Applications of HOX3 gene in improving cotton fiber elongation property

A HOX3, cotton fiber technology, applied in the field of plant genetic engineering and crop genetics, can solve the problems of unsuccessfully obtaining market promotion varieties, hindering the simultaneous improvement of cotton fiber yield and quality, etc.

Inactive Publication Date: 2016-12-07
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, due to the negative correlation between cotton fiber yield and quality, which seriously hinders the simultaneous improvement of cotton fiber yield and quality, it is difficult for traditional breeding methods to break the negative correlation between yield and quality to achieve simultaneous improvement of b...

Method used

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  • Applications of HOX3 gene in improving cotton fiber elongation property
  • Applications of HOX3 gene in improving cotton fiber elongation property
  • Applications of HOX3 gene in improving cotton fiber elongation property

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0141] Isolation of GhHOX3 cDNA and Genomic DNA

[0142] The method described in this embodiment refers to people such as Wang Shui, Control of Plant Trichome Development by a Cotton Fiber MYB Gene (Plant Cell 16,2323-2334.)

[0143] Cotton fiber tissue RNA extraction:

[0144]2 g of material (fibers on the ovule surface of upland cotton R15 9 days after flowering) (upland cotton variety "Jinmian R15", purchased from the Cotton Research Institute of Shanxi Academy of Agricultural Sciences) was ground into powder in liquid nitrogen and transferred to 50ml In the centrifuge tube, add 8ml of extraction buffer (1M Tris HCl, 50mM EDTA, 1% SDS, pH9.0) and an equal volume of water-saturated phenol:chloroform:isoamyl alcohol (25:24:1), shake and mix well, Place on ice for 1 hour and mix every 10 minutes. Centrifuge at 13000g for 20 minutes at 4°C. Repeat phenol: chloroform: isoamyl alcohol extraction 2-4 times, and finally extract once with chloroform: isoamyl alcohol (24:1). Take...

Embodiment 2

[0157] Construction of embodiment 2.GhHOX3 transgenic vector and Agrobacterium transformation

[0158] Use Primer Star DNA polymerase to amplify the target fragment (see Example 1), and determine the restriction enzyme cutting site for the fusion fragment to be inserted into the vector according to the distribution of the polyclonal region of the vector and the restriction enzyme cutting site of the GhHOX3 gene. After linking to the modified pCAMBIA2301 35S promoter, 35S::GhHOX3, 35S::GhHOX3-genomic and 35S::dsGhHOX3 transgene vectors were generated respectively, and verified by sequencing.

[0159] Transformation of Agrobacterium tumefaciens was performed by freeze-thaw method.

[0160] Take a single colony LBA4404 or GV3101 (Invitrogen), and culture it in 3ml LB medium (25μg / ml rifamycin Rif and 50μg / ml Kanamycin Kan and Gentamycin Gen) at 28°C, 220rpm overnight. Add 2ml bacterial liquid to 50ml LB medium (25μg / ml Rif and 50μg / ml Gen), cultivate to OD at 28℃, 220rpm 600 = ...

Embodiment 3

[0161] Example 3. Cotton transgene and screening of transgenic progeny

[0162] After culturing the Agrobacterium containing the vector plasmid on the YEB bacterial medium supplemented with kanamycin 50mg / L, rifampicin 100mg / L and streptomycin 300mg / L for 2-3 days, pick a single colony and inoculate it with the same antibiotic Suspension culture was carried out overnight at 28°C on a shaker at 200 rpm / min in YEB liquid medium. Centrifuge the bacterial solution at 4000rpm / min for 10 minutes, resuspend the pellet with 1 / 2MS liquid medium containing 30g / L glucose and 100μmol / L acetosyringone, and adjust the OD 600 The value is about 0.4-0.6, which is used as an infection solution for later use.

[0163] Cotton R15 (a tetraploid wild-type upland cotton, as the transgenic female parent) seeds were placed in 1 / 2MS0 medium [1 / 2MS salt (purchased from DUCHEFA M0221) + 5g / L glucose + 7g / L agar powder, pH 6.0], germination and culture in the dark, after 5-7 days, the hypocotyls of the...

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Abstract

The invention relates to applications of an HOX3 gene in improving the cotton fiber elongation property, specifically, the invention provides the applications of the cotton fiber elongation gene GhHOX3 gene of cotton and polypeptide of the cotton fiber elongation gene GhHOX3 gene in improving the cotton fiber elongation property. The experiment proves that the genomic gene of the cotton fiber elongation gene GhHOX3 can effectively and exclusively improve the cotton fiber length in the molecular level, and the synchronous improvement of the yield of the cotton fiber is realized.

Description

technical field [0001] The invention relates to the fields of plant genetic engineering and crop genetics, in particular, the invention relates to a cotton fiber elongation gene and application thereof. Background technique [0002] Cotton is an important natural fiber and economic crop, mainly including 4 cultivars: G. hirsutum, G. barbadense, G. herbaceum and G. arboreum. Cotton fiber is the main product of cotton production, and the output and quality of cotton fiber directly determine the output value and benefits of cotton production. [0003] Therefore, improving cotton fiber quality has always been the main goal of cotton breeding. The indicators for evaluating the quality of cotton fiber mainly include color, water content and impurity content in terms of fiber appearance; in terms of fiber intrinsic quality, there are mainly length, fineness, strength, etc. The development of cotton fiber goes through four stages: fiber initiation, cell elongation, secondary wall ...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00C12Q1/68G01N33/68
Inventor 陈晓亚单淳敏关雪莹上官小霞王水王凌健
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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