Plants having improved tolerance to various types of environmental stress, their production, and polyamine metabolism-related enzyme gene

a technology of environmental stress and plant-derived polyamine metabolism, which is applied in the direction of enzymology, biochemistry apparatus and processes, transferases, etc., can solve the problems of no reports on plant-derived polyamine metabolism-related enzyme genes whose expression is induced, and achieve the effects of preventing damage, improving various types of environmental stress tolerance in plants, and reducing the number of plant phenotypes

Inactive Publication Date: 2008-01-10
TOYOBO CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0027] As a result of extensive research to achieve the aforementioned objects, the inventors discovered that various environmental stress tolerance parameters are improved when polyamine metabolism-related enzyme genes involved in polyamine biosynthesis, whose level of expression changes specifically upon exposure to cold stress, are isolated and are introduced and overexpressed in plants so as to bring about changes in the polyamine concentration through the activation of polyamine metabolism.
[0189] The present invention can improve various types of environmental stress tolerance in plants, can prevent damage or alleviate limited growth caused by various types of environmental stress which plants may encounter during their growth, and can be expected to result in more stable cultivation, improve productivity, expand areas of cultivation, and the like.

Problems solved by technology

However, there have been no reports on plant-derived polyamine metabolism-related enzyme genes whose expression is induced and whose level of expression increases upon exposure to cold stress in plant tissue exhibiting cold stress tolerance.

Method used

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  • Plants having improved tolerance to various types of environmental stress, their production, and polyamine metabolism-related enzyme gene
  • Plants having improved tolerance to various types of environmental stress, their production, and polyamine metabolism-related enzyme gene
  • Plants having improved tolerance to various types of environmental stress, their production, and polyamine metabolism-related enzyme gene

Examples

Experimental program
Comparison scheme
Effect test

example 1

Measurement of Polyamine Content in Roots of Cucumbers and Cucurbita Ficifolia Bouche

[0191] (1) Preparation of Samples

[0192]Cucurbita ficifolia Bouche having high cold stress resistance in the roots and cucumber “Suyo” having poor cold stress resistance were planted in glass rooms, and were transplanted to pots filled with commercially available fine soil (Sansan soil, by Takii & Co., Ltd.) at the cotyledon development stage. At the first leaf stage, the plants were placed in artificial ventilation rooms (air temperature: day 26° C. / night 20° C.; relative humidity: day 70% / night 85%; light intensity: 480 μM / m2s; 15 hour long day). Nine each were planted in two cultivation tanks (½-fold Hoagland's solution 1201; night temperature: 23° C.).

[0193] (2) Low Temperature Treatment

[0194] 4 days after planting, the live weight of each was determined, they were replanted, and the night temperature of one cultivation tank was lowered to 14° C.

[0195] (3) Sampling

[0196] Following the low t...

example 2

Cloning of Plant-Induced Polyamine Metabolism-Related Enzyme Gene

[0205] (1) Preparation of Poly(A)+RNA

[0206] Vermiculite was inoculated with Cucurbita ficifolia Bouche, and the plants were transplanted to pots filled with commercially available fine soil (Sansan soil, by Takii & Co., Ltd.) at the cotyledon development stage. The potted Cucurbita ficifolia Bouche was placed in incubators (air temperature: day 26° C. / night 22° C., 13 hour long days) for plant cultivation. At the two leaf stage, the incubator temperature was lowered to day 18° C. / night 14° C. to begin low temperature treatment. After 3 days of low temperature treatment, the plants were divided into roots, stems, and leaves for sampling. The samples were stored in an −80° C. freeze until RNA extraction.

[0207] About 4 g of Cucurbita ficifolia Bouche root tissue was immediately frozen in liquid nitrogen and finely milled in a mortar and pestle in the presence of liquid nitrogen. 10 mL of 0.2 M Tris acetic acid buffer (...

example 3

Preparation of Transgenic Arabidopsis thaliana

[0247] (1) Preparation of Expression Construct

[0248] The FSPD1 polyamine metabolism-related gene given in SEQ ID NO. 1 was cleaved with XhoI in such a way that the entire reading frame of the base sequence was included, and the fragment was purified by the glass milk method. pGEM-7Zf (Promega) was then cleaved with XhoI, and the FSPD1 fragments were subcloned in the sense and antisense directions. The FSPD1 fragments were again cleaved with the XbaI and KpnI restriction enzymes at the multicloning site of pGEM-7Zf, and were subcloned in the sense and antisense direction to the binary vector pBI101-Hm2 to which the 35S promoter had been ligated. The resulting plasmid was designated pBI35S-FSPD1. The structure of this expression construct is given in FIG. 8. Transformed E. coli JM109 was designated Escherichia coli JM109 / pBI35S-FSPD1.

[0249] The polyamine metabolism-related gene FSAM24 given in SEQ ID NO. 3 was cleaved with NotI in such ...

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Abstract

The present invention relates to isolated plant-derived polyamine metabolism-related enzyme genes characterized in that their level of expression changes when exposed to environmental stress; antisense DNA or antisense RNA to such genes; plants and their progeny having improved environmental stress tolerance, in which such genes are retained in a stable manner; methods for producing such plants, and methods for producing calli of such plants; method for selecting transformed plants which grow improved more than plants lacking an exogenous polyamine metabolism-related enzyme gene; and microorganims and plants which are transformed by such genes

Description

TECHNICAL FIELD [0001] The present invention relates to plants having improved environmental stress tolerance, and in particular to plants having improved cold stress tolerance, salt stress tolerance, herbicide stress tolerance, drought stress tolerance, and osmotic stress tolerance. The present invention further relates a method of producing such plants. BACKGROUND ART [0002] Plants adapt to various types of environmental stress such as the temperature and salt of their habitats. However, in terms of temperature stress, for example, plants are susceptible to hot or cold temperatures when exposed to environments over or under the maximum or minimum optimum growth temperature, leading to impairment upon the gradual or sudden loss of the physiological functions of cells. Efforts have been made to expand the temperature adaptability of plants by breeding means such as selection or cross breeding in order to make use of wild plants adapted to various temperature environments for food cr...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A01H1/00A01H5/00C12N1/15C12N1/19C12N1/21C12N9/10C12N15/60C12N15/82
CPCC12N15/8271C12N9/1085
Inventor KASUKABE, YOSHIHISAIHARA, IZUMITACHIBANA, SHOJI
Owner TOYOBO CO LTD
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