Pun1 Gene Isolation from Capsicum frutescens L. cv Cakra Hijau

Elhah Nailul Khasna,Ria Reinnata Juliandari,Dwi Listyorini,Shelly Zairina,Eko Sri Sulasmi

doi:10.18502/kls.v3i4.689

Abstract

Capsicum frutescens L. is one of chili peppers with high pungency. Capsicum frutescens has several cultivars, one of those is C. frutescens cv. Cakra Hijau. This cultivars is known resistance to pests and diseases as well. Pungency is due to the accumulation of capsaicinoids. Pun1 is an important gene responsible for pungency. The full-leght genomic sequence of Pun1 is 1897 bp, containing two exons of 738 bp and 590 bpand one intron of 348 bp in between. This study was aimed to isolate Pun1 gene that free from intron. mRNA was isolated with TriReagentâ furthermore RT-PCR method used Qiagent One-Step RT-PCR and two pairs of primer : F1/R1 (F15’-ATG-GCT-TTT-GCA-TTA-CCA-TCA-3’ / R15’- CTT-AGC-TCG-AAG-TGC-ATC-TA-3’) and F2/R2 (F25’-GAA-GGT-GGC-AGA-AGA-ATC-AG-3’/R25’-TTA-GGC-AAT-GAA-CTC-AAG-GA-3’). The result of this study are isolated 738 bp exon-1 and 590 bp exon 2 of Pun1 gene.<div> Keywords: Capsaicin; Capsicum frutescens L. cv. Cakra Hijau; exon; Pun1 gene </div>

Highlights

IntroductionCapsicum frutescens L. is widely cultivated in tropical and subtropical regions of the world including Indonesia [1, 2]
Publishing services provided by Knowledge ECapsicum frutescens L. is widely cultivated in tropical and subtropical regions of the world including Indonesia [1, 2]
License, which permits unrestricted use and Capsaicinoid was first detected at 20th days after anthesis gradually increased redistribution provided that and reached maximal level around 40 d after anthesis, on the 50 d, the capsaicinoid the original author and source content decreased significantly [6].Capsaicin synthase (CS) is the last enzym in the are credited

Summary

Introduction

Capsicum frutescens L. is widely cultivated in tropical and subtropical regions of the world including Indonesia [1, 2]. Cakra Hijau is one of local cultivar from C. frutescens et al This article is distributed that have economic value, it is resistant to pest and disease as well and has high under the terms of the level of pungency [3]. License, which permits unrestricted use and Capsaicinoid was first detected at 20th days after anthesis gradually increased redistribution provided that and reached maximal level around 40 d after anthesis, on the 50 d, the capsaicinoid the original author and source content decreased significantly [6].Capsaicin synthase (CS) is the last enzym in the are credited. Of Pun gene and a 4 bp deletation in part of exon-1 which distrub translation and so deletion of part of exon-2 including the stop codon [9,10,11]. DNA isolation of Pun gene from C. frutescens cv. This study aimed to isolate coding region from RNA of Pun gene C. frutescens cv.

Reverse Transcriptase-PCR

Result and Discussion