Rapid propagation method in vitro of Lycium barbarum nigra using young seedlings as explant donors

A technology of black fruit wolfberry and explants, which is applied in the field of forestry biology, can solve the problems of vitrification of tissue culture materials, and achieve the effect of convenient storage and low cost

Inactive Publication Date: 2017-12-12
SHENYANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, in order to solve the problem of vitrification of the tissue culture material, the present invention provides a simple method for removing the vitrification of the tissue culture material of Lycium barbarum

Method used

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  • Rapid propagation method in vitro of Lycium barbarum nigra using young seedlings as explant donors
  • Rapid propagation method in vitro of Lycium barbarum nigra using young seedlings as explant donors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] 1. Obtaining sterile seedlings

[0024] Mature seeds of Lycium barbarum were placed in warm water at 35°C for 24 hours. Soak the seeds in 70% (volume / volume) alcohol for 30 seconds on a sterile ultra-clean workbench, then soak them in 0.1% (mass / volume) mercuric chloride aqueous solution for 2 minutes, and rinse them with sterile water 3 times. Seeds were inoculated on MS solid medium without adding any plant growth regulators, and cultured at 20° C. with light for 8 hours per day. 0.43% (mass / volume) solidifying agent agar and 3% (mass / volume) sucrose were added to the MS solid medium used in this step and the following steps. The pH was adjusted to 5.8 with NaOH solution before autoclaving at 121 °C for 15 min.

[0025] 2. Inoculation of Explants

[0026] When the germinated seedlings in step 1 grow to 10 leaves, cut the well-grown young leaves transversely into small pieces with a diameter of 3 mm, and inoculate them on MS solid culture medium containing 6-BA 0.05...

Embodiment 2

[0036] 1. Obtaining sterile seedlings

[0037] Mature seeds of Lycium barbarum were placed in warm water at 37°C for 24 hours. Soak the seeds in 75% (volume / volume) alcohol on a sterile ultra-clean workbench for 1 min, then soak in 2% (mass / volume) sodium hypochlorite solution for 5 minutes, and rinse with sterile water 6 times. The seeds were inoculated on 1 / 2 MS solid medium without adding any plant growth regulator, and cultured at 28° C. with 16 hours of light every day. 0.7% (mass / volume) solidifying agent agar and 4% (mass / volume) sucrose were added to the MS solid medium used in this step and the following steps. Adjust the pH value to 5.4 with KOH solution before autoclaving at 121°C for 20 minutes. Add 2% sucrose to 1 / 2MS solid medium.

[0038] 2. Inoculation of Explants

[0039] When the germinated seedlings in step 1 grow to 20 leaves, cut the well-grown young leaves transversely into small pieces with a diameter of 5 mm, and inoculate them into MS solid culture ...

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Abstract

The invention belongs to the technical field of forest biology, particularly relates to a method for in-vitro rapid propagation of lycium ruthenicum by taking young seedlings as explant donors. The method is completed through obtaining of a sterile seedling, inoculation of an explant, subculture multiplication and acclimatization and transplanting. The method comprises the following steps: taking seedling leaves as explants to carry out inoculation, wherein 1 month after the inoculation, the bud forming rate of the explants is 87.5%; taking stems as explants to carry out inoculation, wherein 1 month after the inoculation, the bud forming rate of the explants is 100%; carrying out starvation and drying treatment, carrying out inoculation, wherein 1 month after the starvation and drying treatment, the vitrification relieving rate is 85.3%. The method is extremely high in regeneration efficiency, along with the increase of the reproduction times, the regeneration efficiency increases exponentially, and both the rooting percentage and the transplanting survival rate are above 80%. According to the method, mature seeds are used as the initial experiment materials, so that the storage is convenient and is not limited by the seasons; by combining a vitrification relieving method, the method is particular suitable for the protection, development and utilization of rare lycium ruthenicum resources; the individual reproduction cost is extremely low, so that the method is suitable for the industrialized seedling production of the lycium ruthenicum.

Description

technical field [0001] The invention belongs to the field of forestry biotechnology, and in particular relates to rapid propagation of Lycium ruthenicum by tissue culture and devitrification of vitrified materials of Lycium ruthenicum. Background technique [0002] Black Goji Berry ( Lycium ruthenicum Murr.) is a perennial thorny shrub of the genus Lycium barbarum in the family Solanaceae. It is one of the important drought-resistant and saline-alkali-resistant desert pioneer tree species. Black fruit wolfberry fruit sugar has the effects of anti-fatigue and prevention and treatment of diabetes. In addition, black fruit wolfberry is known as the king of proanthocyanidins. Its proanthocyanidins have the functions of scavenging free radicals, enhancing immunity, prolonging the lifespan of fruit flies, reducing serum total cholesterol and triglycerides in mice, and increasing serum, liver tissue and total body antioxidant capacity. Lycium barbarum with black fruit integrate...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
CPCA01H4/008
Inventor 王钦美周永斌张志宏崔建国吴月亮张丽杰张群迟峻莱
Owner SHENYANG AGRI UNIV
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