19. In-vitro Analysis of Chenopodium murale extract for resistance against potato virus Y
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Abstract
Potato (Solanum tuberosum) is one of the largely consumed staple food crop of the world, including Pakistan. In Pakistan, potato shares a large percentage in country’s agriculture GDP. In Pakistan majority of good quality virus free seed potato is imported by multinational and local seed companies. This costs a huge burden on country’s economy. Potato Virus Y (PVY) belongs to Potyviridae family, severely damaging leaves of the Solanaceae family including potato. Plant lectins are the potential proteins that specifically bound and cross linked with carbohydrates and target glycans present on the cell surface of viruses through a pathway termed as lectin activation. The present in-vitro study shows the effect of lectin extracted from the seeds of Chenopodium murale against the downregulation of the CP (coat protein) gene of PVY. Stable interaction was found between PVY coat protein and CML in-silico. Full length CP gene from PVY virus was amplified and cloned in mammalian expression vector. CML protein was purified to sub fractions through anion exchange chromatography on SP-sepharose gel. The non-cytotoxic effect and safe limit of CML extract was indicated by CC50 value. The hemagglutination activity of CML against the rabbit erythrocytes was revealed to be 35 µg/ml. The real-time PCR revealed the anti-PVY activity of CML extract. CML extract in concentration of 30, 60 and 90 µg/ml was found effective against PVY CP gene in comparison with control.
Keywords: Anion Chromatography; Coat Protein gene; Haemaglutination assay; HepG2 cancer cells; Lectins extract; Potato Virus Y (PVY); SDS-PAGE