초록 ▼

High throughput VIGS (Virus-Induced Gene Silencing) 기술을 이용하여 식물발달의 기능유전체 연구를 수행하였다. 10,000개의 Nicotina benthamiana sequence 중에서 식물조직발달, 호르몬 작용, 신호전달, 엽록체발달 둥에 관련된 유전자 795개를 선택하여 pTV00 vector로 cloning 하고 Agrobacterium으로 transformation 한 후 각 유전자 당 6개의 Nicotina henthamiana 식물체를 사용하여 Agrobacterium infilt

High throughput VIGS (Virus-Induced Gene Silencing) 기술을 이용하여 식물발달의 기능유전체 연구를 수행하였다. 10,000개의 Nicotina benthamiana sequence 중에서 식물조직발달, 호르몬 작용, 신호전달, 엽록체발달 둥에 관련된 유전자 795개를 선택하여 pTV00 vector로 cloning 하고 Agrobacterium으로 transformation 한 후 각 유전자 당 6개의 Nicotina henthamiana 식물체를 사용하여 Agrobacterium infiltration으로 VIGS를 수행하였다. Infiltration 후 2-3주 후 식물체의 형질 변화를 control과 비교하여 조사하였다. phenotype은 cell death, leaf yellowing, growth arrest, abnormal organs, premature senescence, flower abnormality, apical dominance breakdown등으로 나누어 분석하였다. 795개 유전자 중 65개 유전자가 VIGS 시 심각한 식물체의 생장 및 발달 변화가 유도되었다. 데이터베이스 구축을 위하여 VIGS로 나타나는 형질에 대한 결과를 축적하고 있다. 이 연구를 통하여 식물발달을 조절하는 새로운 유전자들을 발굴하였고 그들 유전자작용 기작을 탐구하므로서 식물 발달조절의 전체 네트워크를 이해하고자 하였다. 식물발달의 주요한 유전자의 기능을 여러 분자생물학적, 생화학적, 세포생물학적인 테크닉을 이용하여 심층분석하여 세계적 저명저널에 논문으로 발표하였다. 또한 발굴된 기능성유전자는 고부가가치 형질전환 식물 개발에 활용하고자 한다.

Abstract ▼

The purpose of this study is to carry out plant functional genomics using virus-induced gene silencing to understand plant developmental reguatory pathways. We used TRV (tobacco rattle virus)-based VIGS system using Nicotiana benthamiana as a host. Three thousand N. benthamiana cDNAs were random-seq

The purpose of this study is to carry out plant functional genomics using virus-induced gene silencing to understand plant developmental reguatory pathways. We used TRV (tobacco rattle virus)-based VIGS system using Nicotiana benthamiana as a host. Three thousand N. benthamiana cDNAs were random-sequenced, and target genes were selected for VIGS-mediated function analysis, The target genes include plant developmental genes, signal transduction genes, and hormone-related genes. Total 795 cDNAs were cloned into pTV00 (VIGS vector), and the recombinant plasmids were transformed into Agrobacterium. Then Agrobacterium suspension was infiltrated into N. benthamiana plants at 3 weeks after germination. After 2-3 weeks after infiltration, the visible phenotypes appeared in the newly emerged plant tissues. Among 795 cDNAs, 65 cDNAs showed visible morphological phenotypes, including stunted growth, cell death, changes in leaf color, wrinkled leaves, changes in plant architecture, and flower abnormality Thus using VIGS, we identified many novel genes that modulate plant developmental porcesses and characterized their action mechanisms in plant development using various molecular and biochemical tools. We also probed the protein-protein interaction in order to understand the global picture of developmental regulatory networks, The detailed results obtained in this project were summarized below.

목차 Contents

• 제 1 장 연구개발과제의 개요...19
• 제 2 장 국내외 기술개발 현황...21
• 제 3 장 연구개발수행 내용 및 결과...25
• 제 1 절: VIGS를 수향한 Ni. benthamiana clone 들의 list...39
• 제 2 절: Mitochondria-associated hexokinases play a role in the control of programmed cell death in Nicotiana benthamiana...46
• 제 3 절: Silencing of NbBTF3 results in developmental defects and disturbed gene expression in chloroplasts and mitochondria of higher plants....61
• 제 4 절: Depletion of UDP-D-apiose/UDP-D-xylose synthases results in rhamnogalacturonan-II deficiency, cell wall thickening, and cell deathin higher plants....72
• 제 5 절: Prohibitin is involved in mitochondrial biogenesis in plants....81
• 제 6 절: Comparative microarray analysis of programmed cell death induced by proteasome malfunction and hypersensitive response in plants....91
• 제 7 절: Functional characterization of NtCDPK1 in tobacco....99
• 제 8 절: Retinoblastoma protein regulates cell proliferation, differentiation, and endoreduplication in plants....105
• 제 9 절: Inactivation of an organellar glutamyl- and seryl-tRNA synthetase leads to a developmental arrest of chloroplasts and mitochondria in higher plants....116
• 제 10 절: Functional characterization of NtCEF1, an AP2/EREBP-type transcriptional activator highly expressed in tobacco callus....131
• 제 11 절: Silencing of NbNAP1 Encoding a Plastidic SufB-like Protein Affects Chloroplast Development in Nicotiana benthamiana...145
• 제 13 절: DNA gyrase is involved in chloroplast nucleoid partitioning....152
• 제 14 절: The shooty callus induced by suppression of tobacco CHRK1 receptor-like kinase is a phenocopy of the tobacco genetic tumor....170
• 제 15 절: Heterologous expression and molecular and cellular characterization of CaPUB1 encoding a hot pepper U-box E3 ubiquitin ligase homolog....177
• 제 16 절: The CaPRP1 gene encoding a putative proline-rich glycoprotein is highly expressed in rapidly elongating early roots and leaves in hot pepper (Capsicum annuumL. cv. Pukang)....195
• 제 17 절: A split enhanced green fluorescent protein-based reporter in yeast two-hybrid system....201
• 제 4 장 목표달성도 및 관련분야에의 기여도...211
• 제 5 장 연구개발결과의 활용계획...215
• 제 6 장 연구개발과정에서 수집한 해외과학기술정보...215
• 제 7 장 참고문헌...217