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.

为了阐明光诱导矮化的机制，采用差异显示法寻找矮化豌豆植株中涉及矮化的特异基因。 Poly(A)^+ RNA 是从光照生长的矮豌豆 (Pisum sativum L.cv. Progress No 9) 和正常豌豆 (cv. Alaska) 幼苗以及两个品种的潮湿生长幼苗中分离出来的。 Poly(A)^+ RNA反转录后，利用72种随机引物，采用差异展示法鉴定特定基因。鉴定出 20 个差异表达带，并选择其中 7 个进行进一步表征。这七个条带被克隆到质粒载体中，确定它们的核苷酸序列并在GenBank中搜索它们的同源物。使用克隆的编码序列作为探针，通过高严格性的Northern印迹进一步表征克隆。结果表明，1个克隆在光生矮豌豆幼苗中特异表达，1个克隆在除光生矮豌豆幼苗外的所有幼苗中表达。因此，这两个新基因克隆可能与光诱导矮化豌豆植株的矮化有关。