Greilhuber Johann
Institute of Botany and Botanical Garden of the University of Vienna, Rennweg 14, A 1030 Vienna, Austria.
Ann Bot. 2005 Jan;95(1):91-8. doi: 10.1093/aob/mci004.
The 6 years since the last Angiosperm Genome Size Discussion Meeting in 1997 have experienced the decline of the then widely held idea of the 'plastic' genome. Several published cases of intra-specific variation in cultivated plants have been questioned on re-investigation with an improved technical approach. At the same time, technical problems caused by staining inhibitors present in the plant material have been recognized. In the accumulation of genome size data more critical methods and rules for best practice are urgently needed. INFRA-SPECIFIC VARIATION RE-VISITED: This review is about (a) the basic requirement for repeatability of results and the need for self-criticism on the part of the investigator and (b) the critical points in the technical procedure, particularly the quantitative Feulgen reaction. Case studies are presented on Dasypyrum villosum (refuting a previously reported 'plastic genome' phenomenon), on Glycine max (refuting previously claimed intraspecific variation) and on Arachis hypogaea and A. duranensis, in which reported C-values are too high by roughly two-fold. In A. hypogaea the reported intraspecific genome size variation could not be confirmed. Furthermore, a claimed negative correlation between altitude and genome size in A. duranensis was shown to be based on an arbitrary omission of data points that did not fit the correlation (although a correlation was found).
The finding of previously published questionable studies was the incentive for a re-consideration of the quantitative Feulgen procedure with regard to best practice in genome size studies. Clarification here of the critical steps of the method should help to improve the data in the literature. It must be stressed that the most important requirement is the need for a self-critical attitude of researchers to their data.
自1997年上次被子植物基因组大小研讨会以来的6年里,当时被广泛接受的“可塑性”基因组概念逐渐式微。一些已发表的栽培植物种内变异案例,经采用改进的技术方法重新调查后受到质疑。与此同时,人们认识到植物材料中存在的染色抑制剂所导致的技术问题。在基因组大小数据的积累过程中,迫切需要更严谨的方法和最佳实践规则。
本综述涉及(a)结果可重复性的基本要求以及研究者自我批评的必要性,(b)技术流程中的关键点,特别是定量福尔根反应。文中给出了关于节节麦(驳斥先前报道的“可塑性基因组”现象)、大豆(驳斥先前声称的种内变异)以及花生和直立花生的案例研究,其中报道的C值大约高出实际值两倍。在花生中,所报道的种内基因组大小变异无法得到证实。此外,有人声称直立花生的海拔与基因组大小之间存在负相关,但结果表明这是基于随意舍弃不符合该相关性的数据点得出的(尽管确实发现了一种相关性)。
对先前已发表的有问题研究的发现促使人们重新审视基因组大小研究中的最佳实践——定量福尔根程序。在此对该方法关键步骤的阐释应有助于改进文献中的数据。必须强调的是,最重要的要求是研究人员对其数据要有自我批评的态度。