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Application of oilseed rape BnC3H gene in improving biological self-luminous intensity of plants

A self-luminous and genetic technology, applied in the biological field, can solve the problems of not making a fundamental breakthrough and not yet put into production, and achieve the effect of improving the intensity of biological self-luminescence

Active Publication Date: 2022-02-08
ZHEJIANG UNIV HANGZHOU GLOBAL SCI & TECH INNOVATION CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, none of these projects have made a fundamental breakthrough and have not yet been put into production.

Method used

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  • Application of oilseed rape BnC3H gene in improving biological self-luminous intensity of plants
  • Application of oilseed rape BnC3H gene in improving biological self-luminous intensity of plants
  • Application of oilseed rape BnC3H gene in improving biological self-luminous intensity of plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Construction process of large fragment bioluminescence module DNA vector pYLTAC380GW-7G / 8G

[0046] In this study, the TransGene Stacking II system was used for multigene assembly. The NPGA (4'-phosphopantetheinyl transferase) gene in the Aspergillus nidulans genome, the H3H (hispidin-3-hydroxylase) gene and the Hisps (Hispidin synthase) gene in the Neonothopanus nambi genome, and the CPH ( Caffey pyruvate hydrolase) gene, rapeseed p-coumaric acid 3-hydroxylase BnC3H (p-coumaric acid 3-hydroxylase) driven by 35S promoter and fungal luciferase (Luz) gene were integrated into the pYLTAC380GW plasmid, and finally passed one step The BP recombination reaction connects and removes the selection marker gene expression cassette element to form a basic vector

[0047] pYLTAC380GW-Hisps-CPH-H3H-NPGA-Luz (pYLTAC380GW-7G) ( figure 1 ), the enhanced carrier

[0048] pYLTAC380GW-Hisps-CPH-H3H-NPGA-Luz-BnC3H (pYLTAC380GW-8G), the enhanced vector contains overexpressed ra...

Embodiment 2

[0059] Example 2 Analysis of Luminescence Intensity After Transient Transformation of Phalaenopsis Petals by Biological Self-luminescence Module

[0060] 1. Streak and activate the EHA105 bacterial solution containing the verified correct vector plasmid pYLTAC380GW-7G / 8G on the LA+Kana+Rif plate, 28°C, 36h, pick colonies from each plate, and transfer them to LB+ Culture in Kana+Rif+15μM As medium, 28℃, 200rpm to OD 600 =0.8-1.0, 4000rpm, 10min to collect the bacterial cells, suspend the Agrobacterium bacterial cells with the infection solution (containing 10mM MgCl, 10mM MES, 150μM As), and let stand at room temperature for 2-3h.

[0061] 2. To verify the transient expression in the Phalaenopsis petals, use a 1mL needle to gently open a small opening on the surface of the Phalaenopsis petals, then use the needle tube with the needle removed to absorb the bacterial liquid, and inject it into the petals from the wound of the Phalaenopsis petals. Indoor normal cultivation for 48...

Embodiment 3

[0063] Example 3 Analysis of Luminescence Intensity of Transgenic Plants of Tobacco Bioself-luminescence Module Mediated by Agrobacterium

[0064]1. Streak the EHA105 bacterial solution containing the verified correct vector plasmid pYLTAC380GW-7G / 8G on the LA+Rif+Kana plate, 28°C, 36h, pick a single clone into 3-5ml LB medium at 200rpm, 28°C , 36h, according to the ratio of 1:100-1:50, expand the culture of 50ml for 3-5h to OD=0.6, then centrifuge the bacterial solution, and suspend the bacteria with MS0 liquid medium (MS+3%Sucrose+PH5.8, 50ml) Body to OD=0.6 for infection;

[0065] 2. Select healthy tobacco leaves (4-5 weeks) that have fully unfolded, cut them into 0.5cm square size with a scalpel (cut off the leaf edge to avoid the main vein), and place the upper surface of the leaf downward on MS1 ​​solid medium (MS+0.5 mg / L IAA+2.0mg / L BA+3% sucrose+0.6-0.8% Phytagel, pH=5.8), culture in dark at 25°C for 2-3 days;

[0066] 3. Add the pre-cultured tobacco leaves to the b...

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Abstract

The invention discloses an application of an oilseed rape BnC3H gene in improving the biological self-luminous intensity of plants, and belongs to the technical field of biology. The CDS sequence of the BnC3H gene is as shown in SEQ ID NO.1. The application comprises the steps: (1) sequentially integrating a Hisps gene, a CPH gene, a H3H gene, a NPGA gene, a Luz gene and a BnC3H gene into a receptor vector by utilizing a multi-gene assembly technology, and constructing to obtain a multi-gene vector; and (2) introducing a target gene segment into a receptor plant by using a transgenic technology to obtain a transgenic plant with enhanced biological self-luminous intensity. The invention discloses co-expression of the BnC3H gene and the Hiss gene, the CPH gene, the H3H gene, the NPGA gene and the Luz gene participating in plant self-luminescence for the first time, and the biological self-luminescence intensity of the plant can be remarkably improved. The invention provides a feasible technical scheme for breeding and production of sustainable luminous plants.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of rapeseed BnC3H gene in improving the intensity of plant bioluminescence. Background technique [0002] Bioluminescence is a natural phenomenon that can be observed in about 10,000 species in 800 genera. Organisms capable of naturally emitting light include bacteria, fungi, algae, invertebrates, and insects, most of which are marine organisms (Haddock et al, 2010). In most cases, the function of bioluminescence is to convey visual signals to warn predators, attract prey or courtship. After further scientific research, it is also expected to develop a self-luminous "tree light" to replace street lights to illuminate the road, so as to reduce the power consumption of urban road lighting and save energy. [0003] There have been related reports on the research on self-illuminating plants. In 2013, synthetic biologists and botanists at Stanford University proposed th...

Claims

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Application Information

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IPC IPC(8): C12N15/53C12N9/02C12N15/84C12N15/64A01H5/00A01H5/02A01H6/82A01H6/62
CPCC12N9/0073C12N15/8205C12N15/8241C12Y114/13036Y02A40/146
Inventor 都浩钟静玲葛杰瑜郑鹏
Owner ZHEJIANG UNIV HANGZHOU GLOBAL SCI & TECH INNOVATION CENT
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