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Application of rice transcription factor Os11g31340.1 gene CDS (Coding Sequence)

A technology of rice transcription factor and transcription factor, which is applied in the field of genetic engineering, can solve the problems of weakening advantages of hybrid breeding and achieve the effect of improving rice grain traits and grain shape

Active Publication Date: 2015-02-11
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current research on increasing rice production is more dependent on limited rice germplasm resources, the advantages of traditional hybrid breeding are gradually weakening, and rice transgenic technology may explore the potential of further increasing rice production

Method used

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  • Application of rice transcription factor Os11g31340.1 gene CDS (Coding Sequence)
  • Application of rice transcription factor Os11g31340.1 gene CDS (Coding Sequence)
  • Application of rice transcription factor Os11g31340.1 gene CDS (Coding Sequence)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Isolation of Os11g31340.1 Gene CDS Sequence and Plant Expression Vector Construction

[0035] Find the Os11g31340.1 gene in the plant transcription factor database (http: / / rice.plantbiology.msu.edu / analyses_search_locus.shtml), design PCR amplification primers according to its CDS sequence, and design PCR amplification primers according to its sequence (F: 5′-CAAAAAAGCAGGCTTC ATGGCCAGCCCTGGAGT-3′ and reverse primer R: 5′-CAAGAAAGCTGGGTCGTTCATGCCATGGCCATAAC-3′).

[0036] The CDS sequence of the Os11g31340.1 gene was obtained by PCR using the total cDNA of wild-type Nipponbare rice as a template, and its nucleotide sequence is shown in SEQ ID No.1.

[0037] 2. Construction of plant expression vectors

[0038] The CDS sequence of the rice transcription factor Os11g31340.1 gene was constructed downstream of the four transcription factor activation motif VP16 coding genes (SEQ ID No.4) through the Gateway system.

[0039] 2.1 Cloning the above PCR product into th...

Embodiment 2

[0049] The acquisition of embodiment 2 transgenic rice plants

[0050] Take the mature seeds of rice 'kitaake', shell them manually or mechanically, select the plump, smooth and spot-free seeds and inoculate them on the induction medium for induction culture after being sterilized. Rice callus with good appearance and good growth was selected as the recipient material, and ubi: VP64-Os11g31340.1 was transferred into the rice callus by the Agrobacterium-mediated method, with 100 μM acetosyringone and O.D. 0.7 AAM culture solution of Agrobacterium. For transformation, the callus soaked in the transformation solution was placed on a co-culture medium for co-cultivation, cultured in the dark at 25°C for 3 days, then placed on a screening medium for about 30 days, and subcultured every 10 days. Then, the screened out resistant calli were transferred to the differentiation medium for differentiation for about 20 days, and subcultured every 10 days. The resistant callus that differ...

Embodiment 3

[0056] Identification of embodiment 3 transgenic positive strains

[0057] In order to detect the overexpression of ubi:VP64-Os11g31340.1 gene in T2 transgenic rice, primers were designed on the vector (forward primer: 5'-GTGGGGACAAGTTTGTACAAAAAAGCAGGCTTC-3' and reverse primer: 5'-GTGGGGACCACTTTGTACAAGAAAGCTGGGTC-3' ) for PCR detection and obtained obvious specific bands, in which WT is wild-type rice 'kitaake', V2401H-12 and V2401H-28 are VP64-Os11g31340.1 transgenic rice lines ( image 3 ). The above primers are designed at the junction of the vector and the target gene, image 3 It shows that the size of the amplified fragment is basically the same as that of the target gene (834bp).

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Abstract

The invention relates to an application of a rice transcription factor Os11g31340.1 gene CDS (Coding Sequence). A transcription factor activation sequence motif VP64 and the rice transcription factor Os11g31340.1 gene CDS are fused and constructed to obtain a constitutive transcription factor, and the constitutive transcription factor is transformed into crops such as rice in order to improve the grain traits of the rice. For example, the lengths of rice grains are increased. The rice transcription factor Os11g31340.1 gene CDS has an important theoretical value in the detailed explanation of the regulation and control of a seed development mechanism. Moreover, the grain shape of the rice can be improved through a transgenic means, so that the rice transcription factor Os11g31340.1 gene CDS has important significance to production practice.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to the application of the rice transcription factor Os11g31340.1 gene CDS sequence. Background technique [0002] Rice (Oryza sativa L.) is one of the three most important food crops in my country and the world, the staple food of more than half of the world's population, and an important model plant for functional gene research. Related genetics and molecular biology studies have been paid much attention by researchers, and the regulation of transcription level is an important way of gene expression regulation. The current research on increasing rice yield is more dependent on limited rice germplasm resources, the advantages of traditional hybrid breeding are gradually weakening, and rice transgenic technology may explore the potential of further increasing rice yield. [0003] In the plant kingdom, plants that can form seeds account for more than two-thirds of the total number ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/82C12N1/21A01H5/00C12N15/11
Inventor 刘斌赵涛李宏宇刘军林辰涛
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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