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1.
ABSTRACT Alternaria citri, a postharvest pathogen, produces endopolygalacturonase (endoPG) and causes black rot on citrus fruit. We previously described that an endoPG-disrupted mutant of Alternaria citri was significantly reduced in its ability to macerate plant tissue and cause black rot symptoms on citrus. In order to investigate colonization of citrus fruit tissues by Alternaria citri, pTEFEGFP carrying a green fluorescent protein (GFP) gene was introduced into wild-type Alternaria citri and its endoPG-disrupted mutant (M60). Green fluorescence was observed in spores, germ tubes, appressoria, and infection hyphae of transformants G1 (derived from wild type) and GM4 (derived from M60). Hyphae of G1 but not GM4 vertically penetrated the peel, but the hyphae of both G1 and GM4 spread equally in the juice sac area of citrus fruit. Green fluorescence of Alternaria citri transformant EPG7 carrying a GFP gene under control of the endoPG gene promoter of Alternaria citri was induced by pectin in the peel during the infection stage, but repressed completely in the juice sac area, likely by carbon catabolite repression by sugars in the juice. 相似文献
2.
Kouhei Ohtani Atsunori Isshiki Hiroshi Katoh Hiroyuki Yamamoto Kazuya Akimitsu 《Journal of General Plant Pathology》2003,69(2):120-125
The Acpg1 gene of Alternaria citri encodes an extracellular endopolygalacturonase that is important for virulence in citrus fruits. Expression of Acpg1 is regulated by substrate induction and carbon catabolite repression. A green fluorescent protein (GFP) gene was employed
as a reporter gene to define 813 bases upstream of the translation start site comprising the Acpg1 promoter. This upstream sequence contains five putative binding sequences of catabolite repressive element A (CreA), a cis-acting zinc finger repressor involved in carbon catabolite repression. We constructed each CreA-binding site-deleted Acpg1 promoters with GFP reporter gene and transformed them to A. citri. The construct PGPDL4 deleted from −401 to −813 showed both substrate induction and catabolite repression, whereas PGPDL5
additionally deleted from −1 to −84, including one putative CreA-binding site, resulted in a loss of catabolite repression
function. Green fluorescence of PGPDL4 was induced by pectin in the peel but was repressed completely in the juice sac area
of citrus fruit. However, green fluorescence of PGPDL5 was induced in both the peel and juice sac area, indicating that repression
of Acpg1 in the juice sac area is likely accomplished by carbon catabolite repression.
Received: October 4, 2002 / Accepted: October 31, 2002
Acknowledgments The authors thank Drs. D. Cullen, A. Van den Wymelenberg, and J. Andrews, University of Wisconsin, for providing pTEFEGFP
containing GFP and Dr. T. Tsuge, Nagoya University, for providing transformation vector pSH75. The nucleotide sequence data
of Acpg1 promoter region in the DDBJ, EMBL, and GenBank sequence databases is under accession number AB047543. This research was supported
in part by grants to K.A. from the Ministry of Education, Culture, Sports, Science, and Technology of Japan. 相似文献
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ABSTRACT Alternaria citri causes Alternaria black rot, a postharvest fruit disease, on a broad range of citrus cultivars. We previously described that an endopolygalacturonase minus mutant of A. citri caused significantly less black rot in citrus fruit. To search for other essential factors causing symptoms in addition to endopolygalacturonase, a random mutation analysis of pathogenicity was performed using restriction enzyme-mediated integration. Three isolates among 1,694 transformants of A. citri had a loss in pathogenicity in a citrus peel assay, and one of these three mutants was a histidine auxotroph. Gene AcIGPD that encodes imidazole glycerol phosphate dehydratase, the sixth enzyme in the histidine biosynthetic pathway, was cloned, and the mutant containing the disrupted target gene, AcIGPD, caused less black rot. 相似文献
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ABSTRACT Phylogenetic analysis revealed that isolates of Alternaria alternata causing black rot of citrus were associated with six well-supported evolutionary lineages. Isolates recovered from brown spot lesions on Minneola tangelo, leaf spot lesions on rough lemon, and healthy citrus tissue and noncitrus hosts were related closely to isolates from black-rotted fruit. Phylogenies estimated independently from DNA sequence data from an endopolygalacturonase gene (endoPG) and two anonymous regions of the genome (OPA1-3 and OPA2-1) had similar topologies, and phylogenetic analysis was performed on the combined data set. In the combined phylogeny, isolates from diverse ecological niches on citrus and noncitrus hosts were distributed in eight clades. Isolates from all lineages, regardless of ecological or host association, caused black rot in fruit inoculation assays, demonstrating that small-spored Alternaria isolates associated with different ecological niches on citrus and other plant hosts are potential black rot pathogens. These data also indicated that the fungi associated with black-rotted fruit do not form a natural evolutionary group distinct from other Alternaria pathogens and saprophytes associated with citrus. The use of the name A. citri to describe fungi associated with citrus black rot is not justified and it is proposed that citrus black rot fungi be referred to as A. alternata. 相似文献
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Nobuya Tashiro Seiji Uematsu Youichi Ide Masafumi Matsuzaki 《Journal of General Plant Pathology》2012,78(3):233-236
A fruit rot, similar to brown rot, occurred on extremely early ripening Satsumas in Saga Prefecture in the early autumn of 1999. A single species of Phytophthora was isolated from the affected fruit. After nonwounding inoculation of healthy fruits of Citrus spp. including Satsuma with the isolated fungus, the fungus was reisolated from fruit with symptoms similar to those in nature. On the basis of its morphology and molecular analysis of the rDNA-internal transcribed spacer regions, the pathogen was identified as P. palmivora. This is the first report of P. palmivora as a causal pathogen of citrus brown rot in Japan. 相似文献
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ABSTRACT Epidemics of citrus brown rot from 1994 to 1997 in the south-central and east-coast citrus areas of Florida were characterized and the causal Phytophthora spp. identified. Two species of Phytophthora, P. palmivora and P. nicotianae, were consistently associated with brown rot. Epidemics caused by P. palmivora appeared to be initiated on immature fruit dropped on the orchard floor. The soilborne fungus infected and sporulated on these fruit and was then disseminated to fruit above 1 m in the canopy. In contrast, infection by P. nicotianae, the common cause of root rot, was confined to the lowest 1 m of the canopy. Fruit infected by P. palmivora produced large amounts of ellipsoidal sporangia available for splash dispersal, whereas those infected by P. nicotianae produced far fewer spherical sporangia. Isolates from brown rot epidemics were compared with P. nicotianae from citrus in Florida and Texas, P. citrophthora in California, P. palmivora, and selected Phytophthora spp. from other hosts. Brown rot symptoms produced by the different pathogenic citrus isolates on inoculated fruit were indistinguishable. Morphology, mating behavior, and isozyme patterns of brown rot isolates from 1988 to 1997 matched P. palmivora from citrus roots, other host plants, and other locations, but were different from characterized isolates of P. citrophthora in California and P. nicotianae in Florida and Texas. Cellulose acetate electrophoresis of the isozyme glucose-6-phosphate isomerase rapidly identified the causal citrus pathogen from infected fruit and soil isolation plates. Although P. palmivora is an aggressive pathogen of citrus roots, bark, and fruit, populations in orchard soils were low compared with P. nicotianae. 相似文献
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近些年链格孢属真菌的分类有了长足的进展,但柑橘褐斑病和黑腐病的病原却还是存在着一些争议。本研究从中国柑橘果实上收集了49株黑腐病菌菌株,并选取了8株从柑橘叶片上分离的具代表性的褐斑病菌菌株,基于Alta1 、endoPG、LSU、OPA10-2、OPAl-3和OPA2-1 等6个基因位点串联序列构建系统发育树。结果表明,柑橘黑腐病和褐斑病都可由不止一种链格孢菌引起,均以Alternaria alternata为主。两种病害的病原菌之间不能通过该系统发育树区分,但在致病性上存在差异,且能通过扩增ACT毒素合成基因进行区分。为了使两种病害的病原更加方便阐述,作者建议以它们的主要类群对其进行命名,柑橘褐斑病病原学名还是遵循前人的称呼,为the tangerine pathotype of A. alternata,即链格孢菌橘致病型,而柑橘黑腐病病原应为A. alternata,即链格孢菌。 相似文献
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中国柑橘黑腐病和褐斑病病原菌的系统发育分析 总被引：4，自引：0，他引：4
近些年链格孢属真菌的分类有了长足的进展,但柑橘褐斑病和黑腐病的病原却还是存在着一些争议。本研究从中国柑橘果实上收集了49株黑腐病菌菌株,并选取了8株从柑橘叶片上分离的具代表性的褐斑病菌菌株,基于Alta1 、endoPG、LSU、OPA10-2、OPAl-3和OPA2-1 等6个基因位点串联序列构建系统发育树。结果表明,柑橘黑腐病和褐斑病都可由不止一种链格孢菌引起,均以Alternaria alternata为主。两种病害的病原菌之间不能通过该系统发育树区分,但在致病性上存在差异,且能通过扩增ACT毒素合成基因进行区分。为了使两种病害的病原更加方便阐述,作者建议以它们的主要类群对其进行命名,柑橘褐斑病病原学名还是遵循前人的称呼,为the tangerine pathotype of A. alternata,即链格孢菌橘致病型,而柑橘黑腐病病原应为A. alternata,即链格孢菌。 相似文献
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Macarisin D Cohen L Eick A Rafael G Belausov E Wisniewski M Droby S 《Phytopathology》2007,97(11):1491-1500
ABSTRACT During the infection of citrus fruit by Penicillium digitatum there is little evidence of a host defense response. This suggests that P. digitatum has the ability to suppress host defenses. The current study demonstrates that P. digitatum suppresses a defense-related hydrogen peroxide (H(2)O(2)) burst in host tissue. In contrast, the nonhost pathogen, Penicillium expansum, triggers production of a significant amount of H(2)O(2) in citrus fruit exocarp. Using laser scanning confocal microscopy, we demonstrated that P. digitatum suppressed an elevation in H(2)O(2) up to 42 h after inoculation. Nevertheless, H(2)O(2) levels around wounds inoculated with P. expansum increased by 63-fold above the control. P. digitatum continued to suppress H(2)O(2) production in citrus fruit exocarp up to 66 h postinoculation and H(2)O(2) levels were actually threefold below that of noninoculated controls. In contrast, the H(2)O(2) level was still about 11-fold above the control value in wound sites inoculated with P. expansum. Studies on the effect of organic acids (as pH modulators) on the response of citrus fruit to compatible and noncompatible pathogens indicated that pathogenicity was enhanced only when host-tissue acidification was accompanied by the suppression of H(2)O(2). Additionally, pathogenicity of both P. digitatum and P. expansum on citrus fruit was significantly enhanced by the H(2)O(2)-scavenging enzyme catalase. Based on our study and previous reports regarding the potential involvement of citric acid and catalase in green mold pathogenesis, we suggest that these compounds are strongly associated with the virulence of P. digitatum. 相似文献
12.
柑桔溃疡病菌PCR快速检验检疫技术研究 总被引：11，自引：0，他引：11
柑桔溃疡病是严重影响全世界柑桔生产的重大检疫性病害,根据柑桔溃疡病菌(Xanthomonas axonopodis pv. citri)新近公布的全基因组中独有的保守蛋白基因序列,设计筛选出一对种特异性引物(JYF5/JYR5),能专一地扩增检出柑桔组织表面所带溃疡病菌的DNA靶带(413 bp)。而柑桔叶面附生的非致病性黄单胞菌、野油菜黄单胞菌近缘种以及健康柑桔样品都不能扩增;靶细菌DNA检测下限1.56 pg/μL,靶细菌悬浮液检测下限10 cfu/μL;在不同PCR仪及各种控温方式下都能稳定地扩增出特征性靶带。这一特异、准确的柑桔溃疡病菌PCR检验技术和研制的预包被固相化PCR检测试剂盒已开始用于我国非疫生产区建设中柑桔苗木、果实的病害检疫检验。 相似文献
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ABSTRACT Asiatic citrus canker (ACC) is a severe disease of several citrus species and hybrids in many tropical and subtropical areas. Populations of Xanthomonas axonopodis pv. citri in leaf and twig lesions are the most important inoculum source for secondary infections. In areas with a marked winter season (e.g., Argentina and Japan), low temperatures induce a decrease of 10(2) to 10(4) in population sizes in lesions, thus creating a discontinuity in the X. axonopodis pv. citri life cycle. The purpose of this study was to evaluate the dynamics of X. axonopodis pv. citri populations in leaf lesions exposed to the mild winter temperatures prevailing in a tropical environment. Internal X. axonopodis pv. citri population levels in Mexican lime leaf lesions reached 10(6) to 10(7) CFU lesion(-1) whatever the lesion size. These densities, however, were not strongly negatively affected by winter temperatures prevailing under experimental conditions. The estimated decrease in internal X. axonopodis pv. citri population sizes was approximately 10-fold. When exposed to 35 mm h(-1) of simulated rainfall, internal population sizes decreased over time by approximately 1 log unit for lesions 1 and 2 months old, but did not for older lesions. A microscopic examination indicated that lignin-like compounds are present in lesions more than 6 months old. The slow decrease over time of X. axonopodis pv. citri population sizes in leaf lesions may be the balanced result of defense reactions by the host at late stages of disease development, and the concomitant multiplication of the pathogen at the margin of old lesions. We conclude that the epidemiological significance of overwintered leaf lesions in the tropics is higher than that reported in other areas. 相似文献
16.
Thomas Thomidis 《Phytoparasitica》2001,29(1):47-49
The relative virulence ofPhytophthora cactorum andP. syringae originating from almond trees, and ofP. citrophthora originating from citrus, to apple, pear, peach, cherry and plum rootstocks, was studiedin vivo andin vitro. Results of the different experiments were in good agreement. All testedPhytophthora isolates showed little virulence to pear rootstocks-causing only minor crown rot symptoms - and no virulence at all to apple
rootstocks. In contrast, they were highly virulent to stone fruit rootstocks, causing crown rot disease. The non-pathogenicity
of these isolates to pome rootstocks could be interpreted as strict host specificity. 相似文献
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Sour rot, green mold and blue mold are postharvest diseases of citrus fruit. Benzimidazole and imidazole fungicides control
molds, and guazatine controls sour rot, but strains ofPenicillium spp. resistant to these chemicals have been reported. Aqueous formulations of fenpropimorph, a morpholine fungicide, are
very active in controlling fungi attacking postharvest citrus fruits. The fungicidal effect of the chemical in the coating
wax was investigated in combined treatments with guazatine in dip or spray applications on various citrus fruit varieties.
Fenpropimorph alone was as good as or a better curative and protective treatment than the current commercial treatments applied
in packinghouses in Israel for decay control. 相似文献
19.
F. Neri L. Cappellin E. Aprea F. Biasioli F. Gasperi A. Spadoni I. Cameldi A. Folchi E. Baraldi 《Plant pathology》2019,68(8):1508-1524
Neofabraea vagabunda, causing bull’s eye rot, produces notable loss during cold storage of apples growing in cool humid regions. The infection initiates in the orchard, but the pathogen lives quiescently in fruit for some months before causing the symptoms of the disease. In vivo and in vitro assays were carried out to gain knowledge on the influence of fruit volatile organic compounds (VOCs) in N. vagabunda development and define volatile markers for pathogen detection, using SPME/GC-MS, PTR-ToF-MS analysis and light microscopic observations. This study reports that: (i) the main VOCs of Cripps Pink apple (highly susceptible to bull’s eye rot) are degraded during the conidial germination of N. vagabunda, stimulating pathogen hyphal growth towards the host; (ii) first disease symptoms appear when fruit releases VOCs related to senescence, which also stimulate pathogen hyphal growth; (iii) VOCs typical of ripe-senescent fruit are also emitted by infected fruit during N. vagabunda quiescence, and methanol and ethanol are the earliest markers of bull’s eye rot; and (iv) the in vitro volatile metabolism of Botrytis cinerea, Penicillium expansum and Colletotrichum fioriniae has similarities with that of N. vagabunda, but the volatile profile of each pathogen is distinguishable. Overall, this study provides novel knowledge on fruit–fungus interaction and insights for the development of tools for early disease detection in packing houses. 相似文献
20.
ABSTRACT Greasy spot, caused by Mycosphaerella citri, is a serious disease of citrus in the Caribbean basin. M. citri is a loculoascomycete and produces pseudothecia in decomposing leaves after intermittent wetting and drying. A new in vitro mating technique was developed for production of pseudothecia on sterilized leaf disks in petri dishes. Of the single-ascospore cultures that were recovered from individual asci, four were one mating type and four were a second mating type (tentatively designated mat+ and mat-), indicating that M. citri probably is heterothallic and bipolar like most other loculoascomycetes. Most populations of ascospores recovered from individual leaves or from leaves from groves of different citrus species and various locations had a 1:1 ratio of mating types consistent with random mating. Cytological studies demonstrated that the ontogeny of pseudothecial development was similar to other loculoascomycetes. The formation of mature pseudothecia required 30 to 45 cycles of wetting and drying of infected, dead leaves which required approximately 60 to 90 days. The in vitro system for pseudothecial production and the knowledge of the mating system in M. citri will facilitate genetic studies of this important pathogen. 相似文献