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Maize gene ZmGRAS37 for regulating and controlling nutrient body largeness, early blossoming and grain weight increment of arabidopsis thaliana at seedling stage and application thereof

A technology of Arabidopsis thaliana and vegetative bodies, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problems of lack of GRAS family members and in-depth research on GRAS family members

Active Publication Date: 2017-08-08
SHANDONG AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the research on GRAS family members in maize is not in-depth, and there is currently a la

Method used

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  • Maize gene ZmGRAS37 for regulating and controlling nutrient body largeness, early blossoming and grain weight increment of arabidopsis thaliana at seedling stage and application thereof
  • Maize gene ZmGRAS37 for regulating and controlling nutrient body largeness, early blossoming and grain weight increment of arabidopsis thaliana at seedling stage and application thereof
  • Maize gene ZmGRAS37 for regulating and controlling nutrient body largeness, early blossoming and grain weight increment of arabidopsis thaliana at seedling stage and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Embodiment 1: Obtaining of transgenic strains

[0050] (1) Corn GRAS37 Gene sequence analysis, cloning and vector construction

[0051] The present invention utilizes MaizeGDB website MaizeGDB to find gene GRAS37 , NCBI analysis shows that this gene, like all GRAS protein family members, has a conserved C-terminal GRAS domain; GRAS37 The full length of the genome sequence is 2258 bp (sequence shown in SEQ ID NO.1), while the CDS sequence is 2001 bp (sequence shown in SEQ ID NO.2), will GRAS37 The amino acid sequence (sequence shown in SEQ ID NO.3) and all members of the GRAS protein family in maize, Arabidopsis, rice and sorghum construct a phylogenetic tree, and the results are as follows figure 1 shown. figure 1 Phylogenetic tree analysis showed that there was no direct homologous gene of this gene in Arabidopsis thaliana, but the gene had a high homology with the Gramineae sorghum, which indicated that the gene was a unique gene of Gramineae.

[0052] base...

Embodiment 2

[0097] Example 2: Transgenic lines have a large vegetative body phenotype at the seedling stage

[0098] (1): Overexpression of Zm GRAS37 Cultivation of transgenic Arabidopsis

[0099] Arabidopsis seeds were sterilized with freshly prepared 70% ethanol for 5 minutes, then sterilized with 2.6% sodium hypochlorite for 10 minutes, and finally rinsed 5-7 times with ddH2O containing Tween-20, and spread the sterilized sterile seeds on 1 / 2MS solid medium. They were treated at 4°C in the dark for 3 days, placed in a long-day incubator at 22°C for 7 days, and then Arabidopsis seedlings were transferred to a substrate containing nutrient solution to continue culturing. The long-day conditions were 16h light / 8h dark at 22°C.

[0100] The experimental results are shown in Figure 5 and Figure 6, Figure 5A , B can show that the transgenic positive plants have a large vegetative body phenotype at the seedling stage. Figure 6A , B, and C can all indicate that the transgenic positive...

Embodiment 3

[0109] Example 3: Transgenic lines have an early flowering phenotype

[0110] (1): Overexpression of Zm GRAS37 Detection of flowering gene expression in transgenic lines

[0111] According to the qRT-PCR primer design requirements, gene-specific primers were designed using software such as Beacon Designer 7 and Primer Premier 5.0. Select the SYBR Green Design option, create the file, enter the sequence, and run BLAST

[0112] After using the search sequence and Template structure search tools, run the Primer search tool to select the optimal primer sequence (first ensure the specificity of the primers and then consider avoiding the influence of the template structure). Primers were synthesized at Shanghai Sangon Bioengineering Service Co., Ltd. and purified by PAGE.

[0113] qRT-PCR analysis was performed using qRT-PCR-specific 96-well plates (Axygen, USA) and high-transmittance parafilm (Axygen, USA), and fluorescent quantitative PCR instrument Icycler real-time PCR syste...

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Abstract

The invention provides a maize gene ZmGRAS37 for regulating and controlling the nutrient body largeness, the early blossoming and the grain weight increment of arabidopsis thaliana at a seedling stage and application thereof. According to the maize gene ZmGRAS37, the gene ZmGRAS37 is separated and cloned from maize; the cDNA (complementary Deoxyribonucleic Acid) sequence of the ZmGRAS37 is connected to an expression vector pPZP211 promoted by a 35S promoter; the arabidopsis thaliana is infected and converted by utilizing an agrobacterium infection method; indicated by an experimental result, the gene can be used for enabling the model-plant arabidopsis thaliana to generate characters of the nutrient body largeness and to further have the early blossoming, the grain weight increment and the like at the seedling stage. Shown by a phylogenetic-tree analysis result, the direct homologous gene of the gene does not exist in the model-plant arabidopsis thaliana; however, the gene has the higher homology with that of sorghum and maize of gramineous plants; the gene is accounted to be the peculiar GRAS (Generally Recognized as Safe) family gene of the graminaceous plants; in consideration of the conservatism of the gene in the graminaceous plants and the phenotypes presented in transgenic arabidopsis thaliana, the gene is proven to have potential application value.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and specifically relates to a corn gene that regulates Arabidopsis thaliana seedling vegetative body size, early flowering and grain weight increase ZmGRAS37 and its application. Background technique [0002] The agricultural production in the Huang-Huai-Hai region of my country is mainly the rotation of wheat and corn. Since the production of maize tends to be directly broadcast by machine after wheat harvest and harvested by machine after maturity, there is an urgent need to breed maize varieties with a short growth period in the breeding work. A large crop vegetative body can significantly increase the photosynthetic efficiency of the crop and increase the yield of the crop for the purpose of obtaining the vegetative body; moderate early flowering can shorten the growth period of the crop and improve the adaptability of the variety, so it is an important agronomic trait of t...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82A01H5/00
CPCC07K14/415C12N15/8261C12N15/827
Inventor 安海龙郑成超王镇黄金光王也孙晋浩孙德慧
Owner SHANDONG AGRICULTURAL UNIVERSITY
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