Testing the infectivity of a Begomovirus by particle bombardment method using a gene gun

A.L Abubakar,M.M Abarshi,M.N Maruthi

doi:10.4314/njb.v35i2.8

Abstract

This study was design to identify the causal agent of Horsegram yellow mosaic disease and to investigate the pathogenicity of Horsegram yellow mosaic viruses (HgYMV) infective clones. The samples were obtained using standard method from the two main horsegram growing areas of Bangalore, Karnataka State of India. The viral DNA from horsegram plants exhibiting severe symptoms was amplified by PCR. An isolate of HgYMV1 and HgYMV2 were associated with severe symptom phenotype from HgYMV. Full-length clones of DNA-A and DNA-B genomic components were constructed and attempts were made to introduce homologous (HgYMV1/HgYMV2) combinations of DNA-A and DNA-B genomic components into Nicotiana benthamiana plants. Inoculation of linearized constructs containing full-length clones or partial head-to-tail dimers of DNA-A and DNAB genomic components resulted in the introduction of DNA-A genomic components into the host plant. However, these combinations of genomic DNA component were not detected in the inoculated plants bombarded using the gene gun. Thus, this study was unable to confirm the pathogenicity of HgYMV infective clones using N. benthamiana as model plant.Keywords: Horsegram, Begomovirus, yellow mosaic viruses, Particle bombardment, Gene gun

Highlights

Begomoviruses are widely distributed in nature and are transmitted through the vector called Bemisia tabaci
Amplification of begomovirus genome was proceeded using a pair of degenerate primers designed for the amplification of the DNA A and DNA B genomic components, Deng-A
Total DNA was extracted from infected leaf tissue and the PCR was used to amplify various Horsegram yellow mosaic viruses (HgYMV) DNA fragments using the primer sequences shown in method section

Summary

Introduction

Begomoviruses are widely distributed in nature and are transmitted through the vector called Bemisia tabaci. They are classified as monopartite with a DNA-A-like component or bipartite containing DNA-A and DNA-B (Brown et al, 2012). Horsegram (Macrotyloma uniflorum) is an pulse crop (legume) that produces seeds, is an annual native to India. It is commonly cultivated in India and serves as good source of protein for rural inhabitants (Kadam and Salunkhe, 1985; Fuller and Harvey, 2006; Fuller and Murphy, 2018)

Methods

Results

Discussion

Conclusion