Application of transcription factor to oryza sativa breeding and oryza sativa breeding method

A technology of transcription factor, rice, applied in the direction of application, chemical instrument and method, botany equipment and method, etc.

Active Publication Date: 2021-05-18
GUIZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention intends to provide a method for rice breeding to solve the technical problem of increasing the number of tiller buds at the beginning of rice and thus increasing rice yield

Method used

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  • Application of transcription factor to oryza sativa breeding and oryza sativa breeding method
  • Application of transcription factor to oryza sativa breeding and oryza sativa breeding method
  • Application of transcription factor to oryza sativa breeding and oryza sativa breeding method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041]Example 1: Construction of OSTCP21 gene super expression plants

[0042]The RNA of Rice Nikami (Ni) was extracted, and it reversed into cDNA, using primer to expand the cDNA of the OSTCP21 gene by PCR (see SEQ ID NO.1, and OSTCP21 gene) by PCR by PCR (protein sequence of OSTCP21 gene). Sequence see SEQ ID No. 2). The primers are F1 and R1, respectively:

[0043]F1: 5'-gagctcggctccggggggggggggcccccatgagctcgcggggcagcaa-3 '(SEQ ID NO.5, enzyme-containing point KPNI);

[0044]R1: 5'-TccaagggcGaattggtcgctgctTgcccttctcctTgatgt-3 '(SEQ ID No.6, enzyme-containing point Bamhi).

[0045]The cDNA of the OSTCP21 gene was then brought into a PCAMBIA-1306 vector (PCAMBIA-1306 vectors from Cambia) by two enzyme-cutting sites KPNI and BamHI, and the super-metallic carrier OSTCP-P1306 of the OSTCP21 gene was constructed. Using Agrobacterium EHA105-mediated genetic conversion method, the super-expression vector is introduced into the normal rice variety in Japan.

[0046]All transgenic seedlings obtained were...

Embodiment 2

[0051]Example 2: Construction of Osgamyb gene super expression plants

[0052]RNAs were extracted in rice, and reverse it into cDNA, using primers to amplify the cDNA of the OSTCP21 gene by PCR (see SEQ ID NO.3, the protein sequence of the Osgamyb gene, see SEQ ID NO.4). The primers are:

[0053]F3: 5'-gagctcggggggggggggggggggatccatgtatcggggtggagagcgagag-3 '(SEQ ID No.9, enzyme-containing point kPni);

[0054]R3: 5'-TccaaggcGAATTGTCGACTTGAATTGAATTGTCGACTTGAATTGAATTGACATTCACAGGCC-3 '(SEQ ID No.10, enzyme-containing point BamHi).

[0055]The super-volume carrier OSGAMYB-P1306 of the Osgamyb gene was constructed by KPNI and BamHi. (PCAMBIA-1306 vectors were purchased from Cambia). Using Agrobacterium EHA105-mediated genetic conversion method, the super-expression vector is introduced into the normal rice variety in Japan.

[0056]All transgenic seedlings obtained were cultured with a normal nutrient solution of gamma maternal, and if the seedlings were able to grow normally, the transgenic plants wer...

experiment example 1

[0071]Experimental Example 1: Hydroponic test

[0072]The seeds of super-expression plants (Example 1), Japan (Ni) and gene knockout plants (Comparative Example 1) were submerged by distilled water with distilled water for 3 days and cultured for 7 days, and the nutrient liquid culture and nutrient formulation were transferred to rice nutrient solution. Referring to the prior art routine international rice formulation, 40 days were cultured, and the water rice form was observed, and the number of rice tillers and biomass (ie, seedlings were fresh), and the phosphor photographed microscope was taken. Tiller bud photo resultsFigure 5 As shown, it can be seen from the figure that the OSTCP21 gene super-expression plants are increased compared to the total number of tillers and tiller buds in the Japanese sunny plants, and achieve significant differences. The OSTCP21 gene knockout mutant plants were reduced by tillement and tillers in compared to Japanese sunny plants, and achieved signifi...

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Abstract

The invention relates to the technical field of oryza sativa molecular breeding, in particular to an application of a transcription factor to oryza sativa breeding and an oryza sativa breeding method. The oryza sativa breeding method comprises the following steps: increasing the protein expression level of an OsGAmyb gene and / or an OsTCP21 gene of an oryza sativa plant. The amino acid sequence of the OsGAmyb gene is as shown in SEQ ID NO. 1, and the amino acid sequence of the OsTCP21 gene is as shown in SEQ ID NO. 3. After the expression level of the protein of the OsGAmyb gene and / or the OsTCP21 gene of the oryza sativa plant is increased, the tiller number, the biomass and the rice quantity of a single rice plant are remarkably increased, so that the yield of the single oryza sativa plant is increased. Overexpression of the OsGAmyb and the OsTCP21 genes can be applied to various aspects of genetic improvement of oryza sativa plant types and yield, and conditions are created for improving the oryza sativa yield and cultivating rice with new plant types.

Description

Technical field[0001]The present invention relates to the field of rice molecular breeding, and more particularly to the application of transcription factors in rice breeding, and rice breeding.Background technique[0002]"People take food as the sky, eat rice as the first", rice (Oryza Sativa L.) is one of the most important food crops in the world, and the high output of its output determines food security in the world. The high and low of rice yield is mainly determined by three agronomicities, namely the number of spikes, number of each ear, and single weight. Among them, the number of spikes in the single plant is closely related to rice in rice. Rice tillering is a special branch of spikes in a single sub-leaf plant. Its formation can be divided into two stages, namely the start of tiller buds, the elongation of Leyser O.Regulation of Shoot Branching by Auxin.2003, 8 (11): 541-545. Typically, the occurrence of rice tiller buds is mainly the armpits of each leaf on the rice main ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8261
Inventor 方中明杭俊楠杨修艳王荣纳
Owner GUIZHOU UNIV
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