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Application of OsGRF6 protein in regulation of plant height

A protein and plant technology, applied in the application field of OsGRF6 protein in the regulation of plant strains, can solve the problems of less research on GRF gene family and unclear biological process, etc.

Inactive Publication Date: 2015-03-25
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there are relatively few studies on the GRF gene family in rice, and the biological processes involved in this gene family are still unclear

Method used

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  • Application of OsGRF6 protein in regulation of plant height
  • Application of OsGRF6 protein in regulation of plant height
  • Application of OsGRF6 protein in regulation of plant height

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1, the cloning of OsGRF6 gene

[0028] Based on the screening results of rice-related genes in the previous laboratory, one of the EST fragments of interest was selected. By utilizing NCBI ( http: / / www.ncbi.nlm.nih.gov ) and TIGR ( http: / / www.tigr.org ) to retrieve the two websites, get the genome sequence corresponding to the EST, and use Gramme ( http: / / www.gramene.org ) database analysis to obtain its complete cDNA sequence.

[0029] Design primers according to the results of database analysis, 5' end primer: 5'-GG GGTACC atgcagggtgcaatggc-3' (the underlined sequence is the Kpn I site), 3' end primer: 5'GA AGATCT caccaggcggatgctcggatg-3' (the underlined sequence is the BglII site), using japonica rice Zhonghua No. 10 three-leaf stage seedling reverse-transcribed cDNA as a template, amplified to 1371bp full-length cDNA by RT-PCR method. The specific operation process is as follows:

[0030] Select the three-leaf stage Zhonghua 10 rice (Oryza sa...

Embodiment 2

[0035] Example 2, Functional identification of OsGRF6 gene

[0036] 1. Acquisition of expression vectors

[0037] 1. Construction of pUN1301 vector

[0038] 1) Cut about 0.2g of corn seedlings (variety name: Zhongzuo-Zhongdan 8, Beijing Zhongnongzuo Technology Development Co., Ltd.) and grind them in liquid nitrogen; then add 800 μL of newly prepared extraction buffer (containing 0.1M Tris -HCl pH8.0, 50mM EDTA, 0.5M NaCl, 1% SDS and 1% β-mercaptoethanol), shake vigorously to suspend it; bathe in 65℃ water for 30 minutes, mix by inversion every 5 minutes; then add 250μL pre- Cold 5M potassium acetate aqueous solution, immediately inverted and mixed, ice bath for 5 minutes; add an equal amount of phenol / chloroform, extract once, centrifuge at 12000rpm for 5 minutes; collect supernatant, add 0.6 times the volume of isopropanol to precipitate DNA, room temperature Place for 40 minutes; centrifuge at 12,000 rpm at 4°C for 15 minutes, discard the supernatant; wash the pellet once...

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Abstract

The invention discloses an application of an OsGRF6 protein in regulation of plant height. According to the application of the OsGRF6 protein in the regulation of the plant height provided by the invention, an amino acid sequence of the OsGRF6 protein is sequence 2 in a sequence table. In the above application, a nucleotide sequence of a coding gene of the OsGRF6 protein is sequence 1 in a sequence table. Experiments show that a gene OsGRF6 of GRF family is cloned in japonica rice (ZH10); and positive transgenic rice lines are obtained by transferring rice ZH10 callus by an agrobacterium-mediated method. By analyzing the transgenic lines and corresponding wild type phenotypes, obvious dwarf characteristics are found in the transgenic rice.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of an OsGRF6 protein in regulating plant strain growth. Background technique [0002] Rice is the main food crop in my country. In the past ten years, with the development of the economy, the area of ​​cultivated land has decreased, and the only way to stabilize and increase the total rice output is to rely on the increase of per unit yield. The improvement of rice plant type plays an important role in increasing the yield per unit area. Rice plant height is an important agronomic trait controlling rice yield, which is mainly regulated by the elongation of rice internodes. The elongation of the uppermost internode of rice can promote the extraction of the young panicle, and then flowering, pollination and grain filling. Plants that are too tall can easily cause lodging and reduce production. Dwarf breeding is a major breakthrough in rice breeding. In the 1950s, Aca...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82C12N15/84A01H5/00
Inventor 种康刘唤唤郭思义徐云远赵原
Owner INST OF BOTANY CHINESE ACAD OF SCI
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