Since traditional taxonomic studies possess intrinsic limitations with plant species identification, combinations of DNA barcodes have been considered a powerful tool to discover undetected genetic variation within species across large geographic areas, providing more precise estimates of biodiversity. However, the lack of efficient and universal markers is often considered a peculiar challenge in molecular taxonomic studies across plant taxa. Similarly, many loci have been proposed for DNA barcodes; still standardizing regions as a DNA barcode is vital for making them efficiently discriminate plant species. In this study, we tested the phylogenetic utility of nuclear (nrDNA) region (ITS2) with chloroplast (cpDNA) regions (matK, psbA-trnH, and rbcL) for efficient discrimination of Triticum species. A total of 109 accessions representing 16 recognized genotypes in the Triticum genus have been sampled to assess the efficiency of barcoding loci to resolve species discrimination. As expected from earlier studies, our results also revealed that a single locus has difficulty in discriminating Triticum species. Species discrimination in Triticum taxa was martially improved by using a combination of gene loci; however, the closely related species, T. aestivum and T. turgidum, had no DNA barcode to separate them. Thus, we recommend further research on finding species-specific SNP using intragenic regions as standard DNA barcode loci in Poaceae.

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Phosphate is one of the major nutrients of growth, development, and reproduction of crop plants and functions in energy metabolism, signal transduction cascades, and regulates enzymatic activities. To understand uptake and usage of this nutrient in Oryza sativa (rice), a model crop plant, global studies on this family is more effective. Here, we conducted phylogenomic analyses of 26 rice and 19 Arabidopsis phosphate transporters (PHT) reported from previous studies, by integrating various meta-expression data to the phylogenic tree context. Subsequently, of four subfamilies, the PHT1 subfamily was a high affinity phosphate transporter, which functioned under low concentrations of phosphorous in soil, while the others (i.e., PHT2, PHT3, and PHT4) were low-affinity phosphate transporter subfamilies. Most members of the PHT1 in rice and Arabidopsis, in contrast to the other transporter subfamilies, showed significant induction under phosphate starvation, and the responses were more obvious in the roots. These results indicated that the functions of PHT1 phosphate transporters in rice and Arabidopsis were well conserved in response to phosphate starvation. We confirmed significant upregulation of seven PHT1 subfamily genes in rice under phosphate starvation, by RT-PCR, indicating that the high affinity phosphate transporters played important roles in the uptake of phosphate under phosphate deficiency. The regulatory network of OsPT4 belonged to the PHT1 subfamily based on RiceNet analysis, suggesting clues for further analyses. Our study showed the significance of at least seven PHT1 subfamily members, which could improve the efficiency of phosphate use in rice, as a model crop plant.

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DNA barcoding is the use of short DNA sequences of the genome for large scale species identification. The Consortium for the Barcode of Life (CBOL) plant-working group recommended a 2-locus combination as the standard plant barcode. The evolutions of the chloroplast regions combined with nuclear gens are sufficiently rapid to allow discrimination between closely related species. We evaluated the efficacy of the proposed plant barcoding loci, matK, along with ITS2 for barcoding the Vigna species. To assess the discriminatory ability of barcoding loci for identifying the Vigna species, we sampled 52 of the taxonomically best known groups in the genus. Topologies of the phylogenetic trees based on ITS2 and matK analyses were similar but a few accessions were placed into distant phylogenetic groups. Neither ITS2 nor matK analyses were able to discriminate some closely related Vigna species. Thus, we used concatenated data to increase the resolving power of ITS2 and used matK as an additional tool for phylogenetic analysis in Vigna because characterization of the nucleotide sequences of the matK region was easier and more cost-effective than that of the ITS region.

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We investigated the species discriminatory efficiency of the proposed plant barcoding loci ITS2 and matK in Vicia species. In 2011, China Plant BOL Group proposed the addition of nuclear ITS2 to matK be accepted as a 2-locus DNA barcode to classify plant species. The matK region was chosen as a DNA barcode because of its effective species discriminating power, high quality sequence recovery, and easy experimental analysis. Integration of matK sequences into Vicia phylogeny could improve phylogenetic reconstruction of this species. To assess the ability of barcoding loci to resolve Vicia species, we sampled 36 of the taxonomically best known groups in the genus. Topologies of the phylogenetic trees based on ITS2 and matK analyses were similar but a few accessions were placed into distant phylogenetic groups. Neither ITS2 nor matK analyses alone could discriminate some closely related Vicia species. Thus, we have proposed a concatenated data approach to increase the resolving power of ITS2 and used matK as an additional tool for phylogenetic analysis in Vicia because characterization of the nucleotide sequences of the matK region was easier to recover and more cost-effective than those of the ITS region.

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