Nuclear Genome Size Determination Of Christia Vespertilionis Via Flow Cytometry

Mohd Razik Midin, M. Fikri, Siti Sarah Zailani
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Abstract

Abstract Christia vespertilionis (butterfly wing plant) is an ornamental plant originated from South East Asia with reported usage in traditional medicine practice and potential as an anticancer and antitumor. This research aims to estimate the genome size of C. vespertilionis via flow cytometry (FCM) method. The research was conducted with the optimisation of nuclear suspension preparation followed by the genome size estimation. Two chopping techniques [manual chopping (MC) and BDTM Medimachine (MM)] and two lysis buffers (Otto and LBO1) were tested. Otto buffer with manual chopping was found to be the most suitable method, generated fine DNA peak with minimum debris background, and coefficient of variation (CV) value less than 3%. Five replicates of the FCM analysis were made for the genome size determination. The estimated genome size of C. vespertilionis was found to be 3.22 pg by using Glycine max cv. Polanka (2C=2.5pg) as an external reference standard. Further comparison with other Christia species was not possible due to the lack of data on genome size. The genome size data of C. vespertilionis can be useful for future morphology and genetics studies of Christia species.
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流式细胞术测定大葱属植物核基因组大小
摘要蝶翅植物是一种原产于东南亚的观赏植物,具有传统医学用途和潜在的抗癌、抗肿瘤作用。本研究旨在利用流式细胞术(FCM)方法估计大褐藻的基因组大小。本研究首先进行了核悬液制备的优化,然后进行了基因组大小的估计。测试了两种切碎技术[手动切碎(MC)和BDTM Medimachine (MM)]和两种裂解缓冲液(Otto和LBO1)。结果表明,人工斩波Otto缓冲是最合适的方法,能以最小的碎片背景生成精细的DNA峰,变异系数(CV)值小于3%。FCM分析进行了5次重复,以确定基因组大小。利用Glycine max cv,估计C. vespertilionis的基因组大小为3.22 pg。波兰卡(2C=2.5pg)作为外部参考标准。由于缺乏基因组大小的数据,无法与其他克里斯蒂亚物种进行进一步比较。该植物的基因组大小数据可为今后植物形态学和遗传学的研究提供参考。
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