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Lilyturf root embryoid induction and culture method

A culture method and embryoid body technology, applied in the field of plant cell engineering, can solve the problems of failing to achieve detoxification of seedlings, purification and rejuvenation, not being a breeding material, and a long culture period, achieving easy regeneration, improving seedling rate, The effect of stable offspring

Inactive Publication Date: 2009-05-06
JIANGSU POLYTECHNIC COLLEGE OF AGRI & FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This kind of culture process uses several kinds of basic medium and hormones together, not only the culture period is long, the cost is high, and it is easy to mutate during the tissue culture process, it is not the best breeding material, let alone achieve the detoxification of seedlings, The role of purification and rejuvenation

Method used

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  • Lilyturf root embryoid induction and culture method
  • Lilyturf root embryoid induction and culture method
  • Lilyturf root embryoid induction and culture method

Examples

Experimental program
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Effect test

Embodiment 1

[0035]Take a healthy plant of dwarf sagegrass, cut off the root and the old stem section at the base, and peel off the old leaves in turn, leaving only 6 young leaves wrapped around the tip of the stem, first rinse it under tap water for about 1-2 hours, and then wash it with Soak in solution for 10 minutes, then rinse several times with tap water, rinse with sterile water 4 to 6 times, disinfect with 2% sodium hypochlorite for 15 minutes, and dry the surface with sterile paper; put the material into 0.1% Disinfect in mercuric chloride solution for 8 minutes, rinse with sterile water for 8 to 10 times, peel off the outer leaves, and inoculate the young explants wrapped in shoot tips in 1 / 2MS+GA ( 5mg / L) + BA (3mg / L) + IBA (0.1mg / L) + sucrose 20g / L + 0.7% agar medium (pH 5.8-6.0, the same below), 3 in each bottle, Aseptic culture, the temperature is 25±2°C during the day and 20±2°C at night, the light intensity is 500-10001x., the light time is 10-14h, and the shoot tip differe...

Embodiment 2

[0037] Take a healthy plant of slender-leafed grass, cut off the roots and old stems at the base, and peel off the old leaves one by one, leaving only 6 young leaves wrapped around the tip of the stem, first rinse it under tap water for 1-2 hours, and then wash it with detergent Soak in the solution for 10 minutes, then rinse several times with tap water, rinse with sterile water 4 to 6 times, disinfect with 2% sodium hypochlorite for 10 minutes, and dry the surface with sterile paper; put the material into 0.1% L Sterilize in mercury solution for 15 minutes, rinse with sterile water for 8 to 10 times, peel off the outer leaves, and inoculate the young explants wrapped in shoot tips in 1 / 2MS+GA (3mg / L) + BA (2mg / L) + IBA (0.1mg / L) + sucrose 20g / L + 0.7% agar medium (pH 5.8-6.0, the same below), 3 pieces per bottle, no For bacterial culture, the temperature is 25±2°C during the day and 20±2°C at night, the light intensity is 500-1000lx., the light time is 10-14h, and the shoot...

Embodiment 3

[0039] Carry out the cultivation of aseptic seedlings by the same embodiment of dwarf sage grass and slender sage grass (substratum 1 / 2MS+GA (4mg / L)+BA (1mg / L)+IBA ( 0.1mg / L)+white sugar 20g / L+0.7% agar), primary embryoid bodies (medium MS+BA(0.1mg / L)+NAA(0.2mg / L)+2.4D(1mg / L)+sucrose 30g / L+0.7% agar) and secondary embryoid bodies (medium MS+BA (4mg / L)+NAA (0.2mg / L)+sucrose 30g / L+0.7% agar) induction and subculture. What needs to be pointed out here is that Grass nigra is a species that is more difficult to regenerate. The inventor improved the medium with SH, B 5 and N 6 The medium was basic medium, and different types of hormones were added to it to induce embryoid bodies, but the ideal embryoid body incidence rate could not be obtained. However, using MS primary embryoid body medium, it takes at least three months for the initial sterile seedling explants to grow primary embryoid bodies, and a large number of embryoid bodies will be produced in the primary embryoid body me...

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Abstract

The invention relates to a dwarf lilyturf embryoid inducing and culturing method which comprises the following steps: the basal buds of the dwarf lilyturf are adopted as explants for culturing aseptic seedlings on an aseptic seedling medium; the aseptic seedlings are transplanted on the medium of a primary embryoid to induce embryogenic calluses; the embryogenic calluses are sub-cultured and differentiated to form the primary embryoid; the primary embryoid is transplanted on a secondary embryoid medium to induce a secondary embryoid; and then the secondary embryoid is transplanted to a seedling medium to generate a complete plant. In the invention, regulation is carried out on the two directions of forming a complete plant and the secondary embryoid after the dwarf lilyturf primary embryoid is formed, which not only can obtain the complete plant in a short time, but also can improve the planting percent to 95 percent through the regulation on the inducing and differentiation on the secondary embryoid; moreover, the steps are simple, the culturing cost is low and the propagation coefficient is high.

Description

technical field [0001] The invention relates to the technical field of plant cell engineering, in particular to a method for inducing and cultivating the embryoid bodies of syringae. Background technique [0002] Ophiopogon japonicus, also known as Ophiopogon japonicus, belongs to the monocot of Liliaceae. The plant is low, evergreen in all seasons, with strong stress resistance, heat resistance, good disease resistance and insect resistance, light-loving, shade-tolerant, and no need for too much pruning. It has become an indispensable landscape in gardens in recent years. Plants and lawn ground cover. Moreover, it has medicinal properties, and it is one of the varieties that can be adjusted by medicine. Due to its wide range of uses, the demand for step grass has been increasing year by year in recent years. [0003] However, the seed-setting rate of stepgrass is low, and seed reproduction is difficult. Usually, the ramets are used to reproduce or apply paclobutrazol to...

Claims

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Application Information

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IPC IPC(8): A01H4/00C12N5/04
Inventor 梁慧敏朱洪生巫建新鲍荣静李晶何任红
Owner JIANGSU POLYTECHNIC COLLEGE OF AGRI & FORESTRY
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