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질산태 질소 공급이 알팔파의 뿌리혹 형성 및 엽중 Nitrate Reductase 활성에 미치는 영향
李錫河,黃石重 韓國作物學會 1993 Korean journal of crop science Vol.38 No.2
알팔파 방목형 품종 Victoria와 건초형 품종 Vernal을 공시하여 실산태 새소 농도 0,2,4,8, 12mM의 다섯 수준으로 급여하여 파종 6주후 초기 생육시기에 지상부 부위별과 뿌리혹 건물증, 엽중 질산태 질소 환원 능력 및 축적 정도를 조사하였으며, 그 결과는 다음과 같이 요약된다. 1. NO3  ̄ 수준이 높아짐에 따라 지상부 건물중이 증가하였으나 NO(NO3 )  ̄ 의 8mM과 12mM간의 차이는 없었다. 2. 두 품종 모두 8mM 이상의 NO3  ̄ 수준에서 뿌리혹 착생이 현저히 억제되었다. 3. 엽 생체중당 nitrate reductase 활성은 NO3  ̄ 수준이 증가함에 따라 높아졌으나 4mM이상에서는 차이가 크지 않았다. 4. 엽중 질산태 질소축적은NO3  ̄ 의 12mM 처리시 크게 증가하였다. A full understanding of the interdependence of leaf nitrate ((No3  ̄) metabolism and symbiotic nitrogen(N2 ) fixation in legume crops is needed to help maximize the use of both N sources as well as to improve forage quality through the inhibition of leaf nitrate accumulation. The present work examines the effects of added nitrate, the level of which are 0,2,4,8 and 12mM, on the nodule formation and leaf nitrate utilization and on the possibility of inducing nitrate-toxicity to livestocks in two alfalfa varieties, ' Vernal ' of grazing type and ' Victoria ' of hay type. Higher level of exogeneous nitrate resulted in the increased above-ground dry weight. Nodulation was inhibited severely when more than 8mM NO3  ̄ was supplied to alfalfa plants, and leaf nitrate reductase reached a maximunm at 4mM nitrate supply. The Vmax of nitrate reductase in leaves of Vernal was similar to that of Victoria, whereas the Km of Vernal was higher than that of Victoria. High accumulation of leaf nitrate, 4~times10-5 g/g leaf fresh weight, was shown at 12mM nitrate supply, which was thought to be not enough to induce nitrate-toxicity to livestocks.icity to livestocks.
이석하 한국작물학회 2007 Journal of crop science and biotechnology Vol.10 No.3
Xanthomonas axonopodis pv. glycines (Xag) is a pathogen that causes bacterial leaf pustule (BLP) disease in soybeans grown in Korea and the southern United States. Typical and early symptoms of the disease are small, yellow to brown lesions with raised pustules that develop into large necrotic lesions leading to a substantial loss in yield due to premature defoliation. After Xag infects PI 96188, only pustules without chlorotic haloes were observed, indicating the different response to Xag. To identify differentially expressed genes prior to and 24 hr after Xag inoculation to PI 96188 and BLP-resistant SS2-2, an oligonucleotide macroarray was constructed with 100 genes related to disease resistance and metabolism from soybean and Arabidopsis. After cDNAs from each genotype were applied on the oligonucleotide macroarrays with three replicates and dye swapping, 36 and 81 genes were expressed as significantly different between 0 hr and 24 hr in PI 96188 and SS2-2, respectively. Six UniGenes, such as the leucine-rich repeat protein precursor or 14-3-3-like protein, were selected because they down-regulated in PI 96188 and up-regulated in SS2-2 after Xag infection, simultaneously. Using tubulin and cDNA of Jangyeobkong (BLP-susceptible) as controls, the oligonucleotide macroarray data concurred with quantitative real-time RT-PCR (QRT RT-PCR) results in most cases, supporting the accuracy of the oligonucleotide macroarray experiments. Also, QRT RT-PCR data suggested six candidate genes that might be involved in a necrotic response to Xag in PI 96188. Xanthomonas axonopodis pv. glycines (Xag) is a pathogen that causes bacterial leaf pustule (BLP) disease in soybeans grown in Korea and the southern United States. Typical and early symptoms of the disease are small, yellow to brown lesions with raised pustules that develop into large necrotic lesions leading to a substantial loss in yield due to premature defoliation. After Xag infects PI 96188, only pustules without chlorotic haloes were observed, indicating the different response to Xag. To identify differentially expressed genes prior to and 24 hr after Xag inoculation to PI 96188 and BLP-resistant SS2-2, an oligonucleotide macroarray was constructed with 100 genes related to disease resistance and metabolism from soybean and Arabidopsis. After cDNAs from each genotype were applied on the oligonucleotide macroarrays with three replicates and dye swapping, 36 and 81 genes were expressed as significantly different between 0 hr and 24 hr in PI 96188 and SS2-2, respectively. Six UniGenes, such as the leucine-rich repeat protein precursor or 14-3-3-like protein, were selected because they down-regulated in PI 96188 and up-regulated in SS2-2 after Xag infection, simultaneously. Using tubulin and cDNA of Jangyeobkong (BLP-susceptible) as controls, the oligonucleotide macroarray data concurred with quantitative real-time RT-PCR (QRT RT-PCR) results in most cases, supporting the accuracy of the oligonucleotide macroarray experiments. Also, QRT RT-PCR data suggested six candidate genes that might be involved in a necrotic response to Xag in PI 96188.
Gene Duplications Revealed during the Process of SNP Discovery in Soybean [Glycine max (L.) Merr.]
이석하,Chun Mei Cai,반규정 한국작물학회 2007 Journal of crop science and biotechnology Vol.10 No.4
Genome duplication (i.e. polyploidy) is a common phenomenon in the evolution of plants. The objective of this study was to achieve a comprehensive understanding of genome duplication for SNP discovery by Thymine/Adenine (TA) cloning for confirmation. Primer pairs were designed from 793 EST contigs expressed in the roots of a supernodulating soybean mutant and screened between 'Pureunkong' and 'Jinpumkong 2' by direct sequencing. Almost 27% of the primer sets were failed to obtain sequence data due to multiple bands on agarose gel or poor quality sequence data from a single band. TA cloning was able to identify duplicate genes and the paralogous sequences were coincident with the nonspecific peaks in direct sequencing. Our study confirmed that heterogeneous products by the co-amplification of a gene family member were the main cause of obtaining multiple bands or poor quality sequence data in direct sequencing. Counts of amplified bands on agarose gel and peaks of sequencing trace suggested that almost 27% of nonrepetitive soybean sequences were present in as many as four copies with an average of 2.33 duplications per segment. Copy numbers would be underestimated because of the presence of long intron between primer binding sites or mutation on priming site. Also, the copy numbers were not accurately estimated due to deletion or tandem duplication in the entire soybean genome.
Genetic Mapping of Novel Symptom in Response to Soybean Bacterial Leaf Pustule in PI 96188
이석하,김길현,박종호,김문영,Sunggi Heu 한국작물학회 2011 Journal of crop science and biotechnology Vol.14 No.2
Soybean bacterial leaf pustule (BLP) is a serious disease caused by Xanthomonas axonopodis pv. glycines. Typical symptoms of BLP are pustules surrounded by small yellow haloes. Interestingly, PI 96188 only exhibits pustules without chlorotic haloes which suggests a resistant response. The objectives of this study are to understand the inheritance mode of the novel symptom to BLP in PI 96188 and to investigate whether or not a gene controlling BLP resistance in PI 96188 is identical to the rxp gene. First, a new BLP resistant genotype, PI 96188 was crossed with the resistant cultivar SS2-2. All F₁ plants showed the same phenotype as SS2-2 and the F₂ population segregated into 75 typical symptoms (haloes presence : 28 novel symptoms (haloes absence) indicating the presence of a single recessive gene. To map the novel symptom to BLP in PI 96188, a population of 88 F_7 recombinant inbred lines was developed from a cross between PI 96188 and the susceptible cultivar Jinjoo1. The BLP resistance gene from PI 96188 was mapped on chromosome (Chr.) 10 (LG O) rather than Chr. 17 (LG D2). This gene was linked with the simple sequence repeat marker, Sat_108at the distal end of Chr. 10. Thus, the BLP resistance gene from PI 96188 was determined to be a new gene.