Combined vector capable of increasing content of coenzyme Q10 in tomatoes as well as construction method and application of combined vector

A construction method and carrier technology, applied in the field of plants, can solve problems such as difficulties in the stable intake of coenzyme Q10

Active Publication Date: 2021-01-29
SHANGHAI CHENSHAN BOTANICAL GARDEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Under the influence of the concept of increasing the proportion of vegetable and fruit diet structure in today's society, maintaining the coenzyme Q in food 10 It becomes extremely difficult to maintain a stable intake of

Method used

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  • Combined vector capable of increasing content of coenzyme Q10 in tomatoes as well as construction method and application of combined vector
  • Combined vector capable of increasing content of coenzyme Q10 in tomatoes as well as construction method and application of combined vector
  • Combined vector capable of increasing content of coenzyme Q10 in tomatoes as well as construction method and application of combined vector

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036]Such asfigure 1 As shown, coenzyme Q10The biosynthetic pathway of the quinone ring mainly includes four processes: (1) the formation of the aromatic ring structure of the quinone ring precursor; (2) the polymerization of isoprene pyrophosphate into a chain; (3) the formation of the aromatic ring structure and the isopentenyl side chain Binding; (4) Modification on the quinone ring after binding. The first three processes directly determine the downstream coenzyme Q10The output. Therefore, promoting the first three steps of the reaction process is the main goal of our technical solution:

[0037](1) Promote the formation of the aromatic ring structure of the precursor of the quinone ring: Coenzyme Q10The precursor of the quinone ring structure is 4-hydroxybenzoic (4-HB). In prokaryotes, 4-HB is formed by chorismate lyase (UbiC) catalyzed by chorismate lyase. In eukaryotes, 4-HB is converted from phenylalanine or tyrosine, but the specific formation process is still unclear. Theref...

Embodiment 2

[0064]This embodiment provides a method for improving coenzyme Q10For the production method of content tomato, the level 2 vector obtained in Example 1 was transformed into Agrobacterium GV3101 by heat shock, and transformed into wild-type tomato MicroTom; after obtaining independent resistant callus, further differentiation and regeneration were obtained to obtain tomato strain HUCD .

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Abstract

The invention relates to a combined vector construction method capable of increasing the content of coenzyme Q10 in tomatoes. The combined vector construction method comprises the following steps: obtaining an E8 promoter sequence, a tobacco Coq2 gene coding sequence, a DPS gene coding sequence, an HMGR2 gene coding sequence, a UbiC gene coding sequence and an hsp terminator sequence; constructingrespective level 0 vectors of the six sequences; constructing respective level 1 vectors of the four genes; and constructing a level 2 vector. The invention also relates to a production method of tomatoes capable of increasing the content of coenzyme Q10, which comprises the following steps: transforming agrobacterium GV3101 by using the combined vector obtained by the construction method throughthermal shock, and transforming wild tomatoes; and after obtaining independent resistant calluses, further differentiating and regenerating to obtain the tomato strain with high coenzyme Q10 content.Through cultivation and screening, the highest yield of T3-generation fruit coenzyme Q10 of the tomato strain HUCD can reach more than 7 times (-0.151 mg / g) of wild tomato fruits, and the tomato strain HUCD can be stably inherited and has no difference in phenotype; namely, the content of the coenzyme Q10 in the tomato strain HUCD is remarkably increased.

Description

Technical field[0001]The present invention relates to the field of plant technology, in particular to a method that can increase coenzyme Q in tomato10Content of the combined carrier and its construction method and application.Background technique[0002]Coenzyme Q10(Coenzyme Q10, Also known as CoQ10The molecular structure of) is composed of benzoquinone parent group and a quinone with decaisopentene side chain. Coenzyme Q10In the human body, it is mainly used as an electron transporter of complex II / III in the oxidative respiratory chain to help cells produce ATP. In addition, it is also an antioxidant on the membrane and is used as a cofactor for many enzymes. In the human body, lack of coenzyme Q10It can cause a variety of serious neurological diseases, such as mitochondrial encephalomyopathy, cerebellar ataxia or Leigh's encephalopathy, and affect the function of multiple organs such as skeletal muscle, heart, and kidney. Under normal circumstances, coenzyme Q10The level will decr...

Claims

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Application Information

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IPC IPC(8): C12N15/66C12N15/84A01H5/00A01H6/82
CPCC12N15/66C12N15/8243C12Y401/0304C12Y301/03047C12N9/1085C12Y205/01039C12N9/16C12N9/88
Inventor 许晶晶范航陈晓亚杨蕾
Owner SHANGHAI CHENSHAN BOTANICAL GARDEN
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