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Tomato lipoxygenase family silent expression vector, and construction method and application thereof

A technology of lipoxygenase and expression vector, applied in the field of genetic engineering, to achieve the effect of improving nutritional value, simple and easy construction method, and improving tomato fruit quality

Inactive Publication Date: 2014-07-02
CHONGQING UNIV
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Problems solved by technology

In recent years, there are relatively many reports on the use of transgenic technology to increase the resistance to diseases and insect pests, herbicides and salt and alkali resistance of tomato, but there is no report on the use of transgenic technology to increase the content of unsaturated fatty acids in tomato fruits.

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  • Tomato lipoxygenase family silent expression vector, and construction method and application thereof
  • Tomato lipoxygenase family silent expression vector, and construction method and application thereof
  • Tomato lipoxygenase family silent expression vector, and construction method and application thereof

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Embodiment Construction

[0033] Preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings. The experimental method that does not indicate specific conditions in the preferred embodiment is usually according to conventional conditions, such as described in the Molecular Cloning Experiment Guide (Third Edition, J. Sambrook et al., translated by Huang Peitang, etc., Science Press, 2002) conditions, or as recommended by the manufacturer.

[0034] The tomato line used in the preferred embodiment is wild-type tomato ( Lycopersicon esculentum Mill. cv Ailsa Craig). For the construction method of plasmid pDHG5.2 (carrying CaMV35S promoter, GFP gene open reading frame and CaMV35S terminator), see literature (Kim SH, Grierson D. Subcellular localization and silencing of ripening-associated membrane protein (TRAMP) in tomato Lycopersicon esculentum Mill. Plant Science, 2005, 169: 1022-1029), obtained by inserting the GFP gene open reading fra...

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Abstract

The invention discloses a tomato lipoxygenase family silent expression vector and its construction method and application. The silent expression vector contains a double-stranded RNA expression unit, including a promoter, a forward segment of the TomloxACD splicing gene, and a reverse segment of the TomloxACD splicing gene and terminator, the TomloxACD splicing gene is spliced ​​by the TomloxA gene fragment (SEQIDNo.1), the TomloxC gene fragment (SEQIDNo.2) and the TomloxD gene fragment (SEQIDNo.3); the silencing expression vector is transferred into by Agrobacterium-mediated The contents of α-linolenic acid and linoleic acid in the fruits of transgenic positive plants obtained from wild-type tomato were significantly higher than those of non-transgenic plants, indicating that the silencing expression vector can be used for breeding tomato plants with high unsaturated fatty acid fruit. It has a good application prospect in improving the fruit quality and improving the nutritional value of tomato fruit.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and relates to a gene silencing expression vector, a construction method of the gene silencing expression vector, and an application of the gene silencing expression vector in plant breeding. Background technique [0002] tomato( Lycopersicon esculentum Mill.) is a kind of important crop of Solanaceae, is one of the most common fruits and vegetables cultivated in the whole world. Essential fatty acids are essential for the life activities of the body, but the body cannot synthesize them or the synthesis speed is too slow to meet the needs of the body. The polyunsaturated fatty acids that must be supplied by food mainly include α-linolenic acid and linoleic acid. Genetic engineering breeding has become an important branch in the field of plant breeding, showing great potential in the directional improvement of plant characteristics. In recent years, there are relatively many reports...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82C12N15/84C12N15/66C12N1/21A01H5/08
Inventor 陈国平胡廷章曾华胡宗利屈霄霄涂昀
Owner CHONGQING UNIV
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