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A kind of method for cork oak callus and adventitious bud induction

A callus induction and callus technology, applied in the field of plant biology, can solve the problems of imperfect asexual reproduction system and low tissue culture proliferation coefficient, and achieve the effect of expanding the propagation coefficient of adventitious buds

Active Publication Date: 2020-09-25
INST OF FORESTRY CHINESE ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The asexual reproduction system of Quercus variabilis is not yet perfect, and there are problems such as low proliferation coefficient in tissue culture

Method used

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  • A kind of method for cork oak callus and adventitious bud induction
  • A kind of method for cork oak callus and adventitious bud induction

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Screening Quercus variabilis callus induction medium

[0020] The mature seeds of a fine single plant of Quercus variabilis were used as explants, and after low-temperature storage at 4°C for 30 days, they were treated with 0.2% HgCl under sterile conditions. 2 Disinfect the surface for 10-15 minutes, rinse with sterile water 3 to 4 times, and put it in MS plus activated carbon 1.5g / L medium for germination culture. The culture temperature is 25±2°C, the light intensity is 800-1000Lux, and the light cycle is 16h / d; when the seedlings of Quercus cork grow 3-6cm high and the root length is 1-1.5cm, the root system is circumcised with a sterile blade. The circumcision depth does not reach the xylem, inoculated in the callus induction medium, the callus induction medium is based on MS, WPM, B5 culture, 6-BA0.5mg / L, 1.0mg / L, 1.50mg / L L, IAA0.1mg / L, 0.3mg / L, 0.8mg / L carry out orthogonal design induction culture (see Table 1), 6 repetitions. The relative humidity o...

Embodiment 2

[0024] Example 2 Screening Quercus variabilis callus proliferation medium

[0025] The induced Quercus variabilis callus was cultured on the basis of WPM, 6-BA0.3mg / L, 0.6mg / L, 0.9mg / L, IAA0.1mg / L, 0.2mg / L, 0.3mg / L for orthogonal Proliferation culture was designed, with 6 repetitions, the relative humidity of the culture room was 70%±5%, the temperature was (24±1)°C, and dark culture was carried out. After 20 days, the proliferation of callus on each culture medium was observed and analyzed.

[0026] It was observed that the culture medium for the proliferation of Quercus variabilis callus is preferably WPM solid medium: 6-BA0.6mg / L, IAA0.2mg / L.

Embodiment 3

[0027] Example 3 Screening callus differentiation induction medium for adventitious buds

[0028] The callus after proliferation was transferred to the differentiation medium for differentiation culture experiment. The medium used WPM as the basic medium, and different concentrations of TDZ, 6-BA, and KT growth regulators were added. See Table 2 for details. Each treatment was inoculated with 6 mediums, and each medium was inoculated with 5 pieces of embryogenic callus. The relative humidity of the culture room was 70%±5%, the temperature was (24±1)°C, cultured under LED light, the photoperiod was 8h·d-1, and the number of adventitious bud regeneration was counted after 35 days of culture.

[0029] Table 2 Differentiation coefficients on different media

[0030]

[0031] Differentiation factor = number of adventitious buds produced / number of inoculated explants

[0032] As can be seen from Table 2, the embryogenic callus has the highest adventitious bud regeneration coef...

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Abstract

The invention discloses a method for callus induction and adventitious bud differentiation of quercus variabilis. The method for callus induction and adventitious bud differentiation of the quercus variabilis adopts the test-tube seedlings obtained by aseptic seeding induction of quercus variabilis seeds as test materials. When the seedlings are 3-6 cm tall and the roots are 1-1.5 cm long, the root systems of the seedlings are subjected to girdling with an aseptic blade and placed in induction mediums for callus induction, then the induced callus is stripped from the root systems, and callus proliferation, adventitious bud differentiation induction and adventitious bud rooting are carried out. The method for callus induction and adventitious bud differentiation of the quercus variabilis adopts the root systems of the girdled seedlings for callus induction culture, and finally obtains the seedlings through the process of callus proliferation, differentiation culture and the like, so that the problems, in the conventional quercus variabilis tissue culture process, of difficult induction of embryogenic callus with stem tips, stem segments and leaves and serious endophyte contaminationof the culture material are solved, the adventitious bud propagation coefficient can be increased, and the method for callus induction and adventitious bud differentiation of the quercus variabilis has practical significance for the large-scale seedling raising of the quercus variabilis.

Description

technical field [0001] The invention belongs to the field of plant biotechnology, and in particular relates to a method for inducing callus of cork oak and adventitious buds. Background technique [0002] Cork oak (Quercus variabilis Bl.) is a plant of the genus Quercus in the Fagaceae family. There are 350 species in this genus, mainly produced in the temperate and subtropical regions of the northern hemisphere. It is a deciduous tree with a height of 30 meters and a diameter at breast height of more than 1 meter. The distribution area of ​​Quercus variabilis is vast in my country, among which, the total area of ​​concentrated distribution area and sub-concentration distribution area of ​​Quercus variabilis is about 1,363,500 hectares. [0003] Seed propagation of Quercus variabilis is not a means of high-efficiency breeding of large-scale improved species, but asexual reproduction can solve the problem of high-efficiency breeding of Quercus variabilis, and it is also an ef...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 毕泉鑫陈梦园李迎超王利兵于海燕
Owner INST OF FORESTRY CHINESE ACAD OF FORESTRY
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