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Kaffir lily in-vitro propagation method using young plants and young blade cut blocks as explants

A technology for explants and plants, which is applied to the field of rapid asexual reproduction in vitro of Clivia, can solve the problems of low regeneration rate, long regeneration period, and limited application of tissue culture technology of Clivia, and achieves short regeneration period, low cost, and high dedifferentiation rate. The effect of high differentiation rate

Inactive Publication Date: 2010-09-08
NORTHEAST NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the Clivia tissue culture experiment done by the predecessors has been initially successful, the application of Clivia tissue culture technology in actual production is limited due to the low regeneration rate and long regeneration cycle.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] 1. The acquisition of explant: get mature seed, put into aseptic ultra-clean table, soak 1min with 75% (volume / volume) alcohol, soak 10 minutes with the mercuric chloride aqueous solution of 0.1% (mass / volume), without Rinse with bacterial water three times, inoculate the seeds on 1 / 2MS semi-solid medium without hormones, and cultivate under light for 8 hours a day at 20°C.

[0015] 2. Explant inoculation and callus induction: Cut the aboveground part (comprising young leaves and young stems) of the plantlet growing 1 leaf in step 1 or the aseptic regenerated plant containing 1 leaf into small pieces with a diameter of 3 mm. Blocks were inoculated on MS semi-solid medium containing 6-BA1mg / L, NAA0.5mg / L, sucrose 3.0% (mass / volume), and irradiated with 40W fluorescent lamp every day at 20°C for 8 hours. Subculture once every 40 days, and induce differentiation when a yellow granular callus grows.

[0016] 3. Induction of clustered buds: the callus is transferred to the ...

Embodiment 2

[0020] 1. The acquisition of explant: get mature seed, put into aseptic ultra-clean table, soak 2min with 75% (volume / volume) alcohol, soak 20 minutes with the mercuric chloride aqueous solution of 0.2% (mass / volume), without Rinse with bacterial water 5 times, inoculate the seeds on 1 / 2MS semi-solid medium without hormones, and cultivate under light at 30°C for 12 hours a day.

[0021] 2. Explant inoculation and callus induction: cut the above-ground part (comprising young leaves and young stems) of the plantlet growing two leaves or the aseptic regenerated plant having two leaves in step 1 into 5mm in diameter Small pieces were inoculated on the MS semi-solid medium containing 6-BA2mg / L, NAA2.0mg / L, sucrose 3.0% (mass / volume), and irradiated with 40W fluorescent lamp for 9 hours every day at 30°C. Subculture once every 50 days, and induce differentiation when a yellow granular callus grows.

[0022] 3. Induction of clustered buds: the callus is transferred to the induction ...

Embodiment 3

[0026] 1. The acquisition of explant: get mature seed, put into aseptic ultra-clean table, soak 40S with 75% (volume / volume) alcohol, soak 15 minutes with the mercuric chloride aqueous solution of 0.15% (mass / volume), without Rinse with bacterial water 4 times, inoculate the seeds on 1 / 2MS semi-solid medium without hormones, and incubate under light at 24°C for 10 hours a day.

[0027] 2. explant inoculation and callus induction: when a leaf grows from the seedling in step 1, the above-ground part of the plantlet (comprising young leaves and young stems) is cut into small pieces with a diameter of 4mm, and inoculated in a seedling containing 6- BA1.5mg / L, NAA1.0mg / L, sucrose 3.0% (mass / volume) MS semi-solid medium, 24 ℃ 40W fluorescent lamp every day for 8.5 hours. Subculture once every 45 days, and induce differentiation when a yellow granular callus grows. The young aerial part of a sterile regenerated plant with one leaf can also be used as an explant.

[0028] 3. Inducti...

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Abstract

The invention relates to a flower propagation method, in particular to a kaffir lily in-vitro propagation method using young plants and young blade cut blocks as explants. The invention adopts the technology such as explant inoculation and healing induction, cluster bud induction, differentiation induction culture, root induction, regeneration plant culture, regeneration plant transplanting and the like for propagating kaffir lily flower. The kaffir lily can propagate plants with the same genetic background and regular external appearance in a large scale and in relatively short time under the condition without influencing the normal development and propagation of the female plants, the material taking is not limited by seasons, the regeneration period is short, the dedifferentiation rate and the differentiation rate are high, the rooting rate is high, the transplanting survival rate is high, and the growth state of the transplanted plants is good. The invention is particularly suitable for fast propagation of rare varieties of the kaffir lily, and is applicable to transgenosis of a leaf disc transformation method of the kaffir lily, the cost of the average single strain regeneration seedlings is very low, and the invention is particularly suitable for factory production.

Description

technical field [0001] The invention relates to a method for flower propagation, in particular to a method for in vitro asexual rapid propagation of Clivia. Background technique [0002] Plant tissue culture is plant aseptic culture technology, which is based on the totipotency of plant cells, using isolated plant organs (such as roots, stems, leaves, stem tips, flowers, fruits, etc.), tissues (such as cambium, epidermis, Cortex, endosperm, etc.) or cells (such as megaspores, microspores, somatic cells, etc.) and protoplasts can induce callus, adventitious buds under sterile and suitable artificial medium and artificial conditions such as light and temperature. , adventitious roots, forming whole plants or techniques for producing other products of economic value. The regenerated plants produced by plant tissue culture can basically maintain the excellent traits of the parents (explant source plants) except for extremely low variation. Since the invention of plant tissue c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 王丽王钦美邹凡雨
Owner NORTHEAST NORMAL UNIVERSITY
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