<div>Background: DNA barcoding is a relatively new method of identifying plant species using short sequences of chloroplast DNA. Although there is a large number of studies using barcoding on various plant species, there are no such studies in the genus Nelumbo.&nbsp; 
 
Methods: Three chloroplast DNA regions (rbcL, matK, trnH-psbA) were tested for their suitability as DNA barcoding regions of thirty three lotus samples which were collected in Thua Thien Hue province, Vietnam. Universal primers were used and sequenced products were analyzed using Minimum Evolution method &nbsp;in the MEGA 7.0 program. 
 
Result: We did not observe high variability in nucleotide sequences within the rbcL region (0.135%). White Nelumbo, while, the most encoding matK (8.013%) and variable trnH-psbA (with different number of repeating regions TAAAA) intergenic regions was the most useful for Nelumbo barcoding. Individual application of the studied regions did not provide the expected results. None of the regions used in the study allowed the division of white and pink lotus varieties of N. nucifera specie according to the adopted classification of the genus Nelumbo. The results confirm that the use of matK, rbcL and trnH-psbA or combine all three regions together is insufficient for DNA barcoding in white and pink lotus varieties of N. nucifera specie and better discrimination within the genus Nelumbo. Our results also indicate the necessity of using a different region. All of the new sequences have been deposited in GeneBank under the following accession numbers: rbcL (MN011708 to MN068956); matK (MN011719 to MN068978) and trnH-psbA (MN011730 to MN086252).&nbsp;</div>

<div>Evaluation of Genetic Diversity by DNA Barcoding of Local Lotus Populations from Thua Thien Hue Province</div>

<div>Background: DNA barcoding, an emerging approach, is being widely used to accurately and quickly identify species using conserved DNA sequences.&nbsp; 
 
Methods: Herein we designed seven universal primers for matK (matK1, matK2, matK3 and matK4), rbcL, psbA-trnH and ITS based on their nucleotide sequences in GenBank to amplify 40 species of leguminosae and grass forages. Sequence alignment was performed using MEGA 5.0 and haplotype and mutation sites were analyzed with DnaSP 5.10. PCR amplification efficiency on using the primers designed for psbA-trnH and ITS was relatively low, making these sequences unsuitable for DNA barcoding. Further, we optimized target fragment amplification conditions for all 40 species analyzed in this study. On purifying, sequencing and analysing amplification products, we selected 5&prime; - and 3&prime; -end conserved fragments in four marked fragments.&nbsp; 
 
Result: Sequences of each maker loci showed that there were 12, 17 and 6 haplotypes of matK1, matK2 and matK3, respectively and 13 of rbcL. Based on these haplotypes of matK1, matK2, matK3 and rbcL, we established a DNA barcoding database for 20 forage species.</div>

Establishment of a DNA Barcoding Database for Legume and Grass Species Identification

<div>Background:&nbsp;The DNA barcoding is currently an effective and widely used tool that enables rapid and accurate identification of plant species.&nbsp; 
 
Methods:&nbsp;DNA barcoding of 9 chloroplast genes (rbcL, matK, trnH-psbA, accD-psaI, ndhA, psbE-petL, Rpl32-trnL, trnW-psaJ, trnSGCU-trnGGCC) were used to provide the theoretical basis for species identification, genetic diversity analysis of lotus population collected in Thua Thien Hue province, Vietnam. Universal primers were used and sequence products were analyzed using the MEGA X program. 
 
Result:&nbsp;The results showed that high levels of haplotype diversity (Hd), ranging from 0.618-0.869 and low levels of nucleotide diversity (Pi), ranging from 0.180 &times; 10-3-3.280 &times; 10-3 base on a total of nine gene regions of chloroplast genome. The neutrality tests show an excess of rare nucleotide position variations in individuals&rsquo; white lotus and derived haplotypes recent expansion. While the evolution of the individuals in the pink lotus may have to decrease. The phylogenetic analyses indicated that combined sequences were not insufficient to make a difference to the DNA barcoding in the individual&rsquo;s lotus of the N. nucifera species this is in the study. The standardized and accurate barcode information of lotus is provided for researchers. It lays the foundation for the conservation, evaluation, innovative utilization and protection of Nelumbonaceae germplasm resources.</div>

Research on Phylogenetic Relationship of Lotus Populations Collected in Thua Thien Hue Province, Vietnam based on the Chloroplast Genome by DNA Barcode