Search for photoreceptor-binding proteins mediating photomorphogenesis and their localization

寻找介导光形态发生的光感受器结合蛋白及其定位

• 批准号: 09440270
• 负责人: WADA Masamitsu
• 金额: $ 7.81万
• 依托单位: TOKYO METROPOLITAN UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09440270/
• 关键词: photomorphogenesis; photoreceptor; phytochrome; Binding protein; Fern; 青色光吸収色素; シダ植物

We screened mutants which are in defect of phototropism and chloroplast photoorientation movement in Adiantum protonemal cells. The mutants were induced by EMS or heavy ion beam of nitrogen. One of the mutants obtained by heavy ion beam irradiation lack AdiPHY3, which is a chimeric gene of phytochrome chromophore binding domain at N-terminus and almost full length of Arabidopsis NPH1at C-terminus, suggesting that the both physiological phenomena are mediated by PHY3.Expression pattern of five Adiantum cryptochromes were studied. CRY1-3 are expressed in all developmental stages tested both in gametophytes and in sporophytes. CRY4 is mainly expressed in spores and dark-grown sporophytes. CRY 5 is mainly in sporophytes grown both in the light and darkness. Competitive PCR shows that CRY4 is down- regulated and CRY5 is up-regulated when spores were irradiated with red light for the induction of spore germination. The expression pattern of fusion proteins of cryptochrome and GUS in onion epidermal cells suggests that CRY3 and 4 are transferred into nuclei in the dark.Candidates of genes of PHY3-binding proteins were screened by two-hybrid method. Now characterization of those genes are underway.

我们在铁线蕨原丝体细胞中筛选出向光性和叶绿体光定向运动缺陷的突变体。诱变剂为EMS或重离子氮束。其中一个重离子束诱变突变体的N端缺少光敏色素结合域基因AdiPHY3，而C端缺少拟南芥NPH 1的几乎全长基因AdiPHY3，表明这两种生理现象都是由PHY3介导的。在配子体和孢子体的所有发育阶段都有表达。EST4主要在孢子和黑暗生长的孢子体中表达。CRY 5主要存在于光照和黑暗条件下生长的孢子体中。竞争性PCR结果表明，红光照射诱导孢子萌发时，E14基因表达下调，E15基因表达上调。隐花色素与GUS融合蛋白在洋葱表皮细胞中的表达模式表明，PHY3和PHY4在黑暗条件下转入细胞核。目前，对这些基因的研究正在进行中。

项目成果

Nozue, K., T.Kanegae, T.Imaizumi, S.Fukuda, H.Okamoto, K.-C.Yeh, J.C.Lagarias and M.Wada: "A phytochorme from the fern Adiantum with features of the putative photoreceptor NPH1." Proc.Natl.Acad.Sci.USA. 95. 15826-15830 (1998)

Nozue, K.、T.Kanegae、T.Imaizumi、S.Fukuda、H.Okamoto、K.-C.Yeh、J.C.Lagarias 和 M.Wada：“一种来自铁线蕨的植物色素，具有假定的光感受器 NPH1 的特征。

K.Nozue: "Isdation of a second : phytochrome cDNA from Adintum capillus-veneris" Plant Physiology. 118. 711 (1998)

K.Nozue：“第二个的确定：来自 Adintum capillus-veneris 的光敏色素 cDNA”植物生理学。

Kadota, A.and M.Wada: "Red light-aphototropic (rap) mutants lack red light-induced chloroplast relocation movement in the fern Adiantum capillus-veneris."Plant Cell Physiol.. 40. 238-247 (1999)

Kadota, A. 和 M.Wada：“红光反光性 (rap) 突变体在蕨类铁线蕨中缺乏红光诱导的叶绿体重新定位运动。”植物细胞生理学.. 40. 238-247 (1999)

T.Kagawa: "Chloroplast-avoidance response induced by high-fluence blue light in prothallial cells of the fern Adiantum capilhes-veneris as analyzed by microbeam irradiation" Plant Physiology. 119. 917-923 (1999)

T.Kakawa：“通过微束照射分析铁线蕨原叶体细胞中高通量蓝光诱导的叶绿体回避反应”《植物生理学》。

Adiantum capillus-veneris (Accession NO.AB016232). Kagawa, T.and M.Wada: "Chloroplast avoidance response induced by high fluence rate of blue light in prothallial cells of fern Adiantum as analyzed by microbeam irradiation." Plant Physiol.119. 917-923 (19

Adiantum capillus-veneris（登记号AB016232）。