A New approach to the practical use of insect-tolerant lines of sugar beet via foreign gene transfer

通过外源基因转移实际应用耐虫甜菜品系的新方法

• 批准号: 11356001
• 负责人: KANAZAWA Akira
• 金额: $ 22.73万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11356001/
• 关键词: sugar beet; insect tolerance; cabbage armyworm; transformation; cry; bioassay; chloroplast; ヨトウガ幼虫; 形質転換; 殺虫性タンパク; cryIC遺伝子; 葉緑体工学; 器官特異性; トランジットペプチド

The larvae of cabbage armyworm (Mamestra brassicae L.) bring heavy damage on the final yield of beet-roots and subsequently, sugar production, by eating stems and leaves of sugar beet plants. In order to establish sugar beet cultivars that are tolerant to insects including cabbage armyworm, we have been developing transgenic sugar beet lines harboring cryIA(b), a gene encoding insecticide protein from Bacillus thuringiensis. In the present study, based on the results so far obtained, we tried to make transgenic sugar beet lines that show increased insecticide activity aiming at practical use of the transgenic lines. First, we introduced cryIC into sugar beet plants and analyzed the effect of its expression. We chose cryIC from various candidates of insecticide protein genes, since its gene product showed highest insecticide activity against cabbage armyworm among various insecticide proteins that were obtained by expressing cloned genes in E. coli. Meanwhile, reports were published that described that the introduction of insecticide protein genes into chloroplast gave increased accumulation of the protein and consequently, gave increased insecticide activity of plants. We, therefore, developed vectors available for chloroplast transformation and selection system of transformed chloroplast in order to introduce insecticide protein genes into sugar beet chloroplasts. We also succeeded in introducing several genes practically useful for sugar beet production and evaluated the effect of gene expression. In addition, we analyzed the stability of the expression of foreign genes in plant cells in order to make sure their experssion.

甘蓝夜蛾（Mamestra brassicae L.）通过食用糖用甜菜植物的茎和叶，对甜菜根的最终产量以及随后的糖生产造成严重损害。为了建立对包括甘蓝夜蛾在内的昆虫具有耐受性的甜菜品种，我们已经开发了携带cryIA（B）的转基因甜菜品系，cryIA（B）是编码来自苏云金芽孢杆菌的杀虫剂蛋白的基因。在本研究中，基于迄今为止所获得的结果，我们试图使转基因甜菜品系显示出增加的杀虫剂活性，旨在实际使用的转基因品系。首先，我们将cryIC基因导入甜菜植株中，并分析其表达效果。我们从多种候选杀虫蛋白基因中选择了cryIC，因为其基因产物在通过在大肠杆菌中表达克隆基因获得的多种杀虫蛋白中对甘蓝粘虫显示出最高的杀虫活性。杆菌同时，有报道称，将杀虫剂蛋白质基因导入叶绿体，增加了蛋白质的积累，从而增加了植物的杀虫剂活性。因此，我们开发了适合于叶绿体转化的载体和转化叶绿体的筛选系统，以便将杀虫蛋白基因导入甜菜叶绿体。我们还成功地引入了几个对甜菜生产实际有用的基因，并评估了基因表达的效果。此外，我们还对外源基因在植物细胞中表达的稳定性进行了分析，以确定外源基因在植物细胞中的表达。

项目成果

Shimamoto Y.: "Resistance to larvae of cabbage armyworm (Mamestra brassicae L.) in ICP gene transformant in sugarbeet"Proc. Japan. Soc. Sugar Beet Technol.. 41. 90-98 (1999)

Shimamoto Y.：“甜菜 ICP 基因转化体对甘蓝粘虫 (Mamestra bassicae L.) 幼虫的抗性”Proc.

Kimoto Y.: "Differences in toxicity to larvae of cabbage armyworm between transgenic sugarbeet lines with cryIA(b) and cryIC"Proc. Japan. Soc. Sugar Beet Technol.. 43. 20-23 (2001)

Kimoto Y.：“带有cryIA(b) 和cryIC 的转基因甜菜品系对甘蓝粘虫幼虫的毒性差异”Proc.

木本裕,島本義也: "cryIC導入テンサイのヨトウガ幼虫耐性"てん菜研究会報. 42号(印刷中). (2000)

Yutaka Kimoto、Yoshiya Shimamoto：“cryIC 引入的甜菜对粘虫幼虫的抵抗力”甜菜研究杂志第 42 期（印刷中）。

Ivashuta S: "Genotype-dependent transcriptional activation of novel repetitive elements during cold acclimation of alfalfa (Medicago sativa)"Plant J.. 31. 615-627 (2002)

Ivashuta S：“苜蓿冷驯化过程中新型重复元件的基因型依赖性转录激活”Plant J.. 31. 615-627 (2002)

Kimoto Y.: "Resistance of transgenic sugarbeet with cryIC to larvae of cabbage armyworms"Proc. Japan. Soc. Sugar Beet Technol.. 42. 8-12 (2000)

Kimoto Y.：“带有cryIC的转基因甜菜对甘蓝粘虫幼虫的抗性”Proc。