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Lilium pumilum somatic embryo direct generation method with effect of significantly shortening induction time

A technology of induction time and somatic embryos, which is applied in the field of plant biology, can solve the problems of complex process, long induction cycle, and limitation of somatic embryo application, and achieve the effect of simple method, high repeatability, and accelerated factory breeding

Active Publication Date: 2018-08-31
SHENYANG AGRI UNIV
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The indirect regeneration method needs to go through multiple steps, the process is complicated, and the induction period is long. It takes at least 45 days from inoculation to obtaining globular embryos (Zhang Jing, 2017), which limits the rapid propagation of somatic embryos in bulbs and genetic transformation. Aspects of application

Method used

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  • Lilium pumilum somatic embryo direct generation method with effect of significantly shortening induction time

Examples

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Comparison scheme
Effect test

example 1

[0033] Example 1 In vitro regeneration technique of somatic embryos of Lily lily

[0034] 1. Test material

[0035] Lily Varieties: Lily Lily ( Lilium pumilum DC. Fisch.), purchased from Liaobei seedling sales office in Guangming Community, Xincheng Street, Kaiyuan City, Liaoning Province.

[0036] 2. Test methods and results

[0037] Step 1: Explant culture stage

[0038] Prepare MS medium first, add 60g of sucrose and 7g of agar powder (strips) to each liter of medium, adjust the pH value to 5.8, and divide into tissue culture bottles. At 121°C and 103.4kPa, autoclave for 20 minutes (medium sterilization method is the same below), observe for three days after sterilization, and inoculate after confirming the sterilization effect.

[0039] On the ultra-clean workbench, inoculate the sterile seedlings of Lilium microphylla in the above-mentioned MS medium. Use an airtight sealing film to ensure that the humidity in the tissue culture bottle reaches 85%. The temperature of...

Embodiment 2

[0049] Example 2 Comparison of in vitro regeneration methods of somatic embryos of Lily lily

[0050] 1. Test material

[0051] Lily Varieties: Lily Lily ( Lilium pumilum DC. Fisch.), purchased from Liaobei seedling sales office in Guangming Community, Xincheng Street, Kaiyuan City, Liaoning Province.

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Abstract

The invention relates to a lilium pumilum somatic embryo direct generation method with effect of significantly shortening induction time. The method comprises the following steps: inoculating asepticseedlings of lilium pumilum into an MS medium with 60g.L<-1> sucrose, culturing the seedlings for 45 days, taking outer scale, inoculating the outer scale into a somatic embryo induction medium, culturing the outer scale in dark for 30 days to obtain globular embryos; transferring a culture containing the globular embryos to a somatic embryo proliferation medium, performing culturing for 25 days to obtain secondary proembryos, and performing multiplication in the form of secondary somatic embryos; transferring the secondary proembryos to the MS medium, performing culturing in light for 20 daysfor germination into seedlings. The lilium pumilum somatic embryos are obtained in a direct regeneration way for the first time, the problem of long embryo induction cycle is effectively solved, obtaining time of the globular embryos and the final tissue culture seedling obtaining time is shortened by 90 days, the method has the advantages of stable explant source, simple and implementable stepsand high repeatability, and an efficient and stable regeneration system can be provided for rapid propagation, germplasm preservation and genetic transformation of the lilium pumilum.

Description

technical field [0001] The invention belongs to the field of plant biotechnology, and in particular relates to a method for direct generation of lily lily somatic embryos which significantly shortens the induction time. Background technique [0002] lily lily ( Lilium pumilum DC. Fisch), Liliaceae (Liliaceae) Liliaceae ( Lilium ) Perennial herbaceous plants, widely distributed in Northeast my country, North China and Northwest China, most of them grow in forest margins, grasslands or hillside grasslands at an altitude of 400-2600 meters. Good parents for resistance breeding (Yang Liping, 2005). The lily lily has a beautiful plant shape, bright colors, petite flowers, revolving and drooping tepals, glossy, fragrant flowers, and high ornamental value. It is often used as a ground cover for flower beds and flower borders. At the same time, the polyphenols in the bulbs of Lilium tenifolia have a certain ability to scavenge various free radicals, so they can also be used as n...

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 孙红梅李宏宇王京王志平
Owner SHENYANG AGRI UNIV
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