Morphological and molecular variability among Indian isolates of <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> causing banded leaf and sheath blight in maize

Rhizoctonia solani, a devastating soil borne fungus inciting banded leaf and sheath blight (BLSB) disease is a constraint in maize production and improvement program. Rhizoctonia isolates collected from seven diverse maize cropping zones of India were examined for morphological and molecular variability. All the tested isolates caused symptoms of BLSB on maize and were also cross infective on rice and sugarcane hosts, but showed significant variability in hyphal diameter, mean hyphal cell size, weight, size and distribution of scleorotia, culture pigmentation, incubation period, pathogenicity and expression of symptoms. Neighbour joining cluster analysis placed the 62 isolates of R. solani into four major groups, A, B, C and D. Group A was more diverse and included isolates of diverse agro-ecological zones. The cluster analysis corresponded well with principle component analysis. Pathogenicity testing of R. solani isolates on maize genotype (CM 501) revealed highly variable virulence pattern of the pathogen population suggesting its high evolutionary potential, and hence adaptability to diverse geographical regions. The study reveals a strong evidence of inherent potential of the R. solani isolates to survive in diverse ecological zones and its probable spread to other maize cultivars across India. Sequence comparisons of the internal transcribed sequence-ribosomal DNA region of 62 isolates did not reveal much diversity among the isolates. Majority of the isolates (n?=?61) clustered together with anastomosis group (AG) AG1-IA used as reference strain in the phylogram, distinct from AG1-IB, AG2–2IIIB and Waitea circinata used as reference strains. BLSB isolates representing distinct geographical locations shared identical sequences indicating long-distance dispersal of the pathogen. The study confirms that the genetic flexibility of the pathogen allows for its adaptation to variable ecological niches and long-distance introduction of new genotypes into the region. The study emphasizes that epidemiological studies may complement the molecular studies.     

Comparison of sequences for the internal transcribed spacer region in <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> AG 1-ID and other subgroups of AG 1

The rDNA-ITS sequence of Rhizoctonia solani AG 1-ID was determined and compared to those of R. solani AG 1-IA, AG 1-IB, and AG 1-IC. The similarity of the isolates from each AG 1 subgroup was almost identical (99%–100%), whereas it was lower between subgroups (91%–95%) than within subgroups. Phylogenetic analysis indicated that isolates of AG 1-ID and other subgroups were separately clustered. Isolates of R. solani AG 1 were clearly separated from R. solani AG 2-1, AG 4, and binucleate Rhizoctonia AG-Bb and AG-K. These results showed that analysis of the rDNA-ITS sequence is an optimal criterion for differentiating R. solani AG 1-ID from other subgroups of R. solani AG 1.     

Identification of Rhizoctonia solani AG 1-IB in Lettuce, AG 4 HG-I in Tomato and Melon, and AG 4 HG-III in Broccoli and Spinach, in Brazil

Fungi isolated in Brazil, from lettuce, broccoli, spinach, melon and tomato, were identified as Rhizoctonia solani. All lettuce isolates anastomosed with both AG 1-IA and IB subgroups and all isolates from broccoli, spinach, melon and tomato anastomosed with AG 4 subgroup HG-I, as well as with subgroups HG-II and HG-III. DNA sequence analyses of ribosomal internal transcribed spacers showed that isolates from lettuce were AG 1-IB, isolates from tomato and melon were AG 4 HG-I, and isolates from broccoli and spinach were AG 4 HG-III. The tomato isolates caused stem rot symptoms, the spinach, broccoli and melon isolates caused hypocotyl and root rot symptoms on the respective host plants and the lettuce isolates caused bottom rot. This is the first report on the occurrence in Brazil of R. solani AG 4 HG-I in tomato and melon, of AG 4 HG-III in broccoli and spinach and of AG 1-IB in lettuce.     

Characterisation of Rhizoctonia solani Anastomosis Groups Causing Bottom Rot in Field-Grown Lettuce in Germany

Bottom rot caused by Rhizoctonia solani is an increasing problem in field-grown lettuce in Germany. During the growing seasons of 1999 and 2000, 95 isolates of R. solani from lettuce plants with bottom rot symptoms were collected from eight locations. The isolates were characterised using hyphal anastomosis, pectic zymograms and morphological characteristics. Ninety-three isolates were identified as anastomosis group (AG) 1-IB, one as AG 1-IC and one as AG 2-1. Optimum hyphal growth was measured over a temperature range of 20–30 °C with an optimum at 25 °C. Aggressiveness of the AG 1-IB isolates varied from weak to strong when tested on detached lettuce leaves. The pathogenic potential of six AG 1-IB isolates was determined on 14 plant species in comparison with lettuce under conditions favourable for the fungus. Radish, broccoli, kohlrabi, spinach and millet seedlings were as severely infected as lettuce seedlings. The same isolates caused little symptoms on maize, tomato and onion. Knowledge about the host range of AGs of R. solani are important for planning an effective crop rotation as part of a control management system.     

Characterization by conventional techniques and PCR of Rhizoctonia solani isolates causing banded leaf sheath blight in maize

Rhizoctonia -diseased specimens were collected from various host species growing in or near maize fields in different geographic regions of the Philippines. A greater range of host species, with varying types of disease symptoms, was found in Mindanao than in Luzon. Fifty-two isolates belonged to anastomosis group AG1-IA and caused banded leaf and sheath blight in maize ( Zea mays ), but they showed considerable variation in virulence. The most and least virulent isolates recovered from maize were both collected from Mindanao. Isolates from necrotic spots/foliar blight of durian and coffee, which were collected from the same region, showed the lowest lesion heights. UPGMA-SAHN clustering analysis from RAPD fingerprint data of 30 haplotypes of R. solani AG1-IA isolates from the Philippines and Japan resolved seven groups of AG1-IA at the 75% similarity level. Variation among isolates from upland crops seemed to be partially correlated with geographical origin and virulence. In the case of paddy rice isolates from Japan and the Philippines, some were closely related, with over 75% similarity, suggesting a common origin. In PCR-RFLP analysis of the rDNA internal transcribed spacer region, no polymorphism was observed among the AG1-IA isolates but they were differentiated from subgroups AG1-IB and AG1-IC using the endonucleases Eco RI, Mbo I and Hin fI.     

Characterization of Rhizoctonia Species Associated with Foliar Necrosis and Leaf Scorch of Clonally-propagated Eucalyptus in Brazil

The objective was to identify and characterize the causal agent of foliar necrosis and leaf scorch of Eucalyptus spp. in Brazil. Nineteen putative isolates of Rhizoctonia obtained from Eucalyptus plants during clonal propagation were compared with isolates from other hosts and with tester strains of anastomosis groups of Rhizoctonia solani. Features compared were morphological characteristics of anamorphs and teleomorphs, numbers of nuclei per cell in the vegetative hyphae, anastomosis of hyphae, and ability to produce necrotic lesions on cuttings and damping-off of E. grandis×E. urophylla hybrid seedlings. Rhizoctonia solani AG1 (AG1-IB like) was the most frequent causal agent isolated from Eucalyptus plants and cuttings with symptoms of leaf scorch and foliar necrosis respectively. These isolates were highly virulent on Eucalyptus cuttings and presented naturally epiphytic growth on Eucalyptus shoots. Binucleate isolates and isolates of R. solani AG4 were also virulent on cuttings and were most virulent on Eucalyptus seedlings causing pre- and post-emergence damping-off. Virulence on Eucalyptus cuttings and seedlings was not restricted to a single species or anastomosis group of Rhizoctonia.     

Population genetic structure of <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> AG1IA from rice field in North India

Rhizoctonia solani AG1IA is an important fungal pathogen causing significant yield and quality losses in rice production. However, little is known about the levels of genetic diversity and structure of this pathogen in North India. Out of 240 samples collected from different rice-growing regions of North India, 112 isolates were identified as R. solani AG1IA subgroups using species-specific primers. All 112 isolates were organized into four groups on the basis of percent disease index (PDI). The majority of the isolates were weakly virulent. Population genetic analysis was performed within and between populations using inter simple sequence repeat (ISSR) markers. A total of 8249 alleles were identified from the 112 isolates of R. solani AG1IA through analysis of the ten inter simple sequence repeat markers. All the ten ISSR markers were polymorphic. The average number of bands per primer was 7.3 which ranged in size from 250 to 1500 bp. Genetic structure of the isolates using inter simple sequence repeat primers showed high degree of polymorphism (PIC ≥0.81). The analysis of molecular variance (AMOVA) indicated that most of the genetic diversity occurred within populations (60%), while the variability among populations and among regions contributed 25 and 15%, respectively. Overall, the present study reveals that a large variation exists among rice-infecting isolates of R. solani AG1IA in North India. Fingerprinting of the isolates using ISSRs along with phenotypic characterization and virulence analysis will help epidemiological studies that can provide new insights into pathogen biology and disease spread.     

Evaluation of Brassica species for resistance to Rhizoctonia solani and binucleate Rhizoctonia (Ceratobasidum spp.) under controlled environment conditions

Isolates of R. solani AG 2–1, AG 8, AG 10 and binucleate Rhizoctonia (Ceratobasidium spp.) were tested for virulence on Brassica crops in growth chamber experiments. Isolate virulence and genotype resistance were determined based on percent of seedling survival, shoot length reduction, and shoot fresh weight. Isolates had significant effects on all tested measurements, compared to the non-inoculated controls. Rhizoctonia solani AG 2–1 appears to be the most aggressive pathogen on all tested genotypes followed by R. solani AG 8, binucleate Rhizoctonia and R. solani AG 10, respectively. Genotype by isolate interaction effects were found to be significant for percent of seedling survival and shoot length reduction. None of the tested genotypes exhibited any level of resistance to R. solani AG 2–1, but three promising genotypes with moderate levels of resistance to R. solani AG 10, R. solani AG 8 and binucleate Rhizoctonia were identified. Moderate heritability (0.57) was observed for the percent of seedling survival in the resistant genotype KS4022.     

Aetiology of Rhizoctonia in sheath blight of maize in Sichuan

Rhizoctonia isolates obtained from maize grown in commercial fields in 33 representative counties (or cities) in Sichuan province in China were characterized according to colony morphology, hyphal anastomosis and pathogenicity. Of 141 isolates, 116 were identified as R. solani , 23 as R. zeae and two as binucleate Rhizoctonia . The isolates of R. solani were assigned to four anastomosis groups (AG): AG-1-IA (101 isolates, accounting for 71.6% of the total), AG-1-IB (2, 1.4%), AG-4 (9, 6.4%) and AG-5 (4, 2.8%). The two isolates of binucleate Rhizoctonia belonged to AG-K. On maize, isolates of AG-1-IA caused typical sheath blight symptoms. Lesions produced by isolates of AG-4, AG-5, AG-1-IB and AG-K were darker than those of AG-1-IA. Rhizoctonia zeae usually caused discontinuous lesions with a dark brown margin and a brown centre on the leaf sheaths, as well as ear rot. Isolates of AG-1-IA were the most virulent to maize, with an average lesion length of approximately 15 cm. Isolates of R. zeae produced lesions approximately 12 cm long, while those of AG-4, AG-5, AG-1-IB and AG-K were progressively shorter. On potato dextrose agar (PDA; pH 6.4), the minimum temperature for mycelial growth of R. zeae isolates was 14–18°C, the maximum 38–40°C and optimum 30°C. Isolates of R. zeae did not grow on PDA (28°C) at pH 2.0, the optimum for growth being pH 6.4.     

First report of white leaf rot on Chinese chives caused by Rhizoctonia solani AG-2-1

Delayed sprouting and white rot of leaf tips were found on Chinese chives in a greenhouse in Hokkaido, Japan, in the spring of 2006 and 2007. The causal fungus was identified as Rhizoctonia solani anastomosis group (AG)-2-1 and discriminated from the leaf rot pathogen R. solani AG-4 HG-I in terms of pathogenicity. Symptoms and the time of year that the disease occurs also apparently differ for the two pathogens. This is the first report of white leaf rot on Chinese chive caused by R. solani AG-2-1.     

Anastomosis Groups, Pathogenicity and Sensitivity to Fungicides of Rhizoctonia solani Isolates Collected on Potato Crops in France

A collection of 241 isolates of Rhizoctonia solani obtained from potato plants grown in different areas in France was characterized for anastomosis grouping, symptomatology on tubers of different cultivars and sensitivity to three fungicides. Most isolates collected belonged to (anastomosis groups (AGs)) AG 3, but 2% and 4% of the isolates were AG 5 and AG 2-1. AG 3 and AG 2-1 isolates were mostly obtained from sclerotia on tubers, but all AG 5, some AG 3 and some AG 2-1 isolates were recovered from superficial tuber alterations, like deformations, corky or scabby lesions. Sclerotia were formed on tubers produced by healthy stem cuttings grown in soil artificially infested with AG 3, but not on tubers grown in soil infested with either AG 5 or AG 2-1. No variation in susceptibility to sclerotial formation was observed among five potato cultivars. In all cases, a large proportion of tubers showed superficial corky lesions, often associated with deformations. The proportion of tubers with lesions and deformations was highest in soil infested with AG 2-1 and significantly lower on cv. Samba in all treatments. All isolates were highly sensitive to flutolanil, iprodione and pencycuron, except the AG 5 isolates, moderately sensitive to pencycuron. These results show that, although AG 3 is the most common R. solani group on potato in France, AG 5 and AG 2-1 may be present. Isolates differed for pathogenicity. In vitro sensitivity to fungicides varied among AGs.     

Characterisation of fungal pathogens causing basal rot of lettuce in Belgian greenhouses

Basal rot is a common disease in lettuce greenhouses. A 3-year study on the diversity of pathogens associated with basal rot in Belgium was carried out. A total of 150 isolates were collected originating from 56 greenhouses. Four pathogens appeared to be involved. Rhizoctonia solani was found to be the causal agent at 23 locations, Sclerotinia spp. at 14, Botrytis cinerea at 17 and Pythium spp. at seven. The isolates of R. solani were further characterised to anastomosis groups and subgroups using morphological characteristics, pectic zymogram and PCR-RFLP. Five anastomosis groups could be distinguished: AG1-1B, AG4 HGI, AG10, AG2-1, AG2-1 Nt and AG3, with isolates of AG4 HGI and AG1-1B being the most prevalent and the most aggressive. Sclerotinia sclerotiorum was found at 13 locations, while S. minor was found at only one location. Based on ITS-sequencing Pythium isolates were assigned to three different species. At 20°C, isolates of all pathogens were able to cause lesions on detached lettuce leaves, except isolates of R. solani AG3 and AG2-1 Nt. A correlation could be found between the occurrence of the pathogens and the growing season. Botrytis cinerea was the most common pathogen in winter, whereas R. solani was most frequently isolated in summer. Sclerotinia spp. and Pythium spp. were isolated in spring, summer and autumn. The information obtained in this study will be most useful in the development of an alternative control strategy for causal agents of basal rot.     

Phylogenetic analysis of <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> subgroups associated with web blight symptoms on common bean based on ITS-5.8S rDNA

Sixty-eight Rhizoctonia solani isolates (31 AG-1, 37 of AG-2-2) associated with web blight (WB) of common bean, Phaseolus vulgaris, were examined for sequence variations in the ITS-5.8S rDNA region. The isolates were collected in bean-growing lowland and mountainous regions in Central and South America. Sequences of these isolates were aligned with other known R. solani sequences from the NCBI GenBank and distance and parsimony analysis were used to obtain phylogenetic trees. WB isolates of AG-1 formed two clades separated from known AG-1 subgroups. WB isolates of AG-2-2 formed one clade separated from known AG-2-2 subgroups. Other isolates belonged to AG-1 IA and AG-1 IB. Based on phylogenetic analysis, we confirmed that at least five genetically different subgroups incite WB of common beans. Three new subgroups of R. solani have been identified and designated as AG-1 IE, AG-1 IF and AG-2-2 WB. DNA sequences of these isolates provided needed information to design taxon-specific primers that can be employed in ecological/epidemiological studies and seed health tests.     

Pathogenicity and Fungicide Sensitivity of <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> and <Emphasis Type="Italic">R. cerealis</Emphasis> Isolates

The Rhizoctonia solani species consists of multinucleate isolates that belong to anastomosis groups AG1–AG3 and differ in virulence and host affinity. R. cerealis is a binucleate species of anastomosis group AG-D which causes sharp eyespot, a common plant disease in Poland. Rhizoctonia spp. is a ubiquitous soil pathogen that poses a significant threat for global crop production due to the absence of effective crop protection products. The aim of this study was to determine the virulence of R. solani and R. cerealis isolates towards Beta vulgaris, Zea mays, Triticum spelta and T. aestivum seedlings, to confirm the presence of endopolygalacturonase genes pg1 and pg5 in the genomes of the tested isolates and to evaluate the tested isolates’ sensitivity to triazole, strobilurin, imidazole and carboxamide fungicides. All tested isolates infected B. vulgaris seedlings. but none of them were virulent against Z. mays plants. R. solani isolates AG4 PL and AG2-2IIIB PL were characterized by the highest virulence (average infestation score of 2.37 and 2.53 points on a scale of 0–3 points) against sugar beet seedlings. The prevalence of infections caused by most of the analysed isolates (in particular R. solani AG4 J—11.8, and R. cerealis RC2—0.78) was higher in spelt than in bread wheat. The virulence of the analysed isolates was not correlated with the presence of pg1 and pg5 genes. The efficacy of the tested fungicides in controlling Rhizoctonia spp. infections was estimated at 100% (propiconazole + cyproconazole), 98.8% (penthiopyrad), 95.4% (tebuconazole) and 78.3% (azoxystrobin).     

The first report of tomato foot rot caused by <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> AG-3 PT and AG-2-Nt and its host range and molecular characterization

Foot rot of mature tomato plants was found in four cities of Hokkaido, Japan, from 2004 to 2007. Six of eight isolates obtained from damaged tissues were identified as Rhizoctonia solani anastomosis group (AG)-3, and the remaining two isolates belonged to AG-2-1. We compared these isolates with nine reference isolates including the different subgroups in AG-3 (PT, TB and TM) and AG-2-Nt (pathogen of tobacco leaf spot) within AG-2-1 in terms of pathogenicity to tomato, tobacco and potato. All eight isolates caused foot rot on tomato. The six AG-3 isolates caused stem rot on young potato plants. While, all reference isolates of AG-3 PT causing stem rot of young potato plants incited foot rot on tomato. The two AG-2-1 isolates and an AG-2-Nt reference isolate caused severe leaf spot on tobacco leaves. The sequences of rDNA- ITS region and rDNA-IGS1 region of the AG-3 isolates showed high similarity to that of AG-3 PT isolates. Phylogenetic tree based on ITS and IGS1 regions of rDNA indicated that the AG-2-1 isolates from tomato formed a single clade with AG-2-Nt isolates and that they were separate from Japanese AG-2-1 isolates (culture type II). Pathogenicity tests and DNA sequence evaluation of the causal fungi revealed that the present isolates of AG-3 and AG-2-1 belonged to AG-3 PT and AG-2-Nt, respectively. This is the first report of tomato foot rot caused by R. solani in Japan.     

Host range and phytotoxicity of <Emphasis Type="Italic">Stemphylium solani</Emphasis>, causing leaf blight of garlic (<Emphasis Type="Italic">Allium sativum</Emphasis>) in China

Since 2004, a new leaf blight disease on garlic of high severity has been observed in Dangyang County, Hubei province, China. Initial symptoms consisted of multiple, small, irregular to oval, white leaf spots, which enlarge to produce sunken purple lesions, sometimes surrounded by a bright yellow margin. As the disease progressed, lesions expanded and merged, resulting in withering of leaf tips. After isolation and pathogenicity testing, the causal agent of leaf blight of garlic was identified as Stemphylium solani from cultural and morphological characteristics, and subsequent analysis of the internal transcribed spacer region of ribosomal DNA. When fungal plugs of two S. solani isolates were inoculated onto 11 garlic cultivars and 20 other crop species, leaf spots appeared on all inoculated plants, but two garlic cultivars (Qingganruanye and Ruanruanye) and three crop species (Capsicum annuum, Brassica napus and Amaranthus mangostanus) showed the smallest leaf spots. In cross-inoculation experiments, no indications of host specificity were observed, but S. solani isolated from garlic was generally the most virulent on five plant species, while the isolate from leek (Allium odorum) was generally the least virulent. Toxicity testing of the crude culture filtrates indicated that garlic isolates produced toxin(s) that were not heat-labile and induced different levels of phytotoxicity toward various garlic cultivars and crops.     

High activities and mRNA expression of pyrophosphate-fructose-6-phosphate-phosphotransferase and 6-phosphofructokinase are induced as a response to Rhizoctonia solani infection in rice leaf sheaths

Rice sheath blight disease caused by Rhizoctonia solani Kuhn results in significant yield and quality losses in rice growing areas worldwide. The glycolytic pathway is important in the resistance response to R. solani infection in rice. This study examined one of the regulatory steps in this pathway catalyzed by pyrophosphate-fructose-6-phosphate-phosphotransferase (PFP) and 6-phosphofructokinase (PFK). PFP and PFK activity in R. solani-infected rice plants increased. The mRNA expression of PFP/PFK isozymes showed that PFK 1, 2, 4 and 5 in the resistant line at 1 dpi were high as compared to the gradual increase observed in the expression of all PFP isozymes. Also PFK 1, PFK 3, PFK 4, PFK 5, PFP 2 and PFP 5 were adaptive to sheath blight disease infection and linked to defence response while, the expressions of PFK 2, PFP 1, PFP 3 and PFP 4 although adaptive, were not specific to R. solani infection. These observations provide evidence that (a) both PFP and PFK have isozymes that play an adaptive role after R. solani infection but while those of PFK are expressed at higher levels within a short time after infection those of PFP are expressed gradually, (b) the adaptive activation of PFP in R. solani-infected rice plants is correlated with the paired expression of its α- and β-subunits as shown by PFP 2 and PFP 5, and (c) the expression of some α-subunits is not specific to R. solani infection as shown by PFP 1, PFP 3 and PFP 4.     

Differentiation of the closely related species,Alternaria solani and A. tomatophila,by molecular and morphological features and aggressiveness

The main causal agent of early blight, the noxious disease of solanaceous crops, is generally considered to be Alternaria solani Sorauer (in a broad sense). However, heterogeneity in many morphological features of this pathogen has been noted suggesting that the disease may not be caused by a single species. Recent research has revealed that several large-spored Alternaria species may cause disease of potato and tomato including A. solani sensu stricto and A. tomatophila. The goal of our research was to compare Russian large-spored Alternaria isolates from tomato and potato to test the hypothesis that early blight of tomato and potato are caused by different species. Cluster analysis of genetic distances estimated from 12 polymorphic molecular markers (universally primed-polymerase chain reaction and randomly amplified polymorphic DNA) revealed two groups of isolates accepted here as A. solani and A. tomatophila that were supported by morphology and host plant association. Differentiation of species was supported by phylogeny derived from the DNA sequences of a portion of the Alt a1, gpd and calmodulin genes. Species-specific primers based on the Alt a1 and calmodulin gene sequences for both species were designed. Under laboratory conditions, A. solani isolates were equally aggressive on both tomato and potato, whereas A. tomatophila was highly aggressive to tomato but only weakly aggressive to potato. In the field, A. solani was isolated from potato, tomato and from several wild potato species including S. schickii, S. papita and S. kurtzianum. The majority (90 %) of A. solani isolates carried the mating type locus 1 (MAT1) idiomorph MAT1-1 while the majority (88 %) of A. tomatophila isolates carried the MAT1-2 idiomorph.     

玉米纹枯病菌细胞核纯化与原生质体转化的探讨

 引起玉米纹枯病的立枯丝核菌(Rhizoctonia solani Kühn)多为多核杂核真菌,也难以通过有性态使其细胞核进行单一化。利用原生质体再生的方法对单核玉米纹枯病菌JN的细胞核进行同核纯化,比较从单个原生质体再生菌获得的rDNA\|ITS序列,发现序列间存在差异,说明其细胞核没有达到预期纯化的效果。继而扩增了双核和多核玉米纹枯病菌的rDNA\|ITS序列,发现它们自身的rDNA\|ITS序列也存在差异,因此认为立枯丝核菌的这种rDNA\|ITS序列的差异可能是长期未经过有性阶段的结果且与其细胞核数目无关。使用转化丝状真菌的载体转化纹枯菌未获得稳定的转化子,进而对载体进行了改造,用纹枯菌(AG\|3)actin基因的启动子和终止子控制潮霉素基因构建了载体pRsA。利用PEG介导的原生质体转化方法,实现了对单核纹枯菌的转化,PCR验证得到3个转化子。但在PDA培养基连续继代5代后,转化子的潮霉素抗性消失。结果对改进纹枯菌的转化方法有借鉴意义。     

Analysis of genetic and pathogenic variation of Alternaria solani from a potato production region

A two-year survey was conducted to investigate the level of genetic variability occurring across growing seasons within natural populations of Alternaria solani, the cause of early blight in potato. Genetic diversity among 151 isolates, taken from a disease resistance breeding trial, was assessed using seven random amplified polymorphic DNA (RAPD) primers and sequence analyses of portions of the internal transcriber spacer (ITS) region and Alt a1 gene. A. solani isolates were grouped into 19 RAPD profiles to examine the distribution patterns of genetically distinct isolates within and between years. Seven RAPD profiles were found spanning both years with profiles 6 and 13 being the most prevalent. Five unique profiles were found only in 2008 and seven were found only in 2009. No variation was observed among isolates of A. solani based on ITS and Alt a1 sequence analyses, but a distinction between A. solani and A. dauci, a close relative outgroup was identified. Pathogenicity was also assessed using a tissue culture plantlet assay on four isolates and two reference cultures. Differences in virulence were observed among the isolates examined.