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Method for cultivating colored cotton

A cultivation method and technology of colored cotton, which is applied in the fields of botany equipment and method, biochemical equipment and method, plant gene improvement, etc., and can solve the problems of monotonous color, lack of color, and low color saturation of colored cotton

Active Publication Date: 2019-01-18
孙玉强
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the lack of germplasm resources of other colors except brown and green in the existing colored cotton, traditional genetic breeding methods cannot solve the problems of monotonous color and low color saturation of colored cotton.

Method used

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  • Method for cultivating colored cotton
  • Method for cultivating colored cotton
  • Method for cultivating colored cotton

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1 Utilizing Upland Cotton Purple Mutant HS2 to Cultivate New Color Cotton

[0060] 1. Obtaining of cotton purple mutant HS2

[0061] The constructed T-DNA expression vector (pBI121) was transferred to Agrobacterium LB4404 strain for expansion and culture, using cotton C312 seedling hypocotyls as explants, and the T-DNA vector-loaded Agrobacterium LB4404 in the basic induction medium ( MSB5 medium, 2,4-D (2,4-dichlorophenoxyacetic acid) 0.1 mg / L + KT (cytokinin) 0.1 mg / L) were co-cultured at 23°C for 36-48 hours. Wash the co-cultured hypocotyls with sterile water containing cephalosporin (500mg / L), and then place the hypocotyls in the induction medium (MSB5 medium, 2,4-D 0.1mg / L+KT 0.1mg / L+kanamycin 50mg / L) cultured at 28°C to induce embryogenic callus. Transfer resistant embryogenic callus to the selection medium (MSB5 medium, IBA (indole acetic acid) 0.5mg / L+KT0.15mg / L+kanamycin 50mg / L) to select successfully transformed embryogenic callus Tissue, differently gr...

Embodiment 2

[0080] Example 2 Utilizing RNAi to interfere with GhCHS, GhANR, and GhLAR genes respectively to cause the color change of colored fibers

[0081] Clone the full-length GhCHS, GhANR, GhLAR genes, select the genes GhCHS (nucleotide sequence shown in SEQ ID NO. 5), GhANR (nucleotide sequence shown in SEQ ID NO. 6), GhLAR (nucleoside The acid sequence is shown in SEQ ID NO. 7) specific sequence fragments to construct interference expression vectors of GhCHS, GhANR, and GhLAR genes by cotyledon osmotic injection method (refer to Fu et al. Acyl-CoA N-acyltransferase influences fertility by regulatinglipid metabolism) and jasmonic acid biogenesis in cotton. Scientific Reports, 2015, 5: 11790) electrotransformation of Agrobacterium GV3101.

[0082] Find the cDNA sequence of the target gene in NCBI, use Primer Express 5.0 software to design virus interference primers for GhPDS, GhCHS, GhANR, and GhLAR. Both ends of the primers are added with SpeⅠ and AscⅠ restriction sites and protective ba...

Embodiment 3

[0094] Example 3 Using RNAi to interfere with the GhOMT1 gene to cultivate red cotton

[0095] (1) The GhOMT1 gene (shown in SEQ ID NO.1) was inserted into the plant expression vector pBI121-35S-NOS in the forward direction, and the CaMV35S promoter was used to promote the expression to construct the plant expression vector pBI21-35S-GhOMT1- containing the GhOMT1 gene. NOS, such as Figure 14 .

[0096] The GhOMT1 gene fragment (shown in SEQ ID NO.1) was inserted into the plant interference expression vector pB7GWIWG2(II), and the CaMV35S promoter was used to start the expression to construct the plant interference expression vector pB7GWIWG2(II)-GhOMT1-F- containing the GhOMT1 gene. T35S, such as Figure 15 .

[0097] The constructed vector was used to transform the competent DH-5α Escherichia coli by heat shock. In this process, the recombinant was cultivated with LB kanamycin selective medium. The plasmid pB7GWIWG2(II)-GhOMT1 extracted from Escherichia coli DH-5α transformed with...

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Abstract

The invention discloses a method for cultivating colored cotton, which comprises the following steps: (1) crossing cotton purple mutant HS2 as parent with different varieties of cotton to obtain colored cotton; (2) knocking out, editing, interfering with or overexpressing the key enzyme genes in the anthocyanin biosynthetic metabolic pathway of cotton to obtain colored cotton; The key enzyme genesinclude PAL, C4H, 4CL-8, CHS, CHI, F3H, F3 'H, F3' 5 'H, DFR, LAR, LDOX, ANR, OMT or GST.

Description

(1) Technical field [0001] The invention relates to a breeding method for cultivating colored cotton by utilizing genetic modification technology of key genes of anthocyanin metabolism pathway and combining conventional hybrid breeding technology. (2) Background technology [0002] Cotton is one of the most important economic crops in the world, and China is the world's largest textile producer and consumer. In order to meet people's demand for colorful apparel, cotton textiles must be chemically dyed before being made into finished products. However, many chemical substances are used in its processing, which may cause harm to human health; at the same time, due to the requirements of printing and dyeing processing technology, printing and dyeing fabrics need to consume a large amount of water during the processing process and discharge water pollutants. According to the production volume of printing and dyeing cloth in my country's printing and dyeing industry in recent years, ...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H1/02A01H5/00A01H6/60
CPCA01H1/02C12N15/8242
Inventor 孙玉强柯丽萍
Owner 孙玉强
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