Agarwal Milee, Shrivastava Neeta, Padh Harish
B. V. Patel Pharmaceutical Education Research and Development Centre, Thaltej-Gandhinagar Highway, Ahmadabad, 380054, India.
Plant Cell Rep. 2008 Apr;27(4):617-31. doi: 10.1007/s00299-008-0507-z. Epub 2008 Feb 2.
Detection and analysis of genetic variation can help us to understand the molecular basis of various biological phenomena in plants. Since the entire plant kingdom cannot be covered under sequencing projects, molecular markers and their correlation to phenotypes provide us with requisite landmarks for elucidation of genetic variation. Genetic or DNA based marker techniques such as RFLP (restriction fragment length polymorphism), RAPD (random amplified polymorphic DNA), SSR (simple sequence repeats) and AFLP (amplified fragment length polymorphism) are routinely being used in ecological, evolutionary, taxonomical, phylogenic and genetic studies of plant sciences. These techniques are well established and their advantages as well as limitations have been realized. In recent years, a new class of advanced techniques has emerged, primarily derived from combination of earlier basic techniques. Advanced marker techniques tend to amalgamate advantageous features of several basic techniques. The newer methods also incorporate modifications in the methodology of basic techniques to increase the sensitivity and resolution to detect genetic discontinuity and distinctiveness. The advanced marker techniques also utilize newer class of DNA elements such as retrotransposons, mitochondrial and chloroplast based microsatellites, thereby revealing genetic variation through increased genome coverage. Techniques such as RAPD and AFLP are also being applied to cDNA-based templates to study patterns of gene expression and uncover the genetic basis of biological responses. The review details account of techniques used in identification of markers and their applicability in plant sciences.
基因变异的检测与分析有助于我们理解植物各种生物学现象的分子基础。由于测序项目无法涵盖整个植物界，分子标记及其与表型的相关性为我们阐明基因变异提供了必要的标识。基于遗传或DNA的标记技术，如限制性片段长度多态性（RFLP）、随机扩增多态性DNA（RAPD）、简单序列重复（SSR）和扩增片段长度多态性（AFLP），经常用于植物科学的生态、进化、分类、系统发育和遗传学研究。这些技术已经成熟，其优点和局限性也已为人所知。近年来，出现了一类新的先进技术，主要是早期基本技术的组合。先进的标记技术倾向于融合几种基本技术的优势特征。新方法还对基本技术的方法进行了改进，以提高检测遗传间断性和独特性的灵敏度和分辨率。先进的标记技术还利用了一类新的DNA元件，如反转录转座子、基于线粒体和叶绿体的微卫星，从而通过增加基因组覆盖范围来揭示基因变异。RAPD和AFLP等技术也被应用于基于cDNA的模板，以研究基因表达模式并揭示生物反应的遗传基础。本文综述了用于鉴定标记的技术及其在植物科学中的适用性。
