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A method for inducing adventitious buds from leaves of Populus trichocarpa and plant regeneration

A technology for inducing poplar and buds, which is applied in the directions of plant regeneration, botanical equipment and methods, horticultural methods, etc., can solve the problems of low induction rate of adventitious buds, difficulty in culturing organs, low plant regeneration rate, etc., and achieves simple operation steps. Feasible, broad application prospects, and the effect of shortening regeneration time

Inactive Publication Date: 2016-08-31
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 difficulty in organogenesis in tissue culture of Populus trichocarpa, low plant regeneration rate, low induction rate of adventitious buds and long period of explants with stems of Populus trichocarpa, and provides a leaf of Populus trichocarpa Method for adventitious bud induction and plant regeneration

Method used

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  • A method for inducing adventitious buds from leaves of Populus trichocarpa and plant regeneration
  • A method for inducing adventitious buds from leaves of Populus trichocarpa and plant regeneration
  • A method for inducing adventitious buds from leaves of Populus trichocarpa and plant regeneration

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specific Embodiment approach 1

[0014] Specific embodiment one: the method for the induction of adventitious buds of leaves of Populus trichocarpa and plant regeneration in the present embodiment is carried out according to the following steps:

[0015] 1. Collection and processing of materials: Select Populus trichocarpa plants that grow robustly and are free of diseases and insect pests cultivated in the greenhouse, collect newly sprouted branches, cut off the leaves on the branches, and cut the remaining branches into lengths of 9 to 11 cm with The stem segments with 3 to 4 bud points are washed with tap water for 1 to 2 hours;

[0016] Two, inoculation culture and the proliferation of aseptic seedlings: after aseptic treatment, the stem section washed with tap water is inoculated into the rooting medium WPM3 and cultivated, and the aseptic seedlings are propagated;

[0017] 3. Induction of adventitious bud differentiation: use the leaves of P. trichocarpa aseptic seedlings cultivated in step 2 as explant...

specific Embodiment approach 2

[0022] Specific embodiment two: the difference between this embodiment and specific embodiment one is: the specific method of inoculation and cultivation and the propagation of aseptic seedlings described in step two is: the stem section washed with tap water is washed once with sterile water , then put the stem section into 70% alcohol for disinfection for 30 sec, take out the stem section and wash it 3 times with sterile water, then disinfect it with 2% sodium hypochlorite for 7 minutes, and wash it again with sterile water for 30 seconds. Finally, use sterile filter paper to dry the water on the surface of the stem section, use sterile scissors to cut off the part of the stem section that has been oxidized by the disinfectant solution, and then cut the remaining stem section into a small section containing only one bud point, and inoculate On the rooting medium WPM3, at a temperature of 23-25°C, with a daily light of 12-16h, and a light intensity of 30-40μmol·m -2 ·s -1 Cu...

specific Embodiment approach 3

[0023] Specific embodiment three: the difference between this embodiment and specific embodiment one or two is: in the step 3, the adventitious bud induction medium WPM1 is made of 400mg / L KNO 3 , 370mg / L MgSO 4 , 96mg / L CaCl 2 , 8.6mg / L ZnSO 4 , 0.25mg / L CuSO 4 , 27.8mg / L FeSO 4 、990mg / L K 2 SO 4 , 170mg / L KH 2 PO 4 , 22.4mg / L MnSO 4 , 6.2mg / L H 3 BO 3 , 0.25mg / L NaMoO 2 2H 2 O, 37.3mg / L Na 2 EDTA, 1.0mg / L thiamine hydrochloride, 0.5mg / L pyridoxine hydrochloride, 0.5mg / L niacin, 2.0mg / L glycine, 100mg / L inositol, 25mg / L sucrose, 5.6mg / L agar, 0.01~1.0mg / L 6-benzyladenine, 0.01~1.0mg / L indole butyric acid, 0.0001~0.01mg / L thiadidiazuron and distilled water, pH value is 5.8. Others are the same as in the first or second embodiment.

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Abstract

A populus trichocarpa Torr.Gray leaf adventitious bud induction and plant regeneration method aims to solve the problems that at present, populus trichocarpa Torr.Gray has difficulty in tissue culture organogenesis and low in plant regeneration rate, and the adventitious bud inductivity is low and the period is long since populus trichocarpa Torr.Gray stems are taken as explants. The method provided by the invention comprises the following steps: 1, material collection and treatment; 2, inoculated culture and aseptic seedling expanding propagation: stems which are flushed by tap water are subject to sterile treatment for inoculated culture to propagate aseptic seedlings; 3, adventitious bud differentiation induction; 4, adventitious bud elongation induction: calluses with adventitious buds are cut into blocks and placed in an adventitious bud elongation culture medium for culture; 5, adventitious bud rootage culture; 6, hardening-seedling and transplanting. According to the invention, populus trichocarpa Torr.Gray aseptic seedling leaves are taken as explants, so that the adventitious bud inductivity and the survival rate are up to 100%, and from explant culture to seedling survival, only about 60 days are required. The populus trichocarpa Torr.Gray leaf adventitious bud induction and plant regeneration method is used for plant tissue culture.

Description

technical field [0001] The invention relates to a method for inducing adventitious buds and plant regeneration of Populus trichocarpa. Background technique [0002] Populus trichocarpa Torr.Gray, also known as Populus trichocarpa Torr.Gray, is a tree species of the genus Populus (Populus L.), and its origin is mainly distributed in the western part of North America. In 2006, the entire genome sequencing of Populus trichocarpa was completed, and the research results were published in the journal "Science" on September 15 of that year (Tuskan et al., 2006). Forest model tree species. However, for the woody model plant Populus trichocarpa whose genome sequencing was completed in 2006, there are very few cases of successful establishment of tissue culture system and genetic transformation. In 2009, Zhang Hongmei et al. In ", the internode stem segments are used as explants, and the buds germinated from the axillary buds are multiplied and cultured. This method is only for the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 李淑娟程玉祥甄成严善春张雅奎
Owner NORTHEAST FORESTRY UNIVERSITY
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