Molecular Cloning of MYMV Genome and Infectivity of Yellow Mosaic Virus in Green Gram Using Different Viral Transmission Tools

Ashwini Talakayala^1,2 , Veerapaneni Bindu Prathyusha¹ , Dhanasekar Divya ¹ , Srinivas Ankanagari²  and Mallikarjuna Garladinne^1*

¹Plant Molecular Biology laboratory, Agri Biotech Foundation, Rajendranagar, Hyderabad-500 030.

²Department of Genetics, Osmania University, Hyderabad-500 007.

Corresponding Author E-mail: galradinnemarjun@gmail.com

DOI : http://dx.doi.org/10.13005/bbra/2932

ABSTRACT: Mungbean yellow mosaic virus (MYMV) causes massive crop losses in green gram. MYMV is a member of begomovirus with bipartite genome comprising DNA-A and DNA-B components, which is transmitted by whiteflies. Cloning and preparation of infectious clone is very much essential for screening germplasm or transgenic material of pulse crops since viruliferous whiteflies may not be available throughout the year. In the current work, we have amplified rolling circle mediated viral genome of MYMV using Φ29 DNA polymerase. The amplified products was digested and cloned into the plant expression vector pCAMBIA2301.The cloned constructs was then transformed into Agrobacterium LBA4404 through freeze thaw method. Further, three viral transmission techniques including mechanical rubbing, Agroinfiltration and Agroinoculation, were employed for assessing the mosaic symptoms in green gram. The molecular confirmation through polymerase chain reaction (PCR) indicated that the yellow mosaic symptoms were formed due to infectivity of MYMV in the green gram.

KEYWORDS: Agroinfiltration; Agroinoculation; Φ29 DNA polymerase; Yellow mosaic disease

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