Effect of activity of the cell wall and vacuole invertase on the sugar accumulation in the strawberry fruit

细胞壁和液泡转化酶活性对草莓果实糖积累的影响

• 批准号: 11660024
• 负责人: OGIWARA Isao
• 金额: $ 2.3万
• 依托单位: Tokyo University of Agriculture and Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11660024/
• 关键词: Strawberry; Sugar accumulation; Invertase; Cloning; Tissue culture

In order to clarify the accumulation mechanism of the sugar in strawberry fruits, the relation between the sugar content in the fruit and inyertase activity was investigated, and isolation and analysis of the invertase gene were carried out. Also the regeneration of shoot from the leaf culture was examined.1. Though glucose and fructose content of 'Reiko' did not increase with the maturation of the fruit, the sucrose content increased at the later stage in the fruit maturation. In 'Toyonoka', glucose and fructose content increased from the early stage, and these sugars continue to increase afterwards.2. The cell wall invertase activity did not differ between the glucose and fructose accumulated 'Toyonoka' variety **d the sucrose accumulated 'Reiko' variety. Whereas the change in the activity of the sucrose accumulated 'Reiko' variety was small, though the activity of the glucose and' fructose accumulated 'Toyonoka' variety, increased, and the tendency was similar to the change of the reducing sugar.3. When the cloning of an invertase gene was performed, two kinds of gene fragments (inv 1 and inv 2) were isolated. From the result of the homogeneous reference, inv 1 was a part of vacuole type invertase mRNA (520bp), and inv 2 was a part of cell wall type in Invertase mRNA (471bp). From the result of western blotting using the antibody made on the basis of inv 1, it was shown that vacuole invertase of the glucose and fructose accumulated 'Toyonoka' variety had mainly appeared at early stage in the fruit maturation.1 As controlling factor the sugar accumulation of the strawberry, the activity of the vacuole type invertase seemed to be greatly concerned.4. When the piece of a leaf was put on 1/2MS medium, which added 2.4D, CPPU, or TDZ and was kept under the dark and 25℃ condition, the rate of the shot formation in 'Nyohou' and 'Reiko' became 60% or more, but the rate of that in ' Toyonoka' was only 5% on this condition.

为了阐明草莓果实中糖的积累机制，研究了草莓果实中糖含量与转化酶活性的关系，并对转化酶基因进行了分离和分析。并对叶片培养的不定芽再生进行了研究.葡萄糖和果糖含量不随果实成熟而增加，但蔗糖含量在果实成熟后期逐渐增加。丰冈品种葡萄糖和果糖的含量从前期开始增加，以后继续增加.积累葡萄糖和果糖的丰香品种和积累蔗糖的玲子品种的细胞壁转化酶活性没有差异。而积累蔗糖的“玲子”品种活性变化较小，积累葡萄糖和“果糖”的“丰香”品种活性升高，其变化趋势与还原糖的变化趋势相似.当进行转化酶基因的克隆时，分离出两种基因片段（inv 1和inv 2）。从同源对照的结果来看，inv 1为液泡型转化酶mRNA的一部分（520 bp），inv 2为细胞壁型转化酶mRNA的一部分（471 bp）。利用在inv 1基础上制备的抗体进行的western blotting结果表明，积累葡萄糖和果糖的丰香品种的液泡型转化酶主要出现在果实成熟的早期。1.作为草莓糖积累的控制因素，液泡型转化酶的活性似乎受到极大的关注。在1/2 MS培养基中添加2.4D、CPPU或TDZ，在25℃避光条件下，'牛宝'和'玲子'的芽形成率可达60%以上，而'丰香'的芽形成率仅为5%。

项目成果

Ogiwara, I. , Y. Ohtsuka, Y. Yoneda, K. Sakurai, N. Hakoda, I. Shiraura: "Extraction method by water followed by microwave heating for analyzing sugars in strawberry fruits"J. Japan. Soc. Hart. Sci.. 68. 949-953 (1999)

Ogiwara, I., Y. Ohtsuka, Y. Yoneda, K. Sakurai, N. Hakoda, I. Shiraura：“水提取法和微波加热分析草莓果实中的糖分”J.

I.Ogiwara: "Extraction method by water followed by microwave heating for analyzing sugars in strawberry fruits"Journal of the Japanese Society for Horticultural Science. Vol.68.N0.5. 949-953 (1999)

I.Ogiwara：“草莓果实中糖分的水提取法和微波加热分析法”日本园艺学会杂志。

萩原勲 他3名: "イチゴ促成栽培における低糖含量果実の発生要因について"園芸学会雑誌. 68. 130-137 (1999)

Isao Hagiwara 等 3 人：“草莓强制栽培中低糖水果发育的因素”日本园艺学会杂志 68. 130-137 (1999)。

Ogiwara 他3名: "Extraction method by water followed by microwave heating for analyzing sugars in strawberry fruits"J. Japan. Soc. Hort. Sci.. 68. 949-953 (1999)

Ogiwara 等 3 人：“草莓果实中糖分的水提取方法”J. Soc. 68. 949-953 (1999)。

Ogiwara, I., M. Shiraishi, N. Hakoda, I. Shimura: "Causes of low sugar levels in forced strawberry fruits"J. Japan. Soc. Hart. Sci.. 68. 130-137 (1999)

Ogiwara, I., M. Shiraishi, N. Hakoda, I. Shimura：“强制草莓果实中糖分含量低的原因”J.