Functional analysis of dwarf genes located in T-DNA of Ri plasmids and its application to plant breeding

Ri质粒T-DNA矮化基因的功能分析及其在植物育种中的应用

• 批准号: 63490005
• 负责人: KAMADA Hiroshi
• 金额: $ 2.75万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1988
• 资助国家: 日本
• 起止时间: 1988 至 1989
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-63490005/
• 关键词: Ri plasmid; Functional analysis; T-DNA; Transgenic plants; Dwarf genes; Gibberellin

It is well known that Cichorium intybus requires low temperature and long day treatments for the flower bub differentiation. However, Cichorium plants transformed with T-DNA of Ri plasmid exhibited flower formation without low temperature treatment under a long day condition. Similar phenomenon of flower bud formation without low temperature treatment was also observed in Cichorium plants transformed with single rol C gene located on the T-DNA of Ri plasmid. When not-transformed (normal) Cichorium plants were treated with some plant growth regulators, such as gibberellin, anti-gibberellin, cytokinin, and ethylene, gibberellin (GA_3) induced the stem elongation, but not the flower bud differentiation. On the other hand, anti-gibberellin (S-3307, a potent inhibitor of gibberellin biosynthesis) induced the flower bud differentiation with low frequency.Moreover, when Cichorium plants transformed with Ri T-DNA were treated with GA_3, flower bud formation could not be observed. These results indicate that a possible physiological function of Ri T-DNA is an inhibition of gibberellin biosynthesis or a suppression of gibberellin activity).Qualitative and quantitative analyses of endogenous gibberellin in Atropa belladonna transformed with Ri T-DNA were carried out by using radio-immunoassay with monoclonal antibodies against some gibberellins. The major gibberellin in the transformed and normal plants was GA_1, and the contents of GA_1 in the transformed plants was less than 1/100 as compared to that of the normal plants. This result suggests that some physiological responses (including dwarfism) observed specifically in plants transformed with Ri T-DNA were induced by the decrease of endosenous gibberellin contents in the transformed plants.

众所周知，菊苣的花芽分化需要低温和长时间的处理。而经Ri质粒T-DNA转化的菊苣在长日照条件下，不经低温处理也能开花。在Ri质粒T-DNA上单个rol - C基因转化的菊苣植株中，也观察到类似的不经低温处理的花芽形成现象。赤霉素、抗赤霉素、细胞分裂素、乙烯等植物生长调节剂对未转化（正常）菊苣的茎伸长有诱导作用，而赤霉素（GA_3）对花芽分化无诱导作用。另一方面，抗赤霉素（S-3307，一种有效的赤霉素合成抑制剂）诱导花芽分化的频率较低。经Ri T-DNA转化的菊苣植株经GA_3处理后，花芽未形成。这些结果表明，Ri T-DNA可能具有抑制赤霉素生物合成或抑制赤霉素活性的生理功能。采用单克隆抗体对部分赤霉素进行放射免疫分析，对经Ri T-DNA转化的颠茄内源赤霉素进行定性和定量分析。转化植株和正常植株的赤霉素主要以GA_1为主，且与正常植株相比，转化植株的GA_1含量不到1/100。这一结果表明，Ri T-DNA转化植株的一些生理反应（包括矮化）是由转化植株内源性赤霉素含量的减少引起的。

项目成果

鎌田博、小野明美: 組織培養. (1989)

Hiroshi Kamata，Akemi Ono：组织培养（1989）。

R.Kato: "Effects of high and very magnetic fields on the growth of hairy roots of Daucus carota and Atropa belladonna." Plant Cell Physiol.30. 605-608 (1989)

R.Kato：“高磁场和强磁场对胡萝卜和颠茄毛状根生长的影响。”

H.Ohkawa: "Effects of gibberellic acid on hairy root growth in Datura innoxia." J.Plant Physiol.134. 633-636 (1989)

H.Ohkawa：“赤霉酸对曼陀罗毛状根生长的影响。”

A.Isogai;N.Fukuchi;M.Hayashi;H.Kamada;H.Harada;A.Suzuki: Agric Biel Chem. 52(12). 3235-3237 (1988)

A.Isogai;N.Fukuchi;M.Hayashi;H.Kamada;H.Harada;A.Suzuki：Agric Biel Chem。

H.Kamada: "Glbberellin Symposium Tokyo 1989(Effenoxia.cts of gibberellic acid on growth and tropane alkaloid synthesis in Ri transformed plant of Datura innoxia.(印刷中)" Springer-Verlag(eds.by N.Takahashi,B.O.Phinney and J.MacMillan),

H. Kamada：“Glbberellin Symposium Tokyo 1989（Effenoxia.cts of赤霉酸对 Ri 转化植物 of Datura innoxia 的生长和托烷生物碱合成。（印刷中）”Springer-Verlag（由 N.Takahashi、B.O.Phinney 和 J 编辑） .麦克米伦），