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Mrna Extraction, Cdna Synthesis and Tillering Specific Gene Isolation from Blb Resistant Rice Br14

Ahmed Hossain*
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Tajrin Farhana Khanom
Department of Environmental Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

P. Jahan
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Fahmida Khatun
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Khondoker Md. Nasiruddin
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Abstract/ Executive Summary:

Tillering or the production of lateral branches (i.e. culms), is an important agronomic trait that determines shoot architecture and grain production in grasses like rice. Due to its agronomic and biological importance, tillering has been widely studied and numerous works demonstrate that the control of AXM initiation, bud development and tillering in the grasses is via a suite of genes, hormones and environmental conditions. The cDNA rice plant was constructed by an improved RT-PCR technique from a template cDNA library derived from the total RNAs extracted from the young plant leaves of a Oryza sativa rice to identify genes pi55(t) involved in the control of rice tillering. The identification of tillering specific gene in BR14 rice germplasm will help in accelerating the production rate of rice in future, including bacterial leaf blight (BLB) disease resistant genes in different varieties. BLB caused by Xanthomonas oryzae pv. oryzae (Xoo) is a major biotic stress in the irrigated rice belts. Genetic resistance is the most effective and economical control for bacterial blight disease. Branching capacity is the most important. Molecular survey was conducted to identify the presence of tillering specific gene in BLB resistant BR14 rice. Reproducible means of BR14 rice at 21 days was designed based on the timing of full expression of the leaf. Gel documentation showed that the size of the synthesized cDNA was 250 bp and tillering specific gene was 201bp long. PCR with specific primers RM5493 for tillering specific gene was used in the study. On the basis of the result of PCR, it was concluded that after PCR, the band that was found in 201bp was indicated tillering specific gene pi55 (t).

Keywords: Bacterial leaf blight, Tillering, PCR, Biotic stress, Rice BR14

Location: Department of Microbiology and Hygiene, Bangladesh Agricultural University (BAU), Mymensingh.

Start Date: 00-07-2013

End Date: 00-12-2013

Program Area: Pest Management

Commodity: Bacteria, Rice

Objectives:

Considering the above facts the present study was designed with the aims to construction of cDNA library from mRNA of BLB resistant rice, and to isolation of tillering specific gene from cDNA of BLB resistant BR14 rice.

Methodology:

The experiment was carried out at Biotechnology laboratory, Department of Biotechnology; and Microbiology laboratory, Department of Microbiology and Hygiene, Bangladesh Agricultural University (BAU), Mymensingh. The study was conducted from July to December 2013 to extract mRNA, synthesis of cDNA and isolate tillering specific gene pi55 (t).

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

This work demonstrated the potential applying of genetic analyses in rice research and it was conducted for isolation and identification of tillering specific gene from BLB resistant rice. The results of this study may be utilized as the local source of gene x21gene for elite molecular breeding program and it can be used as a base for improvement of rice varieties in Bangladesh. More varieties that are resistant will improve yield of the crops and help address the food security issue facing the developing world. Considering the above information, it could be concluded that BR14 rice is the best source sample for isolating and identifying of tillering specific novel recessive gene pi 55 (t) gene. RT-PCR is more effective method for isolating and detecting mRNA, cDNA and tillering specific gene. BR14 rice is the resistant plant against BLB pathogen.

Knowledge Management: Journal