Recombinase Polymerase Amplification assay for rapid detection of a geminivirus associated with potato apical leaf curl disease

Journal of Plant Diseases and Protection - Recombinase Polymerase Amplification (RPA) assay was developed for specific detection of Tomato leaf curl New Delhi virus-potato (ToLCNDV [potato]),...     

Development of simplified and rapid nucleic acid release protocol for PCR based detection of <Emphasis Type="Italic">Potato viruses</Emphasis>

A simple, cost-effective and rapid viral nucleic acid release (NAR) buffer suitable for RT-PCR based diagnostic assay was developed for the detection of potato viruses. The NAR buffer and commercially available RNA isolation kit were compared for RT-PCR based assay, where an amplicon of expected size (~380 bp) targeting PVY was observed in both isolations indicating that it can be used in RT-PCR based diagnostic assays. The same was further validated for its repeatability by running across more than hundred suspected potato leaf samples collected from different sources where, it showed consistent results for the presence of PVY indicating its reliability. The NAR buffer assay was examined for its sensitivity in comparison with the kit based isolation where both the assays were able to detect even up to 10^?5 dilution without affecting the sensitivity. NAR buffer was found stable up to 28 days at -20 °C and for 14 days at 4 °C without losing PCR sensitivity. The assay was also found effective to release the nucleic acid from potato leaves, thrips and aphids for PCR and RT-PCR based detection of DNA viruses like ToLCNDV-potato and other RNA viruses. The developed protocol is simple, less laborious, time-saving (10-15 min) and economical (1/100th of kit) as compared to kit based protocol. The assay can be adopted in diagnostic laboratories for detection of RNA/DNA viruses from potato plants and in thrips as well.     

Conversion of Potato Starch and Peel Waste to High Value Nanocrystals

The present study aimed to convert starch and potato peel waste to nanocrystals. Starch nanocrystals were prepared using two methodologies: direct acid hydrolysis and enzyme pretreatment followed by acid hydrolysis. Direct hydrolysis broke down the starch granules to nanocrystals in 12 days. Enzyme pretreatment with starch hydrolytic enzymes (α-amylase and amyloglucosidase) reduced the time for preparation of starch nanocrystals by 6 days. Starch nanocrystals of optimum size were obtained with both the treatments and the resultant size ranged from 10 to 50 nm. Nanocrystals were disk-like platelets in appearance. Cellulose nanocrystals were derived from cellulosic material in the potato peel. Cellulose was isolated from peel waste with alkali treatment. Further, cellulose nanocrystals from potato peel and cellulose microcrystalline were prepared by acid hydrolysis. Microscopic images revealed that the aqueous suspension of cellulose nanocrystals derived from potato peel were single rod shaped, whereas those derived from cellulose microcrystalline were rod-like nanoparticles, agglomerated in the form of bundles including some of the rods in single units (well separated). The size of potato peel nanocrystals ranged from 40 to 100 nm (length) and cellulose microcrystalline ranged from 4 to 20 nm (diameter) by 110 to 250, given 4 to 20 nm (length), respectively. As starch nanocrystals as well as cellulose nanocrystals are derived from biopolymer, both can be considered safe for humans and the environment. Moreover, the biodegradable nature of these nanocrystals makes them superior over metallic nanoparticles, particularly in the field of nanocomposites.