CN-122012579-A - Application of pinus massoniana geranyl pyrophosphate synthetase gene PmGGPPS14

Abstract

The invention discloses application of a pinus massoniana geranylgeranyl pyrophosphate synthetase gene PmGGPPS and belongs to the technical field of plant genetic engineering. The nucleotide sequence of the Pinus massoniana geranylgeranyl pyrophosphate synthetase gene PmGGPPS is shown as SEQ ID NO. 1. PmGGPPS14, the amino acid sequence of which is shown in SEQ ID NO. 2. The invention discovers PmGGPPS that PmGGPPS can promote the accumulation of photosynthetic pigments, thereby promoting plant development and increasing leaf area. Over-expression PmGGPPS14 can increase the expression of partial genes in the downstream pathway of GGPP, and has important effect on the synthesis of diterpene resin acid. The invention is helpful for the deep research of the gene function of the pinus massoniana PmGGPPS and provides a molecular means and basis for improving the breeding work of pinus massoniana terpenoid synthesis.

Inventors

• JI KONGSHU
• CUI GUANGTONG
• WANG DENGBAO

Assignees

• 南京林业大学

Dates

Publication Date

20260512

Application Date

20260203

Claims (10)

1. 1. A method for cultivating a plant with increased leaf area and/or increased diterpenoid content, characterized in that the activity of a masson pine geranylgeranyl pyrophosphate synthetase gene PmGGPPS is overexpressed or improved in the plant, and the amino acid sequence of a protein encoded by the PmGGPPS gene is shown as SEQ ID No. 2.
2. 2. The method according to claim 1, wherein the plant is a conifer or a model plant, preferably pinus massoniana, tobacco or arabidopsis thaliana.
3. 3. The method according to claim 1 or 2, comprising the steps of: s1, constructing an expression vector containing PmGGPPS genes; S2, transforming the expression vector into plant receptor materials; And S3, screening and cultivating to obtain the transgenic plant with increased leaf area and/or increased diterpenoid compound content.
4. 4. A method for cultivating a model plant with slow needle development and reduced diterpenoid resin acid content, which is characterized in that the expression of a masson pine geranylgeranyl pyrophosphate synthetase gene PmGGPPS is inhibited or silenced in the plant, the amino acid sequence of a protein coded by the PmGGPPS gene is shown as SEQ ID No.2, the inhibiting or silencing method is a virus-induced gene silencing technology, and the plant is masson pine.
5. 5. Use of a model plant obtained by the method of claim 4 for screening or validating key genes of plant terpenoid metabolic pathways.
6. 6. An isolated masson pine geranylgeranyl pyrophosphate synthetase gene PmGGPPS, which is characterized in that the nucleotide sequence is shown in SEQ ID NO. 1.
7. 7. The protein encoded by the gene of claim 6, wherein the amino acid sequence is shown in SEQ ID NO. 2.
8. 8. A biological material comprising the PmGGPPS gene of claim 6, wherein the biological material is an expression cassette, a recombinant vector, a recombinant bacterium, or a transgenic cell line.
9. 9. Use of the gene of claim 6 or the protein of claim 7 for modulating plant response to abiotic stress, wherein the abiotic stress comprises at least one of methyl jasmonate, salicylic acid, abscisic acid, gibberellin, ethephon, hydrogen peroxide, sodium chloride, osmotic stress, and drought.
10. 10. Use of PmGGPPS gene of claim 6 for over-expression in cultivation of pinus massoniana varieties with improved needle development and/or increased diterpene yield.

Description

Application of pinus massoniana geranyl pyrophosphate synthetase gene PmGGPPS14 Technical Field The invention belongs to the technical field of plant genetic engineering, and particularly relates to application of a pinus massoniana geranylgeranyl pyrophosphate synthetase gene PmGGPPS. Background Pinus massoniana (Pinus massoniana lamb.) is a perennial evergreen tree species of Pinaceae Pinus, and is a pioneer tree species for afforestation in barren mountains in southern areas of China, and has the advantages of rapid growth, strong adaptability and drought resistance. The pinus massoniana wood fiber is slender and is very suitable for papermaking, so that pinus massoniana is the main pulp wood tree species in China. In addition, the pinus massoniana is also a main resin-producing tree species, more than 70% of the annual rosin yield in China comes from the pinus massoniana, the rosin can extract turpentine and rosin, and the rosin is a large export commodity of forestry in China, thereby creating great economic and ecological values for China. The low yield of rosin is an important factor for restricting the development of the pinus massoniana rosin industry, so that the selection of high-yield high-quality varieties and the construction of high-yield rosin artificial forests are effective measures for improving the rosin yield. In addition, rosin plays an important role in the anti-biotic stress. The conifer body is rich in terpenoid (Isoprenoids), the biosynthesis of the conifer body can be simply divided into 3 stages, the 1 st stage is the formation of common precursors of the terpenoid, namely isopentenyl pyrophosphate (Isoprene precursor ispentenyl diphosphate, IPP) and dimethylallyl pyrophosphate (DIMETHYLALLYL DIPHOSPHATE, DMAPP), the 2 nd stage is the synthesis of various types of direct terpenoid precursors, such as geranyl pyrophosphate (Geranyl diphosphate, GPP), farnesyl pyrophosphate (Farnesyl diphosphate, FPP) and geranylgeranyl pyrophosphate (GERANYLGERANYL DIPHOSPHATE, GGPP), and the 3 rd stage is the formation of the terpenoid and modification of the terpenoid. Wherein GGPP of the second stage is a key precursor in the terpenoid metabolic pathway. Gibberellin (GA), abscisic acid (Abscisic acid, ABA), strigolactone (Strigolactones, SL), pigments (phytyl side chains of carotenoids and chlorophyll), vitamins (phytyl side chains of carotene vitamin a and vitamin E), and diterpene plant protectors involved in the defense mechanism of plants against pathogenic microorganisms can be synthesized, and these metabolic branches compete together for GGPP, constituting a complex regulatory network. The distribution of GGPP in organisms plays a very important role in the growth and development of the organisms. Therefore, the GGPP related synthetic gene or protein is developed and applied, and an effective molecular means and basis can be provided for the directional breeding work of improving the growth and development of the pinus massoniana leaves and the synthesis of the terpenoid. Disclosure of Invention The invention aims to provide a Pinus massoniana geranyl pyrophosphate synthetase gene PmGGPPS which meets the use requirement, another aim of the invention is to provide an expression protein of a Pinus massoniana geranyl pyrophosphate synthetase gene PmGGPPS, and a further aim of the invention is to provide application of the Pinus massoniana geranyl pyrophosphate synthetase gene PmGGPPS. In order to solve the technical problems, the technical scheme adopted by the invention is as follows: A method for cultivating plants with increased leaf area and/or increased diterpenoid content comprises over-expressing or increasing activity of Pinus massoniana geranylgeranyl pyrophosphate synthetase gene PmGGPPS in plants, wherein the amino acid sequence of protein coded by PmGGPPS gene is shown as SEQ ID NO. 2. Further, the plant is a conifer or model plant, preferably pinus massoniana, tobacco or arabidopsis thaliana. Further, the method comprises the following steps: s1, constructing an expression vector containing PmGGPPS genes; S2, transforming the expression vector into plant receptor materials; And S3, screening and cultivating to obtain the transgenic plant with increased leaf area and/or increased diterpenoid compound content. Further, the nucleotide sequence of the PmGGPPS gene is shown as SEQ ID NO. 1. Further, the method for constructing the expression vector comprises the steps of connecting the PmGGPPS gene with a plant functional promoter to form an expression cassette, and inserting the expression cassette into a vector framework. Further, the plant functional promoter is a cauliflower mosaic virus 35S promoter. Further, the transformation step is achieved by agrobacterium-mediated, biolistic or vacuum infiltration. A method for cultivating a model plant with slow needle development and reduced diterpenoid resin acid content is to inhibit or silence the expression of a Pinus massoniana ger