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Method for efficiently propagating dendrobium by using roots and culture medium of dendrobium

A technology of induction medium and subculture medium, applied in the field of methods and its medium, can solve the problem of not providing callus induction rate, differentiation rate, seedling rate and reproduction coefficient, which cannot be ruled out originating from non-root tissues, Issues such as the origin of clustered buds and protocorms are not clarified, and the effects of low cultivation costs, cost and energy savings, and a simple process of inducing seedlings are achieved

Inactive Publication Date: 2015-04-01
SHANGHAI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this document does not provide data such as callus induction rate, differentiation rate, seedling rate and reproduction coefficient using Dendrobium root as explants. In the past 30 years, no one else has obtained similar results by using the method reported. rarely quoted
[0006] In 2006 and 2007, Zhan Zhonggen and others studied the root tip culture of Dendrobium officinale and the morphogenesis in the culture [Zhan Zhonggen, Xu Cheng, Zhang Ming, Luo Zijuan. Research on Dendrobium officinale Root Tip Regenerated Plants by Somatic Embryos[J] .Journal of Zhejiang University (Agriculture and Life Science Edition) 2007,31(5):579-580; Zhan Zhonggen. Research on root tip induction of Dendrobium officinale[J].Chinese herbal medicine 2006,37(6):928-931], obtained Over-regenerated plants, but the yield is not high, and cannot be industrialized
[0007] In 2008, Qin Tinghao used the root pocket of Dendrobium officinale as explants, inoculated in MS+6-BA0.5mg / L+NAA0.2mg / L medium for light culture, and induced clustered buds and protocorms [Qin Tinghao. Dendrobium officinale Tissue culture and rapid propagation of [J]. Tropical Agricultural Science 2008, 28(1): 25-28], but the origin of these clustered shoots and protocorms has not been clarified, and it cannot be ruled out that they originated from non-root tissues, and only 20% of the root pockets induced protocorms, the multiplication coefficient was 4.28, and the efficiency was not high
[0008] In 2012, in the patent CN102428874 "A Method for Inducing Protocorm with Dendrobium", it was mentioned that the root tip of Dendrobium was used at 1 / 2MS+NAA0.5~2mg / L+6-BA0.3~1mg / L+agarose Protocorms can be induced and seedlings can be induced by dark culture and light culture on 3-4g / L medium, but no specific data and examples are provided
[0009] In addition, the tissue culture of Dendrobium orchid has been troubled by the three major problems of easy browning, easy vitrification and seedling emaciation so far. At present, cumbersome culture techniques are usually used and complicated methods such as adding banana puree, coconut juice, tryptone, etc. to the medium are used. Organic matter to control its harm [Wu Zhigang, Liu Xianwang, Zhang Shouwen. Research progress in tissue culture of Dendrobium plants [J]. Chinese Medicine Research and Information 2005, 7(11): 23-25; Chen Yahong, Hong Lei, Chen Xiongting. Research on Browning in Tissue Culture [J]. Modern Agricultural Science 2009, 16(3): 44-48; Qiao Yongxu, Zhang Yongping. Tissue Culture of Dendrobium [J]. Anhui Agricultural Science 2010, 38(4): 1731- 1732, 1734; He Jia, Peng Feng, Jiang Yumei, Xia Bing, Wang Ren. Establishment and optimization of Dendrobium tissue culture and rapid propagation system [J]. Jiangsu Agricultural Science, 2010, (6): 70-72], As a result, the culture cost is high and the cycle is prolonged, and it is urgent to fundamentally improve it through the innovation of culture methods and culture media.

Method used

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  • Method for efficiently propagating dendrobium by using roots and culture medium of dendrobium
  • Method for efficiently propagating dendrobium by using roots and culture medium of dendrobium
  • Method for efficiently propagating dendrobium by using roots and culture medium of dendrobium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] The roots of Dendrobium officinale test-tube plantlets were used as materials, cut into sections, inoculated, and cultured in dark at a temperature of about 25°C (25±2°C) to induce embryogenic callus. The embryogenic callus induction medium was MS (Murashige and Skoog, 1962)+6-BA (6-benzylaminoadenine) 0.6mg / L+PIC (4-amino-3,5,6-trichloropyridine-2-acid) 3mg / L+sucrose 30000mg / L+agar powder 5000mg / L. After 15 days, the root segment expanded and began to produce embryogenic callus, such as figure 1 As shown, after 60 days, the root segment can be covered with embryogenic callus, and the callus induction rate is about 70%.

[0047] Transfer the obtained embryogenic callus to the medium for protocorm induction to carry out the differentiation and proliferation of the protocorm. The protocorm induction medium is MS+CPPU(N-(2-chloro-4-pyridyl)-N '-phenylurea) 0.1mg / L+NAA (naphthaleneacetic acid) 0.01mg / L+sucrose 30000mg / L+agar powder 5000mg / L. The culture conditions are 16...

Embodiment 2

[0050] Take the root of the test-tube plantlet obtained in Example 1 as material, cut it into sections and inoculate it, culture in dark at a temperature of about 25°C, induce embryogenic callus, and the embryogenic callus induction medium is MS+6-BA0.6mg / L+PIC3mg / L+sucrose 30000mg / L+agar powder 5000mg / L, the callus induction rate is over 80%. Transfer the induced embryogenic callus to the callus subculture medium for dark culture at a temperature of about 25°C, subculture once every 25 days, and the embryogenic callus can proliferate approximately twice after each subculture . The callus subculture medium is MS+6-BA0.5mg / L+PIC2mg / L+sucrose 30000mg / L+agar powder 5000mg / L.

[0051] Transfer the embryogenic callus obtained from two subcultures to the protocorm induction medium for protocorm differentiation and proliferation. The protocorm induction medium is MS+CPPU0.1mg / L+NAA0.01mg / L+sucrose 30000mg / L+agar powder 5000mg / L, the culture conditions are 16 hours of light per day...

Embodiment 3

[0054] The root of the test-tube seedling of Dendrobium huoshanense is used as material, cut into sections and then inoculated, cultured in dark at a temperature of about 25°C, to induce embryogenic callus, and the embryogenic callus induction medium is MS+6-BA0.6mg / L+ PIC3mg / L + sucrose 30000mg / L + agar powder 5000mg / L, the callus induction rate is over 70%. Transfer the induced embryogenic callus to the callus subculture medium for dark culture, subculture for 25 days, and the temperature is about 25°C, the embryogenic callus can proliferate by more than 1 times. The callus subculture medium is MS+6-BA0.5mg / L+PIC2mg / L+sucrose 30000mg / L+agar powder 5000mg / L.

[0055] Transfer the embryogenic callus obtained from the subculture to the protocorm induction medium for protocorm differentiation and proliferation. The protocorm induction medium is MS+CPPU0.1mg / L+NAA0.01mg / L+sucrose 30000mg / L+agar powder 5000mg / L, the culture conditions are 16 hours of light per day, the temperatu...

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Abstract

The invention discloses a method for efficiently propagating dendrobium by using roots and a culture medium of the dendrobium. The method in the invention comprises the following steps of: selecting a root section of the dendrobium, and incubating and inducing the root section to obtain an embryonic callus, wherein the embryonic callus can be greatly multiplied through subculture; transferring the embryonic callus into a protocorm induction culture medium for induction and multiplication to obtain protocorm and bud seedlings; and transferring the protocorm and the bud seedlings into an adult seedling culture medium and culturing a dendrobium rooting plant. According to the method, MS is used as a basic culture medium; 6-BA and PIC are required to be added into the culture medium for the embryonic callus induction or subculture; CPPU and NAA are required to be added into the culture medium for the protocorm induction. The method for culturing regenerated plants by using the dendrobium roots provided by the invention is high in propagation coefficient, high in planting percentage and low in cost; a seedling growing process, a seedling robusting process and a rooting progress of the protocorm can be further finished; the plants are robust, and the roots are robust.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a method for efficiently propagating dendrobium orchids by using roots and a culture medium thereof. Background technique [0002] Dendrobium orchids (D.orchids) is a perennial herbaceous plant of the genus Dendrobium in the family Orchidaceae, and its main uses are ornamental and medicinal [Wu Rui, Li Zesheng, Gao Yan, Geng Xiuying. Research overview and development prospects of Dendrobium[J]. Chinese Tropical Agriculture, 2012(4): 22-23.]. Dendrobium is one of the four most famous orchids in the world with the most ornamental value [Xu Yu, Wang Siqing. Research progress on the sudden emergence of Dendrobium [J]. Journal of North China Agricultural Science, 2005, 20 (Series): 152-157.] , has many excellent characteristics such as various types, bright colors, graceful posture, numerous and dense flowers, and long flowering period. Dendrobium is also a ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 张建军周音陈敏敏谢纪红
Owner SHANGHAI ACAD OF AGRI SCI
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