of P. oryzae were measured by the assay. Sensitivity and accuracy of the assay was improved by choosing the cell fragments of the least cross-reactive strain as the solid-phase antigen. The improved method was successfully applied for sensitive and accurate assay of all ten strains of P. oryzae with the common measuring range between 1 and 100 ng per tube. Other species of microorganisms had little reactivity in this immunoassay indicating that the assay is specific to P. oryzae group microorganisms.     

Comparative Study of Substrates and Inhibitors of Azospirillum lipoferum and Pyricularia oryzae Laccases

Azospirillum lipoferum and Pyricularia oryzae laccases were compared, using several substrates and inhibitors. Sixteen phenolic or nonphenolic compounds were found to be substrates of both fungal and bacterial laccases. In the presence of different phenol oxidase inhibitors, P. oryzae and A. lipoferum laccase activities had similar properties.     

Novel insights into host specificity of Pyricularia oryzae and Pyricularia grisea in the infection of gramineous plant roots

Pyricularia oryzae and Pyricularia grisea are pathogens that cause blast disease in various monocots. It has been reported that P. oryzae infects the leaves and roots of rice via different mechanisms. However, it is unclear to what extent the tissue types affect the host specificities of P. oryzae and P. grisea. Here, we evaluated the tissue‐specific infection strategies of P. oryzae and P. grisea in various gramineous plants. Generally, mycelial plug inoculation caused root browning but the degree of browning did not simply follow the disease index on leaves. Interestingly, the Triticum and Digitaria pathotypes caused strong root growth inhibition in rice, wheat, and barley. Moreover, the Digitaria pathotype inhibited root branching only in rice. Culture filtrate reproduced these inhibitory effects on root, suggesting that some secreted molecules are responsible for the inhibitions. Observation of root sections revealed that most of the infection hyphae penetrated intercellular spaces and further extended into root cells, regardless of pathotype and host plant. The infection hyphae of Digitaria and Triticum pathotypes tended to localize in the outer layer of rice roots, but not in those of wheat and barley roots. The infection hyphae of the Oryza pathotype were distributed in both the intercellular and intracellular spaces of rice root cells. Pathogenesis‐related genes and reactive oxygen species accumulation were induced after root inoculation with all combinations. These results suggest that resistance reactions were induced in the roots of gramineous plants against the infection with Pyricularia isolates but failed to prevent fungal invasion.     

Antimicrobial Activities of Selected Cyathus Species

Twelve selected Cyathus species were tested for their abilities to produce antimicrobial metabolites. Most of them were found to produce secondary exo-metabolites that could induce morphological abnormalities of rice pathogenic fungi Pyricularia oryzae. Some extracts from the cultivated liquid obviously inhibited human pathogenic fungi Aspergillus fumigatus, Candida albicans and Cryptococcus neoformans. Activities against six human pathogenic bacteria were also obtained from some of these extracts.     

Changes in Nucleic Acid Contents in Rice Plants Inoculated with Pyricularia oryzae

RNA contents were reduced in rice leaves after infection by Pyricularia oryzea, Decrease was greater with race IE-2 than with IC-22. No clear cut differences were found between healthy resistant and susceptible plants but differences between susceptible healthy after inoculation plants were significant. A significant increae in DNA contents was observed after inoculation with either race of P.oryzae.     

A diterpene quinone from the bark of Cryptomeria japonica

A diterpene, cryptoquinone, was isolated from the bark of Cryptomeria japonica, the structure, 7,11,14-trioxoabieta-8,12-diene, was established by spectral analyses and X-ray crystallography. This diterpene quinone showed moderate antifungal activities against Pyricularia orizae and Alternaria alternata, and cytotoxic activity against mouse lymphoid neoplasm (P388) cells with an IC(50) of 0.26 microg/ml.     

The Role of Toxin(s) Produced by Germinating Spores of Pyricularia oryzae in Pathogenesis

Pyricularia oryzae produced toxin(s) during spore germination which induced susceptibility to infection by non-pathogenic Alternaria alternata of rice leaves. The induced susceptibility was independent of the compatibility between the races of blast fungi used for obtaining the toxin(s) and the rice cultivars used for bioassay. Susceptibility was also induced in other susceptible species (e.g. barley, Italian ryegrass, perennial ryegrass and wheat), results suggesting that the toxin(s) are host-selective and determine the host specificity at plant species level.     

The Role of Toxin(s) Produced by Germinating Spores of Pyricularia oryzae in Pathogenesis

Pyricularia oryzae produced toxin(s) during spore germination which induced susceptibility to infection by non-pathogenic Alternaria alternata of rice leaves. The induced susceptibility was independent of the compatibility between the races of blast fungi used for obtaining the toxin(s) and the rice cultivars used for bioassay. Susceptibility was also induced in other susceptible species (e.g. barley, Italian ryegrass, perennial ryegrass and wheat), results suggesting that the toxin(s), are host-selective and determine the host specificity at plant species level.     

Participation of Oryza sativa leaf wax in appressorium formation by Pyricularia oryzae

Appressorium formation of Pyricularia oryzae P2 on cover-glass coated with each of the components of rice leaf wax was examined. Wax esters, aldehydes and alcohols, having polar groups and low contact angles, promoted appressorium formation, but alkanes, non-polar molecules having high contact angles, had no effects. Germination of conidia, however, was not aftected with those constituents.     

Comparative analyses of the distribution of various transposable elements in Pyricularia and their activity during and after the sexual cycle

We examined the distribution and activity of six transposable elements found in the blast fungus, Pyricularia spp. Sixty-eight isolates from various gramineous plants were used for the survey, and the elements were plotted on a dendrogram constructed on the basis of their rDNA-ITS2 sequences. MGR586 and Pot2 (Class II elements), Mg-SINE (SINE-like element) and MGR583 (LINE-like retrotransposon) were widely distributed among the Pyricularia isolates, suggesting that they are old elements which arose in, or invaded, the Pyricularia population at very early stages in its evolution. By contrast, the distribution of the LTR-retrotransposons MAGGY and Grasshopper was limited or sporadic, suggesting that they are relatively new elements which recently invaded the Pyricularia population by means of horizontal transfer events. The activity of these elements was evaluated by Southern analysis in progenies derived from a cross between a Setaria isolate and a Triticum isolate. Many new MAGGY signals were observed, which were absent in the parental isolates, at various stages of the sexual cycle and following vegetative growth. In contrast, the other elements yielded few, if any, such signals. Analysis of the sequences flanking the new MAGGY insertions revealed that they were each associated with a 5-bp target-site duplication at both ends of the insertion. These data suggested that MAGGY was the most active of the elements tested for transposition in Pyricularia.     

Characterization of novel <Emphasis Type="Italic">Trichoderma</Emphasis> spp. isolates as a search for effective biocontrollers of fungal diseases of economically important crops in Argentina

Monoconidial cultures of 33 isolates of Trichoderma from Buenos Aires Province, Argentina were characterized on the basis of twenty eight morphological, physiological and biochemical features. All of them were screened for proteinase, endochitinase and β-1,3 glucanase activity. Universally primed PCR (UP-PCR) and inter-simple sequence repeat (ISSR) techniques were used to examine the genetic variability among isolates, which resulted in 127 bands for the total number of isolates. These results were subjected to numerical analysis revealing 20 haplotypes grouped in five clusters. The ability of Trichoderma isolates to antogonize soil-borne fungal plant pathogens using a dual culture assay was done against five fungal species: Alternaria sp., Bipolaris sorokiniana, Fusarium graminearum, F. solani, and Pyricularia oryzae. The highest inhibition values (85% RI) were obtained against B. sorokiniana and P. oryzae. Three isolates of T. harzianum named as FCCT2, FCCT3 and FCCT9 were capable of causing a high growth inhibition on four of the fungal species assayed, which was in agreement with their higher extracellular hydrolytic activity. Our results suggest that these isolates have the potential to be effective agents for biocontrol of cereal and tomato fungal pathogens.     

Genetic and molecular characterization of pathogenic isolates of Pyricularia grisea from wheat (Triticum aestivum Lam.) and triticale (x Triticosecale Wittmack) in the state of Paraná, Brazil

Isolates of Pyricularia grisea from wheat (Triticum aestivum Lam.) and triticale (x Triticosecale Wittmack) spikes with blast symptoms were analyzed by classical (VCG) and molecular (RAPD) techniques. P. grisea mutants, unable to use sodium nitrate (nit) as nitrogen source, were obtained with potassium chlorate. For vegetative compatibility (VCG) tests, genetically complementary nit mutant pairs were inoculated in a medium with sodium nitrate as a single nitrogen source. P. grisea isolates were divided into two vegetative compatibility groups and two RAPD groups. Since vegetative compatible strains may mutually exchange genetic and cytoplasmatic material, the contribution of the parasexual cycle in the genetic variability of Brazilian P. grisea isolates is discussed.     

稻瘟病菌重复顺序PoR6和稻瘟病菌的分型

POR6是一具有高度多态性的稻瘟病菌(Pyricularia oryzae)重复顺序。利用脉冲电泳技术和Southern分析,表明它是非均匀地散布于基因组中的。经测定POR6的拷贝数约为30—40,序列测定未发现在内部有更小的重复单位。用POR6作探针对44株稻瘟病菌进行DNA指纹分析,分析的中国北方地区的22个菌株可根据相似率归并成8个谱系。对一些转管培养中致病型发生变化的菌株用POR6进行指纹分析,发现这些菌株在转管过程中基因组DNA是有变化的。     

In Vitro Effects of Film-Forming Polymers on the Growth and Morphology of Pyrenophora Avenae and Pyricularia Oryzae

Ethokem (Midkem Agrochemicals, Northampton, UK) and Bond (Newman Agrochemicals, Cambridge, UK) completely inhibited linear growth of Pyricularia oryzae at 1 and 2% concentrations of the compounds and strongly inhibited growth of Pyrenophora avenae in vitro . Vapor Gard (Miller Chemical & Fertilizer Corporation, Hannover, PA, USA) was less effective with relatively little inhibition of P. oryzae , but was a more effective inhibitor of P. avenae . All three compounds decreased cell lengths of both pathogens and increased cell diameters of P. avenae . Vapor Gard and Ethokem did not significantly alter cell diameters of P. oryzae , but a highly significant increase was observed with Bond. Gross changes in hyphal morphology were observed including swollen shortened cells, granulation of the cytoplasm, increased branching and collapsed empty cells. Ethokem and Bond decreased growth rate and inhibited biomass production of P. avenae .     

植物病原真菌毒素的研究现状

近几年来,关于寄主专化性毒素(Host-Specific Toxin,HST)的作用机制及分子生物学方面的研究有了新进展;在14种寄主专化性毒素中,以Alternaria属的病原菌产生的专化性毒素的研究更为深入。非寄主专化性毒素(Non-Host-Specific Toxin,NHST)的研究着重在毒素产生的条件、生物活性测定、抗性鉴定以及检测被感染植物体的毒素含量等。本文综述了一些能产生专化性毒素和非专化性毒素的植物病原真菌,有链格孢(Alternaria)、镰孢(Fusarium)、尾孢(Cercospora)、轮枝孢(Verticillium)、梨孢(Pyricularia)、疫霉(Phytophthora)、长蠕孢(Helminthosporium)、黑团孢(Periconia)和核盘菌(sclerotinia)等属的病原菌。从所发表的文献表明真菌毒素在植物病害发展中起着重要作用。