Amplification and Partial Sequencing of Ribosomal Intergenic Spacer (IGS) from the Citrus Fruit Rot Pathogens Penicillium digitatum, P. ulaiense and P. italicum
Penicillium digitatum, P. italicum, and P. ulaiense are difficult to differentiate using morphological methods and the use of molecular techniques is challenged by the presence of a high number of phylogenetically related species. Moreover, these three species have almost identical Internal Transcribed Spacer (ITS) regions. The IGS regions have great potential to differentiate closely related species however their utilization has been limited mainly because of the difficulties related to their amplification and sequencing. In the present work, new universal primers were designed by comparing IGS flanking regions (28S and 18S genes) from a large number of fungi and utilised to amplify the complete IGS regions off. ulaiense, P. digitatum, and P. italicum. Amplicons were of approximately 4 000 bp for P. ulaiense and 3 500 bp for P. digitatum and P. italicum. Amplified regions were cloned with pCR?-XLTOPO? Kit and sequenced for approximately 1 000 nucleotide at both 3 and 5 sides for P. ulaiense and P. digitatum. As opposed, only a very short fragment was effectively sequenced for P. italicum. Alignment of IGS regions from P. digitatum and P. ulaiense with other GenBank available rDNA sequences and BLAST analyses evidenced very high levels of polymorphisms which are valuable for the development of specific molecular markers.
Penicillium digitatum Penicillium italicum Penicillium ulaiense IGS regions Ribosomal DNA IGS universal primers
A. Ligorio L. Schena F. Nigro K. Youssef I. Pentimone A. Ippolito
Department of Plant Protection and Applied Microbiology, University of Bari, Via G. Amendola 165 /A, Department of Management of Agricultural and Forest Systems, Mediterranean University of Reggio Cala
国际会议
11th International Citrus Congress(第11届国际柑橘大会)
武汉
英文
1373-1376
2008-10-01（万方平台首次上网日期，不代表论文的发表时间）