Kim, Jeong-Soon
(Organic Agriculture Division, National Academy of Agricultural Science, Rural Development Administration)
,
Ahn, Sang-Nag
(Depatment of Agronomy, Chungnam National University)
,
Hong, Sung-Jun
(Organic Agriculture Division, National Academy of Agricultural Science, Rural Development Administration)
,
Kwon, Jin-Hyeuk
(Gyeongsangnam-do Agricultural Research and Extension Services)
,
Kim, Yeong-Ki
(Organic Agriculture Division, National Academy of Agricultural Science, Rural Development Administration)
,
Jee, Hyeong-Jin
(Organic Agriculture Division, National Academy of Agricultural Science, Rural Development Administration)
,
Shim, Chang-Ki
(Organic Agriculture Division, National Academy of Agricultural Science, Rural Development Administration)
The objective of this study was to determine the genetic diversities of major rice blast resistance genes among 84 accessions of aromatic rice germplasm. Eighty four accessions were characterized by a dominant 11 set of PCR-based SNP and CAPS marker, which showed the broad spectrum resistance and cl...
The objective of this study was to determine the genetic diversities of major rice blast resistance genes among 84 accessions of aromatic rice germplasm. Eighty four accessions were characterized by a dominant 11 set of PCR-based SNP and CAPS marker, which showed the broad spectrum resistance and closest linkage to seven major rice blast resistance (R) genes, Pia, Pib, Pii, Pi5 (Pi3), Pita (Pita-2), and Pi9 (t). The allele specific PCR markers assay genotype of SCAR and STS markers was applied to estimate the presence or absence of PCR amplicons detected with a pair of PCR markers. One indica accession, Basmati (IT211194), showed the positive amplicons of five major rice blast resistance genes, Pia, Pi5 (Pi3), Pib, Pi-ta (Pi-ta2), and Pik-5 (Pish). Among 48 accessions of the PCR amplicons detected with yca72 marker, only five accessions were identified to Pia gene on chromosome 11. The Pib gene was estimated with the NSb marker and was detected in 65 of 84 accessions. This study showed that nine of 84 accessions contained the Pii gene and owned Pi5 (Pi3) in 42 of 84 accessions by JJ817 and JJ113-T markers, which is coclosest with Pii on chromosome 9. Only six accessions were detected two alleles of the Pita or Pita-2 genes. Three of accessions were identified as the Pi9 (t) gene locus.
The objective of this study was to determine the genetic diversities of major rice blast resistance genes among 84 accessions of aromatic rice germplasm. Eighty four accessions were characterized by a dominant 11 set of PCR-based SNP and CAPS marker, which showed the broad spectrum resistance and closest linkage to seven major rice blast resistance (R) genes, Pia, Pib, Pii, Pi5 (Pi3), Pita (Pita-2), and Pi9 (t). The allele specific PCR markers assay genotype of SCAR and STS markers was applied to estimate the presence or absence of PCR amplicons detected with a pair of PCR markers. One indica accession, Basmati (IT211194), showed the positive amplicons of five major rice blast resistance genes, Pia, Pi5 (Pi3), Pib, Pi-ta (Pi-ta2), and Pik-5 (Pish). Among 48 accessions of the PCR amplicons detected with yca72 marker, only five accessions were identified to Pia gene on chromosome 11. The Pib gene was estimated with the NSb marker and was detected in 65 of 84 accessions. This study showed that nine of 84 accessions contained the Pii gene and owned Pi5 (Pi3) in 42 of 84 accessions by JJ817 and JJ113-T markers, which is coclosest with Pii on chromosome 9. Only six accessions were detected two alleles of the Pita or Pita-2 genes. Three of accessions were identified as the Pi9 (t) gene locus.
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제안 방법
Eighty four accessions of aromatic rice germplasm were screened for the presence of six major rice blast resistance (R) genes, Pia, Pii, Pi5(Pi3), Pib, Pita (Pita-2), and Pi9(t) multiple allele, using eight set of SNP and CAPS markers. All of the eighty four accessions of aromatic rice germplasm possessed more than one blast resistance gene, except for one accession of japonica type ‘Gerdeh’ in Table 2.
For the PCR based screening of Pi-ta (or Pi-ta2) genes on chromosome 12, the experiment showed that six of the 84 accessions under study produced positive bands of 1042 bp with YL155/87 primer set which was tightly linked to the resistant Pi-ta (or Pi-ta2) allele. Six positive accessions of Pi-ta (Pi-ta2) genes possessed four homozygote resistant Pi-ta (or Pi-ta2) alleles (Basmati, Mayataung, Kala Namak, and Masino Basmati) and two heterozygote resistant Pi-ta (or Pi-ta2) alleles (Basmati (IT211194) and Basmati 6113).
In this study, evaluations were made on the molecular markers that can be used reliably in marker-assisted selection (MAS), and eight set of CAPS and SNP markers tightly linked to the six major resistance genes, Pia, Pii, Pi5(Pi3), Pib, Pita (Pita-2), and Pi9(t) multiple allele, from the 84 accessions of aromatic rice germplasm.
이론/모형
The PCR analysis was conducted based on procedures described by Bioneer PCR Pre-mix kit manual (Bioneer Co. Ltd, Korea). The PCR reaction mixture contained 50 ng template DNA, 20 pmol of each primer set, 0.
성능/효과
The data indicated one accessions of domestic tongil type aromatic rice (Hyangmibyeo1ho), five accessions of japonica type (Daebunhyangdo2, Milagrosa Mutant, KungShanWu-Shen-Ken, Daw Dam, and 415 X Ir352) and three accessions of indica type (Goolarath, Seratus Malam, and Aroma) aromatic rice possessed the Pii rice blast resistance gene. Also, five accessions of japonica type (TALLI, Masino Basmai, Kalomasino Basmati, KINANDANG PAT, and Flores) and 37 accessions of indica type aromatic rice germplasm possessed the Pi5 (or Pi3) genes, but two tongil type and three japonica type of domestic aromatic rice did not have Pi5 (or Pi3) gene with PCR based detection of JJ817-Pi5 primer set.
4. Electrophoretical band patterns of 84 aromatic rice germplasms were scored as presence (+) or absence (-) of the rice blast resistance genes, Pi-ta (Pi-ta2), and the rice blast susceptible gene, pi-ta, were shown as a positive DNA fragments were amplified with three of SNP markers, YL155/YL87 (1042 bp) and YL183/YL87 (1042 bp), respectively. M, molecular marker (Bioneer 100 bp ladder plus); lanes 1-84, eighty four accessions of aromatic rice.
From the results, eighty three positive bands of Pi9(t) multi genes were cleaved with Hinf I restriction enzyme and possessed 12 accessions of Piz gene, 52 accessions of Pik-5 and Pish genes, and 14 accessions of Piz-t and Piz-5 genes. However, the Korean japonica and Tongil type varieties were grouped into four types: Koshihikari-type, Piz-t and Piz-t-type, Piz and Pi9-type, and null type, respectively, from two markers, pBA14 and NBS2-O/U (Cho et al.
In conclusion, the approach makes use of the successful identification of eight set of CAPS and SNP markers tightly linked to the six major resistance genes, Pia, Pii, Pi5 (Pi3), Pib, Pita (Pita-2), and Pi9(t) multiple allele, from the 84 accessions of aromatic rice germplasm. It is possible that the marker-assisted selection of rice blast resistance genes will help in the breeding program for the production of multi disease resistant aromatic rice varieties from different genetic resources.
In addition, they demonstrated that the Pi5(t) locus is identical to the Pi3(t) locus. The data indicated one accessions of domestic tongil type aromatic rice (Hyangmibyeo1ho), five accessions of japonica type (Daebunhyangdo2, Milagrosa Mutant, KungShanWu-Shen-Ken, Daw Dam, and 415 X Ir352) and three accessions of indica type (Goolarath, Seratus Malam, and Aroma) aromatic rice possessed the Pii rice blast resistance gene. Also, five accessions of japonica type (TALLI, Masino Basmai, Kalomasino Basmati, KINANDANG PAT, and Flores) and 37 accessions of indica type aromatic rice germplasm possessed the Pi5 (or Pi3) genes, but two tongil type and three japonica type of domestic aromatic rice did not have Pi5 (or Pi3) gene with PCR based detection of JJ817-Pi5 primer set.
This study had successfully detected 48 positive PCR amplicons of the Pia rice blast resistance gene with yca72 marker and identified 48 positive fragments cleaved with Hinf I restriction enzyme to only five accessions (TALLI, Mayataung, Basmati, Basmati 198, and Basmati 5854) as Pia gene. The Pib specific PCR primer set, NSb was developed to produce a 629 bp-positive amplicon in 64 accessions, two accessions of tongil type, 15 accessions of japonica type and 47 accessions of indica type aromatic rice germplasm.
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