Expression Level of DREB Gene of Local Corn Cultivars from Kisar Island-Maluku, Indonesia, Using Quantitative Real Time Polymerase Chain Reaction

Hermalina Sinay,Estri Laras Arumingtyas

doi:10.4081/pb.2018.7005

Abstract

The research objective was to determine the expression level of dehydration responsive element binding (DREB) gene of local corn cultivars from Kisar Island Maluku, Indonesia. The study was designed as randomized block design with single factor consist of six local corn cultivars obtained from farmers in Kisar Island and one reference varieties which has been released as a drought-tolerant varieties and obtained from Cereal Crops Research Institute Maros South Sulawesi. Isolation of total RNA from the second leaf after the flag leaf at the 65 days after planting were carried out according to the protocols of the R and ABlueTM Total RNA Extraction Kit, and was used as a template for cDNA synthesis. Amplification of cDNA from total RNA was carried out according to the protocol of One-Step Reverse Transcriptase PCR Premix Kit. Real Time-PCR was performed usingcDNA from reverse transcription following the procedures of Real MODTM Green Real-Time PCR Master Mix Kit. The real time-PCR data were analyzed using relative quantification method based on the critical point/cycle threshold. The highest DREB gene expression was showed by Deep Yellow local corn cultivar, and the lowest one was showed by Rubby Brown Cob cultivar. The DREB gene expression level of deep yellow local corn cultivar was even higher than Srikandi variety as a reference variety.

Highlights

M Amplification of cDNA from total RNA o was carried out according to the protocol of c One-Step Reverse Transcriptase PCR - Premix Kit
An exploration and documentation of corn germplasm in the Kisar island was conducted and found that there are seven local cultivars of corn that are specific to that Island namely: i) Rubby Brown Cob cultivar, ii) White Brown Cob cultivar, iii) Red Blood, iv) White, v) Waxy, vi) Early Maturing Yellow, and vii) Deep Yellow cultivar
The highest to develop a plant as a superior crop, the properties of plants related to its ability to dehydration responn sive element binding (DREB) gene expression was showed by Deep Yellow local corn cultivar, and the lowest one was showed by Rubby Brown Cob cultivar

Summary

Introduction

N real time-PCR data were analyzed using relprotein DREB which involved in the mechanism of plant adaptation to drought. The purpose of this study was to determine the level of DREB gene expression on local corn cultivars in Kisar Island Maluku.

Objectives

Results

Conclusion