Curcin isoforms in Jatropha (Jatropha curcas L.): sequence analysis, studies on amplicon profile and expression patterns in different tissues

Abstract

As of now, Jatropha (Jatropha curcas L.) stands as the best solution to the problems associated with production of biofuels, as it is well adapted to marginal soil with low nutrient content, and also suitable in different types of soil conditions, rainfall and climates. Disadvantage of Jatropha is the presence of toxic substances, particularly in the seeds, like curcin and phorbol esters which limits their applications. Curcin is the major toxin present in Jatropha. It is mainly found in the endosperm of the seeds of Jatropha. Considerable progress has been made on curcin as revealed in literature survey. Sequence analysis and comparison were made using the available curcin sequences. Apart from sequence relatedness, conserved amino acids were found and some important motifs were predicted which are presented in the comprehensive manner in this report. Hydropathy character of curcin was presented. Multiple sequence alignment was made using different forms of curcin namely curcin precursor, curcin2A, curcin-L precursor, ribosome inactivating protein (RIP) and ribosome-inactivating protein cucurmosin-like, Jatropha curcas. This exercise revealed both sequence identity and divergence between them. Good quality genomic DNA was isolated and purified from field-grown tender leaves using a simple and efficient protocol as introduced in our laboratory. PCR were carried out using curcin2A and curcin precursor cDNA-specific oligonucleotide primers. The amplicons corresponding to the individual PCR were noted in order to analyse and compare between the Jatropha accessions. RNA was isolated and purified from different tissues of plant using a simple and efficient protocol as introduced in our laboratory. RT-PCR approach was adopted to see the expression patterns of curcin2A and curcin precursor in different tissues of Jatropha. This study would be useful for further research on curcin gene in Jatropha.