• 제목/요약/키워드: Transgenic, Antisense

검색결과 40건 처리시간 0.03초

Regulation of methionine biosynthesis in plants; transgenic study

  • Kim, Jungsup;Thomas Leustek
    • 한국식물학회:학술대회논문집
    • /
    • 한국식물학회 2002년도 춘계학술발표대회:발표눈문요지록
    • /
    • pp.73-82
    • /
    • 2002
  • The committing step in Met and S-adenosyl-L-methionine (SAM) synthesis is catalyzed by cystathionine ${\gamma}$ -synthase (CGS). Transgenic Arabidopsis thaliana overexpressing CGS under control of 35S promoter show increased soluble Met and its metabolite S-methylmethionine, but only at specific stages of development. CGS-overexpressing seedlings are resistant to ethionine. Similar results were obtained with transgenic potato plants overexpressing Arabidopsis CGS. Several of the transgenic lines show silencing of CGS resulting in deformed p]ants with a reduced capacity for reproductive growth similar as transgenic plants by antisense RNA (CGS[-]). Exogenous feeding of Met to the CGS[-] and CGS[+] silenced plants partially restores their growth. Similar morphological deformities are observed in plants cosuppressed for SAM synthetase, even though such plants accumulate 250 fold more soluble Met than wild type and they overexpress CGS. The results suggest that the abnormalities associated with CGS and SAM synthetase silencing are due in part to a reduced ability to produce SAM, and that SAM may be a regulator of CGS expression.

  • PDF

Expression of diligent protein and Pinoresinol/Lariciresinol reductase genes of forsythia in transgenic potatoes

  • Chuong, Tran-Van;Kim, Hyun-Soon;Park, Ji-Young;Joung, Jae-Youl;Youm, Jung-Won;Jeon, Jae-Heung
    • Plant Resources
    • /
    • 제4권3호
    • /
    • pp.181-188
    • /
    • 2001
  • We tried to introduce two forsythia genes related in lignan biosynthesis, dirigent protein and pinoresinol/lariciresinol (Ph) reductase, into potatoes for accumulation of lignans in transgenic potatoes. We made binary vectors overexpressing dirigent protein gene and P/L reductase gene driven by a CaMV35S promoter and transformed into potatoes via Agrobacterium mediated transformation. And in order to control the metabolic flux of lignan biosynthesis pathway, we tried to inhibit chalcone synthase genes of potatoes by antisense inhibition technique also. We tried to use PCR screening method for selection of transgenic plants of different vectors. We tried to determine and compare lignan contents from different transgenic potato lines.

  • PDF

Suppression of the ER-Localized AAA ATPase NgCDC48 Inhibits Tobacco Growth and Development

  • Bae, Hansol;Choi, Soo Min;Yang, Seong Wook;Pai, Hyun-Sook;Kim, Woo Taek
    • Molecules and Cells
    • /
    • 제28권1호
    • /
    • pp.57-65
    • /
    • 2009
  • CDC48 is a member of the AAA ATPase superfamily. Yeast CDC48 and its mammalian homolog p97 are implicated in diverse cellular processes, including mitosis, membrane fusion, and ubiquitin-dependent protein degradation. However, the cellular functions of plant CDC48 proteins are largely unknown. In the present study, we performed virus-induced gene silencing (VIGS) screening and found that silencing of a gene encoding a tobacco CDC48 homolog, NgCDC48, resulted in severe abnormalities in leaf and shoot development in tobacco. Furthermore, transgenic tobacco plants (35S:anti-NgCDC48), in which the NgCDC48 gene was suppressed using the antisense RNA method, exhibited severely aberrant development of both vegetative and reproductive organs, resulting in arrested shoot and leaf growth and sterile flowers. Approximately 57-83% of 35S:anti-NgCDC48 plants failed to develop mature organs and died at early stage of development. Scanning electron microscopy showed that both adaxial and abaxial epidermal pavement cells in antisense transgenic leaves were significantly smaller and more numerous than those in wild type leaves. These results indicate that NgCDC48 is critically involved in cell growth and development of tobacco plants. An in vivo targeting experiment revealed that NgCDC48 resides in the endoplasmic reticulum (ER) in tobacco protoplasts. We consider the tantalizing possibility that CDC48-mediated degradation of an as-yet unidentified protein(s) in the ER might be a critical step for cell growth and expansion in tobacco leaves.

PRK1, a Receptor-like Kinase from Petunia inflata, is Essential for Post-meiotic Development of Pollen and Embryo Sac

  • Pai, Hyun-Sook;Karunanandaa, Balasulojini;Gilroy, Simon;Kao, Teh-Hui
    • 한국식물학회:학술대회논문집
    • /
    • 한국식물학회 1996년도 제10회 식물생명공학심포지움 고등식물 발생생물학의 최근 진보
    • /
    • pp.48-60
    • /
    • 1996
  • We previously identified and characterized a predominantly pollen-expressed gene of Petunia inflata that encodes a receptor-like kinase named PRK1. The extracellular domain of PRK1 contains leucine-rich repeats which have been implicated in protein-protein interactions, and the cytoplasmic domain was found to autophosphorylate on serine and tyrosine. To investigate the function PRK1 in pollen development, we transformed P. inflata plants with a construct containing the promoter of a predominantly pollen-expressed gene of tomato, LAT52, fused to an antisense PRK1 cDNA corresponding to part of the extracellular domain of PRK1, There transgenic plants were found to each produce approximately equal amounts of normal and aborted pollen. Analysis of the inheritance of the transgene inserts in two of the transgenic plants, ASRK-13 and ASRK-20, to their progeny revealed that certain transgene inserts cosegregated with the pollen abortion phenotype. Microscopic examination of the aborted pollen grains showed that their outer wall, the exine, was essentially normal, but that their cytoplasm contained only starch-like granules. Staining of the nuclei of the microspores at different stages of uninucleate stage. However, at subsequent stages half of the microspores completed mitosis and developed into normal binucleate pollen, but the other half initially remained uninucleate, then lost their nucleio. Analysis of the amounts of PRK1 mRNA and the antisense PRK1 transcript suggested that the pollen abortion phenotype most likely resulted from down-regulation of the PRK1 gene by the antisense PRK1 transgene. These results suggest that PRK1 plays an essential role in a signal transduction pathway that mediates post-meiotic development of microspores.

  • PDF

웅성불임 유전자의 발현억제를 이용한 임성회복 (Restoration of Fertility by Suppression of Male Sterility- Induced Gene Using an Antisense Construct)

  • 박영두;박범석;김현욱;진용문
    • 원예과학기술지
    • /
    • 제17권4호
    • /
    • pp.473-475
    • /
    • 1999
  • 본 연구는 antisense 유전자를 이용하여 웅성불임 식물체를 유기하는 웅성불임 유기 유전자의 발현을 억제함으로써 임성을 회복하기 위하여 실시하였다. 약특이 promoter(GBAN215-6 promoter)와 antisense 방향의 diphtheria toxin(DTx-A) 유전자로 제작된 pKDA215b로 담배(cv. petit Havana SR1)를 형질전환시키고 형질전환이 확인된 76개의 $R_0$ 세대를 자가수분하여 $R_1$ 세대를 획득하였다. $R_1$ 세대의 유전 분석후 antisene 유전자가 복수로 존재하는 5개의 $R_1$ 계통 (21505, 21507, 21511, 21522, 21525)을 선발하고 동형접합체를 획득하기 위해 자가수분을 하였다. 임성회복을 유도하고 회복친을 선발하기 위하여 antisense 유전자를 가진 $R_2$ 계통과 웅성불임 식물체를 교배하였다. 그 결과 꽃가루가 완전이 회복된 개체, 부분적으로 회복된 개체, 회복되지 않은 개체 등 3종류의 식물체를 획득하였으며 이들 식물체의 화분 이외의 표현형은 정상식물체와 같았고 그 기작 구명을 위해 선발하고 계속 연구가 진행되고 있다.

  • PDF

Expression of dirigent protein and Pinoresinol/Lariciresinol reductase genes of forsythia in transgenic potatoes.

  • Chuong, Tran-Van;Kim, Hyunsoon;Park, Jiyoung;Jaeyoul Joung;Jungwon Youm;Jaeheung Jeon
    • 한국자원식물학회:학술대회논문집
    • /
    • 한국자원식물학회 2001년도 The 8th International Symposium
    • /
    • pp.49-58
    • /
    • 2001
  • We tried to introduce two forsythia genes related in lignan biosynthesis, dirigent protein and pinoresinol/lariciresinol (P/L) reductase, into potatoes for accumulation of lignans in transgenic potatoes. We made binary vectors overexpressing dirigent protein gene and P/L reductase gene driven by a CaMV35S promoter and transformed into potatoes via Agrobacterium mediated transformation. And in order to control the metabolic flux of lignan biosynthesis pathway, we tried to inhibit chalcone synthase genes of potatoes by antisense inhibition technique also. We tried to use PCR screening method for selection of transgenic plants of different vectors. We tried to determine and compare lignan contents from different transgenic potato lines.

  • PDF

Characterization of a Tomato (Lycopersicon esculentum Mill.) Ripening-associated Membrane Protein (TRAMP) Gene Expression and Flavour Volatile Changes in TRAMP Transgenic Plants

  • Kim Seog-Hyung;Ji Hee-Chung;Lim Ki-Byung
    • Journal of Plant Biotechnology
    • /
    • 제7권2호
    • /
    • pp.87-95
    • /
    • 2005
  • The tomato ripening associated membrane protein (TRAMP) (Fray et al., 1994) is a member of the major intrinsic protein (MIP) family, defined as channels facilitating the passage of water and small solutes through membranes. During normal fruit ripening the TRAMP mRNA levels were increased whereas the expression levels of TRAMP in low ethylene ACO1-sense suppressed lines, Nr and rin fruits, were lower than at the breaker stage of wild type fruit. TRAMP mRNA is inhibited by $LaCl_3$, which is an inhibitor of $Ca^{2+}$-stimulated responses, treatment but drought condition did not affect TRAMP expression. The levels of TRAMP mRNA transcripts were substantially higher in the dark treated seedlings and fruits. These suggest that TRAMP function as a water channel may be doubted because of several reasons; no water content was changed during ripening in wild type, antisense and overexpression lines, TRAMP expression under light condition was lower than dark condition and TRAMP expression was not changed in drought condition. Co-suppression plant, 3588 was one of sense suppression lines, which contain CaMV 35S promoter and sense pNY507 cDNA, produced small antisense RNA, approximately 21-25 nucleotides in length, mediated post-transcriptional gene silencing. Therefore, TRAMP expression was inhibited by small antisense and multiple copies might induce gene silencing without any production of double strand RNA. Total seven selected volatile productions, isobutylthiazole, 6-methyl-5-hepten-2-one, hexanal, hexenal methylbutanal, hexenol, and methylbutanol, were highly reduced in sense line whereas total volatile production was increased in TRAMP antisense line. These results suggested TRAMP might change volatile related compounds.