Method for the genetic modulation of the biosynthesis of hemicelluloses, cellulose and uronic acids in plant cells using gene expression cassettes

A technology of gene expression cassettes and plant cells, applied in the direction of using vectors to introduce foreign genetic material, cells modified by introducing foreign genetic material, biochemical equipment and methods, etc.

Inactive Publication Date: 2007-02-07
SUZANO PAPEL E CELULOSE +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, breeding programs that go beyond agronomic traits, such as the development of wood with traits related to paper bleaching and end product characteristics, are rare (Foelkel, C, supra)

Method used

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  • Method for the genetic modulation of the biosynthesis of hemicelluloses, cellulose and uronic acids in plant cells using gene expression cassettes

Examples

Experimental program
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Effect test

Embodiment 1

[0114] total RNA isolation

[0115] In this embodiment, in SALZMAN, R.A, FUJITA, T., ZHU-SALZMAN, K., HASEGAWA, P.M., BRESSAN, R.A. (1999) (An Improved RNAIsolation Method for Plant Tissues Containing High Levels of Phenolic Compounds or Carbohydrates-Plant With the help of the method of Molecular Biology Reporter 17:11-17, 1999), total RNA was obtained.

[0116] Total RNA was isolated from Eucalyptus roots. Eucalyptus roots soaked in liquid nitrogen were transferred to extraction buffer, then extracted 2–3 times with chloroform / isoamyl alcohol at a ratio of 24:1 (v / v), and centrifuged to separate the aqueous and organic phases. Proteins and carbohydrates remaining in the aqueous phase were precipitated with NaCl and ethanol for at least 3 hours at -20°C. After adding phenol:chloroform:isoamyl alcohol (25:24:1) to the supernatant obtained from the above precipitation, centrifugation was performed to separate the aqueous phase from the organic phase. A second precipitation w...

Embodiment 2

[0118] Messenger RNA extraction (mRNA)

[0119] Messenger RNA was isolated using an mRNA purification kit called "Dynabeads" from the American company DYNAL Biotech according to the manufacturer's standard.

Embodiment 3

[0121] specific primer synthesis

[0122] The sequence CACC was ligated immediately in front of the 5' region of the sense primer sequence of the overexpressed genes (UDP-glucose dehydrogenase, UDP-D-glucuronate carboxylase, UDP-glucose pyrophosphorylase), That is, the translation initiation codon is attached.

[0123] The sequence CACC was linked immediately after the 3' region of the antisense primer sequence of the inositol oxygenase gene, ie a translation stop codon.

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Abstract

The present invention refers to gene expression cassettes encoding enzymes involved in the metabolic pathway for the biosynthesis of hemicelluloses, cellulose and / or uronic acids, and to a method for genetic transformation of plant cells through the introduction of one or more gene expression cassettes from the present invention, and the overexpression and repression of these genes in plants. More particularly, the present invention refers to a method for introducing expression cassettes in plants. Additionally, the present invention refers to genetically modified plants. The present invention also refers to the attainment of the genetically modified plant, its derived plants and seeds, as well as the wood, paper and cellulose from this plant.

Description

field of invention [0001] The present invention relates to gene expression cassettes encoding enzymes involved in the biosynthetic metabolic pathways of hemicellulose, cellulose and / or uronic acid, and to the genetic transformation of plant cells by introducing one or more gene expression cassettes into said cells Methods. In addition, the present invention relates to the modification of the metabolite content and / or components of this metabolic pathway in plants or plant parts. [0002] The present invention relates to isolated genes and the respective encoded enzymes involved in the biosynthesis of hemicellulose, cellulose and / or uronic acid. [0003] The invention also relates to methods of obtaining genetically modified plants, plants and seeds derived therefrom, as well as wood, paper and cellulose derived from such plants, and their use. Background technique [0004] It is known that the chemical composition of cellulose pulp, mainly related to the presence of hemice...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/52C12N9/00C12N15/82C12N5/14C12N9/02C12N9/10C12N9/14C12N9/88C12N9/90C12N15/74
CPCC12N9/14C12N9/88D21H19/38C12N9/90D21H19/60C12N15/8243D21H21/52C12N9/0004D21H19/82C12N9/10D21H23/26C12N15/8246D21H23/50
Inventor C·A·拉贝特M·T·V·拉贝特A·L·伯特罗D·D·纳斯曼托G·古特马尼斯M·I·F·法拉尔多
Owner SUZANO PAPEL E CELULOSE
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