Transcription factor for enhancing embryogenesis of plant cell and synthesis of fatty acid as well as encoding gene and application thereof

A technology of embryogenesis and phytogenesis, which is applied in the fields of plant genetic improvement, plant regeneration, botany equipment and methods, etc., can solve the problems such as the unknown transfer pathway of AtMYB transcription factor function signal, and achieve the effect of increasing frequency and promoting transformation

Active Publication Date: 2009-12-09
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, most of the predicted functions of AtMYB transcription f

Method used

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  • Transcription factor for enhancing embryogenesis of plant cell and synthesis of fatty acid as well as encoding gene and application thereof
  • Transcription factor for enhancing embryogenesis of plant cell and synthesis of fatty acid as well as encoding gene and application thereof
  • Transcription factor for enhancing embryogenesis of plant cell and synthesis of fatty acid as well as encoding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1. Cloning of AtMYB115 gene

[0056] Design PCR primers according to the nucleotide sequence of AtMYB115 gene to amplify AtMYB115 gene, the primer sequence is as follows:

[0057] MYB115F (upstream primer): 5′-CCTCGAGGAAGAACATCTACAATAAAATCTCC-3′

[0058] MYB115B (downstream primer): 5'-CAGATCTTCATTCCAACCATTCATGAGCATCC-3'.

[0059] Use TRIZAL reagent (Invitrogen) and refer to the kit instructions to extract total RNA from Arabidopsisthaliana Colombian ecotype (Col-0) fresh siliques, and then use SuperScript TM IIReverse Transcriptase kit (Invitrogen) reverse transcription to synthesize its first strand cDNA, and then use the synthesized cDNA as a template to perform PCR amplification under the guidance of primers P1 and P2. After the reaction is over, perform 1 % Agarose gel electrophoresis detection, use DNA recovery kit (Dingguo Company) to recover the target fragment with a length of about 1000bp, ligate the recovered fragment into the vector pGEM-Teasy (Promega),...

Embodiment 2

[0062] Example 2. Obtainment of AtMYB115 Gene Transformed Arabidopsis

[0063] The plant expression vector pER10-MYB115 constructed in Example 1 was transformed into Agrobacterium GV3101 competent cells by the electric shock method, and coated on the LB resistant plate containing 50 mg / L spectinomycin and 50 mg / L rifampin Cultivate for 12-16 hours at 28℃ and 150rpm, pick the single colony of Agrobacterium that grows and inoculate it in 20mL LB liquid medium containing 50mg / L spectinomycin and 50mg / L rifampicin at 28℃, 150rpm Cultivate for 2 days under shaking, and then inoculate the bacterial solution in 300mL LB liquid medium containing 50mg / L spectinomycin and 50mg / L rifampicin at 1% of the inoculum amount, and shake culture at 28℃ and 150rpm for 16- 18 hours. After the incubation, the cells were collected by centrifugation at 5000 rpm for 20 minutes, and the cells were suspended in 250 mL of Silwetl-77 containing 5% sucrose, and shaken slowly. Finally, the bacterial solution wa...

Embodiment 3

[0065] Example 3 Detection of gene expression in AtMYB115 overexpression plants

[0066] Two different transgenic lines #5 and #6 of the Arabidopsis transgenic lines transgenic with the AtMYB115 gene in Example 2 were selected to detect the expression level of the AtMYB115 gene in Arabidopsis, and the Arabidopsis plants transfected with the pER8 vector were used as controls. The above plant seeds were sown on 1 / 2MS medium (Sigma). Incubate for about 2 weeks at 22°C under 16 hours of light. Then, it was transferred to liquid 1 / 2 MS medium containing 10 μM estradiol and induced for 16 hours. At the same time, it was transferred to liquid 1 / 2 MS medium without estradiol and induced for 16 hours as a control.

[0067] Use TRIZOL reagent (purchased from Invitrogen) and refer to the kit instructions to extract total RNA, and then use Invitrogen's SuperScript TM II Reverse Transcriptase kit and refer to the kit instructions to reverse transcription to synthesize its first strand cDNA, an...

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Abstract

The invention discloses a transcription factor for enhancing the embryogenesis of a plant cell and the synthesis of fatty acid as well as an encoding gene and an application thereof. The transcription factor is the following proteins: (a) a protein formed by an amino acid sequence disclosed by the sequence 3 in a sequence table; (b) relevant proteins derived from (a), enhancing the embryogenesis of the plant cell and obtained through replacing and/or losing and/or adding one or a plurality of amino acids in the amino acid sequence disclosed by the sequence 3 in the sequence table. The invention also discloses the encoding gene of the transcription factor and a recombined expression vector containing the gene, a transgene clone or a recombined germ. The transcription factor for enhancing the embryogenesis of a plant cell and the synthesis of fatty acid and the encoding gene thereof have great application values, such as enhancing the embryogenesis of the plant cell, culturing transgene plants with improved fatty acid content, being used as screening marks of the transgene plates, enabling plants to have asexual reproduction capacity and producing synthetic seed.

Description

Technical field [0001] The invention relates to a transcription factor that promotes plant somatic embryogenesis and fatty acid synthesis, and its coding gene and application. Background technique [0002] The embryo of higher plants is generally the product of fertilization, that is, developed through the combination of sperm and egg. But in some special cases, embryos can also be produced through somatic embryogenesis. The so-called somatic embryogenesis refers to the morphogenesis process in which diploid somatic cells imitate the various stages of sexual zygotic embryogenesis and form a new individual without sex cell fusion. [0003] In theory, any plant cell has the potential to develop into a complete biological individual, but the cell must first return to the meristem state or embryonic cell state in order to express totipotency, and the explants with this reverting ability are subject to The genotype, growth period, physiological state and the condition of the culture m...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21C12N15/11C12N15/82A01H1/00A01H4/00A01H5/10
Inventor 左建儒王兴春腾冲牟金叶
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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