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DNA barcoding as a valuable molecular tool for the certification of planting materials in bamboo.

ABSTRACT: DNA barcodes developed for selected commercially important bamboo species can be utilized for the certification of planting stock in bamboo nurseries in absence of discriminatory features at the juvenile stage. Planting materials such as micropropagated plantlets, rhizome transplants and culm cuttings, generated at nursery level are directly procured for establishment of commercial plantations without any further verification. Very often misidentification and mixing up occur at nursery level and the error is not discovered until several years have passed. The present study evaluated the potentiality of seven Consortium for Barcode of Life (CBOL) recommended standard DNA barcode regions in commercially important bamboo species of India. Among the analyzed barcode regions, multiple sequence alignment (MSA) of psbA-trnH barcode region showed species-specific nucleotide differences in the studied bamboo taxa. The major nucleotide changes observed were transitions/transversions as well as insertions/deletions of nucleotides. Even though species-specific mononucleotide differences could be identified for most of the studied bamboo taxa, a small amount of sequence similarities were found in some of the Dendrocalamus and Bambusa species, which were grouped together in tree-based analysis. In subtribe Melocanninae, Ochlandra travancorica, Melocanna baccifera and M. clarkei showed unique species-specific psbA-trnH barcodes. Similarly, in the genus Oxytenanthera, unique species-specific psbA-trnH barcodes were obtained for O. monadelpha and O. parvifolia. Thus psbA-trnH barcode region generated distinct species-specific barcodes for commercial bamboo species in genera Bambusa, Dendrocalamus, Melocanna, Oxytenanthera as well as Ochlandra. Any national certification agency set up for the purpose can utilize psbA-trnH DNA barcode region to tag species identity and to establish the authenticity of multiplied planting materials in bamboos.

SUBMITTER: Dev SA

PROVIDER: S-EPMC6976193 | biostudies-literature | 2020 Feb

REPOSITORIES: biostudies-literature

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DNA barcoding as a valuable molecular tool for the certification of planting materials in bamboo.

Dev Suma Arun SA Sijimol K K Prathibha P S PS Sreekumar V B VB Muralidharan E M EM

3 Biotech 20200122 2

DNA barcodes developed for selected commercially important bamboo species can be utilized for the certification of planting stock in bamboo nurseries in absence of discriminatory features at the juvenile stage. Planting materials such as micropropagated plantlets, rhizome transplants and culm cuttings, generated at nursery level are directly procured for establishment of commercial plantations without any further verification<i>.</i> Very often misidentification and mixing up occur at nursery le ...[more]

PMID: 32015955

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Project description:BACKGROUND:While DNA barcoding is an important technology for the authentication of the botanical origins of Chinese medicines, the suitable markers for DNA barcoding of the genus Uncaria have not been reported yet. This study aims to determine suitable markers for DNA barcoding of the genus Uncaria (Gouteng). METHODS:Genomic DNA was extracted from the freshly dried leaves of Uncaria plants by a Bioteke's Plant Genomic DNA Extraction Kit. Five candidate DNA barcode sites (ITS2, rbcL, psbA-trnH, ITS, and matK) were amplified by PCR with established primers. The purified PCR products were bidirectionally sequenced with appropriate amplification primers in an ABI-PRISM3730 instrument. The candidate DNA barcodes of 257 accessions of Uncaria in GenBank were aligned by ClustalW. Sequence assembly and consensus sequence generation were performed with CodonCode Aligner 3.7.1. The identification efficiency of the candidate DNA barcodes was evaluated with BLAST and nearest distance methods. The interspecific divergence and intraspecific variation were assessed by the Kimura 2-Parameter model. Genetic distances were computed with Molecular Evolutionary Genetics Analysis 6.0. RESULTS:The accessions of the five candidate DNA barcodes from 11 of 12 species of Uncaria in China and four species from other countries were included in the analysis, while 54 of total accessions were submitted to GenBank. In a comparison of the interspecific genetic distances of the five candidate barcodes, psbA-trnH exhibited the highest interspecific divergence based on interspecific distance, theta prime, and minimum interspecific distance, followed by ITS2. The distribution of the interspecific distance of ITS2 and psbA-trnH was higher than the corresponding intraspecific distance. Additionally, psbA-trnH showed 95.9 % identification efficiency by both the BLAST and nearest distance methods regardless of species or genus level. ITS2 exhibited 92.2 % identification efficiency by the nearest distance method, but 87 % by the BLAST method. CONCLUSION:While psbA-trnH and ITS2 (used alone) were applicable barcodes for species authentication of Uncaria, psbA-trnH was a more suitable barcode for authentication of Uncaria macrophylla.

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DNA barcoding as a valuable molecular tool for the certification of planting materials in bamboo.

Publications

DNA barcoding as a valuable molecular tool for the certification of planting materials in bamboo.

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