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LuMyb gene for regulating and controlling flax cellulose synthesis and application of LuMyb gene

A cellulose and flax technology, applied in the field of genetic engineering, can solve the problem of not being able to determine the direct regulator of secondary wall cellulose synthase, etc.

Pending Publication Date: 2022-04-15
黑龙江省农业科学院经济作物研究所 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented describes an improved way for creating plasmid DNA from bacteria called lymphocytes or phages which can be used to silence genes involved with disease resistance mechanisms such as drug resistances. By utilizing this technique, researchers are able to study how these diseases develop over time without causing harm themselves.

Problems solved by technology

This patents describes how fibers develop during growth stages through various processes like production or transportation from roots into leaves. These changes affect their properties including structure (length), mechanical behavior, etc., which ultimately impacts on its final product quality. Therefore understanding these mechanisms plays an important role in improving agricultural efficiency and reducing costs associated with producing high-quality products.

Method used

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  • LuMyb gene for regulating and controlling flax cellulose synthesis and application of LuMyb gene
  • LuMyb gene for regulating and controlling flax cellulose synthesis and application of LuMyb gene
  • LuMyb gene for regulating and controlling flax cellulose synthesis and application of LuMyb gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 flax LuCesA8 Identification of key regulatory regions in gene promoters

[0026] Download from the Phytozome website (https: / / phytozome-next.jgi.doe.gov / ) LuCesA8 (Lus10029245) The 2000bp promoter sequence upstream of the ORF of the gene, the cis-acting elements in the promoter were analyzed through the PLACE database, and the -426~-52bp upstream of the promoter was predicted to be the key regulatory region of the promoter (as shown in SEQ ID NO.1) . Use the software Primer Premier 5.0 to design specific primers LuCesA8F and LuCesA8R (see Table 1), and use PCR technology to clone in the flax variety "Dazihua" LuCesA8 The sequence between the gene promoter -426~-52bp (as shown in SEQ ID NO.1) obtained the promoter pLuCesA8B (-426~-52bp), and inserted the promoter sequence into the multiple cloning site EcoRI of the pCAMBIA1300-GUS vector Between -XbaI and GUS reporter gene ORF, construct the plant expression recombinant vector p1300LuCesA8 (-426~-52bp)::G...

Embodiment 2

[0028] Example 2 Yeast one-hybrid screening of transcription factor TF upstream of LuCesA8 promoter

[0029] In the early stage, 1 gene was screened by weighted co-expression analysis with LuCesA8 A candidate transcriptional regulator of gene coexpression, LuMyb (Lus10039610). Primers LuMybF and LuMybR were designed to clone the cDNA sequence of LuMyb gene (1158bp, as shown in SEQ ID NO.2), and connected to the pGADT7 vector to construct the prey vector pGADT7-Lus10039610. The primer sequences are shown in Table 1. Will LuCesA8 The gene promoter -426~-52bp sequence (as shown in SEQ ID NO.1) was connected to the pAbAi vector to construct the bait vector pLuCesA8-AbAi. The prey vector pGADT7-Lus10039610 and the bait vector pLuCesA8-AbAi, the two vectors were simultaneously transformed into yeast, and the transformation products were diluted at the ratio of 1:20, 1:50 and 1:100 respectively, and the dilution was prepared in SD / -Leu+ 300ng·mL -1 Cultured on AbA medium, pLuCe...

Embodiment 3

[0031] Example 3 LuMyb Functional verification of genes

[0032] According to the gene sequence of flax LuMyb (Lus10039610) in Phytozome public database, upstream and downstream primers LuVIGSF / LuVIGSR were designed to clone flax LuMyb The target sequence of the gene (441bp, as shown in SEQ ID NO.3), the target sequence fragment was inserted into the pTRV2 vector to construct the VIGS silencing vector pTRV2-LuMyb. flax phytoene dehydrogenase LuPDS (Lus10021967) Gene sequence design upstream and downstream primers LuPDSF / LuPDSR clone flax LuPDS The target sequence of the gene (437bp, as shown in SEQ ID NO.4), the target sequence fragment was inserted into the pTRV2 vector to construct the VIGS silencing vector pTRV2-PDS. The "Dazihua" flax seeds were evenly planted in the nutrient soil, placed in a light incubator, 23°C, 16h light, 18°C, 8h dark cultivation. After normal culture for about 10 days, the seedlings grow 2 cotyledons, and the VIGS test is carried out when the ...

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Abstract

The invention provides a LuMyb gene for regulating and controlling flax cellulose synthesis and application of the LuMyb gene. The cDNA (complementary deoxyribonucleic acid) sequence of the LuMyb gene is as shown in SEQ ID NO.2, and the amino acid sequence of the encoded protein of the LuMyb gene is as shown in SEQ ID NO.5. According to the invention, a virus-induced gene silencing technology is adopted, and a LuMyb gene silencing flax strain SE is obtained through an agrobacterium-mediated method. LuMyb is a direct regulation factor of a secondary wall cellulose synthase LuCesA8 gene, the LuMyb gene has a regulation effect on synthesis of cellulose, hemicellulose and lignin, and the discovery of the gene provides an important gene resource for molecular breeding for improving the quality of flax fibers.

Description

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Claims

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Application Information

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Owner 黑龙江省农业科学院经济作物研究所
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