Method for instantaneously changing orchid color of dendrobium nobile by using DhMYB2 gene
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A Dendrobium orchid technology, applied in the field of molecular biology and biology, can solve the problem of changing flower color and so on
Pending Publication Date: 2022-03-25
TROPICAL CORP STRAIN RESOURCE INST CHINESE ACAD OF TROPICAL AGRI SCI
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[0005] So far, there has been no successful application of VIGS technology in Dendrobium orchids, nor has there been an...
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Embodiment 1
[0050] Embodiment 1, the method for instantaneously changing the color of dendrobium orchid
[0051] 1. Construction of VIGS vector
[0052] 1. DhMYB2-pMD18-T recombinant plasmid
[0053] Using the flower buds of the Dendrobium variety 'Red Pearl' (purchased from Hainan Oriental Canaan Orchid Industry Co., Ltd.) as material, the total RNA was extracted by using the Tiangen Polysaccharide Polyphenol Plant Total RNA Extraction Kit. Thermo Scientific reverse transcription kit was used to synthesize cDNA as a template. Design full-length cDNA amplification primers: MYB2F1: 5'-GCAATGGGAAAAAATCCGTGC-3'; MYB2R1: 5'-CGGGAAAAAAATTCTATGACAAA-3', and use the above templates for amplification.
[0054] The amplification reaction program was 35 cycles of 95°C for 3min, 95°C for 30S, 60°C for 30S, 72°C for 1min for 30S, and 72°C for 10min.
[0055] Amplification reaction system: 10 μL of Vazyme2×Phanta Max Master Mix (purchased from Nanjing Novizyme Biotechnology Co., Ltd.), 0.5 μL of fo...
Embodiment 2
[0116] Example 2, Agrobacterium-mediated transient expression of the pCymMV-pro60 recombinant vector of DhMYB2-1 and DhMYB2-4 in Dendrobium
[0117] 1. Transient expression changes Dendrobium orchid flower color
[0118] The recombinant bacteria 1 (OD) expressing DhMYB2-1 prepared in Example 1 was selected. 600nm value reached 1.0) and recombinant bacteria expressing and DhMYB2-4 4 (OD 600nm The value reached 1.0), and the other two Dendrobium varieties "17002" (parents were hybrids of 'Pomo'×'Swan Orchid' respectively, and the parents were purchased from Hainan Dongfang Canaan Orchid Industry Co., Ltd.) and 'Hua'er '(purchased from Hainan Oncidium Biotechnology Co., Ltd.), the injection method is the same as in Example 1. Seven strains were injected per treatment.
[0119] 30 days after injection, use CIEL under C 2° light source using NF555 colorimeter * a * b * The flower color parameters of petals and sepals of multiple color-changing flowers of Dendrobium orchid det...
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Abstract
The invention discloses a method for instantaneously changing the orchid color of dendrobium nobile by using a DhMYB2 gene. The invention provides application of a substance for inhibiting the biological function of the DhMYB2 protein or a substance for inhibiting the expression of a DhMYB2 protein coding gene in any one of the following phenotypes or cultivation of a plant with any one of the following phenotypes: 1) changing the flower color of dendrobium nobile; (2) the content of anthocyanin in the dendrobium nobile lindl is reduced; (3) increasing the brightness of the flower color of the dendrobe orchid; 4) reducing the brilliance of the dendrobium nobile flowers; (5) enabling the single dendrobium plants to form different flower colors; the method specifically comprises the following steps: constructing a VIGS vector from a segment of sequence of a CDS coding region of a DhMYB2 gene obtained from dendrobium nobile, transferring into agrobacterium, and injecting dendrobium nobile to realize flower color change. According to the method, the flower color changing effect is remarkable, different plants and different flowers are different in expression, and the ornamental value and interestingness are high.
Description
technical field [0001] The invention relates to a method for instantaneously changing the flower color of dendrobium orchids by using DhMYB2 gene, which belongs to the field of molecular biology and biotechnology. Background technique [0002] Dendrobium has more than 1,500 species and many artificial hybrids, mainly grown and planted in tropical and subtropical regions of Asia and eastern Oceania. Dendrobium orchid has bright colors, various flower types, and long fresh-keeping period. It is a flower with high ornamental value. It is also called the fourth in the world together with Phalaenopsis, Oncidium and Cattleya Large ornamental Cayenne orchid is widely used in cut flowers, potted flowers, boutonnieres, bouquets, and table flowers. Flower color is the most important ornamental character of Dendrobium. The main colors of Dendrobium orchids are dark red, purple, pink, yellow, green, and white, and there are rich coloring methods, such as solid color, patterned, and sp...
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