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Application of a sir2 family gene or protein in regulating plant terpenoid production

A gene and family technology, applied in the field of application of Sir2 family genes or proteins in regulating plant terpenoid production

Active Publication Date: 2022-05-13
RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Silence information regulator 2 (silence information regulator Sir2) protein is a family of NAD+-dependent protein deacetylase / ADP ribosyltransferase, and its targets are known to be involved in various cellular proteins of aging, DNA repair and metabolic regulation. However, there is no report on the regulation of Sir2 family proteins or genes in the production of terpenoids in plants.

Method used

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  • Application of a sir2 family gene or protein in regulating plant terpenoid production
  • Application of a sir2 family gene or protein in regulating plant terpenoid production
  • Application of a sir2 family gene or protein in regulating plant terpenoid production

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Cloning and sequence analysis of LcSRT1 and LcSRT2 genes

[0028] The present invention uses the previously obtained transcriptome data of Litsea cubeba (see accessionnumber SRA080286 in the SRA database of NCBI), and screens out two transcripts whose annotation information is Sir2 family protein, and both transcripts have completely open reads frame (ORF). The primers of two genes were designed according to the ORF sequence, and the Sir2 family genes were cloned from Litsea cubeba by RT-PCR. The Sir2 family genes are named as LcSRT1 and LcSRT2, which have 1,359bp (full CDS sequence as the nucleotide sequence shown in Seq ID No: 1) and 1,158bp (full CDS sequence as the nucleotide sequence shown in Seq ID No: 2) respectively. acid sequence), encoding 452 amino acids (amino acid sequence as shown in Seq ID No: 3) and 385 amino acids (amino acid sequence as shown in Seq ID No: 4), respectively.

[0029] Among them, the CDS amplification primer of LcSRT1 gene is...

Embodiment 2

[0035] Example 2 The overexpression of LcSRT1 and LcSRT2 genes can increase the production of plant monoterpenes

[0036] After the LcSRT1 and LcSRT2 genes obtained in Example 1 were heterologously expressed in tobacco (N. benthamiana), the present invention found that in the respective positive transgenic plants, the expression levels of the two genes were respectively increased compared with the SRT gene in tobacco. 3,373 times and 2,512 times, such as figure 1shown. The present invention uses the headspace-SPME-GC-MS method to comparatively analyze the chemical components of the essential oils in LcSRT1ox, LcSRT2ox and wild-type plant WT, and specifically finds that (with figure 2 shown):

[0037] Compared with control wild-type plants, the overexpression of LcSRT1 and LcSRT2 genes can significantly increase the production of (1R)-(+)-α-pinene, D-limonene, cineole, β-ring citral and geranial. Metabolites are important constituents of monoterpenes in essential oils.

[...

Embodiment 3

[0041] Example 3 Overexpression of LcSRT1 and LcSRT2 genes can increase plant vegetative organs

[0042] The LcSRT1 and LcSRT2 genes obtained in Example 1 were heterologously expressed in tobacco (N. benthamiana), and it was also found that the transgenic positive plants of the two genes had a phenotype of plant organ enlargement compared with the wild type, which was reflected in the same developmental period. The phenotypes of enlarged leaves, increased stem diameter (also known as increased stem diameter), increased plant height (also known as increased length), and increased plant fresh weight.

[0043] Specifically: heterologous expression of LcSRT1 and LcSRT2 genes in tobacco. After positive transgenic lines were screened by PCR amplification and sequencing, the present invention compared the length of the transgenic plants and wild-type 1-month-old tobacco seedlings, stem diameter and plant fresh weight (referring to the total weight of all organs of the whole plant, th...

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Abstract

The invention discloses the application of a Sir2 family gene or protein in regulating the output of plant terpenoids; the Sir2 family gene is LcSRT1 gene or LcSRT2 gene; the terpenoid is a monoterpenoid, such as (1R)- (+)‑α‑pinene, D‑limonene, cineole, β‑cyclocitral and / or geranial. In the present invention, by transgenicly overexpressing the Sir2 family gene in plants, it is found that it can increase the production of terpenoids in plants, and provides a new way for the production of terpenoids; in addition, the present invention also expands the Sir2 family gene The application of the protein or the protein provides a new way for the application of the Sir2 family gene or protein.

Description

technical field [0001] The invention relates to the field of plant molecular biology, in particular to the application of a Sir2 family gene or protein in regulating the production of plant terpenoids. Background technique [0002] Terpenoids are the most abundant secondary metabolites in plants, and they play important roles in plants and between plants and other organisms or with the environment. Currently, more than 30,000 terpenoids have been identified from different species and have been extracted for various commercial purposes, such as: perfume, food flavoring, pesticide or pesticide production. Terpenoids are derived from the common C5 precursor isopentenyl diphosphate (IPP) and its allyl isomer dimethylallyl diphosphate (DMAPP), which in higher plants are Produced by two independent pathways located in separate compartments within the cell. In the cytosol, IPP originates from the well-known mevalonate (MVA) pathway, which begins with the condensation of acetyl-Co...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/54C12N9/10C12N15/82A01H5/12A01H5/04A01H5/00A01H6/82
CPCC12N9/1029C12N15/8243C12N15/8261
Inventor 吴立文汪阳东陈益存高暝赵耘霄
Owner RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY