病原菌黑曲霉LKO1核糖体核DNA (rDNA)序列的形态、分子鉴定及系统发育分析

M. George, P. Ramteke
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引用次数: 5

摘要

黑曲霉是果蔬“黑霉病”的致病菌。本研究的目的是在体外对病原菌进行形态学鉴定,并通过rDNA its区序列确定其分子身份和系统发育位置。从土壤中分离出黑曲霉LKO1,分别在PDB和PDA培养基上进行培养。对其生长特征进行了监测,并对其生殖结构进行了分析。生长3 d后,每个菌落直径为1.5 cm,之后菌落以每天1 cm的速度扩大,形成黑色的分生孢子,孢子头呈黑色,孢子头携带大量分生孢子。利用ITS-PCR (GenBank登录号MK696283)对真菌进行分子鉴定。扩增的序列与NCBI GenBank中可用的序列进行比较。该序列与其他分离株的相似性为100%。rDNA ITS区域的分离、形态鉴定和测序将有助于进一步研究该植物病原体的管理和商业利用。
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Morphology, molecular identification and phylogenetic analysis based on internal transcribed spacer (ITS) of the ribosomal nuclear DNA (rDNA) sequence of a pathogenic fungal isolate Aspergillus niger LKO1
Aspergillus niger is the causal organism of “black mold” disease of fruits and vegetables. The objective of the present investigation was to characterize the pathogen morphologically in vitro and to confirm its molecular identity and phylogenetic position by the sequence of ITS region of rDNA. A. niger LKO1 was isolated from soil and cultured in PDB and PDA media. The characteristic of growth was monitored and the reproductive structures were analyzed. After 3 days of growth, the diameter of each colony was 1.5 cm, after which the colony was extended its diameter 1 cm per day and subsequently black conidiophores were formed with black pigmented spore heads carrying numerous conidia. Fungus was also characterized using molecular methods based on ITS-PCR (GenBank accession number MK696283). The amplified sequence was compared with the available sequences in the NCBI GenBank. The sequence showed 100% similarity with other isolates of A. niger. The isolation, morphological characterization and sequencing of ITS region of rDNA will help in further research on the management of this plant pathogen and its commercial utilization.
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