Bennett Matthew S, Triemer Richard E
Michigan State University, Department of Plant Biology, 166 Plant Biology, East Lansing, Michigan 48824, USA.
J Phycol. 2012 Feb;48(1):254-60. doi: 10.1111/j.1529-8817.2011.01101.x. Epub 2012 Jan 10.
One of the foremost issues in the field of algal taxonomy is the inability to acquire, grow, and sequence new taxa. This problem is particularly true in the study of photosynthetic euglenoids where most of the distinct taxa in culture collections have been sequenced, and many other taxa of interest have been resistant to culturing, and thus, sequencing. In an effort to address this problem, we have utilized a new technique, novel to the field of taxonomy, which allows for the sequencing of nuclear genes from a very small number of cells. Through this procedure, a DNA extraction followed by a multiple displacement amplification (MDA), taxa obtained by field collection had their genomic DNA (gDNA) amplified many fold to microgram quantities. The DNA was then used as template DNA for PCR reactions, and multiple nuclear genes were amplified successfully from several different taxa. By applying this procedure, we were able to shed new light on taxa that have been historically difficult to classify, resulting in the assignment of Euglena helicoideus (C. Bernard) M. S. Benn. et Triemer and Phacus horridus (Pochm.) M. S. Benn. et Triemer to the genus Lepocinclis.
藻类分类学领域最主要的问题之一是无法获取、培养新的分类单元并对其进行测序。在光合裸藻的研究中,这个问题尤为突出,因为培养物保藏中心的大多数不同分类单元都已测序,而许多其他感兴趣的分类单元一直难以培养,因此也无法测序。为了解决这个问题,我们采用了一种分类学领域新颖的新技术,该技术能够从极少量细胞中对核基因进行测序。通过这个流程,先进行DNA提取,然后进行多重置换扩增(MDA),野外采集得到的分类单元的基因组DNA(gDNA)被扩增了许多倍,达到微克级数量。然后将该DNA用作PCR反应的模板DNA,并成功地从几个不同的分类单元中扩增出多个核基因。通过应用这个流程,我们能够对历来难以分类的分类单元有新的认识,结果将螺旋裸藻(C. Bernard)M. S. Benn. 及Triemer和粗糙扁裸藻(Pochm.)M. S. Benn. 及Triemer归入鳞孔藻属。