Identification and characterization of suppressor of over expression of constans1 (SOC1) gene in black pepper (Piper nigrum L.)

Abstract

The present study entitled“Identification and characterization of Suppressor of Overexpression of Constans1 gene in Black Pepper (Piper nigrum L.)”was conducted at the Integrated Biotechnology Block, College of Agriculture, Vellayani, during 2015-2016.The study envisagedisolation and sequencing ofSOC1, a flowering integrator gene in black pepper (variety - Karimunda) and functional characterization of the gene by studying the expression patterns. Degenerate primers were designed for the above said gene based on the gene sequences from NCBI database (SOC1 forward and reverse primers) which were used to isolate and identify the gene. Total RNA of black pepper was isolated using modified CTAB method followed by synthesis of cDNA using AMV RT (Avian myeloblastosis virus reverse transcriptase). PCR (Polymerase chain reaction) with degenerate primers was done using cDNA as the template. However no amplifications were observed after the first reactions. Therefore nested PCR reactions were done using the PCR products of the first reaction as the template. Two bands of size 640 bp and 330 bp were produced in the nested reactions. Sequencing of the product yielded four sequences with each of the sequence showing similarity to the SOC1 gene, when done sequence analysis, thus making it the first flowering integrator gene to be identified in black pepper. Microscopy studies were carried out to see the floral characters of black pepper in detail. Microscopy studies were done using FAA fluid as fixative, sectioning the tissues and staining with safranin and fast green were carried out to see the changes occurring in different development stages of spikes from immature spike to complete spike with berries.