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Method for improving cottonseed nutritional quality and application thereof

A cottonseed and nutrition technology, applied in the field of plant genetic engineering, can solve the problems of carotenoid content and improvement in cottonseed that have not yet been seen, and achieve the effects of simple and easy method, enhanced cottonseed nutrition, and increased carotenoid content

Inactive Publication Date: 2016-06-22
SOUTHWEST UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But so far, there is no report about the improvement of cottonseed carotenoid content

Method used

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  • Method for improving cottonseed nutritional quality and application thereof
  • Method for improving cottonseed nutritional quality and application thereof
  • Method for improving cottonseed nutritional quality and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 Acquisition of cotton phytoene synthase gene GhPSY2

[0049] On the Phytozome public database (https: / / phytozome.jgi.doe.gov), using the base sequence of Arabidopsis phytoene synthase gene AtPSY (At5g17230) as bait, BlastN search homologous genes. The results showed that the gene numbered Gorai.006G009400 in Raymond cotton was highly homologous to At5g17230. Using the CDS sequence of Gorai.006G009400 as a reference, design primers (Table 1, GhPSY2-U&GhPSY2-D), use upland cotton cDNA as a template, amplify the obtained fragment, and add BamHI and EcoRI sites at both ends of the fragment. The sequencing results are as follows: SEQ ID NO.1. Cluster analysis showed that this sequence had high homology with known phytoene synthase genes from other species (such as Arabidopsis, tomato, soybean, etc.), so this sequence was considered to be a cotton phytoene synthase gene. Gene encoding erythromycin synthase (GhPSY2).

Embodiment 2

[0050] Example 2 Construction of the GhPSY2 expression vector specific to the seed development stage

[0051] The procedure for constructing GhPSY2 into the plant expression vector pLGN-Nos is shown in figure 2 . The pLGN-Nos vector is a binary plant expression vector transformed from the traditional plant expression vector pBI121. Its T-DNA segment (region between RB and LB, figure 2 ) was replaced by a fusion gene expression cassette controlled by the constitutive CaMV35S promoter (CaMV35S-P) reporting gene GUS and marker gene NPTII, and another expression cassette controlled by CaMV35S-P.

[0052] Use primers (Table 1, pV-F and pV-R) to amplify and clone the promoter pV from the bean genome, and add HindⅢ and BamHI sites at both ends of the promoter. GhPSY2 was excised from the cloning vector with BamHI and EcoRI, and the promoter pV was excised from the cloning vector with HindIII and BamHI. The above two fragments were simultaneously added to the ligation reaction s...

Embodiment 3

[0053] The genetic transformation of embodiment 3 cotton

[0054] The cotton genetic transformation of the above expression vector was carried out by the method mediated by Agrobacterium tumefaciens, and the medium formula used is shown in Table 2. The specific method is as follows: select wild-type Jimian No. 14 cotton seeds (hulled) with full grains, place them in a sterile Erlenmeyer flask, rinse with 75% alcohol for 1 minute, and then rinse with 0.1% HgCl 2 Sterilize for about 10 minutes (shake the Erlenmeyer flask), fully rinse with sterile water 6-8 times, place on a shaker at 30°C (100-110rpm), and wait for the radicle to grow about 1cm (about 24-36h, every 8 hours Change the water once), take it out, and insert the radicle into the seed germination medium. Culture in a dark room at 30°C until the hypocotyls elongate to 2-3cm (about 48h). A single colony of Agrobacterium carrying the expression cassette vector was inoculated in liquid YEB medium containing 50 mg / LKm a...

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Abstract

The invention belongs to the technical field of plant genetic engineering, and particularly relates to a method for improving cottonseed nutritional quality and application thereof. The technical purpose that a new choice is provided for improving the cottonseed carotenoid content and improving cottonseed nutritional quality is achieved. According to the technical scheme, a seed-specific expression promoter is built to drive the plant expression vector of a phytoene synthetase gene PSY, cotton is transformed, the carotenoid content in cottonseeds is improved, and the PSY gene is the cotton GhPSY2 gene. After the target gene is regulated through the method, the carotenoid content in mature cottonseeds is significantly improved. The method is easy and convenient to implement and remarkable in effect, and has the application prospect of producing huge economic benefits.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and in particular relates to a method for improving the nutritional quality of cottonseed and its application. Background technique [0002] Carotenoids belong to isoprenoid compounds and are an important class of plant natural pigments. Carotenoids are a class of fat-soluble pigments with bright colors, mainly yellow, orange and red. Carotenoids are important health-protecting substances for animals and humans. As the only source of vitamin A in humans and animals, vitamin A formed by the transformation of carotenoids is an indispensable nutrient for humans and animals, and can effectively play a role in the health care of the vision and skin systems. In addition, carotenoids have antioxidant effects, and their presence can delay aging and reduce the incidence of cardiovascular diseases and cancer. However, the animal body itself cannot directly synthesize carotenoids, and t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/84A01H5/00
CPCC12N9/1085C12N15/8205C12N15/8243
Inventor 肖月华王毅姚丹李倩罗明侯磊裴炎
Owner SOUTHWEST UNIVERSITY
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