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Method for constructing plant dwarfing and early flowering by using CRY1G380R

A plant and cryptochrome technology, applied in the field of agricultural science, can solve problems such as affecting photolyase/CRY protein function

Inactive Publication Date: 2013-07-03
SHANGHAI JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on how the mutation of the conserved G site affects the function of the corresponding photolyase / CRY protein, and what kind of physiological impact it will have on the organism.

Method used

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  • Method for constructing plant dwarfing and early flowering by using CRY1G380R
  • Method for constructing plant dwarfing and early flowering by using CRY1G380R
  • Method for constructing plant dwarfing and early flowering by using CRY1G380R

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Overexpression of CRY1 in Arabidopsis G380R or OsCRY1b G388R Construction of transgenic plants

[0029] The full-length cDNA sequence of Arabidopsis CRY1 amplified by PCR was cloned between the EcoRI and SacI sites of the pBS-Myc vector to obtain pBS-Myc-CRY1. After sequencing, the 380th glycine of CRY1 was mutated to arginine (G380R) by site-directed mutagenesis PCR technology, and the clone pBS-Myc-CRY1 was obtained G380R . Myc-CRY1 G380R The fragment was digested with XhoI and SacI, and inserted behind the 35S promoter on the pKYL71 vector to overexpress Myc-CRY1 G380R plant expression vector ( figure 1 A), using the obtained plasmid to transform the Agrobacterium GV3101 strain.

[0030] Through amino acid sequence alignment, it was found that G388 of rice OsCRY1b corresponds to G380 of Arabidopsis CRY1 (AtCRY1) ( figure 1 B). The full-length cDNA sequence of rice OsCRY1b amplified by PCR was cloned into the pBS vector between HindIII and SacI si...

Embodiment 2

[0032] Example 2: 35S::CRY1 in Arabidopsis G380R and 35S::OsCRY1b G388R Photomorphogenetic phenotyping of transgenic plants by observing 35S::CRY1 G380R and 35S::OsCRY1b G388R Morphogenic phenotype discovery in transgenic plants, 35S::CRY1 G380R and 35S::OsCRY1b G388R The morphogenetic phenotypes of the seedlings of these two transgenic plants were less affected by the light conditions when they were grown under the main light quality conditions of blue light (BL), red light (RL) and far-red light (FR) and dark (DK) , showing that under various conditions, the elongation of the hypocotyls of the seedlings was significantly inhibited, the cotyledons were fully opened and expanded, and the cotyledons and even part of the hypocotyls of the seedlings were obviously red (respectively figure 2 A-2D, image 3 A-3D). Statistical analysis of hypocotyl length revealed that 35S::CRY1 G380R and 35S::OsCRY1b G388R The transgenic plants were significantly dwarf than WT (respectivel...

Embodiment 3

[0033] Example 3: Transgenic plants 35S::CRY1 in Arabidopsis G380R , 35S::OsCRY1b G388R flowering phenotype analysis

[0034] Observations of the flowering phenotype revealed that, under long-day light (16h white light / 8h dark), 35S::CRY1 G380R , 35S::OsCRY1b G388R Transgenic plants flowered slightly earlier than WT ( Figure 5 A). In addition, Arabidopsis is a long-day plant, and the flowering time of WT will be greatly delayed under short-day (8h white light / 16h dark), while 35S::CRY1 G380R , 35S::OsCRY1b G388R The flowering time of plants under short-day was also significantly earlier than that of WT, which was close to that of WT under long-day (Fig. SB). These results suggest that overexpression of CRY1 G380R , OsCRY1b G388R The flowering time of the plants is not sensitive to the change of photoperiod, and has the ability of early flowering. CRY1 G380R and OsCRY1b G388R The result of early flowering of plants also supports the role of plant CRY1 protein in the...

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Abstract

The invention belongs to the technical field of the agricultural science, and discloses a method for constructing plant dwarfing and early flowering by using CRY1G380R. According to the method, with site-directed mutagenesis of conservative sites of plant cryptochrome CRY1, the constitutive dwarf of the plant can be generated, ie., the light quality and the light intensity generate light influence on the plant dwarfing development; with the method, the plant can significantly and early flowers, and the early flowering phenotype does not depend on photoperiod conditions of plant growing.

Description

technical field [0001] The invention relates to a method in the field of agricultural science and technology, in particular to a mutant protein CRY1 utilizing plant cryptochrome G380R Methods of creating dwarf plants as well as early flowering. Background technique [0002] Light plays an important role in regulating the growth and development of plants and the physiology of animals. In the solar spectrum, the blue light / A region ultraviolet light, red light, and far red light are the main bands that regulate plant growth and development. CRYPTOCHROME (CRY), also known as cryptochrome, is ubiquitous in blue light / A region ultraviolet light photoreceptor in animals and plants. The CRY proteins of animals and plants all have a conserved N-terminal domain, called the PHR domain ( Ph otolyase- R elated Region). These PHR domains have high homology with the ancient DNA photolyase (photolyase) family, belong to a protein superfamily (photolyase / CRY), and have very similar pro...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415
Inventor 杨洪全顾南南
Owner SHANGHAI JIAOTONG UNIV
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