비생물학적 스트레스 관련 벼 Ac/Ds 삽입 변이체의 선발 및 유전자 발현 분석
Selection of (Ac/Ds) insertion mutant lines by abiotic stress and analysis of gene expression pattern of rice (Oryza sativar L.)
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Transposon-mediated insertional mutagenesis is one of powerful strategy for assessing functions of genes in higher plants. In this report, we have selected highly susceptible and tolerance plant by screening about high salt (3% NaCl) and cold stresses (4℃) from F2 seeds of 30,000 Ac/Ds insertional mutagenesis lines in rice (Oryza sativa L. cv. Dongjin). In order to identify the gene tagging, insertion of Ds element was analyzed by Southern blot and these results revealed that 19 lines were matched genotype of selected lines with phenotype from the first selected 212 lines, and 13 lines have one copy of Ds elements. The Franking Sequence Tags (FSTs) of selected mutant lines showed high similarities with the following known function genes: signal transduction and regulation of gene expression (transpoter, protease family protein and apical meristem family protein), osmotic stress response (heat shock protein, O-methyltransferase, glyceraldehyde -3-phosphate dehydrogenase and drought stress induce protein), vesicle trafficking (SYP 5 family protein) and senescence associated protein. The expression pattern of 19 genes were analyzed using RT-PCR under the abiotic stresses of 9 class; 250 mM NaCl, osmotic, drought, 3% H2O2, 100 µM ABA, 100 µM IAA, 0.1 ppm 2,4-D, 4℃ cold and 38℃ high temperature. Isolated knock-out genes showed the positive response about 250 mM NaCl, drought, H2O2, PEG, IAA, 2,4-D, ABA treatment and low (4℃) and high temperature (38℃). The results from this study indicate that function of selected knock-out genes could be useful in improving of tolerance to abiotic stresses as an important transcriptional activators in rice.
Abstract
Transposon-mediated insertional mutagenesis is one of powerful strategy for assessing functions of genes in higher plants. In this report, we have selected highly susceptible and tolerance plant by screening about high salt (3% NaCl) and cold stresses (4℃) from F2 seeds of 30,000 Ac/Ds insertional mutagenesis lines in rice (Oryza sativa L. cv. Dongjin). In order to identify the gene tagging, insertion of Ds element was analyzed by Southern blot and these results revealed that 19 lines were matched genotype of selected lines with phenotype from the first selected 212 lines, and 13 lines have one copy of Ds elements. The Franking Sequence Tags (FSTs) of selected mutant lines showed high similarities with the following known function genes: signal transduction and regulation of gene expression (transpoter, protease family protein and apical meristem family protein), osmotic stress response (heat shock protein, O-methyltransferase, glyceraldehyde -3-phosphate dehydrogenase and drought stress induce protein), vesicle trafficking (SYP 5 family protein) and senescence associated protein. The expression pattern of 19 genes were analyzed using RT-PCR under the abiotic stresses of 9 class; 250 mM NaCl, osmotic, drought, 3% H2O2, 100 µM ABA, 100 µM IAA, 0.1 ppm 2,4-D, 4℃ cold and 38℃ high temperature. Isolated knock-out genes showed the positive response about 250 mM NaCl, drought, H2O2, PEG, IAA, 2,4-D, ABA treatment and low (4℃) and high temperature (38℃). The results from this study indicate that function of selected knock-out genes could be useful in improving of tolerance to abiotic stresses as an important transcriptional activators in rice.
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