Tapia-Tussell Raul, Quijano-Ramayo Andres, Cortes-Velazquez Alberto, Lappe Patricia, Larque-Saavedra Alfonso, Perez-Brito Daisy
Laboratorio GeMBio, Centro de Investigación Científica de Yucatán, Calle 43 # 130, Col. Chuburná de Hidalgo, Merida, Yucatan 97200, Mexico.
Mol Biotechnol. 2008 Nov;40(3):293-8. doi: 10.1007/s12033-008-9093-0. Epub 2008 Aug 1.
Colletotrichum gloeosporioides is the common causal agent of anthracnose in papaya (Carica papaya L.) fruits, and infection by this fungal pathogen results in severe post-harvest losses. In the Yucatán peninsula (Mexico) a different Colletotrichum species was isolated from papaya fruits with atypical anthracnose lesions. The DNAs from a variety of Colletotrichum isolates producing typical and atypical lesions, respectively, were amplified by PCR with C.gloeosporioides-specific primers. All isolates from typical anthracnose lesions yielded a 450 bp PCR product, but DNAs from isolates with atypical lesions failed to produce an amplification product. For further characterization, the rDNA 5.8S-ITS region was amplified by PCR and processed for sequencing and RFLP analysis, respectively, to verify the identity of the papaya anthracnose pathogens. The results revealed unequivocally the existence of two Colletotrichum species causing anthracnose lesions on papaya fruits: C. gloeosporioides and C. capsici. PCR-RFLP using the restriction endonuclease MspI reliably reproduced restriction patterns specific for C. capsici or C. gloeosporioides. The generation of RFLP patterns by MspI (or AluI or RsaI) is a rapid, accurate, and unequivocal method for the detection and differentiation of these two Colletotrichum species.
胶孢炭疽菌是番木瓜(Carica papaya L.)果实炭疽病的常见病原菌,这种真菌病原体的感染会导致严重的采后损失。在尤卡坦半岛(墨西哥),从具有非典型炭疽病病斑的番木瓜果实中分离出了一种不同的炭疽菌物种。分别用胶孢炭疽菌特异性引物通过PCR扩增来自产生典型和非典型病斑的各种炭疽菌分离株的DNA。所有来自典型炭疽病病斑的分离株都产生了一个450 bp的PCR产物,但来自具有非典型病斑的分离株的DNA未能产生扩增产物。为了进一步鉴定,通过PCR扩增rDNA 5.8S-ITS区域,并分别进行测序和RFLP分析,以验证番木瓜炭疽病病原体的身份。结果明确揭示了在番木瓜果实上引起炭疽病病斑的两种炭疽菌物种的存在:胶孢炭疽菌和辣椒炭疽菌。使用限制性内切酶MspI的PCR-RFLP可靠地重现了辣椒炭疽菌或胶孢炭疽菌特有的限制性图谱。MspI(或AluI或RsaI)产生RFLP图谱是一种快速、准确且明确的方法,用于检测和区分这两种炭疽菌物种。