Cloning and Analysis of Molecular Characteristics of Genomic DNA-A MYMIV from Long Bean Plants Affected by Yellow Mosaic Disease

• Main Author: Sapitri, Rima
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/43056
• Institution: Institut Teknologi Bandung

Mungbean Yellow Mosaic India Virus (MYMIV) is a pathogenic virus that causes yellow mosaic in legume plants (Legum Yellow mosaic disease, LYMD) which can cause economic losses. Long beans, Vigna unguiculata is one of the vegetables that is widely cultivated and consumed by people in Indonesia because it has a high protein content. In 2018, there was a reported incident of LYMD associated with long bean plants in Purwakarta and known to use superior plant seeds. Symptoms shown are the appearance of yellow spots on the middle, edges, until almost the entire leaf. The purpose of this study is to characterize the types of viruses that infect and analyze the molecular characteristics of the DNA-A virus genome. The method used is to clone DNA-A completely, the genome that plays an important role in virus replication, into the DH5? bacteria through the pJET1.2 / blunt end vector with the heatshock transformation method. The complete MYMIV DNA-A primer is designed in the coat protein region. Confirmation of successful cloning was done by colony PCR and double digest restrictions. DNA-A is read in its complete nucleotide sequence by the Sanger method by Macrogen, Inc. The sequencing results were analyzed by GenBank®, BioEdit 7.2, and SnapGene® which showed MYMIV in samples of varieties 1 and 2 having a high degree of similarity with MYMIV Brebes 3 isolates (99.21% and 99.17%). Analysis of common region sequences including TATA box, hairpin loop structure, iteron (intergenic region), AG motive, G box and sequence Nanonucleotide TAATAT ? TAC and ORF genome DNA- A were found in both samples of sequential results. The difference between the two is found at several base points in the complete sequence of the DNA-A genome.