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An apple flavonoid glycosyltransferase gene mdgt1 and its application

An apple flavonoid glycosyl and transferase technology, applied in transferase, application, genetic engineering and other directions, can solve problems such as hindering plant photosynthesis and damage to plant cell membranes.

Active Publication Date: 2022-07-15
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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  • An apple flavonoid glycosyltransferase gene mdgt1 and its application
  • An apple flavonoid glycosyltransferase gene mdgt1 and its application
  • An apple flavonoid glycosyltransferase gene mdgt1 and its application

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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) levels: The reactive oxygen species levels in callus were determined with a reactive oxygen species assay kit (Plant ROS ELISA Kit, Beijing Dongge, CK-E0071P), and the assay method was operated according to the instructions.

[0043] 2. Determination of total antioxidant capacity: The total antioxidant capacity of the callus was measured by a total antioxidant capacity detection kit (FRAP method, production batch number S0116), and the measurement 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 was Shimadzu LC-20AT (Shimadzu, Japan). HPLC analysis conditions: the mobile phases of flavon...

Embodiment 1

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

[0048] Using the yellow leaves and green leaves in the prepared materials, these plants were grown on the Beijiadi 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 contents 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 peachtranscriptome landscape. Plant molecular biology, 83, 365-377.). The leaves were ground and blended in triplicate. Anthocyanins and chlorophyll were extracted in 10 mL 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, and library construction and transcriptome sequencing were performed.

[0055] 1. Total RNA extraction:

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

[0057] RNA was analyzed for quantity and quality (purity and integrity) using a nanophotometer spectrophotometer (IMPLEN, Westlake Village, CA, USA) and an Agilent Bioanalyzer 2100 system (Agilent Technologies, CA, USA), respectively.

[0058] 2. Library construction and transcriptome sequencing:

[0059] An equal amount of total RNA in the sample was mixed as a pool. Use magnetic beads with Oligo(dT) to enrich eukaryotic mRNA (if it is prokaryotes, enrich mRNA by removing rRNA by the kit); The mRNA is used as a template, a six-base random primer is used to synthesize one-strand cDNA, then a two-strand reaction system is prepared to synthesize two-strand cDNA, and a kit is used to purify the double-strand...

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Abstract

The invention discloses an apple flavonoid glycosyltransferase gene MdGT1, which is derived from the yellow leaves of Vitex chinensis, the nucleotide sequence of which is shown in SEQ.ID.NO.1, and the amino acid sequence encoded by the nucleotide sequence. As shown in SEQ.ID.NO.2. In vitro enzymatic experiments and in vivo transgenic apple callus experiments proved that the above genes can promote anthocyanin metabolism and enhance the salt resistance of callus when anthocyanin substrates are added. The present invention identifies an apple anthocyanin glycosyltransferase gene MdGT1 for the first time, provides a preliminary explanation for its function, and lays a theoretical and practical foundation for the selection and acquisition of apple germplasm resources in the future.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an apple flavonoid glycosyltransferase gene MdGT1 and an application thereof. Background technique [0002] Flavonoids are polyphenolic compounds that are widely found in various plants, and they are abundant and occupy the top of the list of natural compounds. At present, more than 7,000 flavonoids have been found in nature, mainly including flavonols, flavonoids, isoflavones and anthocyanins. The synthetic pathway of flavonoids is mainly accomplished through this propane metabolic pathway. First, chalcone synthase (CHS) catalyzes 4-coumarin-CoA and malonyl-CoA to form chalcone, and then chalcone undergoes a series of enzymatic reactions to produce dihydroflavonols (including dihydrophenol and dihydrophenol) quercetin), and then under the action of flavonoid synthase (FLS) to generate dihydroflavonols to generate quercetin and kaempferol. Anthocyanins are produced und...

Claims

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Application Information

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Patent Type & Authority Patents(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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