In-vitro preservation method of radix tetrastigme test-tube plantlets

A technology of in vitro preservation and cloverleaf, applied in the field of plant cultivation, can solve the problem that the genetic evaluation of the in vitro preservation technology has not been reported, the normal subculture preservation of tissue culture is frequently transferred, and the planting of cloverleaf germplasm resources is difficult. and other problems, to achieve the effect of preventing species degradation and virus infection, prolonging subculture time, and ensuring purity

Active Publication Date: 2021-04-16
SHANGRAO NORMAL UNIV
View PDF5 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Domestic and foreign studies on Clover mainly focus on clinical application, chemical composition and pharmacological effects. There are a few reports on tissue culture and rapid propagation technology, but there is no in vitro preservation technology and genetic evaluation of its test-tube plantlets. to report
Moreover, it is difficult to plant C. clover germplasm resources in the field, the workload of preservation is heavy, and it is easily affected by the environment, climate and pests and diseases, while the normal subculture of tissue culture is frequently transferred and the cost is high.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] A method for in vitro preservation of green clover test-tube plantlets, comprising the following steps:

[0031] (1) Select the shoot tip of Clover as an explant, soak it in 75% alcohol in mass concentration, wash it twice with sterile water, add dropwise 0.1% mercury liter in mass concentration, rinse it with sterile water for 8 times, and then add it dropwise The mass concentration is 0.1% mercuric chloride, washed 8 times with sterile water, cut into 0.5 cm long and inoculated in the first culture medium; the first culture medium includes 500mg / L NH 4 NO 3 、700mg / LKNO 3 , 150mg / L MgSO 4 ·7H 2 O, 70mg / L KH 2 PO 4 , 0.1mg / L BA, 22g / L sucrose, 8g / L agar; 20±2℃, light intensity 1500lx, light time 16h / d, subculture once every 280d;

[0032] (2) Under aseptic conditions, the isolated stem tip with a diameter of 1mm is cut from the vigorously grown aseptic seedling as material, and the isolated shoot tip is placed in the second medium for cultivation, and the second m...

Embodiment 2

[0039] A method for in vitro preservation of green clover test-tube plantlets, comprising the following steps:

[0040] (1) Select the stem tips of Clover as explants, soak them in 75% alcohol in mass concentration, rinse them twice with sterile water, add dropwise 0.1% mercury liter in mass concentration, rinse them with sterile water for 7 times, and then add them dropwise The mass concentration is 0.1% mercuric chloride, washed 8 times with sterile water, cut into 0.8 cm long and inoculated in the first culture medium; the first culture medium includes 650mg / L NH 4 NO 3 、600mg / LKNO 3 , 120mg / L MgSO 4 ·7H 2 O, 100mg / L KH 2 PO 4 , 0.1mg / L BA, 20g / L sucrose, 10g / L agar; 20±2℃, light intensity 1800lx, light time 16h / d, subculture once every 280d;

[0041] (2) Under aseptic conditions, the isolated stem tip with a diameter of 1mm is cut from the vigorously grown aseptic seedling as material, and the isolated shoot tip is placed in a second medium for cultivation, and the s...

Embodiment 3

[0048] A method for in vitro preservation of green clover test-tube plantlets, comprising the following steps:

[0049] (1) Select the stem tips of A. trifoliate as explants, soak them in 75% alcohol by mass concentration, rinse them twice with sterile water, add 0.1% mercury liter dropwise, rinse them with sterile water for 6 times, and then add them dropwise The mass concentration is 0.1% mercuric chloride, rinsed with sterile water for 6 times, cut into 1.0 cm lengths and inoculated in the first culture medium; the first culture medium includes 600mg / L NH 4 NO 3 、700mg / LKNO 3 , 150mg / L MgSO 4 ·7H 2 O, 60mg / L KH 2 PO 4 , 0.1mg / L BA, 20g / L sucrose, 5g / L agar; 20±2℃, light intensity 1800lx, light time 16h / d, subculture once every 300d;

[0050] (2) Under aseptic conditions, the isolated stem tip with a diameter of 1mm is cut from the vigorously grown aseptic seedling as material, and the isolated shoot tip is placed in a second medium for cultivation, and the second medi...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses an in-vitro preservation method of radix tetrastigme test-tube plantlets, and belongs to the technical field of plant cultivation. The in-vitro preservation method comprises the following steps: selecting a radix tetrastigme stem tip as an explant, carrying out pre-culture, and then carrying out secondary culture; embedding balls in the stem tips cultured for the second time by using sodium alginate; taking out the embedding balls containing the stem tips, placing the embedding balls on sterile filter paper on a sterile culture dish, drying for 1-2 h in sterile air flow, after drying the sterile air flow, putting the embedding balls into freezing pipes, and putting 10-20 embedding balls into each freezing pipe; and putting the freezing pipe filled with the embedding balls into a freezing pipe frame or a sandbag, and then putting into liquid nitrogen for ultralow-temperature storage. According to the in-vitro preservation method, no callus tissue is formed, genetic variation is avoided, the genetic stability is good, the regenerated seedlings recover and grow well, and the plant regeneration rate can reach 99% or above.

Description

technical field [0001] The invention relates to the technical field of plant cultivation, in particular to a method for in vitro preservation of test-tube plantlets of A. trifoliate. Background technique [0002] Clover (Tetrastigma hemsleyanum Diels et Gilg), also known as three-leaf cliff climbing vine, golden thread hanging gourd, belongs to Rhamnaceae, a herbaceous vine of Vitis vinaceae. Herbaceous vines with three leaves, slender branchlets, longitudinally ribbed, glabrous or sparsely pilose. Tendrils unbranched, opposite to leaves with intervals of 2 nodes. The leaves are 3 leaflets, the leaflets are lanceolate, oblong-lanceolate or egg-lanceolate, 3-10cm long, 1.5-3cm wide, apex acuminate, sparsely acute, base cuneate or round, lateral leaflet base not Symmetrical, nearly round, with 4-6 serrations on each side of the edge, thin or sometimes thicker, green above, light green below, glabrous on both sides; 5-6 pairs of lateral veins, not obvious on both sides of the...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 洪森荣周聪张子鑫余洁姚尧尹明华
Owner SHANGRAO NORMAL UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap