阅读说明：本技术 一种控制杨树不定根形成和株高发育的关键基因PeSCE1及其应用 (Key gene PeSCE1 for controlling adventitious root formation and plant height development of poplar and application thereof ) 是由 刘思安 胥猛 祁浩然 吴玲 于 2020-05-21 设计创作，主要内容包括：本发明公开了一种控制杨树不定根形成和株高发育的关键基因PeSCE1及其应用,关键基因PeSCE1的核苷酸序列如SEQ ID NO.1所示,其表达蛋白的氨基酸序列如SEQ NO.2所示。本发明以“南林895”杨不定根为材料,通过RACE技术克隆了PeSCE1基因,利用Gateway技术构建过量表达载体35S::PeSCE1并转入杨树中。PeSCE1转基因杨树的不定根数目明显减少,株高也明显降低。这表明PeSCE1基因是调控杨树不定根形成和株高发育的关键基因,在林木基因工程和无性系林业领域有重要应用价值。(The invention discloses a key gene PeSCE1 for controlling adventitious root formation and plant height development of poplar and application thereof, wherein the nucleotide sequence of the key gene PeSCE1 is shown as SEQ ID NO.1, and the amino acid sequence of an expression protein thereof is shown as SEQ NO. 2. The invention takes Nanlin 895 poplar adventitious roots as a material, clones PeSCE1 gene by RACE technology, and constructs an overexpression vector 35S by Gateway technology, namely PeSCE1 and transfers the overexpression vector into poplar. The number of the indefinite roots of the transgenic poplar PeSCE1 is obviously reduced, and the plant height is also obviously reduced. The result shows that the PeSCE1 gene is a key gene for regulating the formation of adventitious roots and the plant height development of poplar trees and has important application value in the fields of forest genetic engineering and clone forestry.)

1. A key gene PeSCE1 for controlling the formation of adventitious roots and the plant height development of poplar trees has a nucleotide sequence shown in SEQ NO. 1.

2. The expression protein of the key gene PeSCE1 for controlling the adventitious root formation and plant height development of poplar according to claim 1, wherein the amino acid sequence of the expression protein is shown as SEQ NO. 2.

3. A vector containing PeSCE1, which is the key gene for controlling the formation of adventitious roots and the plant height development of poplar according to claim 1.

4. The carrier of claim 3, wherein: the vector is assembled and combined into a strong expression promoter P35S at the 5' end of the PeSCE1 gene.

5. The carrier of claim 3, wherein: the vector has strong terminator NOS assembled at the 3' end of the PeSCE1 gene.

6. The carrier of claim 3, wherein: the vector is assembled into an NPT II gene expression cassette which is used as a screening marker of transgenic poplar.

7. A host cell comprising PeSCE1, which is the key gene for controlling adventitious root formation and plant height development of poplar according to claim 1 or the vector according to claim 3.

8. The use of the key gene PeSCE1 for controlling the adventitious root formation and plant height development of poplar according to claim 1 in improving the adventitious root number and plant height of poplar.

Technical Field

The invention belongs to the technical field of plant genetic engineering, and particularly relates to a key gene PeSCE1 for controlling adventitious root formation and plant height development of poplar and application thereof.

Background

The poplar is one of the important fast-growing industrial wood species and greening afforestation species, and plays an important role in the sustainable development of clonal forestry. In the process of seedling culture, the poplar is generally subjected to cutting seedling culture, so the difficulty of rooting of cutting slips directly influences the survival rate of afforestation, and the adaptability and the stress resistance of a clone are related. Although most poplar trees are easier to root by cutting, many elite clones have difficulty rooting by cutting. The importance of tree height to tree species of timber is self-evident, and is an important trait for breeders to improve varieties. With the development of forest genetic improvement work and genetic engineering technology, people have new understanding on the importance of poplar cutting propagation, and are aware of the necessity and the urgency of strengthening the research on the cutting rooting mechanism in theory and practice.

Ubiquitin is a small peptide related to protein degradation, and a small ubiquitin-like modifier (SUMO) is one of numerous ubiquitin-like modifications with structural conformation and connection mechanism similar to the ubiquitin system. SUMO is an important post-translational modification, and there are three major enzymes involved in SUMO: SUMO activating enzyme (E1 activating enzyme, SAE), SUMO binding enzyme (E2 binding enzyme, SCE1) and SUMO ligase (E3 ligase, SIZ and MMS21/HPY 2). The three enzymes are activated, combined and linked to carry out the SUMO process. In plants, SUMO plays an essential role in the growth and development of plants and in stress response. In Arabidopsis, SUMO plays an important role in abiotic stresses such as drought and low temperature, and in growth and development. In rice, SUMO is involved in cold stress, ABA stress response and growth development regulation. However, the key enzyme gene SCE1 in the SUMO process is still less researched at present, and the function research of SCE in forest trees is rarely reported.

No research report of PeSCE1 exists in the poplar, and the cloning and research of the function of the gene are helpful for understanding the regulation and control mechanism of the gene on the formation of adventitious roots and the height of plants of the poplar, can also promote the development process of forest molecule breeding, and have immeasurable value on the clonal afforestation of the poplar and the increase of the wood yield.

Disclosure of Invention

The purpose of the invention is as follows: aiming at the defects in the prior art, the invention aims to provide a key gene PeSCE1 for controlling the formation of adventitious roots and the plant height development of poplar. The invention also aims to provide the application of the key gene PeSCE1 for controlling the formation of adventitious roots and the plant height development of the poplar in improving the number and the plant height of the adventitious roots of the poplar.

The technical scheme is as follows: in order to achieve the purpose of the invention, the technical scheme adopted by the invention is as follows:

a key gene PeSCE1 for controlling the formation of adventitious roots and the plant height development of poplar trees has the nucleotide sequence shown in SEQ NO. 1.

The PeSCE1 expression protein for controlling the formation of adventitious roots and the plant height development of poplar trees has the amino acid sequence shown in SEQ NO. 2.

The carrier containing the key gene PeSCE1 for controlling the adventitious root formation and the plant height development of the poplar.

As an embodiment:

the vector is assembled and assembled to form a strong expression promoter P35S at the 5' end of the PeSCE1 gene, and the PeSCE1 gene can be efficiently expressed in a poplar body.

The carrier is provided with a strong terminator NOS at the 3' end of the PeSCE1 gene, and can effectively terminate the transcription of the PeSCE1 gene.

The vector is assembled with an NPT II gene expression cassette, and the NPT II gene expression cassette is used as a screening marker of transgenic poplar, and can be used for screening transgenic poplar by using kanamycin.

The vector assembles LB and RB sequences, and facilitates the integration of the PeSCE1 gene expression frame and the screening marker gene NPT II assembled between the sequences into the chromosome of the poplar receptor cell.

The host cell containing the key gene PeSCE1 for controlling the adventitious root formation and the plant height development of the poplar or the vector.

The invention finally provides the application of the key gene PeSCE1 for controlling the formation of the adventitious roots and the plant height development of the poplar in improving the number and the plant height of the adventitious roots of the poplar.

The invention takes Nanlin 895 poplar adventitious roots as a material, and clones the PeSCE1 gene by RACE technology. Meanwhile, a Gateway cloning technology is adopted to construct a poplar over-expression vector 35S, namely PeSCE1, the gene is positioned behind a promoter P35S, and the PeSCE1 can be efficiently expressed in a poplar body under the drive of the promoter P35S, so that the formation of adventitious roots and the plant height development of the poplar can be regulated and controlled.

Has the advantages that: according to the invention, the PeSCE1 gene is transferred into the poplar, the number of adventitious roots of the transgenic poplar with the overexpression PeSCE1 gene is obviously reduced, and the plant height is also obviously reduced, which indicates that PeSCE1 is a key gene for controlling the formation of adventitious roots and the plant height development of the poplar. The poplar with different adventitious root numbers and plant heights can be obtained by regulating the expression level of the gene, so that the gene has important application value in the fields of forest genetic engineering and clonal forestry.

Drawings

FIG. 1 is a schematic structural diagram of a constructed plant expression vector 35S, namely PeSCE 1;

FIG. 2 is the molecule detection of transgenic poplar with over-expression of PeSCE1 gene;

FIG. 3 is a graph comparing the overall morphology of transgenic poplar overexpressing the PeSCE1 gene with that of non-transgenic poplar; CK in the figure is not transgenic poplar;

FIG. 4 is the statistics of the number of adventitious roots of transgenic poplars and non-transgenic poplars overexpressing the PeSCE1 gene.

Detailed Description

The technical solution of the present invention is further described in detail by the following specific examples.