Application of cotton transcription factor GaMAN1 to plant grease metabolism regulation and control

A technology of vegetable oil and transgenic plants, which is applied in the application field of cotton transcription factor GaMAN1 in the regulation of plant oil metabolism, and can solve the problems of increasing oil content and so on

Active Publication Date: 2019-01-15
INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to increase the yield of seed oil, so far, some key enzymes involved in oil biosynthesis have been cloned and used for improvement, but the results show that simply changing a gene cannot significantly increase oil content (Song, Li et al.2013)

Method used

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  • Application of cotton transcription factor GaMAN1 to plant grease metabolism regulation and control
  • Application of cotton transcription factor GaMAN1 to plant grease metabolism regulation and control
  • Application of cotton transcription factor GaMAN1 to plant grease metabolism regulation and control

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Embodiment 1, the expression analysis of GaMAN1 in different organs of cotton

[0075] 1. Synthesis of cDNA

[0076] Total RNA was extracted from the roots, stems, leaves, flowers and ovules of Asian Cotton Rock No. 1 (provided by Cotton, Chinese Academy of Agricultural Sciences), and cDNA was synthesized by reverse transcription with reverse transcriptase.

[0077] 2. Real Time-PCR

[0078] Using the cDNA in step 1 as a template, Real Time-PCR was performed using primer F: 5'-CATGGTGAAGGCAATTGGGC-3' and primer R: 5'-ATCAAAGCAACGAACCGCAC-3'. The cotton His3 gene was used as an internal reference, and the internal reference gene primers were Primer-F: 5'-TCAAGACTGATTTGCGTTTCCA-3' and Primer-R: 5'-GCGCAAAGGTTGGTGTCTTC-3'. At the same time, wild-type Arabidopsis was used as a control.

[0079] The result is as figure 1 As shown, the results showed that the transcription of GaMAN1 gene was almost undetectable in roots, stems, leaves and flowers, but the expression leve...

Embodiment 2

[0080] Example 2, the acquisition of GaMAN1 Arabidopsis and the analysis of its total oil and fatty acid content

[0081] 1. Obtaining GaMAN1-transformed Arabidopsis

[0082] 1. Acquisition of transcription factor GaMAN1

[0083] 1) Synthesis of cDNA

[0084] Total RNA was extracted from the ovules of Asian Cotton Rock No. 1 (provided by the Cotton Institute of the Chinese Academy of Agricultural Sciences), and the RNA was reverse-transcribed with reverse transcriptase to synthesize cDNA.

[0085] 2) PCR amplification and sequencing

[0086] According to the information of GaMAN1 full-length cDNA sequence in the cotton genome sequence of CottonFGD, the following primer sequences were designed: GaMAN1-F: 5'-GGGGTACCATGTCAGAAGAAATGAATCTATC-3' and GaMAN1-R: 5'-ACGTCGACTTATACCGACAGGTGGCACAAG-3'. Using the cDNA obtained in step 1) as a template, PCR amplification was performed using GaMAN1-F / R primers.

[0087] The PCR reaction system and reaction procedure are as follows:

[...

Embodiment 3

[0122] Embodiment 3, the acquisition of GaMAN1 cotton and its oil content analysis

[0123] 1. Obtaining of trans-GaMAN1 cotton

[0124] 1. Acquisition of transcription factor GaMAN1

[0125] 1) Synthesis of cDNA

[0126] Total RNA was extracted from the ovule of Mianmianshixiya No. 1 (provided by the Cotton Institute of Chinese Academy of Agricultural Sciences), and the RNA was reverse-transcribed with reverse transcriptase to synthesize cDNA.

[0127] 2) PCR amplification

[0128] Using the cDNA obtained in step 1) as a template, PCR amplification was performed using GaMAN1-F' / R' primers to obtain a PCR product. The primer sequences are as follows:

[0129] GaMAN1-F': 5'-CGGAATTCATGTCAGAAGAAATGAATCTATC-3';

[0130] GaMAN1-R': 5'-ACGTCGACTTATACCGACAGGTGGCACAAG-3'.

[0131] 2. Construction of pCAMBIA2301-GaMAN1 overexpression vector

[0132]The PCR product obtained in step 1 and the carrier pCAMBIA2301 (purchased from Beijing Huayueyang Biotechnology Co., Ltd.) were dou...

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Abstract

The invention discloses application of a cotton transcription factor GaMAN1 to plant grease metabolism regulation and control. The transcription factor GaMAN1 is the protein shown as a) or b) or c) ord): (a) protein with the amino acid sequence shown as the sequence 2; b) fusion protein obtained by connecting tags onto the end N and / or the end C of the protein shown as the sequence 2; c) proteinwith the same function obtained by performing substitution and / or deletion and / or addition of one or several amino acid residues on the amino acid sequence shown by as sequence 2; d) protein with 75 percent or 75 percent or above of homology with the amino acid sequence shown as the sequence 2 and with the same function. Experiments prove that the cotton transcription factor GaMAN1 can improve thecrop grease content and the fatty acid content. The gene has the important theoretical and practical significance on crop grease ingredient increase and improvement, particularly on cotton seed grease ingredient improvement and high-grease variety culture.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of cotton transcription factor GaMAN1 in the regulation and control of vegetable oil metabolism. Background technique [0002] Cotton is an important economic crop. In addition to lint as a textile raw material, its cottonseed is also an important oil resource in my country. According to the data of the National Grain and Oils Information Center, among the major oil-producing crops in the world, the total output of the eight major oil crops in my country in 2015 was 57.244 million tons, of which the output of cottonseed was 10.089 million tons, accounting for 17.62%, ranking fourth in my country. Large oil crops. [0003] Vegetable oils are higher fatty acid glycerides synthesized from fatty acids and glycerol, stored in the form of triacylglycerol (TAG). During seed development, 16 or 18 carbon saturated fatty acids and oleic acid (18:1) are first synthe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N5/10A01H5/00A01H6/20A01H6/60
CPCC07K14/415C12N15/8247
Inventor 马磊时萌方丹徐双娇田新权
Owner INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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