Transformation of a very chilling-sensitive orchid, "Phalenopusis" with Agrobacreium carrying spinach fatty acid desaturase

用携带菠菜脂肪酸去饱和酶的农杆菌转化一种对寒冷非常敏感的兰花“蝴蝶兰”

• 批准号: 03806005
• 负责人: MATSUO Tomoaki
• 金额: $ 1.54万
• 依托单位: Kagoshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-03806005/
• 关键词: Chilling-resistant plant; Transformation; Chilling injury; Stearoyl ACP desaturase; Agrobacterium tumefaciens; Orchid, Phalenopusis; 低温傷害(障害); 形質転換植物; 低温耐性; ラン; 植物形質転換; 不飽和化酵素

Temperature is a major environmental factor governing the distribution of both wild and cultivated plant species. Many economically important plants having tropical or subtropical origins, for example maize, banana, citrus and squash, are injured by exposure to temperatures slightly above the freezing point of their tissues. Lyons and Raison have proposed that chilling injury is primarily caused by the physical response of membrane lipids to low temperature. Recently, Murata proposed that, in chloroplasts of higher plants, phosphatidylglycerol (PG) might be the molecule resposible for the primary event in chilling injury by virtue of its content of molecular species containing saturated fatty acids.Properties of two enzymes, Glycerol-3-phosphate acyltransferase and Stearoyl ACP desaturase (SAD), have been recognized to be very important on synthesis of unsaturated PG species. SAD introduces the first double bond into C18 fatty acids in higher plants and, threfore, it is involved in cotrolling the extent of unsaturation of membrane lipids. We have reported the isolation and characterization of a cDNA clone that encodes a precursor to SAD of spinach young leaf, chilling resistant plant. Then, we tried transformation of chilling sensitive plants, petunia and Phalenopusis, with Agrobacterium tumefaciens carrying pBl 121/SAD which was inserted the spinach cDNA clone. Several transformants of petunia obtained thus have been continued to compare the fatty acid composition of PG and the chilling sensitivity. No transformant of Phalenopusis is obtained at present time.

温度是决定野生和栽培植物物种分布的主要环境因素。许多具有重要经济意义的热带或亚热带植物，例如玉米、香蕉、柑橘和南瓜，由于暴露在略高于其组织冰点的温度下而受到伤害。Lyons和Raison提出低温损伤主要是由膜脂对低温的物理反应引起的。最近，Murata提出，在高等植物叶绿体中，磷脂酰甘油（PG）可能是负责冷害主要事件的分子，因为它含有饱和脂肪酸的分子种类。甘油-3-磷酸酰基转移酶（Glycerol-3-phosphate acyltransferase）和硬脂酰ACP去饱和酶（Stearoyl ACP desaturase， SAD）的性质对不饱和PG的合成有重要影响。SAD在高等植物中引入C18脂肪酸的第一个双键，因此，它参与控制膜脂的不饱和程度。我们已经报道了一个编码菠菜幼叶SAD前体的cDNA克隆的分离和鉴定。然后，我们尝试将携带pBl 121/SAD的农杆菌插入菠菜cDNA克隆，转化冷敏植物矮牵牛和蝴蝶兰。由此获得的几个矮牵牛花转化体已被继续比较PG的脂肪酸组成和冷却敏感性。目前还没有获得蝴蝶兰的转化。

项目成果

I.Nishida,T.Beppu,T.Matsuo,N.Murata: "Nucleotide sequence of a cDNA clone encoding a precursor to stearoylー(ecylーcarrienーprotein)desaatnrase from spinach"

I.Nishida、T.Beppu、T.Matsuo、N.Murata：“编码菠菜硬脂酰－（ecyl－载体－蛋白）脱氨酶前体的 cDNA 克隆的核苷酸序列”

Nishida I.,et al.: "Nucleotide seguence of a cDNA clone oncuding a precursor to stearoyl-Cacyl-carrier-protein)desaturase from spinach" Plant Molecular Biology. 19. 711-713 (1992)

Nishida I.,et al.：“来自菠菜的硬脂酰基-载体蛋白)去饱和酶前体的 cDNA 克隆的核苷酸序列”植物分子生物学。