Application of cymbidium goeringii miR159a in accelerating plant life cycle

A life cycle, technology of plants, applied in the field of plant genetic engineering

Active Publication Date: 2020-04-10
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, although miR159 plays an important role in the growth and development of plants, its specific regulatory mechanism has not yet been clarified. Therefore, using genetic engineering technology, it will be cloned from Chunlan miR159a The transfer of precursor genes into other plants is of great significance for the study of their functions and has great application prospects

Method used

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  • Application of cymbidium goeringii miR159a in accelerating plant life cycle
  • Application of cymbidium goeringii miR159a in accelerating plant life cycle
  • Application of cymbidium goeringii miR159a in accelerating plant life cycle

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1 Functional Prediction of Genes

[0017] Material 1 used in this embodiment is the roots, stems, and leaves of Chunlan "Song Mei" in the vegetative growth period and reproductive growth period, and material 2 is the young leaves of Chunlan "Song Mei" in the childhood period and the maturity of the vegetative growth period in a year. The leaves and the leaves in the reproductive growth period were quickly frozen in liquid nitrogen after harvest, and stored in an ultra-low temperature refrigerator (-80°C).

[0018] 1) Extraction of Total Small RNA from Chunlan tissues

[0019] According to the instructions of the TaKaRa plant total RNA extraction kit, the specific operation is as follows:

[0020] The ultra-low temperature cryopreserved Chunlan tissues were quickly transferred to a mortar pre-cooled with liquid nitrogen, and the tissues were ground with a pestle, during which liquid nitrogen was continuously added until they were ground into powders; the powdere...

Embodiment 2

[0031] Example 2 Cloning and Transformation of Genes

[0032] The plant material used in this embodiment is the fresh leaf of Chunlan "Song Mei", Arabidopsis thaliana ( Arabidopsis thaliana ) is of the "Columbia" type. The Escherichia coli strain used is Trans5α, which is used for gene cloning and overexpression vector construction. The vector construction is as follows: figure 2 Shown; the Agrobacterium strain is GV3101, which is used to transform Arabidopsis; the plant expression vector used in the experiment is pBI121. The strains used were purchased from Qingke Biotechnology Co., Ltd. and Price Biotechnology Co., Ltd., respectively.

[0033] 1) Extraction of gDNA from Chunlan leaves

[0034] Follow the instructions of the TaKaRa Plant Genomic DNA Extraction Kit, the specific operations are:

[0035] Transfer the fresh leaves of Chunlan "Songmei" to a liquid nitrogen precooled mortar, grind the tissue with a pestle, and add liquid nitrogen continuously until it is gro...

Embodiment 3

[0076] Example 3 Chunlan miR159a Accelerated functional identification of plant life cycles

[0077] Acquisition of transgenic homozygous plants: Harvested transgenic T1 generation seeds were sterilized, screened and cultivated, and then transplanted in nutrient soil, cultured at 22°C, 16 h light / 8 h dark; after the detection, the initially confirmed transgenic plants were retained and waited for Harvest the seeds of the T1 generation after maturity, number them, and obtain the T2 generation; like the T1 generation, sterilize the T2 generation seeds and spread them on the screening medium containing antibiotics, place them at 22°C under continuous light; about 10 days, The survival rate of T2 generation seeds with different numbers was counted, and the plants with a survival ratio of 75% were selected for transplantation and cultured in nutrient soil at 22°C, 16 h light / 8 h dark, and the leaves were taken for positive detection; positive T2 generation plants continued Numberi...

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Abstract

The invention discloses a cymbidium goeringii miR159a gene and an application of the cymbidium goeringii miR159a gene in accelerating plant life cycle. An miR159a precursor sequence fragment coding gene is obtained from cymbidium goeringii cultivar ''Song Mei'', expression analysis is performed in the cymbidium goeringii, then the gene is constructed into an overexpression vector and introduced into a target plant to verify the function, and results find that overexpression miR159a gene arabidopsis thaliana plants have a shortened juvenile period, an advanced flowering period, advanced fruit ripening and other phenotypes that accelerate the life cycle, so that the gene has wide applications in the production and breeding of orchids and other horticultural plants.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, in particular to a kind of Chunlan non-coding RNA (ribonucleic acid) miR159a Application in accelerating plant life cycle. Background technique [0002] Orchidaceae (Orchidaceae) is one of the largest families of flowering plants, with more than 25,000 species in the world, accounting for about 10% of all flowering plants. Chunlan ( Cymbidium goeringii ) belongs to the small-flowered ground orchid species in the Orchidaceae genus. It has a peculiar flower shape, elegant flower color, quiet flower fragrance, beautiful leaf appearance, and high ornamental and economic value. However, it takes a long time for Chunlan to bloom from seedlings. Wild Chunlan seedlings need about ten years to bloom, and artificial tissue culture seedlings need at least three years to bloom under the condition of proper maintenance. Therefore, it is of great significance to study the molecular mechan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/113C12N15/82A01H5/00A01H6/20
CPCC12N15/113C12N15/8261C12N15/827C12Q1/6895C12Q2600/13C12Q2600/158C12Q2600/178
Inventor 刘倩徐子涵胡凤荣张鸽香
Owner NANJING FORESTRY UNIV
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