Unlock instant, AI-driven research and patent intelligence for your innovation.

ghlim2 protein and its gene and its application in improving cotton fiber quality

A cotton fiber and protein technology, applied in the field of genetic engineering

Active Publication Date: 2016-01-13
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, studies in recent years have found that there is a phenylpropane metabolic pathway (GuoJY, WangLJ, ChenSP, HuWL, ChenXY (2007) Gene expression and metabolite profiles of cotton fiber during cell elongation and secondary cell wall synthesis. CellRes. 17: 422-434) in the secondary cell wall synthesis of cotton fiber. Confirmed to contain phenylpropanoid lignin (FanL, ShiWJ, HuWR, HaoXY, WangDM, HuiYuan and Hong-YingYan (2009) Molecular and Biochemical Evidence for Phenylpropanoid Synthesis and Presence of Wall-linkedPhenolics in Cotton Fibers.JIntegrPlantBiol.51:626-637) in mature cotton fibers, and found that its existence may The structure of the fiber cell wall and the growth and development of the fiber cells have an important influence

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • ghlim2 protein and its gene and its application in improving cotton fiber quality
  • ghlim2 protein and its gene and its application in improving cotton fiber quality
  • ghlim2 protein and its gene and its application in improving cotton fiber quality

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Embodiment 1, the cloning of its coding gene of cotton GhLIM2 protein

[0049] 1. Cloning of the gene encoding the cotton GhLIM2 protein

[0050] Analysis of transgenic cotton with silk fibroin light chain gene from upland cotton R15 (Gossypium hirsutum, Shangguan Xiaoxia, Wang Lingjian, Li Yane, etc.) at the elongation stage 12 days after flowering (12DPA). Acta Crops, 2007, 33(5): 697 -702; the public can obtain from the Institute of Microbiology, Chinese Academy of Sciences) isolated a cDNA encoding LIM protein family gene from the fiber, named the gene GhLIM2, and its nucleotide base is sequence 1 in the sequence table, the gene is open for reading The frame (ORF) is the 104th to 676th nucleotides from the 5' end of sequence 1 in the sequence listing. The protein encoded by this gene is named GhLIM2, and the amino acid sequence of the protein is sequence 2 in the sequence listing, and sequence 2 in the sequence listing Consists of 190 amino acid residues.

[0051]...

Embodiment 2

[0054] Example 2, construction and functional identification of transgenic GhLIM2 cotton

[0055] 1. Construction of transgenic GhLIM2 cotton

[0056] 1. Construction of recombinant expression vector

[0057] RNA was extracted from upland cotton R15 (Gossypium hirsutum) fibers. The cDNA obtained by reverse transcription was used as a template, 5'-GATCGGATCCATGGCATTTGCAGGAACAAC-3' was used as an upstream primer (with a BamHI restriction site) and 5'-GCTACCGCGGGAGCTCAGATTCAGCAGCAACTTCAGTTCCA-3' was used as a downstream primer (with a SacII and SacI restriction site ) to obtain a 570bp PCR product (after sequencing, sequence 1 with sequence listing from 104th to 673rd nucleotides at the 5' end).

[0058] Construction of pPZP-GhLIM2: Digest the above PCR product with BamHI and SacI, and combine the obtained restriction product with the pPZP vector (HajdukiewiczP, SvabZ, MaligaP (1994) The small, versatilePZPfamilyofAgrobacteriumbinaryvectorsforplant transformation. PlantMolBiol2...

Embodiment 3

[0089] Example 3, Analysis of subcellular localization of GhLIM2

[0090] The pPZP-GFP-GhLIM2 plasmid was introduced into Agrobacterium EHA105 (Biovector, catalog number: 197343) to obtain recombinant bacteria. The extracted plasmid was sent for sequencing. The plasmid was pPZP-GFP-GhLIM2, and the recombinant bacteria containing the plasmid was named pPZP-GFP-GhLIM2 / EHA105.

[0091] The pPZP-GFP-GhLIM2 / EHA105 and tobacco suspension cell BY-2 (NagataT, NemotoY, HasezawaS (1992) TobaccoBY-2 cell line as the 'HeLa' cell in the cell biology of higher plants. IntRevCytol132: 1-30; public available from the Institute of Microbiology, Chinese Academy of Sciences) were mixed statically Placed for 4 hours, 28°C, 110rpm and co-cultivated for 28-36 hours, and screened on kanamycin medium to obtain a cell line transfected with pPZP-GFP-GhLIM2. The BY-2 cells transfected with pPZP-GFP-GhLIM2 were observed with a confocal fluorescence microscope, and the excitation wavelength of GFP was 48...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
lengthaaaaaaaaaa
Login to View More

Abstract

The invention discloses GhLIM2 protein and its gene and its application in improvement of cotton fiber quality. The GhLIM2 protein is as following (a) or (b): (a) is a protein comprising an amino acid sequence shown as sequence 2 of a sequence table; and (b) is a protein which is obtained by substitution and / or deletion and / or addition of one or more amino acid residues of the amino acid sequence shown as the sequence 2 of the sequence table, and has homologous function and is associated, and derived from sequence 1. Experiments prove that the invention founds the new protein GhLIM2, when its gene is introduced into cotton, the cellulose length, fineness and strength are increased, the lignin content is also increased, the protein and its coding gene lay the foundation of the function in the cotton fiber development process, and a new target gene for improvement of the cotton fiber quality by use of a gene engineering method is probably provided.

Description

technical field [0001] The invention belongs to the field of genetic engineering, in particular to GhLIM2 protein and its gene and its application in improving cotton fiber quality. Background technique [0002] Cotton fiber is a major source of natural textile fibers and plays an important role in global economic development. With the improvement of human living standards, the requirements for high-grade fiber textiles are getting higher and higher, so improving the quality of cotton fibers has very important practical significance. However, in the process of cotton development, cotton fiber quality traits (fiber length, specific strength, micronaire value, etc.) are affected by many factors, and it is difficult to improve cotton fiber quality traits, especially simultaneous improvement. [0003] Cotton fiber is a single cell formed by the epidermal cells of the integument of the ovule through four developmental stages: differentiation, protrusion, elongation, secondary wa...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/21C12N15/11C12N15/82C12N15/84A01H5/00
CPCC07K14/415C12N15/8261
Inventor 夏桂先韩利波焦改丽李元宝王海云
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI