Aiyaz Mohammed, Divakara Shetty Thimmappa, Mudili Venkataramana, Moore Geromy George, Gupta Vijai Kumar, Yli-Mattila Tapani, Nayaka Siddaiah Chandra, Niranjana Siddapura Ramachandrappa
Department of Studies in Biotechnology, University of Mysore, Mysore-570006, India;
DRDO-BU-Centre for Life Sciences, Bharathiar University campus, Coimbatore, Tamil Nadu-640046, India.
Curr Genomics. 2016 Apr;17(2):132-44. doi: 10.2174/1389202917666151116213056.
A total of 106 maize seed samples were collected from different agro-climatic regions of India. Sixty-two Fusarium isolates were recovered, 90% of which were identified as Fusarium verticillioides based on morphological and molecular characters. Use of the tef-1α gene corrected/refined the morphological species identifications of 11 isolates, and confirmed those of the remaining isolates. Genetic diversity among the Fusarium isolates involved multilocus fingerprinting profiles by Inter Simple Sequence Repeats (ISSR) UPGMA and tef-1α gene phenetic analyses; for which, we observed no significant differences among the isolates based on geographic origin or fumonisin production; most of the subdivision related to species. Genotyping was performed on the F. verticillioides isolates, using 12 primer sets from the fumonisin pathway, to elucidate the molec-ular basis of fumonisin production or non-production. One fumonisin-negative isolate, UOMMF-16, was unable to amplify nine of the 12 fumonisin cluster genes tested. We also used the CD-ELISA method to confirm fumonisin production for our 62 Fusarium isolates. Only 15 isolates were found to be fumonisin-negative. Interestingly, genotypic characterization re-vealed six isolates with various gene deletion patterns that also tested positive for the production of fumonisins via CD-ELISA. Our findings confirm the importance of molecular studies for species delimitation, and for observing genetic and phenotypic diversity, among the Fusaria.
从印度不同农业气候区共采集了106份玉米种子样本。分离出62株镰刀菌,其中90%根据形态和分子特征被鉴定为轮枝镰孢菌。tef-1α基因的使用校正/完善了11株分离株的形态学物种鉴定,并确认了其余分离株的鉴定结果。镰刀菌分离株之间的遗传多样性涉及通过简单重复序列区间(ISSR)UPGMA和tef-1α基因聚类分析的多位点指纹图谱;为此,我们观察到基于地理来源或伏马菌素产生,分离株之间没有显著差异;大多数分类与物种有关。对轮枝镰孢菌分离株进行基因分型,使用来自伏马菌素途径的12组引物,以阐明伏马菌素产生或不产生的分子基础。一株伏马菌素阴性分离株UOMMF-16,在所测试的12个伏马菌素簇基因中有9个无法扩增。我们还使用竞争性酶联免疫吸附测定(CD-ELISA)方法来确认我们的62株镰刀菌分离株的伏马菌素产生情况。仅发现15株分离株为伏马菌素阴性。有趣的是,基因型特征揭示了6株具有不同基因缺失模式的分离株,它们通过CD-ELISA检测伏马菌素产生也呈阳性。我们的研究结果证实了分子研究对于镰刀菌物种界定以及观察其遗传和表型多样性的重要性。