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( Myeong Il Mun ),( Hankuil Yi ),( Ill Sup Nou ),( Yoon Kang Hur ) 한국육종학회 2015 Plant Breeding and Biotechnology Vol.3 No.1
Grafting has widely used in the agriculture of fruit-bearing crops and trees because rootstocks have a profound influence on many aspects of scion development and scion responses to the environmental changes including biotic and abiotic stress tolerance. These effects appear to result from the change of gene expression on scion, but only limited numbers of papers have been published demonstrating it. To identify altered expression of genes in Chinese cabbage, Chiifu(Brassica rapa ssp. pekinensis, inbred line) shoot was grafted on three Brassica rootstocks: mustard, turnip and broccoli. After head formation, Br300K microarray experiment was conducted using total RNAs from scion leaves collected in two different seasons, spring (June) and fall (October). A large number of differentially expressed genes (DEGs) were identified both in two seasonal samples, but DEGs were more notable in June sample than in October sample. However, the number of DEGs by three rootstocks were high in October with respect to up-regulation, but high in June for down-regulation. Categories of DEGs included metal ion binding, response to hormonal stimuli, response to endogenous stimuli, regulation of transcription, oxidation reduction and response to stress. Up-regulated genes in both June and October samples were similar in mustard and turnip rootstocks, but different in broccoli rootstock. Two genes were found to respond to all experimental conditions: Brapa_ESTC049008 (hypothetical protein) as an up-regulated gene and Brapa_ESTC016027 (CNGC12) as a down-regulated gene. Together with the previous reports, these results suggest that grafting-induced gene expression depends on the species involved.
Evaluation of Genetic Diversity among Korean Wild Codonopsis lanceolata by Using RAPD
Jin-Hee Park,Young-Ho Kim,Ill-Sup Nou,Jae-Eul Choi,Il-Yong Shim,Kwon-Kyoo Kang 한국자원식물학회 1997 한국자원식물학회지 Vol.10 No.3
The introduction of molecular biology methodologies to plant improvement programs offers an invaluable opportunity for extensive germplasm characterization. We have applied the developed technique of random amplification of polymorphic DNA(RAPD)to the analysis of evaluating genetic diversity among Korean wild Codonopsis lanceolata. A total of 340 polymorpic hands were gernerated on agarose- and polyacrylamide-gel by 19 primers of abitrary sequence. grouped by cluster analysis using sample matching coefficients of similarity. Among of the samples. the minimum genetic distance value was obtained between sample no. 1(Girisan) and no. 2(Girisan), and the largest value between sample no. 11(Sulaksan) and no. 17(Sulaksan).In separate cluster dendrograms based on agareose - and polyacryamide-gel. some differences were observed; In the case of agarose gel,41 samples could be devided into 7 groups at below about 0.44 level of distance. However they were divided into 6 gourps at below about 0.40 level of distance in the case of polyacrylamide gel. These results showed that polymophic data in agrose were not grouped to wild plant selected from each mountainous district except for wild plants selected from Sulaksan and Chiaksan. We believe that polyacrylamide-RAPD is a superior method for detecting DNA polymorphism compared to agarose-RAPD method.
Kim, Daeun,Jin, Bingkui,Je, Byoung Il,Choi, Youngmi,Kim, Byung Sup,Jung, Hee-Jeong,Nou, Ill-Sup,Park, Younghoon,Archambault, A. Canadian Science Publishing 2018 Genome Vol.61 No.10
<P> Reductions in growth and quality due to powdery mildew (PM) disease cause significant economic losses in tomato production. Oidium neolycopersici was identified as the fungal species responsible for tomato PM disease in South Korea in the present study, based on morphological and internal transcribed spacer DNA sequence analyses of PM samples collected from two remote regions (Muju and Miryang). The genes involved in resistance to this pathogen in the tomato accession ‘KNU-12’ (Solanum lycopersicum var. cerasiforme) were evaluated, and the inheritance of PM resistance in ‘KNU-12’ was found to be conferred via simple Mendelian inheritance of a mutant allele of the PM susceptibility locus Ol-2 (SlMlo1). Full-length cDNA analysis of this newly identified mutant allele (Slmlo1.1) showed that a 1-bp deletion in its coding region led to a frameshift mutation possibly resulting in SlMlo1 loss-of-function. An alternatively spliced transcript of Slmlo1.1 was observed in the cDNA sequences of ‘KNU-12’, but its direct influence on PM resistance is unclear. A derived cleaved amplified polymorphic sequence (dCAPS) and a high-resolution melting (HRM) marker were developed based on the 1-bp deletion in Slmlo1.1, and could be used for efficient marker-assisted selection (MAS) using ‘KNU-12’ as the source for durable and broad-spectrum resistance to PM. </P>
Kwon Kyoo Kang,Young Ho Kim,Il Sup Nou,Il Yong Shim 한국자원식물학회 1997 한국자원식물학회지 Vol.10 No.3
The relationship of nine Medicago species belonging to four subgenera were analyzed by using SDS-PAGE and restriction fragment length polymorphism (RELP) methodologies. Sixty-eight bands of alcohol and salt soluble proteins and 85-133 RFLP markers were used to estimate the genetic distance among the species. These species were clustered together at around 0.1 to 0.4 level of distance for both kind of markers, indicating that Medicago species have a large genetic similarity. A combined cluster diagram, at a dissimilarity level of 0.3, differentiated nine species in four groups: group 1, M. littoralis , M. truncatulam, M.scutellata and M. rigidula; group 2, M. sativa ; group 3, M. lupulina ; group 4, M. orbicularis, M. radiata and M. minima. All of them, but except for M. minima. corrensponded to the existing four subgenera of the genus Medicago classified by Lesins and Lesins(1979).The most similar species were M. littoralis and M. trucatula and the most dissimilar one was M. lupulina. In separate cluster diagrams based on RFLP and protein markers, some differences were observed. In the case of RFLP or DNA markers, M. sativa (alfalfa) was distantly clustered with other Medicago species. But in the case of protein markers, M. sativa was closely clustered with M. scutellata, M. littorulis and M. truncatula.
제주산 파인애플 유래 Bromelain관련 유전자 (BL1)를 이용반 형질전환 상추의 특성
정유진,김기훈,최장선,이순열,노일섭,박진희,강권규,Jung, Yu-Jin,Kim, Gi-Hyun,Choi, Jang-Sun,Lee, Soon-Youl,Nou, Il-Sup,Park, Jin-Heui,Kang, Kwon-Kyoo 한국식물생명공학회 2006 식물생명공학회지 Vol.33 No.1
파인애플 (Ananas comosus) 줄기에서 얻어지는 bromelain은 단백질 분해효소 중 cysteine protease의 복합체로 알려져 있다. 본 연구에서는 제주산 파인애플 줄기를 이용하여 bromelain 관련 유전자를 분리하였다. 분리된 BL1 유전자는 총 933개의 염기서열로 311개의 아미노산을 coding 하였다. 지금 까지 알려진 식물 유래 bromelain 관련 유전자와의 alignment 분석한 결과 BAA21929 유전자와 94%, T10516 유전자와 93% 및 P14518 유전자와 81%의 상동성을 보였다. BL1 유전자를 상추 게놈내에 도입하고자 NPTII 유전자 와 BL1 유전자로 제작한 pBI 121 BL 벡터를 Agrobacterium tumefacience LBA4404에 도입한 후, 상추잎 절편에 감염시켜 embryogenic callus 및 재분화 식물체를 육성하였다. 이들식물체로부터 T1세대를 육성하여 PCR 분석을 통해 왜래유전자의 도입 여부를 확인하였다. 또한 형질전환체의 발현여부는 Nothern blot분석 및 eno protease활성을 통해 형질전환체에서 BL1유전자가 안정적으로 상추세포내에서 발현되고 있음을 확인하였다. 따라서 본 실험에서 육성된 bromelain 관련 BL1 유전자가 도입한 형질전환 상추를 육종소재르 활용한다면 상업적으로 유용한 단백질을 분해하는 가수분해효소로써 건강 보조제, 사료첨가제 등에 널리 사용할 수 있을 것으로 생각되어진다. To clarify the roles of bromelain in plants, we isolated BL1 gene encoding bromelain from pineapple stem tissues and sequenced. The full length cDNA is 933 bp and encodes a polypeptide of 311 amino acid residues. The cDNA is most similar 94% at the amino acid level to bromelain previously isolated from pineapple (BAA21929). Explants of Lactuca sativa were co-cultivated with Agrobacterium tume-faciences LBA 4404 strains containing nptII and BL1 gene for transformation. Through initial selection of regenerated explants by culturing on a kanamycin and carbenicillin containing MS medium, multiple shoots were obtained after 2 months of culture. For a complementary step of selection, putative transgenic shoots were transferred to 1/2 Ms basal medium supplemented with 100 mg/L kanamycin and 500 mg/L carbenicillin. The selected shoots were obtained T1 generation seeds with emasculation, and tested with PCR analysis using 35S promoter and BL1 specific primers whether BL1 gene was introduced to genome of the plants. These results confirmed that produced the specific PCR bands in the putative transgenic lines. Additionally the Northern blot and endo protease activity showed that transcripts of BL1 gene were detected in transgenic lines. Theses results suggest that BL1 gene be successfully integrated and transcripted in the transgenic lettuce plants.