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Method for obtaining anti-wilting hydrangea macrophylla flowers

A hydrangea and hydrangea technology, applied in the field of gardens, can solve the problems of easy degradation of traits, difficult maintenance, long flowering period, etc., and achieve the effect of increasing the deposition of secondary cell walls

Inactive Publication Date: 2018-05-15
范瑶飞
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Deciduous shrub, clustered, 1-4m high, corymb inflorescence, terminal in spherical shape, white when it first blooms, then turns blue, and pink when it fades, flowering period from May to July, long flowering period, large and plump flowers, now widespread Cultivation, the market demand is very large, only relying on cuttings is far from meeting the market demand, and due to improper cultivation management measures, the traits are prone to degeneration
[0003] Fresh-cut hydrangea flowers are beautiful and have beautiful meanings, but they are difficult to maintain. Especially in the dry environment in the north, fresh-cut hydrangea flowers will wilt to varying degrees and irreversible almost the next day after they are separated from the body. To solve this problem, the present invention provides a How to Get Wilting Resistant Hydrangeas

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Embodiment 1: obtain the nucleotide sequence of cellulose overexpression protein

[0019] The amino acid sequence (754 amino acids) of Xre cellulose overexpression protein, as shown in SEQ ID NO: 1 in the sequence listing; The nucleotide sequence (2262 cores) of coding is corresponding to the amino acid sequence of described Xre cellulose overexpression protein nucleotide), as shown in SEQ ID NO:2 in the sequence listing.

[0020] The nucleotide sequence of the Xre cellulose overexpressed protein (shown as SEQ ID NO: 2 in the sequence listing) was synthesized by Nanjing GenScript Biotechnology Co., Ltd. The 5' end of the synthetic Xre cellulose overexpression protein nucleotide sequence (SEQ IDNO:2) is also connected with an NcoI restriction site, and the Xre cellulose overexpression protein nucleotide sequence (SEQ IDNO :2) The 3' end is also connected with a SwaI restriction site.

Embodiment 2

[0021] Example 2: Construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0022] 1. Construction of recombinant cloning vector containing Xre cellulose overexpression protein

[0023] The nucleotide sequence of the synthetic Xre cellulose overexpressed protein was connected to the cloning vector pGEM-T (Promega, Madison, USA, CAT: A3600), and the operation steps were carried out according to the instructions of the pGEM-T vector produced by Promega Company to obtain a recombinant clone Vector Xre01-T.

[0024] Then the recombinant cloning vector Xre01-T was transformed into Escherichia coli T1 competent cells (Transgen, Beijing, China, CAT: CD501) by heat shock method, and the heat shock conditions were: 50 μl Escherichia coli T1 competent cells, 10 μl plasmid DNA, 42 ℃ water bath for 30 seconds; 37 ℃ shaking culture for 1 hour (shaking at 100rpm speed), coated with IPTG (isopropylthio-β-D-galactoside) and X-gal (...

Embodiment 3

[0029] Embodiment 3: the acquisition of transgenic hydrangea plants

[0030] Cut the tender stems about 2.0-3.0cm long from the hydrangea plant, remove the leaves, rinse with tap water first, then rub the decontamination powder with a brush to scrub repeatedly, and then rinse with pure water. Under aseptic conditions, soak in 75% alcohol for 10-20 seconds, soak in 0.1% mercuric solution for 6-8 minutes, rinse with sterile water for 5-6 times, and finally cut the treated tender stems into about 0.5-1.0 long According to the conventionally adopted Agrobacterium infection method, co-cultivate with the Agrobacterium transformed by the recombinant expression vector described in Example 2 and 3, inoculate on the pre-starting medium, and the pre-starting medium adopts MS culture 6-BA3mg / L and NAA0.1mg / L were added to MS medium. Two weeks after inoculation of the explants, the axillary buds swelled, sprouted and grew leaves. Transfer the sterile buds obtained above to the differenti...

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Abstract

The invention provides a method for obtaining anti-wilting hydrangea macrophylla flowers, and relates to the technical field of gardening. The method comprises the steps that an Xre cellulose overexpression gene is introduced into a hydrangea macrophylla plant, so that the hydrangea macrophylla plant expresses Xre cellulose overexpression protein; the hydrangea macrophylla flowers with anti-wilting capability are obtained. The plant is changed through genetic engineering; the plant has lignin depositions or xylan depositions basically concentrated on xylem tissue ducts of the plant, so that the increased secondary cell wall depositions or the increased wax / cutin depositions are obtained. The plant subjected to engineering modification can be used for bioenergy production; the anti-lodgingand anti-wilting goals are achieved by improving the density and cellulose content of biomass from the plant.

Description

technical field [0001] The invention relates to the technical field of gardens, in particular to a method for obtaining wilting-resistant hydrangeas. Background technique [0002] Hydrangea macrophylla, also known as Hydrangea, belongs to the deciduous shrub of the family Saxifragaceae. It is native to the central and southern parts of my country, and is now cultivated in the north and south. It likes a semi-shady, warm, and humid environment. It requires acidic soil rich in humus, moist, and well-drained, with a pH of 4.0 to 4.5. Its flower color is closely related to the pH of the soil. Deciduous shrubs, clumps, 1-4m high, corymbs, spherical inflorescences, white at the beginning, then turn blue, and pink when withering, flowering period from May to July, long flowering period, large and full flowers, now widespread Cultivation, the market demand is very large, and only relying on cuttings is far from meeting the market demand. At the same time, due to improper cultivati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/29A01H5/00A01H6/80
CPCC07K14/415C12N15/8261
Inventor 范瑶飞
Owner 范瑶飞
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