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Overexpression of a gene encoding LRR protein improves rice resistance against Magnaporthe oryzae


Azizi, Parisa and Yusop, Mohd Rafii and Abdullah, Siti Nor Akmar and Musa, Mohamed Hanafi and Sahebi, Mahbod (2017) Overexpression of a gene encoding LRR protein improves rice resistance against Magnaporthe oryzae. In: International Conference on Big Data Applications in Agriculture (ICBAA2017), 5-6 Dec. 2017, Auditorium Putra, TNCPI Building, Universiti Putra Malaysia. (pp. 134-140).

Abstract / Synopsis

Magnaporthe oryzae is a rice blast fungus and plant pathogen that causes a serious rice disease and, therefore, poses a threat to the world’s second most important food security crop. Plant transformation technology has become an adaptable system for cultivar improvement and to functionally analyze genes in plants. The objective of this study was to determine the effects (through over-expressing and using the CaMV 35S promoter) of Pikh on MR219 resistance because it is a rice variety that is susceptible to the blast fungus pathotype P7.2. Thus, a full coding DNA sequence (CDS) of the Pikh gene, 1206 bp in length, was obtained through amplifying the cDNA template from a PH9-resistant rice variety using a specific primer. Agrobacterium-mediated transformation technology was also used to introduce the Pikh gene into the MR219 callus. Subsequently, transgenic plants were evaluated from the DNA to protein stages using polymerase chain reaction (PCR), semi-quantitative RT-PCR, real-time quantitative PCR and high-performance liquid chromatography (HPLC). Transgenic plants were also compared with a control using a real-time quantification technique (to quantify the pathogen population), and transgenic and control plants were challenged with the local most virulent M. oryzae pathotype, P7.2. The Pikh gene was up-regulated in the transgenic plants compared with the control plants. The quantity of the amino acid leucine in the transgenic rice plants increased significantly from 17.131 in the wild-type to 47.865 mg g-1 in transgenic plants. The M. oryzae population was constant at 31, 48 and 72 h after inoculation in transgenic plants, while it was increased in the inoculated control plants. This study successfully clarified that over-expression of the Pikh gene in transgenic plants can improve their blast resistance against the M. oryzae pathotype P7.2.

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Additional Metadata

Item Type: Conference or Workshop Item (Paper)
Divisions: Faculty of Agriculture
Institute of Plantation Studies
Institute of Tropical Agriculture and Food Security
Publisher: Institute of Plantation Studies, Universiti Putra Malaysia
Keywords: Plant pathogen; Coding DNA sequence; Agrobacterium-mediated transformation; Real-time quantitative PCR; High performance liquid chromatography
Depositing User: Nabilah Mustapa
Date Deposited: 07 Feb 2018 17:24
Last Modified: 07 Feb 2018 17:24
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