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1.
During a series of sampling in 2008 and 2009, stem rot disease was detected in Hylocereus polyrhizus plantations in Malaysia, with symptom appeared as circular, brown sunken lesion with orange sporodochia and white mycelium formation on the lesion surface. Eighty‐three isolates of Fusarium were isolated from 20 plantations and were morphologically identified as F. proliferatum based on the variability of colony appearance, pigmentation, growth rate, length of chains, production of bluish sclerotia, concentric ring aerial mycelium and sporodochia. Three species‐specific primers, namely ITS1/proITS‐R, PRO1/2 and Fp3‐F/4‐R successfully produced PCR products and confirmed that the isolates from stem rot of H. polyrhizus were F. proliferatum isolates. From BLAST search of translation elongation factor 1‐alpha (TEF1‐α) sequences, the isolates showed 99–100% similarity with F. proliferatum deposited in GenBank which further confirmed that the isolates were F. proliferatum. The results from amplification of MAT‐allele specific primers indicated that 14.5% of F. proliferatum isolates carried MAT‐1 allele and 85.5% carried MAT‐2. Crossing results showed that all 83 F. proliferatum isolates were male fertile showing positive crosses with the tester strains of MATD‐1 and MATD‐2. Perithecia oozing ascospore were produced. Forty isolates as representative were evaluated for pathogenicity test, produced rot symptoms similar to those observed in the fields which confirmed the isolates as the causal agent of stem rot of H. polyrhizus. To our knowledge, this is the first report of stem rot of H. polyrhizus caused by F. proliferatum in Malaysia.  相似文献   

2.
Pineapple (Ananas comosus) is one the important fruit crops planted in Malaysia, and this study was conducted to determine Fusarium spp. associated with diseases of the fruit crop as Fusarium is prevalent in tropical countries. Our objective was to identify and characterize Fusarium spp. associated with pineapple fruit rot and leaf spot mainly found on the fruits and leaves in Peninsular Malaysia. Fusarium isolates (n = 108) associated with pineapple fruit rot and leaf spot were characterized by morphological, molecular and phylogenetic analyses, a mating study and pathogenicity testing. TEF‐1α sequence analysis identified Fusarium proliferatum, Fusarium verticillioides, Fusarium sacchari and Fusarium sp. Mating was successful only between tester strains of F. proliferatum and F. verticillioides. Sexual crosses with standard tester strains showed that 82 isolates of F. proliferatum produced fertile crosses with mating population D (Gibberella intermedia) and three isolates of F. verticillioides were fertile with the tester strain of mating population A (Gibberella moniliformis). All isolates were pathogenic, causing pineapple fruit rot and leaf spot, thus fulfilling Koch's postulates.  相似文献   

3.
During the year 2008 to 2009, a new disease of stem canker was noticed in most red‐fleshed dragon fruit (Hylocereus polyrhizus) plantations in Malaysia. The symptoms observed were small circular sunken orange spot, black pycnidia and rotted stem. This study was conducted to determine the occurrence of the stem canker on H. polyrhizus in Malaysia, subsequently to isolate, identify and characterize the fungal pathogen based on morphology and molecular characteristics and pathogenicity test. From the surveyed 20 plantations in Malaysia, stem canker was detected in all the plantations. A total of 40 isolates of Scytalidium‐like fungus were isolated and identified as Neoscytalidium dimidiatum based on morphological characteristics and ITS region sequences, which showed 99% similarity to N. dimidiatum (FJ648577). From the phylogenetic analysis using maximum‐likelihood tree, isolates of N. dimidiatum from stem canker of H. polyrhizus were grouped together and did not show any sequence variation. From pathogenicity test, all 40 isolates of N. dimidiatum were pathogenic causing stem canker on H. polyrhizus. To our knowledge, this is the first report of stem canker of H. polyrhizus caused by N. dimidiatum in Malaysia.  相似文献   

4.
During August 2010 and January 2011, 10 isolates of Colletotrichum were recovered from stem anthracnose lesions of Hylocereus polyrhizus in the states of Kedah and Penang, Malaysia. Based on the morphological characteristics of colony colour and appearance, and shapes of conidia as well as sequences of internal transcribed spacer regions (ITS), β‐tubulin, actin (ACT) and glyceraldehyde 3‐phosphate dehydrogenase (GAPDH), the fungus was identified as Colletotrichum truncatum. Pathogenicity test showed that C. truncatum isolates were pathogenic to the artificially inoculated H. polyrhizus stem. This is the first report of C. truncatum causing anthracnose on H. polyrhizus stems in Malaysia.  相似文献   

5.
Isolates from the Fusarium fujikuroi species complex, mainly F. sacchari, have been reported to be the causal agents of pokkah boeng in sugarcane in Brazil. However, inadequate information was available on the occurrence and genetic diversity of F. sacchari in Northeast Brazil, which is a limiting factor on management. Thus, isolates of F. subglutinans sensu lato from sugarcane plants with symptoms of pokkah boeng were evaluated using the sexual cross-fertility to determine species. All the isolates produced black perithecia when they were crossed with the test isolates of F. sacchari. Three weeks after the crossing, the formation of fertile ascospores cirri was observed. Thirty-four isolates were self-sterile hermaphrodites, while 21 were fertile only as males. Five isolates were homothallic. The effective size [Ne(f)] of the population as a function of the frequency of hermaphrodites and female sterile strains was 95.5%. The F. sacchari isolates were separated into four genetic groups independent of geographic location. The mean of polymorphism among all populations was 79%, and the average unbiased genetic diversity (uh) was considered moderate (0.31). This study in addition to confirming that F. sacchari as the main species associated with pokkah boeng in sugarcane in Northeast Brazil, reveals the relationship of mating type and genetic diversity of F. sacchari. The unrestricted gene flow between regions is probably the best explanation for the low geographic correlation. This knowledge will help in the adoption of management measures with fungicides or resistant cultivars.  相似文献   

6.
Isolates of Fusarium subglutinans mating population E are usually found on maize. This fungus forms part of the so-called Gibberella fujikuroi species complex. Previously, F. subglutinans has been associated with two additional mating populations (B and H) and a variety of plant hosts. This was mainly due to a lack of diagnostic morphological characters, but the use of DNA sequence information showed that the strains making up mating populations B, E and H, as well as those associated with the different plant hosts, represent separate species. Recently, another putative mating population has been reported on the wild teosinte relatives of maize. Based on sexual compatibility studies, these isolates were apparently closely related to the pitch canker fungus, F. subglutinans f. sp. pini (= F. circinatum;G. fujikuroi mating population H). The aim of the current study was to determine whether the population of F. subglutinans from teosinte constitutes a new or an existing lineage within the G. fujikuroi complex. For this purpose, portions of the mitochondrial small subunit ribosomal DNA, calmodulin and β-tubulin genes from the fungi were sequenced. Phylogenetic analyses and comparison with sequences from public domain databases indicated that the F. subglutinans isolates from teosinte are most closely related to strains of G. fujikuroi mating population E. These results were confirmed using sexual compatibility studies. The putative mating population from the wild relatives of maize therefore forms part of the existing E-mating population and does not constitute a new lineage in the G. fujikuroi species complex.  相似文献   

7.
Fusarium pseudograminearum is one of the major pathogens causing crown rot of wheat in the semi‐arid and arid areas in Tunisia. In this study, the molecular diversity of 74 isolates of F. pseudograminearum representing three populations from Tunisia and a set of isolates from the world collection was investigated. The potential mycotoxin‐producing ability was tested by PCR using primer pairs specific for the Tri3, Tri7 and Tri13 genes. Results indicated that all the isolates are potentially DON and/or 3‐AcDON producers. The mating‐type idiomorphs were identified using diagnostic PCR primer for MAT1‐1 and MAT1‐2. Both mating types were recovered from the same region and in some cases from the same field. Restriction analysis of the nuclear ribosomal DNA (nrDNA) intergenic spacer region (IGS) revealed 11 haplotypes, five of which were identified in the world collection. The analysis of population structure using the combined IGS and MAT data revealed that the total gene diversity (HT = 0.108) was mostly attributable to diversity within populations (HS = 0.102) and that the genetic differentiation among the four populations was low (GST = 0.09). The analysis of molecular variance (amova ) showed that 15% of the variability was between the Tunisian populations and the world collection. These findings indicate that quarantine measures should be in place to limit the introduction of new populations of F. pseudograminearum into Tunisia.  相似文献   

8.
The distribution of mating types of Ascochyta rabiei (teleomorph: Didymella rabiei) was determined in Tunisia using a MAT‐specific PCR assay. Among 123 isolates tested, 80% were MAT1‐1 and 20%MAT1‐2. Only MAT1‐1 isolates were present in the Beja and Bizerte regions of Tunisia, whereas both mating types were present in Nabeul, Kef and Jendouba. In the latter three regions, the hypothesis of random mating could not be rejected based on chi‐squared tests of mating‐type ratios (P > 0.05). The lower frequency of the MAT1‐2 coupled with the restricted distribution of this mating type in Tunisia may indicate a recent introduction of MAT1‐2 in Tunisia. This speculation is consistent with the recent (2001) observation of D. rabiei pseudothecia on chickpea debris in Tunisia. Forty isolates representative of the five regions were genetically analysed using 10 random amplified polymorphic DNA (RAPD) primers to provide a preliminary estimate of genetic diversity of the pathogen in Tunisia. Among 129 putative RAPD loci amplified, 81% were polymorphic and 32 unique RAPD fingerprints were detected. A high level of genetic differentiation was detected among subpopulations (GST = 0.33). Cluster analyses revealed that isolates from Bizerte, Beja and Jendouba were genetically similar and distinct from isolates sampled in Nabeul and Kef. MAT1‐1 isolates were clustered separately from MAT1‐2 isolates in Jendouba and Nabeul suggesting that recombination may not yet be occurring in these regions despite the occurrence of both mating types in equal frequency in these regions. This lack of recombination between MAT1‐1 and MAT1‐2 also supports the hypothesis of a recent introduction of MAT1‐2 into Tunisia.  相似文献   

9.
Northern corn leaf spot, a foliar disease caused by Cochliobolus carbonum, has become prevalent in southwestern China, especially in the Yunnan Province. Races and mating types were identified for 169 isolates collected from 13 prefectures of Yunnan by artificial inoculation using six hybrid corns as differential hosts and by crossing with three standard mating strains: CC092 (MAT1‐2), CC120 (MAT1‐1) and CC026 (MAT1‐1). Results showed the existence of three races: CCR1 (one isolate), CCR2 (43 isolates) and CCR3 (125 isolates). Most isolates were moderately or weakly virulent with only five being highly virulent. CCR3 was widely distributed and significantly more virulent than CCR2 that coexisted with CCR3 in many locations. On Sach's nutrient agar, 20.71% of the Yunnan isolates self‐mated, forming sterile perithecia. Fully developed perithecia could be formed between isolates of different geographic origins, but only 15.98% strains mated successfully with CC092 and 5.33% formed mature perithecia with 4–6 ascospores per asus. Similar results were obtained in crossing with CC026 or CC120. Mating could also occur between CCR3 and CCR2. Both mating types were found in Yunnan with 84 MAT1‐1 strains (one CCR1, 10 CCR2 and 73 CCR3) and 85 MAT1‐2 strains (33 CCR2 and 52 CCR3) and they coexisted in most areas. To identify the mating type rapidly, three specific primers were successfully developed and employed to amplify the mating‐type genes, with stable patterns of 1627 and 876 bp fragments obtained from MAT1‐1 and MAT1‐2 isolates, respectively. The ratio between MAT1‐1 and MAT1‐2 was 1 : 1, indicating that the mating‐type genes segregated randomly in the field naturally.  相似文献   

10.
Fungus strains designated asFusarium sambucinum, F. torulosum, orFusarium sp. nov. were crossed withMAT1-1 andMAT1–2 tester strains ofGibberella pulicaris. Of the 40 field strains that were crossed with the tester strains, 13 strains produced fertile crosses and 27 strains did not produce fertile crosses. One strain designated asF. torulosum was fertile with a tester strain ofG. pulicaris, suggesting that this is an intraspecies cross and that the strain isG. pulicaris, and, consequently,F. sambucinum rather thanF. torulosum. The lack of fertile crosses between tester strains and 27 of the 40 field strains suggests that these strains are notG. pulicaris. Although the ability to form a fully fertile cross with a tester strain can determine the species of a fertile strain, it is more problematic to exclude a strain only because it is infertile.  相似文献   

11.
Cryptococcus neoformans var. grubii (Cng) is the most common cause of fungal meningitis, and its prevalence is highest in sub‐Saharan Africa. Patients become infected by inhaling airborne spores or desiccated yeast cells from the environment, where the fungus thrives in avian droppings, trees and soil. To investigate the prevalence and population structure of Cng in southern Africa, we analysed isolates from 77 environmental samples and 64 patients. We detected significant genetic diversity among isolates and strong evidence of geographic structure at the local level. High proportions of isolates with the rare MAT a allele were observed in both clinical and environmental isolates; however, the mating‐type alleles were unevenly distributed among different subpopulations. Nearly equal proportions of the MAT a and MATα mating types were observed among all clinical isolates and in one environmental subpopulation from the eastern part of Botswana. As previously reported, there was evidence of both clonality and recombination in different geographic areas. These results provide a foundation for subsequent genomewide association studies to identify genes and genotypes linked to pathogenicity in humans.  相似文献   

12.
Chickpea fields were surveyed in nine major chickpea‐growing provinces of Syria in 2008 and 2009 to determine the prevalence and severity of Ascochyta blight, and the distribution of Didymella rabiei mating types (MATs) in the country. A total of 133 Ascochyta rabiei isolates were assayed for mating type, including isolates from older collections that date back to 1982. Multiplex MAT‐specific PCR with three primers was used for MAT analysis. Out of the 133 tested isolates, 64% were MAT1‐1 and 36% were MAT1‐2. Both MATs were found in six provinces but MAT1‐1 alone was found in three provinces. Chi‐squared analysis was used to test for a 1 : 1 ratio of MAT frequencies in all samples. The MAT ratios in the six provinces were not significantly different from 1 : 1, suggesting that there is random mating of the pathogen population under natural conditions. The presence of the two MATs is expected to play a role in the evolution of novel virulence genes that could threaten currently resistant chickpea varieties. Overall analysis of the 133 isolates showed a significant deviation from the 1 : 1 ratio with almost twice as many MAT1‐1 isolates than MAT1‐2 isolates, which indicates a competitive advantage associated with MAT1‐1 in Syria. However, the overall picture of an unequal frequency in MATs indicates that there may be limited sexual recombination occurring in the Syrian population.  相似文献   

13.
Previous research indicated that black and yellow leaf spots on Cymbidium, Ondontioda, Dendrobium and Cattleya could be caused by Fusarium proliferatum worldwide. However, the agent causing leaf spot on Cymbidium spp. plants is still obscure in Taiwan. Thirty‐five F. fujikuroi species complex (FFSC)‐like isolates were collected from Cymbidium leaf spot from different greenhouses in Taiwan. All isolates were identified as F. proliferatum based on morphological characteristics and molecular analysis. Sequence of translation elongation factor 1‐alpha gene showed 99%–100% homology with F. proliferatum. In addition, two assay techniques using either detached leaves or seedlings were used to evaluate the pathogenicity and host range of the isolates and consequently their effects on Cymbidium and other orchid plants. Pathogenicity assays revealed that all isolates induced black and necrotic spots on detached leaves of Cymbidium, showing 9.4%–29.5% severity on seedlings of Cymbidium. Results of host specificity tests on detached leaves of different plants indicated that the F. proliferatum isolates collected from Cymbidium plants caused severe black spots on Oncidium, Cymbidium, Dendrobium and Cattleya plants. The symptoms on Phalaenopsis plants were relatively mild. Results of host specificity tests on plant seedlings indicated that the F. proliferatum isolates of Cymbidium origin were also pathogenic to Oncidium, Cymbidium and Dendrobium, but not to Cattleya and Phalaenopsis. Phylogenetic analysis of the translation elongation factor (TEF) gene among all fungal isolates using maximum parsimony (MP) and Bayesian inference (BI) methods revealed that the isolates of F. proliferatum from Cymbidium spp. could be separated from other FFSC‐like species with high phylogenetic support.  相似文献   

14.
The distribution was examined of the mating type idiomorphs (MAT‐1 and ‐2) of Phaeosphaeria (anamorph Stagonospora) nodorum using DNA from 49 isolates collected from commercial and experimental fields in 2003 and 2004 in Central Asia. MAT‐1 and ‐2 isolates were present in the Kazakh and Russian origins of P. nodorum, but no MAT‐2 isolates were found in Tajikistan. The possibility of a skewed Tajik population cannot be excluded, considering that the sampled region in Tajikistan was geographically isolated from Kazakhstan and Russia.  相似文献   

15.
This study was designed to identify and compare the Fusarium species of the Gibberella fujikuroi complex on pearl millet (Pennisetum glaucum (L.) R. Br) and corn (Zea mays L.) crops grown in southern Georgia, and to determine their influence on potential fumonisin production. Pearl millet and corn samples were collected in Georgia in 1996, 1997 and 1998. Three percent of the pearl millet seeds had fungi similar to the Fusarium species of the G. fujikuroi species complex. One hundred and nineteen representative isolates visually similar to the G. fujikuroi species complex from pearl millet were paired with mating population A (Fusarium verticillioides (Sacc.) Nirenberg), mating population D (F. proliferatum (Matsushima) Nirenberg) and mating population F (F. thapsinum (Klittich, Leslie, Nelson and Marasas) tester strains. Successful crosses were obtained with 50.4%, 10.1% and 0.0% of these isolates with the A, D and F tester strains, while 39.5 of the isolates did not form perithecia with any tester strains. Two of the typical infertile isolates were characterized by DNA sequence comparisons and were identified as Fusarium pseudonygamai (Nirenberg and ODonnell), which is the first known isolation of this species in the United States. Based on the pattern of cross-compatibility, conidiogenesis, colony characteristics and media pigmentation, a majority of the infertile isolates belong to this species. Fumonisins FB1 and FB2 were not detected in any of the 81 pearl millet samples analyzed. The species of the G. fujikuroi species complex were dominant in corn and were isolated from 84%, 74% and 65% of the seed in 1996, 1997 and 1998, respectively. Representative species of the G. fujikuroi species complex were isolated from 1996 to 1998 Georgia corn survey (162, 104 and 111 isolates, respectively) and tested for mating compatibility. The incidence of isolates belonging to mating population A (F. verticillioides) ranged from 70.2% to 89.5%. Corn survey samples were assayed for fumonisins, and 63% to 91% of the 1996, 1997 and 1998 samples were contaminated. The total amount of fumonisins in the corn samples ranged from 0.6 to 33.3 g/g.  相似文献   

16.
Isolates of Magnaporthe grisea causing gray leaf spot on rice were collected in Argentina and analyzed for mating distribution and fertility. One hundred and twenty-five isolates of M. grisea were collected from rice plants between 2000 and 2003. Each isolate was tested for mating type through a polymerase chain reaction based assay. All M. grisea isolates from Argentina belonged to a single mating type, MAT1.1. The fertility status of isolates was determined using controlled crosses in vitro, pairing each isolate with GUY11 and KA9 (MAT1.2 standard hermaphroditic testers). Production of perithecia was scarce among isolates of the blast pathogen since a low percentage of them (7.2%) developed perithecia with only one of the fertile tester (KA9); all crosses failed with the other tester strain. Asci and ascospores were not observed. The presence of only one mating type and the absence of female fertile isolates indicate that sexual reproduction is rare or absent in M. grisea populations associated with rice in Argentina.  相似文献   

17.
Rice is the staple food crop of more than 60% of the population of the world. This crop suffers from blast disease caused by Magnaporthe oryzae. Information on the mating-type allele distribution and diversity of the pathogen population for the state of Karnataka, India is scanty. With this background, a total of 72 isolates of M. oryzae from rice in different districts of Karnataka were examined for identifying sexual mating alleles MAT1, MAT2 and understanding the genetic diversity based on DNA fingerprint of pot2, an inverted repeat transposon. Among 72 isolates, 44 isolates belonged to MAT1 type (male fertile) and 28 isolates were of MAT2 (female fertile) and there were no hermaphrodite isolates. In a given geographical location, only one mating type was identified. Results revealed that the isolates obtained from these regions are not sexually fertile showing predominant asexual reproduction. Hence, genetic variation observed in the pathogen may be mainly because of high copy number of transposons. A high copy number transposon, namely Pot2, was selected in our study to detect genetic diversity of the pathogen. Pot2 rep-PCR DNA fingerprinting profile showed 27 polymorphic bands with bands ranging in size from 0.65 to 4.0 kb and an average of 10 to 14 bands per isolate. Five distinct clusters were formed with two major, two minor, and one outlier. Clusters 4 and 5 are further subdivided into three sub-clusters. Some of the isolates belonging to clusters 3, 4, and 5 are interlinked as these locations are close to one another sharing common geographical parameters and boundaries. This knowledge on the sexual behavior and genetic diversity of M. oryzae is important with respect to breeding for disease resistance.  相似文献   

18.
用标准菌株对1997~1999年在江苏省吴江市,宜兴市、通州市、高邮市和赣榆县采集的325个猪瘟病菌单孢分离菌株的可育性和交配型进行了测定,结果表明江苏省稻瘟病菌菌株的育性较低,可交配率为22.77%,可育率仅为7.08%。不同年份、不同地区采集的猪瘟病菌茵株的性亲和力和交配型有较大的差异,三年的交配率分别为26.61%、8.26%和33.64%;温州地区和赣榆地区菌株的交配率相对较高,分别为26.15%和25.42%,宜兴地区菌株的交配率较低,只有15.38%。江苏省稻瘟病菌菌株的交配型在不同年份亦出现很大差别,1997年29个可交配菌株中有21个菌株表现为MAT1-2 交配型,而1999年36个可交配茵株均为MAT1-1交配型。用江苏省稻瘟病菌的可育菌株进行互交,25个组合中只有6个组合能产生子囊壳和子囊,但均不产生子囊孢子,提示江苏省稻瘟病菌在田间产生健康有性后代的几率不大。对杂交后代的遗传学分析表明,菌株的交配型是受单基因控制的。  相似文献   

19.
Moroccan isolates of Pyricularia grisea (Magnaporthe grisea) from Oryza sativa and Stenotaphrum secundatum were crossed with standard fertile isolates (mating type Mat 1.1 and Mat 1.2) by using the three‐points culture method on oatmeal agar. Only Mat 1.1 isolates from rice were identified and considered to have only the ability to function as males, whereas no isolate from S. secundatum mated with Mat 1.1 or Mat 1.2. Cross‐inoculation studies using Pyricularia grisea (M. grisea) isolates from Oryza sativa and S. secundatum show that rice varieties cultivated in Morocco were susceptible to isolates originating from S. secundatum. Similarly, S. secundatum was susceptible to some isolates from O. sativa.  相似文献   

20.
Several isolates of three Fusarium species associated with the Gibberella fujikuroi species complex were characterized for their ability to synthesize gibberellins (GAs): Fusarium sacchari (mating population B), Fusarium konzum (mating population I) and Fusarium subglutinans (mating population E). Of these, F. sacchari is phylogenetically related to Fusarium fujikuroi and is grouped in the Asian clade of the complex, while F. konzum and F. subglutinans are only distantly related to Fusarium fujikuroi and belong to the American clade. Variability was found between the different F. sacchari strains tested. Five isolates (B-12756; B-1732, B-7610, B-1721 and B-1797) were active in GA biosynthesis and accumulated GA3 in the culture fluid (2.76–28.4 μg/mL), while two others (B-3828 and B-1725) were inactive. GA3 levels in strain B-12756 increased by 2.9 times upon complementation with ggs2 and cps-ks genes from F. fujikuroi. Of six F. konzum isolates tested, three (I-10653; I-11616; I-11893) synthesized GAs, mainly GA1, at a low level (less than 0.1 μg/mL). Non-producing F. konzum strains contained no GA oxidase activities as found for the two F. subglutinans strains tested. These results indicate that the ability to produce GAs is present in other species of the G. fujikuroi complex beside F. fujikuroi, but might differ significantly in different isolates of the same species.  相似文献   

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