MYB (Myeloblastosis) gene PeMYB103 (Petal Epidermis Myeloblastosis 103) related to Euramerican populus anther development and RNAi (Ribose Nucleic Acid interfere) vector thereof

A pemyb103, carrier technology, applied in genetic engineering, plant genetic improvement, recombinant DNA technology, etc., can solve the problems of transgenic plant safety, female plant flying flocculation, male plant loose powder, etc., and achieve the effect of great research value and application potential.

Inactive Publication Date: 2011-09-14
LUDONG UNIVERSITY
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  • Abstract
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Problems solved by technology

[0005] Poplar is a model plant for the study of tree genetics, development and genetic engineering improvement. It is also an important economic tree species and is widely planted in

Method used

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  • MYB (Myeloblastosis) gene PeMYB103 (Petal Epidermis Myeloblastosis 103) related to Euramerican populus anther development and RNAi (Ribose Nucleic Acid interfere) vector thereof
  • MYB (Myeloblastosis) gene PeMYB103 (Petal Epidermis Myeloblastosis 103) related to Euramerican populus anther development and RNAi (Ribose Nucleic Acid interfere) vector thereof
  • MYB (Myeloblastosis) gene PeMYB103 (Petal Epidermis Myeloblastosis 103) related to Euramerican populus anther development and RNAi (Ribose Nucleic Acid interfere) vector thereof

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specific Embodiment approach

[0024] The specific embodiment: the present invention will be described in further detail below in conjunction with accompanying drawing:

[0025] The methods used in the following examples are conventional methods unless otherwise specified.

[0026] experiment material

[0027] 1. Strains and vectors: Escherichia coli strain DHα and Agrobacterium strain LBA4404 were purchased from Huamei Biological Company; expression vector pCambia1300 vector was purchased from Cambia Company; expression vector pBI121 was purchased from Clontech Company.

[0028] 2. Main reagents: Taq DNA polymerase was purchased from Beijing Quanshijin Company, DNA standard molecular weight was purchased from Takara Company; restriction endonuclease, T4 DNA ligase, M-MLV reverse transcriptase were purchased from Promega Company; RNeasy Plant Mini Kit was purchased from Qiagen; BDSMART TM RACE cDNA Amplification Kit was purchased from Clontech. All other chemical reagents were of domestic analytical gra...

Embodiment 1

[0048] Example 1. Cloning and identification of Populus americana MYB gene PeMYB103

[0049] 1.1 Isolation of the 3′ end of PeMYB103

[0050] 1.1.1 Amplification of the 3′ end of PeMYB103

[0051] The inflorescences of Yang Xiong in Europe and the United States were used as materials, and the total RNA was extracted using the RNeasy Plant Mini Kit. With BD SMART TM The RACE cDNA Amplification Kit kit was used for reverse transcription to synthesize the first strand of cDNA. The above specific operations were performed according to the kit instructions. According to the amino acid sequence RCGKSCR of the conserved region and a degenerate primer PMYBf, 3' RACE PCR was carried out using cDNA as a template to obtain a specific fragment. The conditions of PCR amplification were as follows: 94°C for 3 minutes after pre-denaturation, 94°C for 1 minute, 50°C for 1 minute, 72°C for 90s, a total of 35 cycles.

[0052] 1.1.2 Ligation, transformation, sequencing

[0053] Ligation: A...

Embodiment 2

[0060] Example 2, Sequence analysis and homology comparison of Populus americana PeMYB103 gene

[0061] Using DNAstar and Primer Premier 5.0 software to analyze the amino acid sequence encoded by PeMYB103 cDNA, the results show that PeMYB103 is a protein composed of 311 amino acids, with a molecular weight of 35KD and an isoelectric point of 5.6. It has a typical R2R3MYB domain: R2 (12 -amino acid 64) and R3 (amino acids 65-115). Starting from the 17th amino acid in the R2 domain, there is a conserved hydrophobic amino acid residue tryptophan every 19 amino acids, a total of 3 Ws; the first W in the R3 domain is phenylalanine (F, Amino acid at position 70) is substituted, and there is a W residue every 18 amino acids from L. The above analysis results indicated that PeMYB103 is a typical R2R3 MYB transcription factor family member. Using the Blast software in the NCBI database to compare PeMYB103 with the sequence in GenBank and search for similarity, it was found that the h...

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Abstract

The invention discloses an MYB (Myeloblastosis) gene PeMYB103 (Petal Epidermis Myeloblastosis 103) related to Euramerican populus (Populus X euramericana) anther development and an RNAi (Ribose Nucleic Acid interfere) vector thereof. In the invention, an anther specific MYB gene PeMYB103 is obtained from the Euramerican populus for the first time; and a nuclear localization transcription factor consisting of 311 amino acids is encoded by the gene. The homology between the PeMYB103 and an important transcripton AtMYB103 for controlling the development of an Arabidopsis anther is 52%. In the invention, an RNAi vector of the PeMYB103 is constructed and used for reducing or suppressing the expression of the PeMYB103 to obtain a transgenic plant with pollen abortion. The gene disclosed by the invention can be potentially used in creating a male sterile line in a genetic engineering way, in particular breeding a new variety of the populus.

Description

Technical field: [0001] The invention relates to a MYB gene PeMYB103 related to the anther development of Populus x euramericana and its RNAi carrier, belonging to the field of molecular biology and biotechnology. Background technique: [0002] Anthers are the male reproductive organs of plants, where the male gametophytes - pollen - develop. Studying the mechanism of anther development is of great significance for understanding the genetic mechanism of plant reproduction, improving breeding methods for agricultural and forestry crops, and preventing transgene drift. [0003] MYB is one of the largest families of plant transcription factors, and the members of this family have a common feature of containing MYB domain at the N-terminus. According to the number of MYB domains, MYB transcription factors can be divided into three types: MYB1R, R2R3-MYB, and MYB3R, which contain 1, 2 and 3 MYB domains, respectively. Each MYB domain usually consists of 51-52 amino acids. In ter...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/63C12N1/21C12R1/19C12R1/01
Inventor 宿红艳王磊冯培勇张娟刘林德
Owner LUDONG UNIVERSITY
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