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Method for carrying out in-vitro culturing on adventitious buds of juglans mandshurica maxim and inducing plant regeneration

A technology of in vitro culture and walnut catalpa, applied in plant regeneration, botany equipment and methods, horticultural methods, etc., can solve problems such as difficulty in cultivating organs, browning, low induction rate and rooting rate, and achieve wide application value, The effect of reducing the degree of browning and simple and feasible operation steps

Active Publication Date: 2017-03-08
FORESTRY RES INST OF HEILONGJIANG PROVINCE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention aims to solve the problems of the existing tissue culture organs of walnut catalpa difficult to produce, serious browning phenomenon, low induction rate and rooting rate, and provides a method for in vitro culture of adventitious buds of walnut catalpa to induce plant regeneration

Method used

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  • Method for carrying out in-vitro culturing on adventitious buds of juglans mandshurica maxim and inducing plant regeneration
  • Method for carrying out in-vitro culturing on adventitious buds of juglans mandshurica maxim and inducing plant regeneration

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specific Embodiment approach 1

[0021]Specific embodiment one: the method for plant regeneration induced by in vitro culture of adventitious buds of walnut catalpa in the present embodiment comprises the following steps:

[0022] One, carry out pretreatment to walnut catalpa zygotic embryo;

[0023] 2. Take the walnut catalpa zygote embryos pretreated in step 1 and inoculate them with 1.0-1.5 mg / L thiadizuron, 0.5-1.0 mg / L 6-BA, 350 mg / L glutamine, 500 mg / L hydrolyzed casein, 0.5~1.0g / LNa 2 S 2 o 3 , 0.5~1.0g / LPVP, 6.0g / L agar and 30g / L sucrose MS modified medium, cultured in dark until callus appeared;

[0024] 3. Inoculate the callus tissue in a medium containing 0.5~1.0mg / L 6-BA, 0.5~1.0g / L Na 2 S 2 o 3 , 0.5~1.0g / L PVP and 6.0g / L agar MS modified medium, under the light intensity of 32~36μmol·m -2 ·s -1 , in an environment with a temperature of 23-25°C, alternately cultivate light and dark until adventitious buds grow;

[0025] 4. Cut the callus with adventitious buds into pieces and transfer th...

specific Embodiment approach 2

[0028] Specific embodiment two: what this embodiment is different from specific embodiment one is: the method for carrying out pretreatment to the walnut catalpa zygote embryo described in the step one is specifically: draw materials in May, scrub the walnut catalpa fruit surface with washing powder water earlier, Then rinse with running water for 1-2 hours, then rinse with sterile water for 3 times, place it at 2-4°C for 12-24 hours, then move it to an ultra-clean bench, use ultraviolet light to sterilize for 15-30 minutes, and use a mass concentration of Soak in 75% ethanol solution twice, each time for 30s, and then use 0.1% HgCl 2 Soak in the solution for 5-10 minutes, then rinse with sterile water for 3-5 times, and set aside. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0029] Embodiment 3: This embodiment differs from Embodiment 1 or Embodiment 2 in that: the temperature of the dark culture in step 2 is 22-25°C. Others are the same as in the first or second embodiment.

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Abstract

The invention discloses a method for carrying out in-vitro culturing on adventitious buds of juglans mandshurica maxim and inducing plant regeneration, relates to a plant regeneration method of the juglans mandshurica maxim and aims to solve the problems of difficulty, serious browning phenomenon, low inductivity and low rooting rate of existing juglans mandshurica maxim tissue culture organogenesis. The method comprises the following steps: 1, pretreating zygotic embryos of the juglans mandshurica maxim; 2, grafting the pretreated zygotic embryos of the juglans mandshurica maxim to a culture medium for culturing until appearing callus; 3, grafting the callus tissues to a culture medium, and carrying out light and darkness alternation culture until growing adventitious buds; 4, cutting the callus tissues with the adventitious buds into cubes for illumination culture, carrying out elongation of the adventitious buds, and transferring the adventitious buds in a culture medium for carrying out stem drawing of the adventitious buds; 5, transferring the adventitious buds after stem drawing into a culture medium for rooting of the adventitious buds; 6, transplanting the adventitious buds to a greenhouse for culture after hardenining seedlings. According to the method disclosed by the invention, the callu inductivity and the adventitious bud inductivity are up to 100 percent, the stem drawing rate and the rooting rate of the adventitious buds are higher, and the browning rate can be remarkably reduced. The method is applicable to the field of tissue culture of the juglans mandshurica maxim.

Description

technical field [0001] The invention relates to a plant regeneration method of walnut catalpa. Background technique [0002] Juglans mandshurica Maxim is a plant of the genus Juglans walnut, also known as Juglans mandshurica Maxim, commonly known as hickory. Produced in Northeast my country and North China, due to severe damage and a large reduction in natural resources, it is listed as a national second-class rare tree species. Walnut catalpa is fine and tough in texture, elegant in color and dense in texture. It is an excellent material for furniture and construction. Its fruit, hickory nut, contains more than 70% fat, unique flavor and strong fragrance. In my country, the young fruit, bark, and root bark of Chinese catalpa have been used as traditional Chinese medicinal materials for clearing away heat, detoxification, and anti-inflammatory. The green bark of Chinese catalpa contains juglone, flavonoids, terpenes and other active ingredients, which are beneficial to the...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/008
Inventor 张海峰袁显磊殷东生黄海娇张建瑛周志军王福德翁海龙赵彦龙
Owner FORESTRY RES INST OF HEILONGJIANG PROVINCE
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