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.

我们筛选了在adiantum质子细胞中光质和叶绿体光方向运动缺陷的突变体。突变体是由EMS或氮的重离子束诱导的。由重离子束辐照获得的突变体缺乏脂肪含量3，它是N末端处的植物色素发色团结合结构域的嵌合基因，几乎全长的拟南芥nph1at c-terminus，这表明两种生理现象都是由phy3.3.33.的表达模式介导的五个Adiantunmum cryoct crypomes crypoct of phy3.3. CRY1-3在在配子体和孢子体中测试的所有发育阶段表达。 Cry4主要在孢子和深色生长的孢子体中表达。 Cry 5主要是在光明和黑暗中生长的孢子菌。竞争性PCR表明，当用红光照射孢子以诱导孢子发芽时，CRY4受到下调，Cry5被上调。洋葱表皮细胞中加密色素和GUS融合蛋白的表达模式表明，在黑暗中，Cry3和4转移到核中。通过两种杂交方法筛选了Phy3结合蛋白的基因的can剂。现在，这些基因的表征正在进行中。

项目成果

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》。

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。

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。

T.Kanegae: "Isolation and characterization of homologues of plant blue-lighit photorecaptor (cryptochrome) genes from fern Adiantum capillus-veneris." Molecular Genes and Genetics. 259. 345-353 (1998)

T.Kanegae：“从蕨类铁线蕨中分离和鉴定植物蓝光光感受器（隐花色素）基因的同源物。”

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）。