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.
BackgroundEnset (Ensete ventricosum (Welw.) Cheesman; Musaceae) is a multipurpose drought-tolerant food security crop with high conservation and improvement concern in Ethiopia, where it supplements the human calorie requirements of around 20 million people. The crop also has an enormous potential in other regions of Sub-Saharan Africa, where it is known only as a wild plant. Despite its potential, genetic and genomic studies supporting breeding programs and conservation efforts are very limited. Molecular methods would substantially improve current conventional approaches. Here we report the development of the first set of SSR markers from enset, their cross-transferability to Musa spp., and their application in genetic diversity, relationship and structure assessments in wild and cultivated enset germplasm.ResultsSSR markers specific to E. ventricosum were developed through pyrosequencing of an enriched genomic library. Primer pairs were designed for 217 microsatellites with a repeat size > 20 bp from 900 candidates. Primers were validated in parallel by in silico and in vitro PCR approaches. A total of 67 primer pairs successfully amplified specific loci and 59 showed polymorphism. A subset of 34 polymorphic SSR markers were used to study 70 both wild and cultivated enset accessions. A large number of alleles were detected along with a moderate to high level of genetic diversity. AMOVA revealed that intra-population allelic variations contributed more to genetic diversity than inter-population variations. UPGMA based phylogenetic analysis and Discriminant Analysis of Principal Components show that wild enset is clearly separated from cultivated enset and is more closely related to the out-group Musa spp. No cluster pattern associated with the geographical regions, where this crop is grown, was observed for enset landraces. Our results reaffirm the long tradition of extensive seed-sucker exchange between enset cultivating communities in Southern Ethiopia.ConclusionThe first set of genomic SSR markers were developed in enset. A large proportion of these markers were polymorphic and some were also transferable to related species of the genus Musa. This study demonstrated the usefulness of the markers in assessing genetic diversity and structure in enset germplasm, and provides potentially useful information for developing conservation and breeding strategies in enset.Electronic supplementary materialThe online version of this article (doi:10.1186/s12863-015-0250-8) contains supplementary material, which is available to authorized users.
Three rates of phosphate (0, 25, and 60 kg/ha P 2 O 5 ) were applied to phosphorus-deficient native grassland at Tel Hadya, in northern Syria, and biomass productivity, botanical composition and number of legume seeds in the soil were monitored for five seasons (1984/85-1988/89). The experiment was grazed at low (0-8 sheep/ha per year) and high (1-7 sheep/ha per year) stocking rates from the second to the fourth seasons of the experiment; in the fifth season, the low and high stocking rates were increased to 11 and 2-3 sheep/ha per year, respectively. The experimental site was typical of native grassland within the cereal zone of west Asia, where cropping is not possible because of shallow, stony soils and steep slopes.The results showed that annual applications of phosphorus, even as low as 25 kg P 2 O 6 /ha, alleviated the deficiency in soil P and resulted in improved pasture production, even in dry years. Legume production showed the greatest response to P, increasing by 0-3-3 times the production of the control treatments. By the fifth season, legume seed mass had increased threefold and number of seeds sixfold in the P-treated plots, compared with the first season, while in the control plots there was little change. Rain-use efficiency on the P-treated plots was more than double that of the controls by the fourth and fifth seasons.Practical application of the results depends on whether (i) legumes are as frequent in native grasslands, as a whole, as they are at Tel Hadya, (ii) the P deficiency observed at Tel Hadya is widespread, and (iii) grazing of communally owned grasslands can be controlled. It is suggested that all three criteria will often be fulfilled and, therefore, that grassland productivity in west Asia could be substantially increased. Furthermore, the results suggest that above-ground cover and soil organic matter will also increase after P application, both of which will help to reduce soil erosion and thereby increase the sustainability of livestock production in west Asia.
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