不同辣椒炭疽病菌对唑菌酯的敏感性差异

辣椒炭疽病是辣椒生产上的重要病害,严重制约辣椒生产,唑菌酯防治辣椒炭疽病的相关研究未见报道。为此,本研究利用菌丝生长速率法测定了4种不同炭疽病菌对唑菌酯的敏感性,并对其作用靶标基因细胞色素b基因(Cytb)进行比较分析。结果表明:唑菌酯对Colletotrichum brevisporum YYGXZ07,C.truncatum CZHP03,C.acutatum HHBY48和C.gloeosporioides CSLL11的EC50分别为0.098、3.363、10.156和31.982μg/mL,而且供试菌株在含药培养基上的生长速率排序为C.truncatum CZHP03C.brevisporum YYGXZ07C.acutatum HHBY48C.gloeosporioides CSLL11;不同菌株的作用靶标基因细胞色素b氨基酸比对分析发现C.acutatum HHBY48、C.truncatum CZHP03和C.brevisporum YYGXZ07与C.gloeosporioides CSLL11的Cytb氨基酸序列同源性分别为82.98%、88.30%和87.77%,推测6个氨基酸位点可能与菌株对唑菌酯的敏感性存在关联。     

Characterization and pathogenicity of Colletotrichum species associated with anthracnose on chilli (Capsicum spp.) in Thailand

Fungal isolates from chilli ( Capsicum spp.) fruits in Thailand that showed typical anthracnose symptoms were identified as Colletotrichum acutatum , C . capsici and C . gloeosporioides . Phylogenetic analyses from DNA sequence data of ITS rDNA and β-tubulin ( tub 2) gene regions revealed three major clusters representing these three species. Among the morphological characters examined, colony growth rate and conidium shape in culture were directly correlated with the phylogenetic groupings. Comparison with isolates of C . gloeosporioides from mango and C . acutatum from strawberry showed that host was not important for phylogenetic grouping. Pathogenicity tests validated that all three species isolated from chilli were causal agents for chilli anthracnose when inoculated onto fruits of the susceptible Thai elite cultivar Capsicum annuum cv. Bangchang. Cross-infection potential was shown by C . acutatum isolates originating from strawberry, which produced anthracnose on Bangchang. Interestingly, only C . acutatum isolates from chilli were able to infect and produce anthracnose on PBC 932, a resistant genotype of Capsicum chinense . This result has important implications for Thai chilli breeding programmes in which PBC 932 is being hybridized with Bangchang to incorporate anthracnose resistance into chilli cultivars.     

Defence responses of pepper (Capsicum annuum L.) infected with incompatible and compatible strains of Phytophthora capsici

In this study, we investigated the activities of β-1,3-glucanase and peroxidase enzymes in the leaves of pepper cultivar A3 infected with the incompatible strain PC and the compatible strain HX-9 of Phytophthora capsici. The activities of β-1,3-glucanase and peroxidase enzymes substantially increased in the incompatible interactions compared to the compatible interactions. We also analysed the expression patterns of four defence-related genes, including CABPR1, CABGLU, CAPO1 and CaRGA1, in the leaves and roots of pepper inoculated with different strains of P. capsici. All gene expression levels were higher in the leaves than in the roots. Markedly different expression patterns were observed between incompatible and compatible host-pathogen interactions. In the incompatible interactions, the expression levels of CABPR1, CABGLU and CAPO1 genes in leaves increased by a maximum of 17.2-, 13.2- and 20.5-fold at 24, 12 and 12 h, respectively, whereas the CaRGA1 gene expression level increased to a lesser degree, 6.0-fold at 24 h. However, in the compatible interactions, the expression levels of the four defence-related genes increased by a maximum of 11.2-, 8.6-, 7.9- and 2.0-fold at 48, 24, 48 and 72 h, respectively. Compared to the leaves, the expression levels of the four defence-related genes were much lower in the roots. The highest levels of mRNA were those of the CABPR1 gene, which increased 5.1-fold at 24 h in the incompatible and 3.2-fold at 48 h in the compatible interactions. The other three genes exhibited lower expression levels in the incompatible and compatible interactions. These results further confirmed that defence-related genes might be involved in the defence response of pepper to P. capsici attack.     

Development of conventional and real-time PCR protocols for specific and sensitive detection of Colletotrichum capsici in chilli (Capsicum annuum L.)

Anthracnose, caused by Colletotrichum capsici, is a major disease of chilli (Capsicum annuum L.) affecting both fruit and seed quality. The pathogen is both internally and externally seedborne. However, a rapid and sensitive method for detection of this pathogen in seeds is currently limited. In this study, a polymerase chain reaction (PCR) method based on sequence characterized amplified region (SCAR) marker was developed for specific and sensitive detection of C. capsici in chilli seeds and fruits. The developed SCAR primers were highly specific to C. capsici and resulted in the amplification of an expected 250-bp fragment from genomic DNA of all seven of the C. capsici isolates tested. No amplification occurred when the SCAR primers were tested with genomic DNA from three other fungal isolates and four other Colletotrichum species. The SCAR primers successfully amplified similar sized fragments from DNA derived from C. capsici-infected chilli fruits. The molecular detection sensitivity of C. capsici was 1 pg of purified C. capsici DNA template and 25 ng of DNA from C. capsici-infected chilli fruits. A real-time PCR assay was also developed using SYBR Green chemistry for detection of C. capsici in chilli fruits and seeds. The standard curve obtained showed a linear correlation between copy number of the cloned target DNA sequence of C. capsici and cycle threshold (Ct) values, with R² of 0.98. These PCR-based assays may be highly useful in detection of this important pathogen in chilli seeds and fruits in plant quarantine laboratories.     

Serratia marcescens associated with bell pepper (Capsicum annuum L.) soft-rot disease under greenhouse conditions

Soft-rotting bacteria affecting bell peppers crops represent an economically destructive disease of growing importance worldwide. In Venezuela since 2006, soft-rot symptoms have been occasionally observed in bell pepper fruits grown under greenhouse conditions. Affected fruits presented water-soaked lesions that progressed to complete fruit maceration. Bacteria were isolated from water-soaked lesions in order to identify the causal agent of this disease. Of 13 bacterial isolates recovered from affected fruits, only isolate AGPim1A was able to produce a hypersensitive reaction in tobacco plants and to reproduce soft-rot symptoms in bell peppers fruits. Several methods, including classical bacteriological tests and carbon utilization profiling, alongside with sequence analysis of 16S rRNA and housekeeping genes gyrB and groES-groEL, allowed identification of the soft-rotting bacterium as Serratia marcescens. To our knowledge, this is the first report showing a S. marcescens strain associated with soft-rot disease in bell pepper fruits.     

RAPD and virulence analyses of Colletotrichum capski isolates from chilli (Capsicum annuum)

Journal of Plant Diseases and Protection - Chilli (Capsicum annuum) fruits showing symptoms of an-thracnose disease were collected from farmer’s fields in different chilli-growing areas of...     

Identification of the anthracnose fungus of walnut (Juglans spp.) and resistance evaluation through physiological responses of resistant vs. susceptible hosts

Walnut anthracnose is one of the most serious diseases of walnut globally. In this study, the pathogen was identified as Colletotrichum gloeosporioides by morphological and molecular approaches. Then, the resistance of 18 walnut genotypes was evaluated by field investigation as well as field and detached leaflet inoculation. A field inoculation assay was used as a benchmark for comparing field evaluation and the detached leaflet inoculation assay. Seven genotypes, Panhe1 and clones 71 and 200 of Juglans sigillata, Yanyuanzao and Qingxiang of J. regia, and Shujiang1 and Chuanzao1 of J. regia × J. sigillata, displayed high resistance in all assays, suggesting that they may be good candidates for further evaluation. Subsequently, two genotypes (Panhe1 with high resistance, and clone 199 with high susceptibility) were selected to analyse the different physiological responses between highly resistant and susceptible genotypes after inoculation. C. gloeosporioides infection induced the activities of superoxide dismutase (SOD), peroxidase (POD), polyphenol oxidase (PPO), and chitinase; they were all significantly increased in both Panhe1 and clone 199. However, the POD activity and its change rate in Panhe1 were significantly higher than those in clone 199. The change rates of PPO activity, and total phenol and proline content in Panhe1 were significantly higher than those in clone 199. Moreover, the change rates of SOD activity, malondialdehyde, and soluble sugar content in clone 199 were significantly higher than those in Panhe1. These findings not only enhance our understanding of the resistance of walnut to anthracnose, but also lay the foundation for breeding anthracnose-resistant walnuts in the future.     

Identification and development of molecular markers linked to Phytophthora root rot resistance in pepper (Capsicum annuum L.)

Phytopthora root rot in pepper (C. annuum) is caused by Phytophthora capsici L., which exhibits a high level of pathogenic diversity. Resistance to this disease is conditioned by a number of quantitative trait loci. Pyramiding resistance alleles is desirable and could be simplified by the use of molecular markers tightly linked to the resistance genes. The purpose of this study was development of molecular markers linked to Phytophthora root rot resistance. An F₈ recombinant inbred line (RIL) population derived from a cross between YCM334 and a susceptible cultivar ‘Tean’ was used in combination with bulk segregant analysis utilizing RAPD and conversion of AFLP markers linked to Phytophtora root rot resistance into sequence-characterized amplified region (SCAR) markers. In conversion: one marker was successfully converted into a co-dominant SCAR marker SA133_4 linked to the trait. In bulked segregant analysis (BSA): three RAPD primers (UBC484, 504, and 553) produced polymorphisms between DNA pools among 400 primers screened. Genetic linkage analysis showed that the SCAR and RAPD markers were located on chromosome 5 of pepper. Quantitative trait locus (QTL) analysis showed that the SA133_4 and UBC553 were linked to Phytophtora root rot resistance. These markers were correctly identified as resistant or susceptible in nine promising commercial pepper varieties. These markers will be beneficial for marker-assisted selection in pepper breeding.     

Molecular and Pathogenicity Characteristics of Phytophthora nicotianae Responsible for Root Necrosis and Wilting of Pepper (Capsicum annuum L.) in Tunisia

Isolates of Phytophthora from pepper, produced in Tunisia, were characterised according to molecular and pathogenicity criteria. Polymerase chain reaction amplification of the ITS1 region in the ribosomal DNA resulted in different sized fragments. The pepper isolates and P. nicotianae yielded a fragment of 310bp that distinguished it from P. capsici with a fragment of 270bp. The ribosomal RNA gene amplicons of both internal transcribed spacers and the 5.8 S of the pepper Phytophthora and P. nicotianae were digested with 8 endonucleases. The patterns generated, with the 2 enzymes that cut, were identical for both taxa. This molecular analysis corroborated the morphological and biological characteristics and suggests strongly that the isolates of Phytophthora from pepper belong to the species P. nicotianae. Inoculation of pepper, tomato, eggplant and tobacco plants with the isolates of P. nicotianae from pepper showed they were highly pathogenic on pepper but not on tobacco, while their pathogenicity was weak on tomato and eggplant and was associated with atypical symptoms not observed in the field. These pathogenicity tests suggest that pepper isolates of P. nicotianae are particularly adapted to their host and may thus constitute a forma specialis of P. nicotianae.     

Genetic diversity analysis of severely infesting invasive thrips,Thrips parvispinus (Karny) in chilli (Capsicum annuum L.) in India

Phytoparasitica - Thrips are economically important pests of crops and vectors of viral diseases worldwide. Invasion and outbreak of thrips have recently caused havoc in agricultural and...     

Effect of chitin on biological control activity of Bacillus spp. and Trichoderma harzianum against root rot disease in pepper (Capsicum annuum) plants

Two bacterial isolates and one strain of Trichoderma harzianum were tested alone and in combination with chitin for efficacy in control of root rot disease caused by Phytophthora capsici and Rhizoctonia solani in pepper plants under greenhouse conditions. These bacteria (Bacillus subtilis HS93 and B. licheniformis LS674) were isolated from repeatedly washed roots of pepper plants. In in vitro assays, HS93, LS674 and T. harzianum were antagonistic against P. capsici and R. solani and produced high levels of chitinase. Seed treatment and root drenching with bacterial suspensions of HS93 with 0.5% chitin was more effective against Phytophthora and Rhizoctonia root rot than addition of the organisms without chitin. LS674 and T. harzianum reduced Rhizoctonia but not Phytophthora root rot. In two greenhouse tests, seed treatment and root drenching with HS93 amended with chitin enhanced its biocontrol activity against P. capsici but not on R. solani. The effects of LS674 and T. harzianum against R. solani were significantly enhanced when they were used as suspensions with 0.5% chitin for root drenching, but this had no effect on P. capsici. In both greenhouse experiments, the use of 0.5% chitin alone for root drenching reduced Rhizoctonia root rot. Reduction of root rot disease was accompanied by increased yield. These results show that the antagonistic activity of HS93, LS674 and T. harzianum may be stimulated by chitin resulting in significant improvements in their effectiveness against pathogens.     

不同药剂进行土壤消毒对辣椒生长和根际土壤微生物的影响

土传病害和连作障碍是保护地蔬菜生产上的突出问题。本研究选用棉隆、二甲基二硫、多菌灵3种药剂进行土壤消毒, 通过测定土壤酶活性、微生物数量及辣椒农艺性状和产量等指标的变化, 明确不同药剂处理对辣椒生长和根际土壤微生物的影响。结果表明:3种药剂处理后土壤脲酶活性、蔗糖酶活性和过氧化氢酶活性均呈现出先抑制后激活的变化趋势; 消毒后土壤中真菌和细菌的数量均受到抑制, 随着恢复培养时间的增加, 抑制效果逐渐减弱; 3种药剂进行土壤消毒均可以增加辣椒的产量, 其中二甲基二硫处理对辣椒增产效果最为明显, 增产幅度达55.19%。     

Phylogenetic and morphological identification of Colletotrichum godetiae,a novel pathogen causing anthracnose on loquat fruits (Eriobotrya japonica)

Journal of Plant Diseases and Protection - Loquat (Eriobotrya japonica) has potential to be produced in the central part of Mexico. During summer 2017, anthracnose symptoms were observed in Tetela,...     

Phenotypic and genetic characterization of Erwinia carotovora from mulberry (Morus spp.)

Phenotypic and genetic characteristics of nine bacterial strains isolated from mulberry ( Morus spp.), which were originally described as Erwinia carotovora ssp. carotovora (Ecc), were investigated. Based on the results of biochemical tests, these bacterial strains were divided into two different types, type 1 and type 2. Two strains of type 1 were similar to Ecc, whereas seven strains of type 2 were distinct from Ecc. A polyphasic study that included serological assay, specific PCR assay for E. carotovora ssp. atroseptica (Eca), PCR-RFLP of a pectate lyase ( pel ) gene and RAPD-PCR was performed on the type 2 strains, and the data were compared with those of related E. carotovora subspecies. The results of serological and specific PCR assays for Eca showed that the type 2 strains were distinct from Eca. In RFLP analysis of the pel gene using Sau 3AI, the type 2 strains showed a unique RFLP pattern. On the basis of RAPD analysis, similarity of RAPD patterns within the type 2 strains was very high. A unique RAPD fragment was isolated from the type 2 strains and used as a probe for Southern hybridization. This probe hybridized only with PCR products from the type 2 strains. Based on phenotypic, serological and genetic characteristics, the type 2 strains isolated from mulberry may belong to a distinct E. carotovora subspecies other than Eca or Ecc.     

Morphological and molecular diversity of Colletotrichum spp. causing pepper spot and anthracnose of lychee (Litchi chinensis) in Australia

Since the 1980s a new disease has been affecting Australian lychee. Pepper spot appears as small, black superficial lesions on fruit, leaves, petioles and pedicels and is caused by Colletotrichum gloeosporioides, the same fungus that causes postharvest anthracnose of lychee fruit. The aim of this study was to determine if a new genotype of C. gloeosporioides is responsible for the pepper spot symptom. Morphological assessments, arbitrarily‐primed PCR (ap‐PCR) and DNA sequencing studies did not differentiate isolates of C. gloeosporioides from anthracnose and pepper spot lesions. The ap‐PCR identified 21 different genotypes of C. gloeosporioides, three of which were predominant. A specific genotype identified using ap‐PCR was associated with the production of the teleomorph in culture. Analysis of sequence data of ITS and β‐tubulin regions of representative isolates did not group the lychee isolates into a monophyletic clade; however, given the majority of the isolates were from one of three genotypes found using ap‐PCR, the possibility of a lychee specific group of C. gloeosporioides is discussed.     

Biochemical defense responses of black pepper (Piper nigrum L.) lines to Phytophthora capsici

A study on biochemical factors involved in black pepper defense response against Phytophthora capsici, was carried out in P. capsici susceptible (Sreekara) and resistant (04-P24, shows root resistance to the pathogen) black pepper lines. Seven important factors – change in membrane conductance, total phenols, orthodihydroxy (OD) phenols, lignin and defense related enzymes (peroxidase, β-1,3 glucanase and β-1,4 glucanase) – were studied under uninoculated and pathogen (P. capsici, isolate 06-04) inoculated condition to know the preformed and induced responses. The pathogen was inoculated (soil inoculation) and plants were observed for changes, at 24 h intervals for 10 days. On 8th day after inoculation symptoms started appearing on Sreekara and increased the severity till 10th day. Both root and stem samples were subjected for biochemical analysis. Of the factors analyzed, it was found that membrane conductance, OD phenol, lignin and peroxidase activity play significant role in root resistance to P. capsici in 04-P24. Light microscopy of the portion of root – where pathogen found attached – was also done.     

Molecular identification and characterization of ‘Candidatus Phytoplasma asteris’-related strain associated with pepper (Capsicum annuum L.) yellow crinkle disease in China

Journal of General Plant Pathology - Pepper yellow crinkle disease was first reported in China. PCR amplification using primer pairs specific for 16S rRNA, groEL, tuf or secA gene of phytoplasma...