Abstract

This paper is an overview of the diverse, predominantly molecular techniques, used in assessing plant genetic diversity. In recent years, there has been a significant increase in the application of molecular genetic methods for assessing the conservation and use of plant genetic resources. Molecular techniques have been applied in the analysis of specific genes, as well as to increase understanding of gene action, generate genetic maps and assist in the development of gene transfer technologies. Molecular techniques have also had critical roles in studies of phylogeny and species evolution, and have been applied to increase our understanding of the distribution and extent of genetic variation within and between species. These techniques are well established and their advantages as well as limitations have been realized and described in this work. Recently, a new class of advanced techniques has emerged, primarily derived from a combination of earlier, more basic techniques. Advanced marker techniques tend to amalgamate advantageous features of several basic techniques, in order to increase the sensitivity and resolution to detect genetic discontinuity and distinctiveness. Some of the advanced marker techniques utilize newer classes 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 most important and recent advances made in molecular marker techniques are discussed in this review, along with their applications, advantages and limitations applied to plant sciences.

Highlights

  • Understanding the molecular basis of the essential biological phenomena in plants is crucial for the effective conservation, management, and efficient utilization of plant genetic resources (PGR)

  • Molecular markers represent a class of molecular tools that are sensitive to new genome-based discoveries and technical advancements and are, subject to continuous evolution

  • Most molecular marker techniques are employed in the evaluatation of genetic diversity and constructruction of genetic and physical maps

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Summary

Introduction

Understanding the molecular basis of the essential biological phenomena in plants is crucial for the effective conservation, management, and efficient utilization of plant genetic resources (PGR). Genetic or DNA based marker techniques such as Restriction Fragment Length Polymorphism (RFLP), Random Amplified Polymorphic DNA (RAPD), Simple Sequence Repeats (SSR) and Amplified Fragment Length Polymorphism (AFLP) are in common use for ecological, evolutionary, taxonomical, phylogenic and genetic studies of plant sciences These techniques are well established and their advantages and limitations have been documented [2,3,4]. The advanced marker techniques utilize newer classes of DNA elements such as retrotransposons, mitochondrial and chloroplast based microsatellites, allowing increased genome coverage Techniques such as RAPD and AFLP are being applied to cDNA-based templates (i.e., sequences of complementary DNA obtained by mRNA retrotranscription) to study patterns of gene expression and uncover the genetic basis of biological responses. The present review details the molecular techniques of genetic variability and their application to plant sciences

Molecular Assessment of Genetic Diversity
Biochemical Markers
Molecular Markers
Restriction-Hybridization Techniques
Sequence Specific PCR Based Markers
Microsatellite-Based Marker Technique
Markers Based on Other DNA than Genomic DNA
Transposable Elements-Based Molecular Markers
RNA-Based Molecular Markers
Real-Time PCR
11. New Generation of Sequencing Technology
12. Conclusions

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