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Method for inducing hybrid larch plant regeneration through in vitro culture of adventitious buds

A technology of in vitro culture and larch, applied in plant regeneration, botany equipment and methods, horticultural methods, etc., can solve the problems of difficulty in culturing organs, low rate of adventitious bud induction, low rate of plant regeneration, etc., and achieves wide application value effect

Inactive Publication Date: 2015-08-19
NORTHEAST FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The present invention aims to solve the problems of existing hybrid larch tissue culture organ generation difficulty, low plant regeneration rate and low adventitious bud induction rate, and provides a method for in vitro cultured adventitious bud induction of hybrid larch

Method used

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  • Method for inducing hybrid larch plant regeneration through in vitro culture of adventitious buds
  • Method for inducing hybrid larch plant regeneration through in vitro culture of adventitious buds
  • Method for inducing hybrid larch plant regeneration through in vitro culture of adventitious buds

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specific Embodiment approach 1

[0014] Embodiment 1: The method for in vitro culturing adventitious buds of hybrid larch to induce plant regeneration in this embodiment is carried out according to the following steps:

[0015] 1. Take pretreated hybrid larch zygote embryos and inoculate them on agar containing 2.0-2.5mg / L thidiazuron, 1g / L inositol, 450mg / L glutamine, 500mg / L hydrolyzed casein, 6.5g / L agar and 30g / L sucrose in the BM modified medium, carry out dark culture or light culture until callus appears;

[0016] 2. Inoculate the callus in the BM modified medium containing 2.0-2.5 mg / L thidiazuron at a light intensity of 33-36 μmol·m -2 ·s -1 , the temperature is 23 ~ 25 ℃ environment for light cultivation, to grow adventitious buds;

[0017] 3. Cut the callus with adventitious buds into pieces and transfer them to BM modified medium containing 0.1-1.0mg / L TDZ and 0-0.04mg / L NAA for cultivation under the light intensity of 32-38μmol·m -2 ·s -1 , in an environment with a temperature of 22-24°C for ...

specific Embodiment approach 2

[0020] Embodiment 2: The difference between this embodiment and Embodiment 1 is that the hybrid larch in step 1 is Japanese larch × Xing'an larch. Others are the same as the first embodiment.

specific Embodiment approach 3

[0021] Embodiment 3: This embodiment is different from Embodiment 1 or 2 in that: the method for obtaining the pretreated hybrid larch zygote embryos described in step 1: peeling off the seed coat from the mature and plump hybrid larch seeds , rinsed with running water for 2 days; then in the ultra-clean bench, the seeds with the seed coat were sterilized with alcohol with a mass concentration of 70% for 1 minute, rinsed with sterile water for 3 times, and then the seeds were transferred to a mass concentration of 4% sodium hypochlorite In the solution, add 2 to 4 drops of Tween 20 for 15 minutes of shaking and disinfection, then wash with sterile water for 6 times, strip off the endosperm and disinfect with 0.5% sodium hypochlorite for 2 minutes, and wash the embryos with sterile water for 3 times to obtain Pretreated hybrid larch zygotic embryos. Others are the same as in the first or second embodiment.

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Abstract

The invention discloses a method for inducing hybrid larch plant regeneration through in vitro culture of adventitious buds, relates to a method for inducing pine plant regeneration through in vitro culture of adventitious buds, and aims at solving the problems that in the existing tissue culture method of hybrid larch, organogenesis is difficult, plant regeneration rate is low, and adventitious bud induction rate is low. The method comprises the following steps: 1 inoculating pretreated zygotic embryos of the hybrid larch into a culture medium, and carrying out dark cultivation or light cultivation until callus tissue is generated; 2 inoculating the callus tissue into the culture medium, carrying out light cultivation until an adventitious buds grow; 3 cutting the callus tissue with the adventitious buds into blocks, transferring into the culture medium, carrying out subculture for once after 3 weeks, carrying out adventitious bud elongation, transferring into the culture medium, and carrying out stooling of the adventitious buds; and 4 transferring the stooled adventitious buds into the culture medium, and carrying out light cultivation to realize rooting of the adventitious buds. According to the method disclosed by the invention, the induction rate of the adventitious buds is 87.73%; the stooling rate is 75.96%; the rooting rate is 45%; and the method is applied for inducing hybrid larch plant regeneration through in vitro culture of adventitious buds.

Description

technical field [0001] The invention relates to a method for in vitro culturing adventitious buds of pine trees to induce plant regeneration. Background technique [0002] Larch is the main timber species in my country and occupies an important position in forestry production. It has the characteristics of wide distribution, strong adaptability, rapid growth and strong cold resistance. Hybrid Larch (Hybrid Larch) is a very important fast-growing tree species in the genus Larch. Its genetic gain is large, and it has the characteristics of straight and round trunk, hard wood and corrosion resistance. It is also one of the important wood species in Northeast my country. Due to the long growth cycle of hybrid larch, the conventional breeding technology takes a long time to select new varieties, and the effect is slow. At the same time, there are also constraints such as lack of gene source and incompatibility of hybridization. The target gene is directly introduced into the rec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 张磊张莉张含国
Owner NORTHEAST FORESTRY UNIVERSITY
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