한국콩연구회

품종 개량을 위한 콩의 dicarboxylater transporter의 기능 분석

Functional analysis of dicarboxylate transporter from soybean for the crop improvement

사업명 : 농업생명공학기술개발

주관연구기관 : 서울대학교

연구책임자 : 안정선

발행년월 : 2008-02

주관부처 : 농촌진흥청

원문

http://www.ndsl.kr/ndsl/commons/util/ndslOriginalView.do?dbt=TRKO&cn=TRKO201100016187&rn=&url=&pageCode=PG18

초록

For the overall understanding of the symbiotic interaction between legumes and Rhizobis, it is important to identify the root nodule- specific genes and investigate their functions. Although it has been suggested that the transporter carrying dicarboxylates from host plants to the symbiotic bacteroid is present on peribacteroid membrane of the nodules, the molecular information on the on the transporter is limited. Therefore, we have isolated a full length cloneCl,755nt) fot the dicarboxylate transporter-like gene from Glycine max, GmDCAT. Deduced amino acid sequence of GmDCA T showed high sequence similarity with that of peptide transporter from Arebidopsis tbsliene, which belongs to the proton-dependenet oligopeptide transporter(POT) family with twelve transmembrane domains and PTR2 domain. However, GmDCAT has only ten transmembrane domains. To analyze the expression pattern of GmDCAT in soybean, we also performed RT- PCR and in situ hybridization. As a result, it was highly expressed in the nodules and specifically in most part of the infected cells, suggesting its role for the symbiotic nitrogen fixation. We are carrying out yeast complementation and patch clamp experiment in Xenopus oocyte expressing GmDCAT to verify which specific substrates GmDCAT protein actually transport. Furthermore, we built the hairy root transformation experiment to verify how GmDCAT affects on nitrogen fixation in nodules. We already constructed a GmDCAT overexpressing vector and a RNAi vector to transform specific soybean lines. Through this hairy root transformation technique, many other researchers would find out functions of specific legumes' genes in plant level. Therefore, we could create transgenic high quality super beans and contribute to food supply.

1. 콩의 dicarboxylate transporter 기능분석

1) 콩의 dicarboxylate transporter 유전자 분리

2) 유전자 발현분석 및 전기생리학적 연구

3) 기질 분석을 포함한 단백질 기능분석

4) 프로모터 분석

2. 작물개량에의 응용

1) Hairy root 형질전환 및 표현형 분석

2) 콩 형질전환체 제작 및 분석