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Sailila E Abdula,Hye-Jung Lee,Moo-Geun Jee,Dae-Won Jang,Marjohn Nino,Yong-Gu Cho,Kwon Kyoo Kang 한국육종학회 2012 한국육종학회 심포지엄 Vol.2012 No.07
The latest report on draft genome of Brassica rapa sequence has been published. To elucidate the functions of a large population of these genes and to search efficiently for agriculturally useful genes, the Full-length cDNA Over-eXpressor (FOX) gene hunting system was used. The FOX library was transformed into rice by Agrobacteriummediated transformation. Approximately 1,150 FOX-rice lines were generated. Genomic PCR analysis indicated that the average length of FL-cDNAs was 900∼1,200 bp with functional annotation of many unknown function (35.5%). Most of the randomly selected transgenic rice lines showed overexpression (92%) and barely mRNA expression in the wild type Gopum. Moreover, 94% of the 850 transgenic rice lines were moderately tolerant (slightly yellow) to cold and 9 lines were tolerant (seedling light green). For the salinity evaluation, most of the transgenic lines (85%) were highly susceptible whereas seven lines were tolerant. In addition, morphological evaluation of rice lines showed minimal phenotypic alteration (12%). About 25.1 and 22% were earlier in terms of days to heading and chlorophyll contents, respectively. Further, 18% of FOX rice lines showed lower chlorophyll contents. Filled grains, number of tillers, panicle length, culm and plant height were relatively less variable among the lines. These results provided useful genes for functional analyses in the mechanisms of identified transgenic tolerant lines.
Sailila E. Abdula,이혜정,Reneeliza J. Melgar,Mingmao Sun,강권규,조용구 한국식물생명공학회 2011 식물생명공학회지 Vol.38 No.1
Alcohol dehydrogenase (E.C.1.1.1.1) is an enzyme present in higher plants involved in the anaerobic fermentation pathway that catalyzes the reduction of pyruvate to ethanol, resulting in continuous NAD^(+) regeneration. It also plays an important role in many plant developments including tolerance to anoxia condition. Here, a cDNA clone encoding alcohol dehydrogenase (ADH) was isolated from Chinese cabbage (Brassica rapa) seedlings. The gene named Bradh1 had a total length of 1,326 bp that contains a single open reading frame of 1,140 bp. The predicted protein consists of 379 amino acid residues with a calculated molecular mass of 41.17 kDa. Expression pattern analysis revealed a tissue-specific expressing gene in different tissues and strongly expressed in the shoot, roots and seeds of Chinese cabbage. Agrobacterium transformation of full-length cDNA Bradh1 into rice Gopumbyeo showed high efficiency. Furthermore, induction of ADH in transgenic rice enhanced tolerance to anaerobiosis stresses and elevated mRNA transcripts. The overexpression of Bradh1 in rice increases germination under anaerobiosis stresses, implying the possibility of developing new varieties suited for direct seeding or flood-prone rice field.
Abdula, Sailila E.,Lee, Hye-Jung,Ryu, Hojin,Kang, Kwon Kyoo,Nou, Illsup,Sorrells, Mark E.,Cho, Yong-Gu Springer-Verlag 2016 Plant molecular biology reporter Vol.34 No.2
<P>The calcineurin B-like protein (CBL)-CBL-interacting protein kinase (CIPK) pathway is emerging as a major signaling pathway in plants. To understand the function of CIPK, the gene named BrCIPK1 from Brassica rapa were introduced into rice. Characterization of BrCIPK1 gene showed a 1982 bp, containing 1509 bp coding region and 502 amino acids. Green fluorescent protein (GFP)-tagged BrCIPK1 was observed exclusively in the cytoplasmic and peripheral regions in the plant cell. Gene expression showed that its messenger RNA (mRNA) transcription in B. rapa was differentially accumulated in the presence of cold, salinity, and drought, indicating its biological roles in multiple stress response pathways in plants. Furthermore, Ubi-1::BrCIPK1 rice lines showed significantly higher biomass, water content, and proline and free sugar content relative to those in the wild-type Gopum. The BrCIPK1 interacted with rice calcineurin B-like protein 1 and 5 (OsCBL1, OsCBL5), suggesting that it is activated by Ca2+-bound CBLs in the cytosol by calcium spiking and regulates its downstream target proteins in these regions to increase abiotic stress tolerance. The results imply that BrCIPK1 gene may be involved in stress adaptations through the activation of pyrroline-5-carboxylate synthase in the proline biosynthetic pathway. In this paper, a hypothetical mechanism of elevated tolerance to cold, drought, and salinity is presented.</P>
Sailila E Abdula,Hye-Jung Lee,Moo-Geun Jee,Dae-Won Jang,Franz Nogoy,Kwon Kyoo Kang,Yong-Gu Cho 한국육종학회 2012 한국육종학회 심포지엄 Vol.2012 No.07
UDP-glucose 4-epimerase (UGE) catalyzes the reversible conversion of UDP-glucose to UDP-galactose. The gene, named BrUGE1, isolated from a Chinese cabbage had a total length of 1,328 bp that contains a single open reading frame (ORF) of 1,056 bp which encodes a polypeptide of 351 amino acid residues with a calculated mass of 39.0 kDa. Sequence analysis of BrUGE1 protein has the characteristic of an active site tetrad and NAD-binding motif (typically TGXXGXXG) of the extended short chain dehydrogenase/ reductase (SRD) superfamily. Expression analysis showed that BrUGE1 is tissue specific and highly expressed in stem of rice plant. Interestingly, BrUGE1 mRNA was highly accumulated by drought stress with significantly higher amount of soluble sugar. Morphological evaluation showed an increase in yield by 27%. Panicle length, number of productive tillers/hill, and filled spikelets were significantly increased by 17~20% compared to the wild type Gopum. Moreover, the growth of the wild type Gopum seedlings on galactose was increasingly inhibited with a decrease in UDP-glc epimerase 1 expression compared to the transgenic rice lines. In the Ubi-1::BrUGE1 lines, the increase of UDP-glc epimerase 1 expression was apparently sufficient to overcome the toxic effects of galactose. Taken together, the Ubi-1::BrGUE1 rice lines increased yield probably by increasing the rate of filled grains. The enhanced drought tolerance may be due to the induction of soluble sugar which may act as osmolyte to compensate dehydration during drought stress.
Abdula, Sailila E.,Lee, Hye-Jung,Melgar, Reneeliza J.,Sun, Mingmao,Kang, Kwon-Kyoo,Cho, Yong-Gu The Korean Society of Plant Biotechnology 2011 식물생명공학회지 Vol.38 No.1
Alcohol dehydrogenase (E.C.1.1.1.1) is an enzyme present in higher plants involved in the anaerobic fermentation pathway that catalyzes the reduction of pyruvate to ethanol, resulting in continuous $NAD^+$ regeneration. It also plays an important role in many plant developments including tolerance to anoxia condition. Here, a cDNA clone encoding alcohol dehydrogenase (ADH) was isolated from Chinese cabbage (Brassica rapa) seedlings. The gene named Bradh1 had a total length of 1,326 bp that contains a single open reading frame of 1,140 bp. The predicted protein consists of 379 amino acid residues with a calculated molecular mass of 41.17 kDa. Expression pattern analysis revealed a tissue-specific expressing gene in different tissues and strongly expressed in the shoot, roots and seeds of Chinese cabbage. Agrobacterium transformation of full-length cDNA Bradh1 into rice Gopumbyeo showed high efficiency. Furthermore, induction of ADH in transgenic rice enhanced tolerance to anaerobiosis stresses and elevated mRNA transcripts. The overexpression of Bradh1 in rice increases germination under anaerobiosis stresses, implying the possibility of developing new varieties suited for direct seeding or flood-prone rice field.
( Sailila E. Abdula ),( Hye Jung Lee ),( Moo Geun Jee ),( Yu Jin Jung ),( Kwon Kyoo Kang ),( Ill Sup Nou ),( Sang Bok Lee ),( Won Ha Yang ),( Yong Gu Cho ) 한국육종학회 2013 Plant Breeding and Biotechnology Vol.1 No.1
The latest report on the draft genome of Brassica rapa sequence has been published. To elucidate the functions of these genes and to efficiently search for agriculturally useful genes, a Full-length cDNA Over-eXpressor (FOX) gene hunting system was used. The FOX library from Chinese cabbage was introduced into rice via Agrobacterium-mediated transformation. Approximately 1,150 FOX-rice lines were generated. Genomic PCR analysis indicated that the average length of FL-cDNAs introduced into individual lines was 900~1,200 bp. Basic Local Alignment System Tool (BLAST) analysis of the FL-cDNA genes revealed that 35.5% have unknown function. Most of the randomly selected transgenic rice lines showed overexpression (92%) of these genes relative to the wild-type Gopum. Moreover, 94% of the 850 transgenic rice lines were moderately tolerant (slightly yellow) to cold and 9 lines were tolerant (seedlings were light green). Morphological evaluation of the transgenic rice lines showed minimal phenotypic alteration (12%). Approximately 25.1% and 22% of the plants were significantly ahead in the days to heading and had elevated chlorophyll content, respectively. Other agronomic traits such as filled grains, number of tiller, panicle length, and culm and plant height were relatively less variable among the transgenic lines. These results provide a resource for defining genes that are associated with tolerance in transgenic rice lines.
Development and abiotic stress screening of transgenic rice with FL-cDNA derived from Brassica rapa
Sailila E. Abdula,Sung Hee Kim,Hye Jung Lee,Rae Young Suk,Sang Hyuk Lee,Ming Mao Sun,Kwon Kyu Kang,Bo Kyeong Kim,Dong Sub Kim,Sun Hee Woo,Hong Sig Kim,Yong Gu Cho 한국육종학회 2010 한국육종학회 학술발표회 발표요지 Vol.42 No.1
Marjohn Nino,Sailila E. Abdula,Hye-Jung Lee,Dal-A Yu,Mi-Sun Kim,Sanguk Byeon,Franz Nogoy,Kwon-Kyoo Kang,Illsup Nou,Yong-Gu Cho 한국육종학회 2013 한국육종학회 심포지엄 Vol.2013 No.07
Bacterial blight is a serious problem of rice in irrigated and rainfed lowlands. It is caused by Xanthomonas oryzae pv. oryzae (Xoo) which is represented by many pathotypes, making it difficult to control. Plant proteases are important players in immunity acting either in the execution of attack, in signaling cascade or in perception of invader. This study demonstrates the response of cysteine protease (CP) upon interaction with the pathogen. The cysteine protease encoding full-length cDNA was identified and characterized using web-based tools. Conserved domain of the gene revealed its affinity to Peptidase_CIA family. The full-length cDNA of CP in Brassica rapa was then cloned and overexpressed in rice. Insertion of gene was verified in the transformants through PCR assay. Spatiotemporal expression of the gene was performed in transgenic rice. To evaluate the resistance of CP-overexpression lines to Xoo, transgenic plants were inoculated with two races of Xoo. In planta analysis of enzymatic activity of CP was also performed before and after infection by the pathogen.
Marjohn Nino,Sailila E. Abdula,Hye-Jung Lee,Joonki Kim,Dal-A Yu,Sanguk Byeon,Franz Nogoy,Kwon-Kyoo Kang,Illsup Nou,Yong-Gu Cho 한국육종학회 2014 한국육종학회 심포지엄 Vol.2014 No.07
In spite of the overwhelming number of cysteine proteases in plants, only a few were substantially investigated. Papain-like cysteine proteases (PLCPs) are commonly implicated to disease immunity in some key pathosystems in plants, such as in tomato – Cladosporium fulvum, potato/tomato – phytopthora infestans, and Arabidopsis – Ralstonia solanacearum, among the few others. This study demonstrates the function of cysteine protease gene cloned form Brassica rapa (BrCP) related to resistance to Xanthomonas oryzae pv. oryzae in transgenic rice lines. The cysteine protease-encoding full-length cDNA was identified and characterized using web-based tools. The gene is 2,267 bp in size with an open reading frame of 1,365 bp that encodes predicted polypeptide of 455 amino acids. Blast analysis of the conserved domain of the gene confirmed its affinity to Peptidase_CIA family. Full-length cDNA of PLCP in Brassica rapa was then cloned and co-overexpressed in rice with HPT marker. Introgression of the gene was confirmed in the transformants through genomic PCR assay. RT-PCR analysis showed that the gene was constitutively expressed and present in all tissues. The overexpression rice lines exhibited an enhanced resistance when screened with four Korean Xoo isolates.