[논문]Screening of Rice Blast Resistance Genes from Aromatic Rice Germplasms with SNP Markers

Kim, Jeong-Soon; Ahn, Sang-Nag; Kim, Chung-Kon; Shim, Chang-Ki

doi:10.5423/ppj.2010.26.1.070

Screening of Rice Blast Resistance Genes from Aromatic Rice Germplasms with SNP Markers 원문보기

The plant pathology journal, v.26 no.1, 2010년, pp.70 - 79

Kim, Jeong-Soon (National Agrobiodiversity Center, Rural Development Administration) , Ahn, Sang-Nag (Department of Agronomy, Chungnam National University) , Kim, Chung-Kon (National Agrobiodiversity Center, Rural Development Administration) , Shim, Chang-Ki (National Agrobiodiversity Center, Rural Development Administration)

Abstract ▼ AI-Helper

Rice blast is one of the serious devastating diseases. This study was carried out to determine the genetic diversities of blast resistance (R) genes form 86 accessions of aromatic rice with eight SNP markers, z4792, zt4792, z60510, zt6057, k6415, k6411, k39575 and t256, which showed the close-set linkage to 6 major genes, Piz, Piz-t, Pik, Pik-m, Pik-p, and Pit. Four accessions of indica type, Mayataung, Yekywin Yinkya Hmwe, Basmati9-93, and Basmati5854, showed the positive amplicons of six major genes. Among 86 accessions, 83 accessions were detected both or one of Piz and Piz-t genes. Seventy three accessions contained the Piz gene with z4792 marker. In addition, 30 and 71 accessions possessed Piz-t gene with zt4792 and zt6057 markers, respectively. Ten accessions showed the positive bands for the Piz-t gene with both zt4792 and zt6057 markers. Only one accession, Khau Nua Keo, was not amplified for both Piz and Piz-t gene. But japonica type, Gerdeh, possessed only Piz gene between Piz and Piz-t. Fifty two accessions showed the three of Pik multiple genes and Pit gene. Four accessions, Iari7447, Daebunhyangdo2, Shiyayuuine, and Basmati 6129 possessed a Pik-p gene. Especially, Pit gene on chromosome 1 was detected with t256 marker in all of 83 accessions, exception of A-2, one accession of japonica type.

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문제 정의

In this study, the marker-assisted selection of rice blast resistance genes will help in the breeding program in multi diseases resistant aromatic rice varieties in genetic resources of aromatic rice. Some of these germplasma may have special properties that are important to breeding program whereas others may possess proprietary technology.

제안 방법

0, and 30 mM KCl), and 1 unit of Taq DNA polymerase in 20 ul PCR reaction volume. PCR amplifications were carried out in a MyGenie96 Thermal cycler (Bioneer Co. Ltd, Korea), and programmed that template DNA was initially denatured at 95℃ for 4 min, followed by 30 cycles of PCR amplification steps with the following parameters; a 30 sec denaturation at 94℃, 30 sec primer annealing at from 42 to 62℃, and a 60 sec primer extension at 72℃ allowed for completion of primer extension, with a final extension at 72℃ for 10 min. Initially 4 μl of the amplified products were electrophoretically resolved on a 1.
This study was carried out to acquire information for genetic diversities of resistance genes against rice blast disease in aromatic rice germplasms for improving aromatic rice breeding efficiency using PCR-based markers including several candidate SNP (single nucleotide polymorphisms) markers.

대상 데이터

Application of molecular markers will enhance the efficiency of rice evaluation and improvement program. Eighty four accessions of aromatic rice germplasm were screened for the presence of 6 major rice blast resistance (R) genes, Piz, Piz-t, Pik, Pik-m, Pik-p, and Pit, using the eight of SNP markers listed in Table 1. An allele specific PCR marker assays the genotype of SNP marker by estimating the presence or absence of PCR amplicon from a pair of PCR reactions.

성능/효과

Piz-t gene on chromosome 6 was detected in eighty three accessions of aromatic rice germplasm with together or any one of zt4792 and zt6057 SNP marker. Among the eighty three, thirty accessions and seventy one accessions were amplified with each one of allele specific SNP marker, zt4792 and zt6057, respectively. Eighteen accessions including six of japonica type and twelve of indica type showed the positive bands with both zt4792 and zt6057 SNP markers (Fig.
Three accessions of japonica type (Milagrosa mutant, Khau Nua Keo, and Khau Tan Luong) and two accessions of indica type (Basmati6311 and Dinorado) were showed the positive ampilcon for Pik-m of three Pik multi genes with k6441 SNP marker. Each of the seven accessions of japonica type (Hyangnambyeo, Arnahyangchalbyeo, Mihayangbyeo, Jahyangna86-1, Gerdeh, Flores, and Kung-Shan Wu-Shen-Ken) and indica type (Goolarath, Arc6011, Azucena, Milfor6, Khao Dawk Mali 105, Aroma, and Ds20) aromatic rice germplasms was possessed the positive bands of Pik-m and Pik-p genes. Allele specific resistance genes of Pik and Pik-p gene were detected in three accessions of japonica type (Hyangmibyeo1ho, Inaguhu, and 415xIr352) and four accessions of indica type (Tarana Deshi, Iranbyeopssi, Basmati Dhan, and Basmati), respectively (Fig.
Estimation of PCR results for the Piz and Piz-t rice blast resistance genes were determined by visualization of amplicons on 340 bp, 240 bp, and 360 bp of positive fragments using two set of SNP primer, set 1 (z4792 and zt4792) and set2 (z60510 and zt6057) (Fig. 1). Piz gene on chromosome 6 was scored to 73 accessions of aromatic rice germplasm, two accessions of two accessions of tongil type, twenty four accessions of japonica type, and forty seven accessions of indica type (Fig.
Pit gene was founded in an indica rice variety K59. Fortunately in our results, Pit gene was detected all of 83 accessions of aromatic rice germplasms, two tongil types, twenty five japonica types, and fifty six indica types, and expected one accession of japonica type, A-2, and originated from Bhutan.
Our results, the genetic frequency of the six major rice blast resistance genes, Piz, Piz-t, Pik, Pik-m, Pik-p, and Pit, ranged from 30% to 99%. Significantly, eighty three accessions (99%) were detected with both or one of Piz and Pizt genes.
The amplified fragments using SNP markers, z4792 (A), zt4792 (B), z60510 (C), and zt6057 (D), were scored as presence (+) or absence (−) of amplicons linked to Piz gene.

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