MOLECULAR BIOLOGICAL STUDY ON THE MECHANISM OF LIGHTINDUCED DWARFISM (LIGHT-INDUCED GENE EXPRESSION IN DWARF PEA PLANTS)

光致矮化机制的分子生物学研究（矮化豌豆植株光致基因表达）

• 批准号: 09660353
• 负责人: KATO Hisashi
• 金额: $ 1.73万
• 依托单位: KAGAWA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 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-09660353/
• 关键词: Dwarf gene; Light-induced growth inhibition; Dwarf pea; Transgenic plant; Differential display method; RT-PCR; mRNA; Molecular biology

In order to clarify the mechanism of light-induced dwarf ism, a search for specific gene involved the dwarfism in dwarf pea plants was undertaken using differential display method. Poly(A)^+ RNA was isolated from light-grown dwarf pea (Pisum sativum L.cv. Progress No 9) and normal pea (cv. Alaska) seedlings and dank-grown seedlings of both cultivars. After reverse transcription of the Poly(A) ^+ RNA, the differential display method was employed to identify specific genes using 72 kinds of random primers. Differentially expressed 20 bands were identified and seven of them were chosen for further characterization. These seven bands were cloned into plasmid vectors and their nucleotide sequences were determined and searched for their homologues in GenBank. Using the cloned coding sequence as probes, the clones were characterized further by Northern blotting at high stringency. The result shown that one clone was specifically expressed in the light-grown dwarf pea seedlings and one was expressed in all seedlings expect the light-grown dwarf pea seedlings. Thus, the two clones, which are novel genes, may be involved in the light-induced dwarfism of the dwarf pea plants.

为了明确光致矮化的发生机制，采用差异展示法对矮化豌豆植株进行了特异基因的研究。聚(A)^+ RNA是从轻矮豆（Pisum sativum L.cv）中分离得到的。进展9号)和普通豌豆(cv。阿拉斯加)两种品种的幼苗和潮湿生长的幼苗。将Poly(A) ^+ RNA逆转录后，采用差异展示法，利用72种随机引物对特定基因进行鉴定。鉴定出差异表达的20条条带，选择其中7条进行进一步鉴定。将这7条条带克隆到质粒载体上，测定其核苷酸序列，并在GenBank中查找其同源物。以克隆的编码序列为探针，利用Northern blotting对克隆进行高强度鉴定。结果表明，1个克隆在轻矮豆苗中特异表达，1个克隆在除轻矮豆苗外的所有苗中均有表达。因此，这两个新基因克隆可能参与了矮豌豆植物的光致矮化。