Agrobacterium-mediated co-inoculation of okra plants with cloned okra enation leaf curl virus DNA and bhendi yellow vein mosaic beta-satellite DNA furthers Koch’s postulates for enation leaf curl disease

Abstract

The enation leaf curl disease (ELCuD) is one of the several viral diseases affecting the cultivation of okra (Abelmoschus esculentus L.) in the Indian subcontinent. Several begomoviruses and satellites are associated with ELCuD. However, to date, there are no reports of the re-introduction of any cloned ELCuD-associated viral DNA back into okra to cause ELCuD-like symptoms. Okra enation leaf curl virus (OELCuV) and various satellites, which includes bhendi yellow vein mosaic beta-satellite (BYVMB) have earlier been reported to be associated with ELCuD and with other okra diseases such as bhendi yellow vein mosaic disease. In this report, it is shown that agrobacterium-mediated inoculation of a cloned DNA of OELCuV and BYVMB to the shoot apex of virus-free okra plants led to symptoms resembling ELCuD. The OELCuV and the BYVMB DNAs could be PCR- amplified from the symptomatic leaves of the agro-inoculated plants. Full-length OELCuV DNA could also be amplified from the same symptomatic leaves, part of whose DNA sequence matched with that of the DNA which was inoculated. Hence, this work is an important step towards the fulfilment of Koch’s postulates for ELCuD.