Development of Techniques for Detection of Latent Infection and Molecular Identification of Mango Anthracnose Fungi
Date Issued
2005
Date
2005
Author(s)
Tsai, Hui-Ling
DOI
zh-TW
Abstract
In Taiwan, Colletotrichum gloeosporioides are important plant pathogens which cause leaf spotting, fruit staining and rot on Mangifera indica (mango). The detection and diagnosis of Colletotrichum spp. are usually difficult, because of the latent infection and highly morphological variations in the culture conditions. To improve the efficiency of detection and identification of Colletotrichum, we used freezing process, chemical treatment and molecular techniques. In this study, a more safe and convenient of freezing treatment process was investigated and developed as a substitute for Paraquat and Ethephon in the detection of latent infection of Colletotrichum spp. on mango plants. Besides, we designed two primer pairs within ribosomal RNA internal transcribed spacer (ITS) to detect C. gloeosporioides by Nested polymerase chain reaction (Nested PCR). However, due to the sequence conservation of the ITS region between C. gloeosporioides and C. musae, we failed to discriminate by Nested PCR. To improve the species-specificity detection techniques, degenerate primers were designed bared on the mitochondrial cytochrome c oxidase subunitⅠ(COXⅠ) and subunit Ⅱ (COXⅡ) of Colletotrichum spp. The results indicated that an expected fragment genes in C. actatum, but not C. gloeosporioides and C. musae was amplified only. Furthermore, the sequences of the fragments from C. actatum were similar to those COXⅠand COXⅡ reported in Ascomycota. In addition, Amplified fragment length polymorphism (AFLP) was utilized to analyze the difference between C. gloeosporioides and C. musae at the molecular level. Sequence analyses of the differentially amplified fragment are needed to further develop more efficient PCR detection technique for C. gloeosporioides and C. musae.
Subjects
芒果
炭疽病
潛伏感染
內轉錄間隔區
粒線體
mango
anthracnose
latent infection
ITS
mitochondrial
Type
other
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