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Method for improving carotenoid content of tomato

A carotene and tomato technology, applied in the fields of botanical equipment and methods, biochemical equipment and methods, measuring devices, etc., to achieve the effects of high yield, simple method and easy production

Inactive Publication Date: 2011-02-23
山东省农业科学院高新技术研究中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in the retrievable prior art, the method of increasing tomato carotenoid content by expressing cryptochrome gene has not been reported yet.

Method used

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  • Method for improving carotenoid content of tomato
  • Method for improving carotenoid content of tomato
  • Method for improving carotenoid content of tomato

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Construction of cryptochrome gene 1 (cry1), cryptochrome gene 1 carboxyl end (GUS::CCT1) and cryptochrome gene 2 carboxyl end (GUS::CCT2) overexpression vector.

[0046] The primers were designed according to the Arabidopsis cryptochrome genes cry1 (gene accession number: AY124863.1) and cry2 (gene accession number: NM_100320.3) in NCBI (National Center for Biotechnology Information). The total RNA of Arabidopsis was extracted, and the sequences of cry1, CCT1 and CCT2 were amplified from Arabidopsis by RT-PCR method (see sequence table). Such as figure 1 As shown, the plant expression vector pBI121 was modified, the β-glucuronidase (GUS) was replaced with the green fluorescent protein tag (GFP tag) gene, and the CRY1 gene was forwardly linked to the cauliflower mosaic virus (CaMV) 35S promoter Later, it was constructed into the plant expression vector pBI121 to obtain the plant expression vector GFP::CRY1; the carboxyl end of the CRY1 gene and the carboxyl end of...

Embodiment 2

[0047] Example 2 Transformation of plants

[0048] 1. Establishment of tomato tissue culture regeneration system

[0049] Sterilize tomato seeds with sodium hypochlorite for 10 minutes, wash them with sterile water 5 times, plant them on 1 / 2MS medium, and germinate in a culture room at 25°C for 6 days. When the cotyledons of the seedlings are fully extended and the true leaves just appear , Cut the cotyledons or hypocotyls of the seedlings and place them on the regeneration medium for cultivation. After 1 month, when obvious small shoots grow on the explants, move the explants together with the small shoots to the elongation medium. When the regenerated shoots grow to about 1 cm, the small shoots are cut off and placed on a rooting medium to induce rooting. When the root length is about 1 cm, move the regenerated seedlings to the soil.

[0050] Through the optimization of the culture medium and the cultivation conditions and the selection of the explants' age, the regeneration fre...

Embodiment 3

[0070] Example 3 Obtaining transgenic tomato fruit

[0071] Harvest tomato seeds, plant and obtain second-generation seeds, extract DNA for PCR detection and Western hybridization, verify transgenic plants, and harvest homozygous fruits of T2 generation.

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Abstract

The invention discloses a method for improving carotenoid content of a tomato. Overexpression vectors of cryptochrome gene 1, the c-terminus of the cryptochrome gene 1 and the c-terminus of cryptochrome gene 2 are constructed by a gene engineering means to transform the tomato so as to obtain the transgenic tomato. By the method, the lycopene content and total carotenoid content in the transgenic tomato fruit are greatly improved by 3.6 times and 3.4 times respectively, so that the extraction of the natural lycopene from the tomato is more efficient, and the additional value of the tomato product is greatly improved.

Description

Technical field [0001] The invention relates to a method for the content of tomato carotenoids, in particular to a method for increasing the content of tomato carotenoids by expressing cryptochrome genes, and belongs to the field of biotechnology. Background technique [0002] Carotenoids are the general term for a class of important natural pigments, and their main components are isopentenoid polymers containing 40 carbons (C), namely tetraterpenoids. Carotenoids are commonly found in yellow, orange-red or red pigments in animals, higher plants, fungi, algae and bacteria. [0003] Carotenoids can inhibit the body's aging-related diseases and reduce the incidence of certain cancers [1] . Eating more foods rich in carotenoids such as fruits and vegetables can reduce the incidence of aging-related diseases such as coronary heart disease (Coronary heart disease), age-related macular degeneration (AMD), cataracts and other diseases . Studies have shown that if the body's intake of c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/53A01H5/08G01N30/02C12N15/82
Inventor 王兴军姜娜娜洛佩斯·哈维尔赵传志
Owner 山东省农业科学院高新技术研究中心
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