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Application of paddy rice BG1 proteins and encoding genes of paddy rice BG1 proteins to adjusting growth and development of plants

A technology for encoding genes and transgenic plants is applied in the application field of rice BG1 protein and its encoding gene in regulating plant growth and development, which can solve the problems of limited application and little understanding of molecular mechanism research.

Active Publication Date: 2013-11-27
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, little is known about the molecular mechanisms underlying the cloned genes controlling rice grain size
In addition, most of these genes are involved in the process of negative regulation of seed development, which limits the application in agricultural production to a certain extent.

Method used

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  • Application of paddy rice BG1 proteins and encoding genes of paddy rice BG1 proteins to adjusting growth and development of plants
  • Application of paddy rice BG1 proteins and encoding genes of paddy rice BG1 proteins to adjusting growth and development of plants
  • Application of paddy rice BG1 proteins and encoding genes of paddy rice BG1 proteins to adjusting growth and development of plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0100] Example 1, Obtaining and Functional Identification of BG1 Overexpressed Rice

[0101] The gene involved in this example is derived from rice (Oryza. BG1 protein). Sequence 1 consists of 993 nucleotides, and Sequence 2 consists of 310 amino acids.

[0102] 1. Obtaining of rice overexpressing BG1

[0103] 1. Construction of recombinant expression vector

[0104] (1) Extraction of rice genomic DNA

[0105] Rice genomic DNA was extracted using the CTAB method. The specific process is as follows: Put an appropriate amount of rice (Oryza. , 100mM Tris-HCl, 1.4M NaCl), mix well and incubate at 65°C for 30min; after cooling, add 800μl chloroform to each tube, mix well and let stand for 5min, then centrifuge at 12,000rpm for 5min at room temperature; take 600μl supernatant in 1.5ml Add 600 μl of pre-cooled isopropanol to the centrifuge tube, mix thoroughly and precipitate at -20°C for 30 minutes; centrifuge at 12,000 rpm at room temperature for 5 minutes; wash the DNA pell...

Embodiment 2

[0158] Embodiment 2, the acquisition and functional identification of RNAi rice of BG1 gene

[0159] 1. Obtaining the RNAi rice of BG1 gene

[0160] 1. Construction of BG1RNAi vector

[0161] (1) Extraction of rice total RNA and acquisition of cDNA

[0162] Same as Step 1 of Example 1.

[0163] (2) Construction of BG1RNAi vector

[0164] Using the cDNA obtained in step (1) as a template, PCR amplification was performed with primers 3 and 4, and the product was purified after the reaction, which showed that a fragment of about 440bp was amplified, and sequencing showed that the fragment had the sequence 1 in the sequence list. The coding sequence (ORF) of the BG1 gene is shown.

[0165] Primer 3: 5'- CTCGAG CAGCCGTCTTTCTCTCATCCAC-3' (the underlined part is the recognition site of Xho I, and the following sequence is the 61-80th position of sequence 1)

[0166] Primer 4: 5'- GGATCC GCGAAGATGGAGTTGAGGAA-3' (the underlined part is the recognition site of BamH I, and the f...

Embodiment 3

[0188] Example 3, the acquisition of BG1 transgenic Arabidopsis and its functional identification

[0189] 1. Obtaining BG1 transgenic Arabidopsis

[0190] 1. Construction of recombinant expression vector pCAMBIA2300-35S-BG1-GFP

[0191] (1) Extraction of rice total RNA and acquisition of cDNA

[0192] Same as Step 1 of Example 1.

[0193] (2) Construction of recombinant expression vector pCAMBIA2300-35S-BG1-GFP

[0194] Using the cDNA obtained in step (1) as a template, PCR amplification was carried out with primers 5 and 6, and the product was purified after the reaction, which showed that a fragment of about 945bp was amplified, and sequencing showed that the fragment had the sequence 1 in the sequence list The coding sequence (ORF) of the BG1 gene is shown.

[0195] Primer 5: 5'- TCTAGA ATGGAGAGGTGGGCGGCGCCCAAG-3' (the underlined part is the recognition site of Xba I, and the following sequence is the 1st-24th position of sequence 1)

[0196] Primer 6: 5'- ACTAGT ...

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Abstract

The invention discloses application of paddy rice BG1 proteins and encoding genes of the paddy rice BG1 proteins to adjusting growth and development of plants and particularly provides application of proteins consisting of the amino acid sequences represented by Sequence 2 in a sequence table or the encoding genes of the proteins to adjusting and controlling growth and development of plants. Experimental results show that overexpression of the BG1 genes in wild Nipponbare causes increase of the sizes of paddy rice grains, the thousand seed weight, the single-plant yield and biomass, and imply the possible application potential of the BG1 genes in agricultural production. Meanwhile, overexpression of the BG1 genes in arabidopsis can enlarge organs (seeds and leaves), which shows that the BG1 genes probably have a common conservative mechanism in adjusting and controlling the sizes of grains (seeds) in monocotyledons and dicotyledons. The BG1 proteins and the encoding genes have a wide market and application prospect in the fields of crop genetic improvement and the like.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to the application of rice BG1 protein and its coding gene in regulating plant growth and development. Background technique [0002] Rice is one of the most important food crops in the world and is widely grown in Asia. Increasing rice yield is an important goal of breeding as the world population grows. At the same time, rice is a model plant of monocots, and the analysis of its molecular mechanism of yield can provide a theoretical basis for the increase of yield of other crops. The yield of rice is affected by many factors, mainly including environmental factors and genetic factors. Environmental factors mainly include temperature, light, soil, biotic stress and abiotic stress. From the perspective of genetic factors, rice yield traits are mainly quantitative traits controlled by multiple genes, so continuous variation and environmental influences greatly increase the difficulty of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/113C12N15/82A01H5/00
CPCC07K14/415C12N15/8261Y02A40/146C12Q1/6895C12Q2600/13C12Q2600/156
Inventor 储成才刘林川童红宁胡斌梁成真车荣会徐凡
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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