CN-122012527-A - Korean pine PkDOF5.6 gene and application thereof in regulation and control of Korean pine somatic embryogenesis

Abstract

The invention relates to the technical field of plant genetic engineering, in particular to a Korean pine PkDOF5.6 gene and application thereof in regulating and controlling Korean pine somatic embryogenesis. The nucleotide sequence of the Korean pine PkDOf5.6 gene is shown as SEQ ID No. 1. The Korean pine PkDOF5.6 gene provided by the invention regulates and controls Korean pine somatic embryogenesis, and the overexpression of the Korean pine PkDOF5.6 gene remarkably improves the yield and quality of Korean pine somatic embryos and promotes the growth of Korean pine somatic embryo seedlings. The invention has important significance for optimizing the embryogenesis system of Korean pine and realizing efficient large-scale propagation.

Inventors

• YANG LING
• HE XIN
• Tang Liran

Assignees

• 北京林业大学

Dates

Publication Date

20260512

Application Date

20260331

Claims (10)

1. 1. The Korean pine PkDOF5.6 gene is characterized in that the nucleotide sequence of the Korean pine PkDOF5.6 gene is shown as SEQ ID No. 1.
2. 2. The red pine PkDOF5.6 gene according to claim 1, wherein the amino acid sequence encoded by the red pine PkDOF5.6 gene is shown in SEQ ID No. 2.
3. 3. Use of the red pine pkdof5.6 gene of claim 1 for regulating embryogenesis of red pine.
4. 4. The use according to claim 3, wherein overexpression of the red pine PkDOF5.6 gene positively modulates red pine somatic embryogenesis.
5. 5. Use according to claim 3, characterized in that overexpression of the red pine pkdof5.6 gene increases the yield and quality of red pine embryos.
6. 6. The use according to claim 3, wherein overexpression of the red pine PkDOF5.6 gene promotes red pine somatic embryo growth.
7. 7. Use according to claim 3, characterized in that overexpression of the red pine pkdof5.6 gene increases the soluble sugar and soluble protein content in the red pine embryo.
8. 8. An over-expression vector, characterized in that the red pine PkDOF5.6 gene of claim 1 is connected to pCambia1300-35s-sGFP vector to obtain the over-expression vector.
9. 9. The overexpression vector according to claim 8, characterized in that the nucleotide sequence of the pCambia1300-35s-sGFP vector is shown in SEQ ID No. 3.
10. 10. A method for improving the yield and quality of Korean pine embryos, comprising the steps of: 1) Transforming the over-expression vector of claim 8 or 9 into agrobacterium to obtain a transformed bacterium; 2) Infecting the embryogenic callus of Korean pine with the transformant in the step 1).

Description

Korean pine PkDOF5.6 gene and application thereof in regulation and control of Korean pine somatic embryogenesis Technical Field The invention relates to the technical field of plant genetic engineering, in particular to a Korean pine PkDOF5.6 gene and application thereof in regulating and controlling Korean pine somatic embryogenesis. Background Korean pine is an important economic and ecological tree species, and the somatic embryogenesis technology of Korean pine has important significance for preservation, propagation and genetic improvement of germplasm resources. Along with the rapid development of biotechnology, the transgenic technology provides a new research means and application prospect for Korean pine somatic embryogenesis. However, the transformation efficiency of conifer seeds is low, the conventional breeding and molecular breeding are not tightly combined, the popularization and application of transgenic plants are not much, the transgenic technology is effectively combined, key genes and molecular regulation mechanisms in the development process of conifer seed embryo are researched, and other leading edge technologies such as gene editing and the like are utilized to cultivate new varieties with rapid growth, high yield and strong resistance. The application of the transgenic technology in the embryogenesis regulation of Korean pine has important value, can effectively shorten the breeding period and improve the breeding efficiency by accurately regulating and controlling the expression of target genes, and has key significance for promoting the efficient and large-scale propagation of Korean pine excellent germplasm resources. The Dof transcription factor can regulate the growth and development of plants, is widely involved in plant biological and abiotic stress responses and other processes, and plays an important role in the growth and development process of plants. Disclosure of Invention In order to solve the problems, the invention provides a Korean pine PkDOF5.6 gene and application thereof in regulating and controlling Korean pine somatic embryogenesis, and the Korean pine PkDOF5.6 gene provided by the invention regulates and controls Korean pine somatic embryogenesis, and over-expresses the Korean pine PkDOF5.6 gene to obviously improve the yield and quality of Korean pine somatic embryos and promote the growth of Korean pine somatic embryo seedlings. In order to achieve the above object, the present invention provides the following technical solutions: the invention provides a Korean pine PkDOF5.6 gene, wherein the nucleotide sequence of the Korean pine PkDOF5.6 gene is shown as SEQ ID No. 1. Preferably, the coded amino acid sequence of the Korean pine PkDOF5.6 gene is shown as SEQ ID No. 2. The invention also provides an application of the Korean pine PkDOF5.6 gene in regulating and controlling Korean pine somatic embryogenesis. Preferably, overexpression of the Korean pine PkDOF5.6 gene positively regulates Korean pine somatic embryogenesis. Preferably, overexpression of the Korean pine PkDOF5.6 gene improves yield and quality of Korean pine somatic embryos. Preferably, overexpression of the Korean pine PkDOF5.6 gene promotes the growth of Korean pine somatic embryos. Preferably, overexpression of the Korean pine PkDOF5.6 gene increases the soluble sugar and soluble protein content in Korean pine somatic embryos. The invention also provides an over-expression vector, which is obtained by connecting the Korean pine PkDof5.6 gene in the technical scheme into the pCambia1300-35s-sGFP vector. Preferably, the nucleotide sequence of the pCambia1300-35s-sGFP vector is shown as SEQ ID No. 3. The invention also provides a method for improving the yield and quality of the Korean pine somatic embryos, which comprises the following steps: 1) The over-expression vector in the technical scheme is transformed into agrobacterium to obtain transformed bacteria; 2) Infecting the embryogenic callus of Korean pine with the transformant in the step 1). The invention has the beneficial effects that: The Korean pine PkDOF5.6 gene provided by the invention regulates and controls Korean pine somatic embryogenesis, and the overexpression of the Korean pine PkDOF5.6 gene remarkably improves the yield and quality of Korean pine somatic embryos and promotes the growth of Korean pine somatic embryo seedlings. The invention has important significance for optimizing the embryogenesis system of Korean pine and realizing efficient large-scale propagation. Drawings In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required to be used in the embodiments will be briefly described below. FIG. 1 is an electrophoretogram showing cloning of the PkDOF5.6 gene, M: DL 2000 DNA Marker;1: amplified PkDOF5.6 sequence; FIG. 2 is a diagram showing pCambia1300-35s-sGFP plasmid; FIG. 3 is an electrophoretogram showing construction of