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Method of lettuce isolated-culture high-frequency regeneration

An in vitro culture and high-frequency regeneration technology, which is applied in the field of plant cell tissue culture, can solve the problems of difficult hybridization technology, unstable character separation, and small flower organs, so as to save experiment and labor costs and reduce the link of inducing budding , the effect of short seedling time

Inactive Publication Date: 2019-11-15
SHANGHAI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is almost no obvious heterosis in lettuce. Even if there is a certain degree of heterosis, it is difficult to apply it to production because of its small flower organs and difficult hybridization technology.
Therefore, the first generation of hybridization is rarely used in the varieties promoted in production, and the breeding of new lettuce varieties through traditional hybrid breeding methods faces a series of problems such as time-consuming and labor-consuming, and unstable trait separation. In order to shorten the breeding period and improve more targeted The existing "Shenxuan series" of lettuce for the purpose of obtaining better quality new varieties must rely on cell and genetic engineering technology to assist traditional breeding, and the establishment of the "Shenxuan series" lettuce different genotypes in vitro culture and high-efficiency regeneration system is a further development. The premise and foundation of lettuce cell engineering and transgenic research and new variety breeding

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0026] A method for high-frequency regeneration of lettuce culture in vitro, the specific steps are as follows:

[0027] (1) Soak the dried lettuce seeds in warm water at 30°C for 3 hours until the seeds are fully imbibed;

[0028] (2) Sterile inoculation: transfer the seeds into a sterile bottle, use 1% sodium hypochlorite as a surface disinfectant, vibrate up and down for 10 minutes, discard the disinfectant and rinse the seeds twice with sterile water, each time for 5 minutes; drain Inoculate on MS medium after hydration, and cultivate in a light culture room with a temperature of 24°C, a light intensity of 2000lx, and a light duration of 16h / d;

[0029] (3) Induction of callus and germination: After the aseptic seedlings grow for 5 days, cut 3mm hypocotyls and 5mm cotyledons as explants and place them in the germination medium to induce callus and bud differentiation. Pay attention to cutting the cotyledons Face up to ensure that the incision is fully in contact with the ...

Embodiment 2

[0033] A method for high-frequency regeneration of lettuce culture in vitro, the specific steps are as follows:

[0034] (1) Soak the dried lettuce seeds in warm water at 40°C for 4 hours until the seeds are fully imbibed;

[0035] (2) Aseptic inoculation: transfer the seeds into a sterile bottle, use 2% sodium hypochlorite as a surface disinfectant, shake up and down for 15 minutes, drain the disinfectant and rinse the seeds with sterile water for 3 times, each time for 8 minutes; drain Inoculate on MS medium after hydration, and cultivate in a light culture room with a temperature of 26°C, a light intensity of 2000lx, and a light duration of 16h / d;

[0036] (3) Induction of callus and germination: After the sterile seedlings grow for 7 days, cut 6mm hypocotyls and 8mm cotyledons as explants and place them in the germination medium to induce callus and bud differentiation. Pay attention to cutting the cotyledons Face up to ensure that the incision is fully in contact with th...

Embodiment 3

[0040] A method for high-frequency regeneration of lettuce culture in vitro, the specific steps are as follows:

[0041] (1) Soak the dried lettuce seeds in warm water at 35°C for 3.5 hours until the seeds are fully imbibed;

[0042] (2) Aseptic inoculation: transfer the seeds into a sterile bottle, use 1.5% sodium hypochlorite as a surface disinfectant, shake up and down for 12 minutes, drain the disinfectant and rinse the seeds twice with sterile water, each time for 6 minutes; drain Inoculate on MS medium after hydration, and cultivate in a light culture room with a temperature of 25°C, a light intensity of 2000lx, and a light duration of 16h / d;

[0043] (3) Induction of callus and germination: After the sterile seedlings grow for 6 days, cut 4mm hypocotyls and 7mm cotyledons as explants and place them in the germination medium to induce callus and bud differentiation. Pay attention to cutting the cotyledons Face up to ensure that the incision is in full contact with the m...

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Abstract

The invention relates to a method of lettuce isolated-culture high-frequency regeneration. The method includes the steps that cotyledons and hypocotyls of aseptic lettuce seedlings serve as explants to be subjected to isolated culture and are induced to bud on a budding culture medium, and then, buds are transplanted to a rooting culture medium to form a mass of seedlings. Main components of the budding culture medium include 0.2 mg / L of an MS basic medium, 0.5 mg / L of NAA, 30 g / L of 6-BA and 8 g / L of agar; main components of the rooting culture medium include 0.1 mg / L of an MS basic medium, 30 mg / L of IAA, 30 g / L of sucrose and 8 g / L of agar. According to the method of lettuce isolated-culture high-frequency regeneration, the specific hormone-proportioned media are adopted, the culture period is shortened by 7-10 days compared with current regeneration systems, the differentiation frequency of the explants can achieve 100%, it can be ensured that the seedling regeneration rate reaches95% or above, and experiment and labor costs are saved.

Description

technical field [0001] The invention belongs to the technical field of plant cell tissue culture, and in particular relates to a high-frequency regeneration method for in vitro culture of lettuce. Background technique [0002] Plant cell engineering is a biotechnology that uses in vitro cultured cells for genetic manipulation to improve plant varieties. As the basic means of cell engineering and genetic engineering research, plant cell tissue in vitro culture technology has the characteristics of controllable experimental conditions, strong repeatability, and consistent growth of test-tube plantlets. It is also the technical basis for obtaining genetically modified plants. Lettuce is a variety of Lactuca genus in Compositae. It is rich in vitamin C and various mineral elements. It is a vegetable group with high edible value. At the same time, this group has short growth cycle, wide suitable cultivation area and simple cultivation conditions. characteristics; moreover, exoge...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 王颖王虹阎君宋云鹏朱为民
Owner SHANGHAI ACAD OF AGRI SCI
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