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CN-119709796-B - Paeonia lactiflora PlDGAT gene and application thereof in increasing total fatty acid content and/or regulating fatty acid composition in plant seeds

CN119709796BCN 119709796 BCN119709796 BCN 119709796BCN-119709796-B

Abstract

The invention discloses a peony PlDGAT gene and application thereof in increasing total fatty acid content and/or regulating and controlling fatty acid composition in plant seeds, belonging to plant biotechnology, wherein the nucleotide sequence of the peony PlDGAT gene is shown as SEQ ID NO. 1. The paeonia lactiflora PlDGAT gene can increase the total fatty acid content after being transferred into plant seeds, and meanwhile, the proportion of each component of fatty acid is changed. Provides a new idea for creating and controlling the germplasm with strong fatty acid accumulation capability in plant seeds.

Inventors

  • MENG JIASONG
  • MA DI
  • XU HUAJIE
  • ZHAO DAQIU
  • TAO JUN

Assignees

  • 扬州大学

Dates

Publication Date
20260508
Application Date
20241227

Claims (4)

  1. 1. The application of PlDGAT protein expressed by SEQ ID NO.2 in the aspect of regulating the fatty acid composition in plant seeds is characterized in that the regulation of the fatty acid composition in plant seeds refers to the improvement of the content of palmitic acid and/or alpha-linolenic acid in the seeds, and the plant is tobacco.
  2. 2. The application of the expression cassette, the recombinant vector, the recombinant cell or the recombinant strain containing PlDGAT gene shown in SEQ ID NO.1 in regulating the fatty acid composition in plant seeds is characterized in that the regulation of the fatty acid composition in plant seeds refers to the improvement of the content of palmitic acid and/or alpha-linolenic acid in the seeds, and the plant is tobacco.
  3. 3. The use according to claim 2, wherein the recombinant vector is obtained by introducing the ORF sequence of paeonia lactiflora PlDGAT gene into a plant binary expression vector.
  4. 4. The use according to claim 3, wherein the primer sequences for amplifying the ORF sequence of paeonia lactiflora PlDGAT gene are shown as SEQ ID NO.3 and SEQ ID NO. 4.

Description

Paeonia lactiflora PlDGAT gene and application thereof in increasing total fatty acid content and/or regulating fatty acid composition in plant seeds Technical Field The invention relates to a paeonia lactiflora PlDGAT gene and application thereof in increasing total fatty acid content and/or regulating fatty acid composition in plant seeds, belonging to plant biotechnology. Background Paeonia lactiflora is known for its important ornamental value in China. In recent years, as peony belonging to the genus Paeonia is identified as a novel resource food, the oil use value of Paeonia has also come to be paid attention. In the agronomic traits of oil crops, the oil content of seeds and the type of fatty acid are important research hotspots, the effective increase of the fat content of seeds is one of the main targets of the current breeding of various oil crops, and in the traditional breeding method, the new variety is usually selected and hybridized for many years, so that the progress is slow. The biotechnology can accelerate the cultivation process of high-seed oil content and high-quality fatty acid component varieties, change the anabolic pathway of crop oil by utilizing the biotechnology means, increase the content of unsaturated fatty acid in the oil and realize the creation of the character of the high unsaturated fatty acid. Members of the diacylglycerol acyltransferase (DGATs) gene family were first characterized over twenty years ago, and the enzymes encoded by these genes play a central role in plant oil production, metabolic regulation and stress response, and are believed to be key enzymes for the conversion of Diglycerides (DAGs) to Triglycerides (TAGs), leading to their extensive research in a range of plant species. They are also the only rate-limiting enzymes involved in the Kennedy pathway that regulates de novo TAG biosynthesis. Class 4 DGAT genes, including DGAT1, DGAT2, DGAT3 and WSD/DGAT, have been found in plants, which are plant-specific and tissue-specific and which exert different effects on lipid synthesis and accumulation. Numerous studies have shown that DGAT plays a crucial role in the synthesis of TAG. In plants, DGAT determines its different enzymatic activities due to the different gene codes, which makes its role in TAG synthesis and accumulation in different tissues of the plant also different. DGAT2 is typically between 315 and 373 amino acids in length, and DGAT2 may also have substrate preference for different fatty acids in different species. During the development of castor seeds, the DGAT2 has higher expression level than DGAT1, and contributes to the accumulation of ricinoleic acid (18:1). DGAT2 in diatom algae increases palmitic acid (16:0) and oleic acid (18:1) when expressed in yeast systems. DGAT2 is mainly expressed in the late-maturing stage of soybean seeds and may be involved in the accumulation of oil in late-maturing seeds. Peony DGAT2 shows unique substrate preference in the accumulation of linolenic acid (18:3). In the peony, no report of DGAT2 in fatty acid accumulation of peony seeds is seen. Disclosure of Invention The invention aims to provide a paeonia lactiflora PlDGAT gene and application thereof in increasing total fatty acid content and/or regulating and controlling fatty acid composition in plant seeds. The invention provides a peony PlDGAT gene, wherein the nucleotide sequence of the peony PlDGAT gene is shown as SEQ ID NO. 1. The invention also provides the protein encoded by the gene, and the amino acid sequence of the protein is shown as SEQ ID NO. 2. The invention also provides an amplification primer for cloning the full-length sequence of the peony PlDGAT gene cDNA, and the sequence of the amplification primer is shown as SEQ ID NO.9 and SEQ ID NO. 10. The invention also provides an application of the expression cassette, the recombinant vector, the recombinant cell or the recombinant strain containing the paeonia lactiflora PlDGAT gene in increasing the total fatty acid content and/or regulating the fatty acid composition in plant seeds. Further, the regulation of fatty acid composition in plant seeds refers to increasing the content of palmitic acid and/or alpha-linolenic acid in the seed. Further, the plant comprises tobacco. Further, the recombinant vector is obtained by introducing the ORF sequence of the peony PlDGAT gene into a plant binary expression vector. Further, the primer sequences for amplifying the ORF sequence of the peony PlDGAT gene are shown as SEQ ID NO.3 and SEQ ID NO. 4. The invention also provides a method for identifying high fatty acid plant seeds, which comprises the following steps: 1) Identifying whether the plant seeds contain peony PlDGAT protein with the amino acid sequence shown as SEQ ID NO. 2; 2) Identifying whether the plant seeds contain peony PlDGAT gene with the nucleotide sequence shown as SEQ ID NO. 1. Further, the primer sequences used for PCR identification are shown as SEQ ID NO.3 and SEQ ID NO.4