Molecular characterization of large subunit ribulose-1-5-bisphosphate carboxylase/oxygenase (rbcL) gene in three napier grass(Pennisetum purpureum Shumach.) genotypes

Date

6-2016

Degree

Bachelor of Science in Agricultural Biotechnology

College

College of Agriculture and Food Science (CAFS)

Adviser/Committee Chair

Antonio C. Laurena

Restrictions

Restricted: Not available to the general public. Access is available only after consultation with author/thesis adviser and only to those bound by the confidentiality agreement.

Abstract

Genomic DNA from Napier grass (Pennisetum purpureum) cultivars Pakchong, Bana, and Common was extracted using a modified CTAB extraction method. PCR-based approaches were employed to clone and sequence the 599-bp barcoding region of the rbcL gene encoding for the large subunit of Rubisco using published gene-based primers rbcLa-F and rbcLa-R. The pGEM®-T Easy Cloning Vector System was used to clone the amplified rbcL region and were sent to AIT Biotech, Singapore for sequencing. Alignment of the rbcL sequences and corresponding amino acid sequences of the three cultivars of P. purpureum showed that the sequences are almost identical except for a few single nucleotide differences. The location of the amplified rbcL barcoding region was found to be the first 599 bases of the full length of the rbcL coding sequence. Multiple sequence alignment showed that the amplified region of the gene was still able to differentiate different plant species due to nucleotide sequence variations. However, it was not able to differentiate among the Pakchong, Bana and Common genotypes. Hence, another barcode with higher discriminatory power or a combination of barcodes may be required to differentiate the three Napier grass genotypes. The rbcL gene sequences of the three cultivars used in this study also shows 100 % homology to rbcL gene sequences of other cultivars of P. purpureum, namely Dwarf, Florida, and Malaysia. BLASTn analysis revealed that the rbcL gene region amplified is most homologous to Setaria chondrachne, an, annual robust grass, belonging to the same family (Poacea) indicating possible close evolutionary relatedness of the two species. This study is the first to clone, sequence, and characterize the barcoding region of the rbcL gene of P. purpureum cultivars Pakchong, Bana, and Common.

Language

English

Location

UPLB Main Library Special Collections Section (USCS)

Call Number

LG 993.5 2016 A127 /M36

Notes

Major in Crop Biotechnology

Document Type

Thesis

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