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1.
ABSTRACT Genetic diversity was investigated among California populations of Botryosphaeria dothidea, causal agent of Botryosphaeria panicle and shoot blight of pistachio, with random amplified polymorphic DNA (RAPD) and microsatellite-primed polymerase chain reaction (MP-PCR). We surveyed 120 isolates, 112 of which originated from the California Central Valley and included pistachio isolates (n = 52) and isolates from other plant species (n = 60). Out-group isolates (n = 8) were obtained from pistachio in Greece. There was a strong correlation (r = 0.99; P < 0.0001) between the RAPD- and MP-PCR dissimilarity data sets. Little genetic variation (haplotypic diversity [Hs] < 0.002) was detected among B. dothidea isolates collected from central and southern California pistachio plantings. We observed relatively high diversity for isolates from a northern California pistachio orchard (Hs = 0.0146), where the disease was first diagnosed, and from the Chico U.S. Department of Agriculture Germ Plasm Repository (Hs = 0.0726), where the first pistachio trees were planted in California in 1929. Isolates obtained from other hosts, especially those associated with the rare occurrence of the sexual stage of this fungus, showed the highest levels of diversity (Hs = 0.1689). Thirty-eight pistachio isolates (73.1%) had DNA fingerprints identical to 28 pycnidiospore-derived isolates (56.0%) obtained from other host species. Greenhouse inoculations demonstrated that all isolates obtained from other hosts were capable of infecting pistachio and produced characteristic disease symptomology. Thus, California populations of B. dothidea from pistachio are, for the most part, genetically uniform, with the sexual stage rare to absent. However, the rare occurrence of the sexual stage of B. dothidea on other hosts, and more importantly, the capacity of these isolates to infect pistachio, indicate that other host species may serve as sources of inoculum and genetic variation.  相似文献   

2.
Ma Z  Luo Y  Michailides TJ 《Phytopathology》2004,94(4):326-332
ABSTRACT Spatiotemporal changes in the population structure of Botryosphaeria dothidea, causal agent of panicle and shoot blight of pistachio, were analyzed by using microsatellite-primed polymerase chain reaction (MPPCR), partial sequences of the RNA polymerase II (RPB2) gene, and vegetative compatibility groups (VCGs). We examined 390 isolates, 378 recovered from pistachio in seven counties of California from 1990 to 2001 and 12 recovered from peach, apple, and sycamore in Georgia, North Carolina, South Carolina, Illinois, and Pennsylvania. Six microsatellite primers generated 116 polymorphic bands. Based on MP-PCR data, we observed very high (>98%) levels of genetic identity among populations of B. dothidea collected from the commercial pistachio orchards in California. The near identity of these populations was supported by VCGs and partial sequences of the RBP2 gene. These findings suggest that populations of B. dothidea from commercial pistachio orchards are spatially and temporally stable, at least in the past 5 years.  相似文献   

3.
Pistachio represents an emerging nut crop across the Mediterranean basin. In Spain, pistachio has been traditionally cultivated in marginal-dry areas with unfavourable climatic conditions for plant diseases. Consequently, little attention has been given to research on pistachio diseases until recently. Symptoms of branch dieback and cankers, and shoot and panicle blight have been recently observed in commercial pistachio orchards across southern Spain. In this study, 10 commercial pistachio orchards showing disease symptoms were surveyed between 2017 and 2018. Botryosphaeriaceae fungi were consistently isolated from affected shoots, among other fungal families with minor relevance. Representative isolates of each family were characterized based on colony and conidial morphology, optimum growth temperature, and the comparison of DNA sequence data (ITS, LSU, EF, TUB2, and ACT genomic regions). Detached and attached shoots, and attached panicles of pistachio cv. Kerman were inoculated with mycelial plugs or conidial suspensions to demonstrate the pathogenicity of the selected isolates. Botryosphaeria dothidea, Lasiodiplodia pseudotheobromae, Neofusicoccum mediterraneum, N. parvum, Diaporthe neotheicola, Diaporthe sp., Eutypa lata, Eutypa sp., Cytospora sp., and Phaeoacremonium minimum were identified. P. minimum had the highest optimum growth temperature (29.6 °C) and Cytospora sp. the lowest (21–22 °C). Botryosphaeriaceae isolates showed the largest lesions on inoculated shoots, with L. pseudotheobromae being the most aggressive, followed by Neofusicoccum species. Panicles inoculated with N. mediterraneum showed blight symptoms and canker formation 6 weeks after inoculation, without significant differences in aggressiveness between isolates. This work reports relevant information about this emerging disease in the novel Spanish pistachio-growing areas.  相似文献   

4.
ABSTRACT Didymella bryoniae (anamorph Phoma cucurbitacearum) is the causal agent of gummy stem blight, although other Phoma species are often isolated from cucurbit plants exhibiting symptoms of the disease. The molecular and phylogenetic relationships between D. bryoniae and these Phoma species are unknown. Isolates of D. bryoniae and Phoma obtained from cucurbits grown at various geographical locations in the United States were subjected to random amplified polymorphic DNA (RAPD) analysis and internal transcribed spacer (ITS) sequence analysis (ITS-1 and ITS-2) to determine the molecular and phylogenetic relationships within and between these fungi. Using RAPD fingerprinting, 59 isolates were placed into four phylogenetic groups, designated RAPD group (RG) I, RG II, RG III, and RG IV. D. bryoniae isolates clustered in either RG I (33 isolates), RG II (12 isolates), or RG IV (one isolate), whereas all 13 Phoma isolates clustered to RG III. There was greater than 99% sequence identity in the ITS-1 and ITS-2 regions between isolates in RG I and RG II, whereas isolates in RG III, P. medicaginis ATCC 64481, and P. exigua ATCC 14728 clustered separately. On muskmelon seedlings, a subset of RG I isolates were highly virulent (mean disease severity was 71%), RG II and RG IV isolates were slightly virulent (mean disease severity was 4%), and RG III isolates were nonpathogenic (disease severity was 0% for all isolates). The ITS sequences indicate that RG I and RG II are both D. bryoniae, but RAPD fingerprints and pathogenicity indicate that they represent two different molecular and virulence subgroups.  相似文献   

5.
ABSTRACT Laboratory and field experiments were conducted to study the dispersal of Botryosphaeria dothidea conidia using single-drop impactions and natural and simulated precipitations. For laboratory studies, 200 single drops were released from a height of 1 m on infected pistachio nuts. On pieces of photographic film, 50% of the droplets were collected within 20 mm (average droplet travel distance) of the target area, and the droplets ranged from 0.041 to 3.19 mm in diameter, with an average of 0.3 mm. Each droplet carried an average of 23 B. dothidea conidia. In 3 years of field experiments, rainwater was collected in funnels connected to bottles positioned at different heights inside the tree canopy and at different distances away from the edge of tree canopy in three commercial pistachio orchards in San Joaquin, Yolo, and Glenn counties in California. Numbers of conidia in rainwater varied among and within sampling seasons by sampling dates and orchards. Up to 67,000 conidia/ml were obtained in rainwater samples collected from an orchard in Yolo County. Rainwater from orchards in Yolo and Glenn counties contained a consistently higher number of conidia than rainwater collected from the orchard in San Joaquin County. Variation in numbers of conidia also existed among heights where bottles were located. There were significantly more conidia in rainwater collected inside than outside tree canopies. Inside tree canopies, bottles located at 100 and 150 cm above ground collected more B. dothidea conidia than those placed at 50 and 200 cm. Conidia were collected as far as 1 m from the tree canopy edge. Based on data from the Glenn County orchard, a linear relationship between number of conidia (Y) and rainfall amount (X) in millimeters was determined as Y = 240X - 3,867, with r(2) = 0.91, which meant that a minimum of 16.1 mm of rain was needed to disperse conidia of B. dothidea. The power law model best described the dispersal gradients of B. dothidea propagules in the 1999-2000 and 2001-02 sampling seasons, with r(2) values of >/=0.73, whereas the exponential law model fit best for the 2000-01 data, with r(2) values of >/=0.81. In a rain simulation experiment, the intensity of the rain generated by a nozzle at 138 kPa of pressure inside the tree canopy was approximately five times higher than rain recorded outside the tree canopy. Rain removed up to 65% of conidia from infected fruit. These results confirmed that B. dothidea is a splash-dispersed pathogen with relatively short distances of spore dispersal within pistachio orchards. Only pycnidia are present in pistachio orchards; therefore, the results also indicate that inoculum of B. dothidea should be entirely splashed dispersed.  相似文献   

6.
 随着桉树人工纯林的面积扩大,桉树病害也日趋加重,严重影响和制约桉树产业的发展。2008年至今,在云南省石林县和红河州桉树种植区发生一种病害,主要为害嫩枝、枝条和主干,发病初期,感病枝条和主干上产生红褐色圆或椭圆形坏死斑,以后逐渐扩大成椭圆或不规则溃疡斑,病变扩展到木质部后,常造成枝条生长畸形,易引起刮风断裂。主干树皮下明显呈黑褐色坏死,并造成感染面周围树皮开裂,溃疡处一般可看到流胶,严重时造成整株干枯死亡。调查发现,该病发病率通常在 10%~25%,重者可达 40% 以上,引起桉树溃疡和枯萎死亡。本文通过病原菌分离、致病性测定及传统、分子鉴定明确了该病的病原,以期为病害防治提供理论依据。  相似文献   

7.
ABSTRACT The population structure of Guignardia citricarpa sensu lato (anamorph: Phyllosticta citricarpa), a fungus of which strains pathogenic to citrus are subject to phytosanitary legislation in the European Union and the United States, was investigated. Internal transcribed spacer sequences revealed two phylogenetically distinct groups in G. citricarpa. This distinction was supported by amplified fragment length polymorphism analysis that also supported the exclusion of two isolates that had apparently been misclassified as G. citricarpa. On cherry decoction agar, but not on other media, growth rates of group I isolates were lower than those of group II isolates. Conidial dimensions were similar, but group I isolates formed conidia with barely visible mucoid sheaths, whereas those of group II formed conidia with thick sheaths. Cultures of isolates belonging to group I produced rare infertile perithecia, whereas fertile perithecia were formed by most isolates of group II. Colonies of isolates belonging to group I were less dark than those of group II, with a wider translucent outer zone and a lobate rather than entire margin. On oatmeal agar, exclusively group I isolates formed a yellow pigment. Group I harbored strains from citrus fruits with classical black spot lesions (1 to 10 mm in diameter) usually containing pycnidia. Group II harbored endophytic strains from a wide range of host species, as well as strains from symptomless citrus fruits or fruits with minute spots (<2-mm diameter) without pycnidia. These observations support the historic distinction between slowly growing pathogenic isolates and morphologically similar fast-growing, nonpathogenic isolates of G. citricarpa. The latter proved to belong to G. mangiferae (P. capitalensis), a ubiquitous endophyte of woody plants with numerous probable synonyms including G. endophyllicola, G. psidii, P. anacardiacearum, and P. theacearum. G. mangiferae occurs in the European Union and the United States on many host species including citrus, and does not cause symptoms of citrus black spot, justifying its exclusion from quarantine measures.  相似文献   

8.
Species of Botryosphaeria are well-recognized pathogens of pome and stone fruit trees. The taxonomy of these fungi, however, has been confused in the past. Recent taxonomic changes to the Botryosphaeriaceae further influence the literature pertaining to these fungi. This study reviews the taxonomic status of Botryosphaeriaceae associated with fruit tree diseases, identifies them in South Africa and elsewhere, and develops a reliable identification technique for them. Comparisons were made using DNA sequence data from the nuclear ITS rRNA operon and anamorph morphology. These analyses distinguished six clades amongst isolates associated with fruit tree diseases, corresponding to Neofusicoccum ribis (=  B. ribis ), N. parvum (=  B. parva ), N. australe (=  B. australis ), B. dothidea , Diplodia mutila (=  B. stevensii ) and ' Botryosphaeria ' obtusa (the genus Botryosphaeria is no longer available for the fungus known as B. obtusa , but a new name has not been proposed yet). Isolates from fruit trees in South Africa were grouped in the N. australe and ' Botryosphaeria' obtusa clades. This is the first report of N. australe from fruit trees. PCR-RFLP analysis using the restriction endonucleases Cfo I and Hae III distinguished the major clades. However, two groups of closely related species, N. ribis and N. parvum , and N. australe and N. luteum (=  B. lutea ), had identical RFLP profiles. Using RFLP, it was shown that ' Botryosphaeria ' obtusa is the dominant species on fruit trees in the Western Cape Province of South Africa. These results and methods will be useful in future epidemiological studies and disease management of Botryosphaeriaceae from fruit trees.  相似文献   

9.
Ninety-six strains of Pantoea ananatis were isolated from 14 plant species including melon, rice, tea and other crops of economic importance. They were classified into three groups (group I, II, III) based on a welsh onion stabbing assay, tobacco infiltration test, and polymerase chain reaction to detect indole acetic acid (IAA) biosynthesis genes (iaaM and iaaH) and a cytokinin biosynthesis gene (etz). Group Ι strains were characterized as causing significant blight symptom on welsh onion and inducing a hypersensitive response (HR)-like reaction on tobacco leaves after 36–48 h and encompassed 20 isolates from foxtail millet, hydrangea, pineapple, river water and rice. These 20 isolates did not possess iaaM, iaaH, or etz genes. Group II, consisting of 34 melon isolates, harbored iaaM, iaaH and etz genes, but did not cause either blight on welsh onion or HR-like reaction on tobacco. Group III strains did not have the iaaM, iaaH, and etz genes, nor did they cause any reaction on welsh onion or tobacco. The 42 strains in group III were isolated from bamboo grass, Chinese silver grass, citrus, dogwood, melon, mugwort, silk tree, sweet corn, tea and welsh onion. Representative strains of the three groups were tested for pathogenicity on melon and rice. Group Ι strains caused palea browning on rice but not internal fruit rot on melon. On the contrary, group II strains did not cause disease on rice but caused internal fruit rot on melon. Group III strains were not pathogenic on rice or melon. These results suggested that the host range of P. ananatis may be predicted by the reactions of welsh onion and tobacco and detection of iaaM, iaaH and etz genes. These tools may serve as rapid tests to identify the pathogenicity groups of P. ananatis.  相似文献   

10.
During 2015 and 2016, we detected blighted leaves of pistachio (Pistacia vera) trees in different orchards in Arizona (USA). A Phoma-like species was isolated from pycnidia that appeared embedded in the leaf tissue. The pathogen was identified by means of morphological characteristics and DNA analysis (by sequencing of the ITS, BT, and EF regions) as Didymella glomerata. The optimum temperature for mycelial growth of the pathogen was around 25 °C. Inoculation tests were conducted on healthy and wounded pistachio leaves, fruits, and shoots of the cv. Kerman (female) and the cv. Peters (male). Overall, the pathogen was highly virulent on leaves of both pistachio cultivars and did not need injuries for infection. Conversely, the pathogen did not cause any macroscopic symptoms on the inoculated fruits and shoots but showed a certain endophytic behavior in the shoots. Also, data on the ability of different fungicides to inhibit the in vitro mycelial growth of the pathogen are presented.  相似文献   

11.
红瑞木溃疡病病原菌的鉴定   总被引:1,自引:0,他引:1  
罗卿权  徐颖  刘美 《植物保护》2012,38(4):115-117
为了明确近年来引起上海辰山植物园红瑞木上枝干溃疡病的病原菌种类,首先对病原菌分离并依据柯赫氏法则进行了致病性测定和病原菌验证,然后进行了形态学特征观察、rDNA-ITS序列分析和系统发育树构建。病原菌形态学特征与葡萄座腔菌(Botryosphaeria dothidea)一致,其rDNA-ITS序列(已提交GenBank,登录号:JN638454)与GenBank中葡萄座腔菌相似性达到99%,在系统发育树上与葡萄座腔菌处于同一分支。将辰山植物园红瑞木枝干溃疡病的病原菌鉴定为葡萄座腔菌(B.dothidea)。  相似文献   

12.
测定了甲基硫菌灵、百菌清、腐霉利、戊唑醇、啶酰菌胺、苯醚甲环唑和醚菌酯对山核桃干腐病菌Botryosphaeria dothidea的离体抑菌活性。结果表明,上述7种杀菌剂抑制该病原菌菌丝生长的活性依次为苯醚甲环唑>戊唑醇>腐霉利>甲基硫菌灵>百菌清>醚菌酯>啶酰菌胺。进一步研究表明,苯醚甲环唑对采自安徽和浙江两省从未使用过甾醇脱甲基抑制剂类(DMIs)杀菌剂的150株山核桃干腐病菌菌丝生长的平均EC50值为0.43( ±0.11) μ g/mL,可作为其敏感基线用于以后的抗药性监测。  相似文献   

13.
Cai G  Schneider RW 《Phytopathology》2008,98(7):823-829
Random amplified polymorphic DNA (RAPD) and microsatellite-primed polymerase chain reaction (MP-PCR) were used to characterize 164 isolates of Cercospora kikuchii, most of which were collected from Louisiana. Plant tissue (seeds versus leaves), but not host cultivar, had a significant impact on pathogen population differentiation. Cluster analysis showed that the Louisiana population was dominated by a primary lineage (group I) with only a few Louisiana isolates belonging to the minor lineage that also included the non-Louisiana isolates (group II). A previous study showed that isolates could be differentiated according to vegetative compatibility groups (VCGs). However, RAPD and MP-PCR data demonstrated that isolates of C. kikuchii were not generally clustered according to these VCGs. Furthermore, genetic relationships within and between VCGs were examined using sequences of the intergenic spacer region of rDNA. These analyses showed that VCG is not an indicator of evolutionary lineage in this fungus. Our results suggest the likely existence of a cryptically functioning sexual stage in some portion of the C. kikuchii population.  相似文献   

14.
利用苹果轮纹病菌敏感菌株TS1和抗戊唑醇突变体UV-TS1-10进行了苯丙氨酸解氨酶(PAL)、过氧化物酶(POD)活力、可溶性蛋白含量及酯酶同工酶图谱等生理生化特性的测定。经不同浓度药剂处理0~24h,抗性突变体UV-TS1-10体内PAL活性一直高于敏感菌株TS1,两者都呈现先上升后下降的趋势,并在1.5h达到最高值;抗药突变体UV-TS1-10体内POD活力也高于TS1,一直处于上升趋势,24h达到最高值时两菌株活力差异最大;UV-TS1-10体内可溶性蛋白含量为TS1的1.3倍,同一菌株在不同浓度戊唑醇处理后可溶性蛋白含量差异不大,稳定性好;抗药突变体UV-TS1-10比敏感菌株TS1的酯酶同工酶图谱少了一条Rf=0.33的谱带,多了Rf=0.14的特征性谱带,表明抗戊唑醇突变体UV-TS1-10其生理生化特性发生了较大变化。本文以研究敏、抗菌株间生理生化特性的差异为基础,探讨苹果轮纹病菌对戊唑醇可能的抗性机制,为科学地指导生产用药提供理论依据。  相似文献   

15.
The inhibitory effect of isoprothiolane(diisopropyl 1,3-dithiolan-2-ylidenemalonate), a fungicide for rice blast control, on the metabolism of dibutyl N-methyl-N-phenylphosphoramidate (BPA) by 20 isolates of Pyricularia oryzae was examined in relation to sensitivity of the isolates to the reference fungicide IBP(S-benzyl diisopropylphosphorothiolate). The isolates were divided into five groups based on the modes of BPA metabolism and the inhibition of BPA metabolism by isoprothiolane. Every isolate in groups I and II, which was either a field isolate or a stock culture, decomposed BPA rapidly and produced both hydroxylated and N-demethylated BPA as metabolites. BPA decomposition by these isolates was strongly inhibited by isoprothiolane, resulting in the decreased production of both metabolites in group I and of the hydroxylated metabolite in group II. These isolates were almost equally sensitive to isoprothiolane. Isolates in groups III, IV, and V were all obtained from selection of the fungus mutants found growing on media containing isoprothiolane. Isolates in group III, derived by plating large numbers of conidia, did not decompose BPA to any extent. Mutants of groups IV and V were obtained from fast-growing sectors on agar containing isoprothiolane. Both these groups decomposed BPA, but isolates belonging to group IV produced copious amount of N-demethylated BPA whereas isolates in group V did not. BPA metabolism by these in vitro mutants in groups III, IV, and V was not inhibited by isoprothiolane. Thus, the inhibitory effect of isoprothiolane on BPA metabolism was correlated with sensitivity of an isolate to isoprothiolane. The inhibitory effect of IBP on BPA metabolism was not always correlated with the sensitivity of an isolate to IBP.  相似文献   

16.
Genetic Diversity Within Colletotrichum acutatum sensu Simmonds   总被引:1,自引:0,他引:1  
ABSTRACT Isolates of Colletotrichum acutatum from several hosts were characterized by various molecular methods in comparison with morphological identification. Species-specific primer analysis was reliable for grouping C. acutatum isolates to their designated species. Arbitrarily primed polymerase chain reaction and A+T-rich DNA analyses identified four subgroups within C. acutatum. Subgroup I contained U.S. isolates from almond, apple, peach, and pecan, subgroup II contained isolates from anemone, olive, and strawberry, subgroup III contained isolates from almond (Israel) and strawberry (Spain), and subgroup IV contained a single isolate from anemone (the Netherlands). Likewise, sequence analysis of the internal transcribed spacer (ITS) 2 region alone or the complete ITS (ITS 1-5.8S-ITS 2) region grouped the isolates into the same four subgroups. Percent similarity of the complete ITS region within each cluster ranged from 99.6 to 100.0, 99.8 to 100.0, and 98.6% among subgroups I, II, and III, respectively. DNA sequence analysis of the ITS 2 region alone or the entire ITS 1-2 region was more informative than that of the ITS 1 region, which could only group the isolates into two main clusters. The molecular methods employed for studying genetic variation in populations of C. acutatum determined that this species is diverse, indicating that isolates within populations of each subgroup are not host specific.  相似文献   

17.
Molecular analysis of the major Phytophthora species on cocoa   总被引:1,自引:0,他引:1  
The internally transcribed spacer (ITS) regions of the ribosomal RNA (rRNA) gene cluster of 161 isolates of Phytophthora species involved in pod rot, stem canker and leaf blight of cocoa were analysed to determine inter- and intraspecific variation in this disease complex. The species P. palmivora , P. megakarya , P. capsici , P. citrophthora and P. nicotianae could all be clearly distinguished by PCR amplification of the ITS region followed by restriction analysis with Hae III, Hinf I, Pvu II and Alu I. This method provided a relatively rapid identification procedure for these species, and was able to distinguish isolates that had previously been misidentified by morphological methods. Sequence analysis showed that the four main cocoa-associated species formed two distinct groups, one comprising P. capsici and P. citrophthora , and the other P. palmivora and P. megakarya . Detailed sequence analysis and comparison with published literature suggested that P. capsici isolates from cocoa may be closely related to P. tropicalis , a species recently described from Cyclamen and Dianthus .  相似文献   

18.
Early blight caused by Alternaria solani is a highly destructive disease of potatoes. Control of early blight mainly relies on the use of preventive fungicide treatments. Because of their high efficacy, azoxystrobin and other quinone outside inhibitors (QoIs) are commonly used to manage early blight. However, loss of sensitivity to QoIs has previously been reported for A. solani in the United States. Two hundred and three A. solani field isolates collected from 81 locations in Germany between 2005 and 2011 were screened for the presence of the F129L mutation in the cytochrome b gene; of these, 74 contained the F129L mutation. Sequence analysis revealed the occurrence of two structurally different cytb genes, which differed in the presence (genotype I) or absence (genotype II) of an intron, with genotype I being the most prevalent (63% of isolates). The F129L mutation was detected only in genotype II isolates, where it occurred in 97%. Sensitivity to azoxystrobin and pyraclostrobin was determined in conidial germination assays. All isolates possessing the F129L mutation had reduced sensitivity to azoxystrobin and, to a lesser extent, to pyraclostrobin. Early blight disease severity on plants treated with azoxystrobin was significantly higher for A. solani isolates with reduced fungicide sensitivity in the conidial germination assay compared with sensitive isolates. Data suggest an accumulation of F129L isolates in the German A. solani population over the years 2009–2011. It is assumed that the application of QoIs has selected for the occurrence of F129L mutations, which may contribute to loss of fungicide efficacy.  相似文献   

19.
Alternaria late blight caused by Alternaria spp. in the alternata, tenuissima, and arborescens species-groups is one of the most common fungal diseases of pistachio in California. In this study, a field iprodione-resistant (FIR) isolate of the arborescens species-group and a laboratory-induced iprodione-resistant (LIIR) isolate of the alternata species-group were characterized by fungicide and osmotic sensitivity, virulence on detached pistachio leaflets, and sequence of the coiled-coil region (six repeats of approximately 90-amino-acid domain) of the two-component histidine kinase (HK) gene. Both FIR and LIIR isolates were sensitive to azoxystrobin and tebuconazole, and azoxystrobin-resistant isolates were sensitive to iprodione and tebuconazole. The LIIR isolate showed more sensitivity to osmotic stress than its wild-type parent. However, the FIR isolate did not show higher osmotic sensitivity compared to field iprodione-sensitive (FIS) isolates. Laboratory inoculation tests showed that both FIR and LIIR isolates remained highly virulent on pistachio. Analysis of DNA sequences of the HK coiled-coil region showed that there were no differences in deduced amino acid sequence of this region from the LIIR, FIR, and FIS Alternaria isolates from pistachio in California.  相似文献   

20.
Thirty-eight isolates of Alternaria alternata from pistachio orchards with a history of Pristine (pyraclostrobin + boscalid) applications and displaying high levels of resistance to boscalid fungicide (mean EC(50) values >500 microg/ml) were identified following mycelial growth tests. A cross-resistance study revealed that the same isolates were also resistant to carboxin, a known inhibitor of succinate dehydrogenase (Sdh). To determine the genetic basis of boscalid resistance in A. alternata the entire iron sulphur gene (AaSdhB) was isolated from a fungicide-sensitive isolate. The deduced amino-acid sequence showed high similarity with iron sulphur proteins (Ip) from other organisms. Comparison of AaSdhB full sequences from sensitive and resistant isolates revealed that a highly conserved histidine residue (codon CAC in sensitive isolates) was converted to either tyrosine (codon TAC, type I mutants) or arginine (codon CGC, type II mutants) at position 277. In other fungal species this residue is involved in carboxamide resistance. In this study, 10 and 5 mutants were of type I and type II respectively, while 23 other resistant isolates (type III mutants) had no mutation in the histidine codon. The point mutation detected in type I mutants was used to design a pair of allele-specific polymerase chain reaction (PCR) primers to facilitate rapid detection. A PCR-restriction fragment length polymorphism (RFLP) assay in which amplified gene fragments were digested with AciI was successfully employed for the diagnosis of type II mutants. The relevance of these modifications in A. alternata AaSdhB sequence in conferring boscalid resistance is discussed.  相似文献   

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