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Apple flavone glycosyl transferase gene MdGT1 and application thereof

A technology of apple flavonoid glycosyl and transferase, which is applied in the direction of transferase, application, genetic engineering, etc., can solve the problems of plant cell membrane damage and hindrance of plant photosynthesis

Active Publication Date: 2021-02-23
QINGDAO ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, under salt stress conditions, high concentrations of Na+ ions hinder plant photosynthesis and cause plant cell membrane damage

Method used

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  • Apple flavone glycosyl transferase gene MdGT1 and application thereof
  • Apple flavone glycosyl transferase gene MdGT1 and application thereof
  • Apple flavone glycosyl transferase gene MdGT1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment approach

[0041] In the specific embodiment, various experimental methods are as follows:

[0042] 1. Detection of reactive oxygen species (ROS) level: The reactive oxygen species level of the callus was measured with a reactive oxygen species assay kit (Plant ROS ELISA Kit, Beijing Dongge, CK-E0071P), and the determination method was operated according to the instructions.

[0043] 2. Determination of total antioxidant capacity: the total antioxidant capacity of the callus was measured with a total antioxidant capacity detection kit (FRAP method, production batch number S0116), and the determination method was operated according to the instructions.

[0044] 3. Determination of proline content: refer to the international general method. Determination of soluble sugar content: refer to the international general method anthrone method.

[0045] 4. HPLC detection: The high performance liquid chromatograph is Shimadzu LC-20AT (Shimadzu, Japan). HPLC analysis conditions: the mobile phase ...

Embodiment 1

[0047] Determination of anthocyanin and chlorophyll contents in apple yellow leaf mutants and green leaves.

[0048] Using yellow and green leaves from the prepared materials, these plants were grown in the North Homestead of Qingdao Academy of Agricultural Sciences. Leaves were collected on April 15, 2016, flash frozen in liquid nitrogen, and stored at -80°C until further processing.

[0049] The total anthocyanin and chlorophyll content in leaves were determined by the method proposed by Wang et al. (Wang, L., S. Zhao, C. Gu, Y. Zhou, H. Zhou, J. Ma, J. Cheng & Y. Han ( 2013) Deep RNA-Seq uncovers the peach transcriptome landscape. Plant molecular biology, 83, 365-377.). The leaves were ground and blended in triplicate. Anthocyanins and chlorophyll were extracted in 10 ml of methanol (containing 1% hydrochloric acid) for 2 hours at 4°C in the dark. The anthocyanin and chlorophyll contents were measured at 553 and 600 nm, and the experimental results are shown in Table 1. ...

Embodiment 2

[0054] Total RNA was extracted for library construction and transcriptome sequencing.

[0055] 1. Total RNA extraction:

[0056] The total RNA in the green leaves (GL) and yellow leaves (YL) of the above apples were extracted by the modified CTAB method.

[0057] Quantity and quality (purity and integrity) of RNA were analyzed using Nanophotometer Spectrometer (IMPLEN, Westlake Village, CA, USA) and Agilent Bioanalyzer 2100 System (Agilent Technologies, CA, USA), respectively.

[0058] 2. Library construction and transcriptome sequencing:

[0059] Equal amounts of total RNA in the sample were mixed as a pool. Use magnetic beads with Oligo(dT) to enrich eukaryotic mRNA (if it is prokaryotic, use the kit to remove rRNA to enrich mRNA); mRNA as a template, use six-base random primers to synthesize one-strand cDNA, then prepare two strands into a reaction system to synthesize two-strand cDNA, and use the kit to purify the double-strand cDNA; the purified double-strand cDNA is t...

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Abstract

The invention discloses an apple flavone glycosyl transferase gene MdGT1. The apple flavone glycosyl transferase gene MdGT1 is derived from yellow leaves of malus baccata, the nucleotide sequence of the apple flavone glycosyl transferase gene MdGT1 is shown in SEQ. Id. NO.1, and the amino acid sequence coded by the nucleotide sequence of the apple flavone glycosyl transferase gene MdGT1 is shown in SEQ. Id. NO. 2. In-vitro enzymatic experiments and in-vivo transgenic apple callus experiments prove that when an anthocyanin substrate is added into the gene, anthocyanin metabolism is promoted, and the salt resistance of callus can be enhanced. The apple anthocyanin glycosyl transferase gene MdGT1 is identified for the first time, preliminary explanation is provided for the function of the apple anthocyanin glycosyl transferase gene MdGT1, and a theoretical and practical foundation is laid for selection and acquisition of apple germplasm resources in future.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an apple flavone glycosyltransferase gene MdGT1 and its application. Background technique [0002] Flavonoids are polyphenolic compounds widely present in various plants, they are numerous and occupy the top of the list of natural compounds. At present, more than 7000 kinds of flavonoids have been found in nature, mainly including flavonols, flavones, isoflavones and anthocyanins. The synthetic pathway of flavonoids is mainly completed through this propane metabolic pathway. First, chalcone synthase (CHS) catalyzes 4-coumarin-CoA and malonyl CoA to generate chalcone, and then chalcone undergoes a series of enzymatic reactions to generate dihydroflavonols (including dihydrophenol and dihydro quercetin), and then under the action of flavonoid synthase (FLS) to generate dihydroflavonols to generate quercetin and kaempferol. Anthocyanins are produced under the action of hy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/54C12N9/10A01H4/00A01H5/00A01H6/74
CPCC12N9/1048C12N15/825A01H4/00
Inventor 葛红娟沙广利宋永骏李攀张世忠黄粤马荣群张蕊芬孙红涛孙吉录展恩军
Owner QINGDAO ACAD OF AGRI SCI
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