Specific PCR‐based detection of Phomopsis heveicola the cause of leaf blight of Coffee (Coffea arabica L.) in China. (18th December 2020)

• Record Type: Journal Article
• Authors: Lu, Y.; Wu, W.H.; He, C.P.; Liang, Y.Q.; Huang, X.; Zheng, J.L.; Xi, J.G.; Tang, S.B.; Yi, K.X.
• Is Part Of: Letters in applied microbiology. Volume 72:Number 4(2021)
• Page Start: 438
• Page End: 444
• Publication Date: 2020-12-18
• Subjects: leaf blight of Coffee -- Phomopsis heveicola -- species‐specific PCR; Microbiology -- Periodicals; 660.62
• DOI: 10.1111/lam.13396
• Languages: English
• ISSNs: 0266-8254

Abstract

Abstract: Coffee ( Coffea arabica L.) is currently grown in many tropical and subtropical areas countries and is a major traded commodity for the developing world. Coffee leaf blight, caused by Phomopsis heveicola, is one of the most important fungal diseases dangerous to coffee crops in China. This study aimed to develop a PCR‐based diagnostic method for detecting P. heveicola in planta. Specific primers (CPHF/CPHR) were designed based on sequence data of region of internal transcribed spacer (ITS1 and ITS4) of P. heveicola . The efficiency and specificity of CPHF/CPHR were established by PCR analysis of DNA from P. heveicola strains isolated from China and fungal isolates of other genera. A single amplification product of 318 bp was detected from DNA P. heveicola isolates. No amplification product was observed with any of the other fungal isolates tested. The specific primers designed and employed in PCR detected P. heveicola up to 3 pg from DNA isolated. This is the first report on the development of a species‐specific PCR assay for identification and detection of P. heveicola . Thus, the PCR‐based assay developed was very specific, rapid and sensitive tool for the detection of pathogen P. heveicola . Abstract : Significance and impact of the study: Phomopsis heveicola is an important pathogenic fungus responsible for leaf blight of coffee in China. Disease incidence ranged from 10% to 20% in the affected coffee garden, and in extreme cases, 45% of the trees were affected can be a major limiting factor for coffee production. Causes severe losses to coffee production. The conventional detection methods including culture and morphological identification are time consuming, laborious and potentially unreliable. A PCR‐based diagnostic method was developed with specific primers based on sequence data of ITS1 and ITS4 of nuclear ribosomal RNA gene repeats of P. heveicola .