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Method for inducing proliferation of bud cluster of protocorm of cremastra appendiculata

A technology of protocorm buds and protocorms, which is applied in the field of inducing protocorm bud clusters of Rhododendron orchids to proliferate, can solve the problems of low reproduction coefficient, abortion of seed embryos, and inability to realize large-scale production, and achieve the effect of low reproduction coefficient

Active Publication Date: 2016-12-07
GUIZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Rhododendron orchids naturally have difficulty in sexual reproduction. During the long-term evolution process, they have adapted to the vegetative reproduction of pseudobulbs. The pseudobulbs produced year by year form pseudobulb strings, but each year only the latest pseudobulbs germinate and grow into new plants, forming a new pseudobulb. , so the reproductive coefficient is extremely low
In artificial planting, the pseudobulb clusters can be propagated by division, although the reproduction coefficient has been improved to a certain extent, but it is still unable to realize its large-scale production.
Although seeds can be obtained through artificial pollination, the embryos are aborted and difficult to germinate naturally into seedlings

Method used

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  • Method for inducing proliferation of bud cluster of protocorm of cremastra appendiculata
  • Method for inducing proliferation of bud cluster of protocorm of cremastra appendiculata

Examples

Experimental program
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Effect test

Embodiment 1

[0016] Embodiment 1: protocorm proliferation culture

[0017] The plant material Rhododendron orchid [Cremastra appendiculata (D.Don.) Makino] in the present invention is collected from Qiandongnan Prefecture, Guizhou Province. After potting, the seeds are obtained by artificial pollination, and the seeds are aseptically germinated on the culture medium to obtain protocorms.

[0018] Protocorms germinated from seeds were used as explants and inoculated in 1 / 2 MS basic medium + 6-BA 1.0mg·L -1 +IBA1.0mg·L -1 +Activated carbon 0.5g·L -1 + sucrose 30g·L -1 +Agar 5.5g·L -1 cultured on proliferation medium. The culture period was 40 days, and the growth of the protocorms was observed and recorded every 10 days, and the culture temperature was 15°C. After about 15 days of culture, the protocorms changed from white to yellow-green. After 40 days of culture, the protocorms multiplied obviously, forming clusters of protocorms with many white hairs on the surface of the protocorms....

Embodiment 2

[0019] Embodiment 2: Bud cluster induction culture

[0020] Select clustered protocorms with good growth potential in Example 1, inoculate them with different types and concentrations of plant growth regulating substances, different concentrations of sucrose and different basic medium 1 / 4MS, 1 / 2MS or MS, using TDZ, NAA, Sucrose, basic medium 4 factors 3 levels of orthogonal experiments, except that the culture temperature is (25 ± 2) ℃, other culture conditions are the same as embodiment 1. After 30 days, the bud cluster induced proliferation coefficient is counted (wherein, the bud cluster proliferation coefficient=inoculation The number of buds after inoculation / the number of buds before inoculation) was used to investigate the effects of different treatments on the formation of bud clusters induced by protocorms. Conclusion: Protocorm induction bud cluster medium is 1 / 4MS+TDZ 1.5mg·L -1 +NAA0.3mg·L -1 + sucrose 20g·L -1 +Agar 5.5g·L -1 The effect is better (Table 1). In...

Embodiment 3

[0024] Embodiment 3: bud cluster proliferation subculture

[0025] The bud cluster induced in Example 2 is divided into two parts, one part is the undifferentiated protocorm in the bud cluster, which is inserted into the bud cluster induction medium in the above-mentioned embodiment 2 to continue the bud cluster induction formation, and after 30 days, no The base of the differentiated protocorm germinates new buds, and some of the new bud leaves begin to stretch, which can be subcultured again; the other part is a protocorm with obviously enlarged pseudobulbs and a tendency to differentiate into seedlings. Rooting medium 1 / 4 MS+6-BA2.0mg·L -1 +NAA 0.1mg·L -1 +Activated carbon 0.5g·L -1 + sucrose 30g·L -1 +Agar 5.5g·L -1 Carry out rooting culture in the middle, after 15d, blade unfolds, and root emerges, grows 3~5 after 30d and is the strong rhododendron orchid seedling of 2~3cm root, and culture condition is the same as embodiment 2.

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Abstract

The invention discloses a method for inducing proliferation of a bud cluster of a protocorm of cremastra appendiculata, wherein the method comprises the following steps: 1) taking the protocorm obtained through germ-free germination of a hand-pollinate seed of the cremastra appendiculata as an explant; 2) putting the protocorm in a minimal medium 1 / 2MS for proliferation culture; 3) putting the proliferated protocorm into a minimal medium 1 / 4MS to induce the bud cluster; 4) cutting an undifferentiated part and a differentiated part of the bud cluster, putting the undifferentiated part and the differentiated part of the bud cluster into different mediums to proliferate the bud cluster and differentiate to emerge a seedling and culture; 5) putting a plant with the emerged seedling in a minimal medium 1 / 4MS for rooting culture. The tissue culture seedling of the cremastra appendiculata is hardened and transplanted to a medium suitable for growth until the seedling grows into a strong cremastra appendiculata plant. The method effectively improves the propagation coefficient of the cremastra appendiculata seedling, thereby laying a foundation for the large-scale production.

Description

technical field [0001] The invention relates to a method for plant tissue culture and rapid propagation, in particular to a method for inducing the proliferation of rhododendron orchid protocorm clusters. Background technique [0002] Rhododendron orchid [Cremastra appendiculata (D.Don) Makino] is a wild rare medicinal plant of the Orchidaceae Rhododendron orchid genus, and its dried pseudobulb is used as medicine. Rhododendron orchid pseudobulbs contain alkaloids, flavanones, aglycones, phenanthrenes and other ingredients. Internal use has anti-liver cancer, breast cancer, uterine cancer and other effects, and external use can cure sores, snake bites, skin scalds or burns Wait. [0003] Rhododendron orchid is naturally difficult to reproduce sexually. During the long-term evolution process, it has adapted to the vegetative reproduction of pseudobulbs. The pseudobulbs produced year by year form pseudobulb strings, but each year only the latest pseudobulbs germinate and grow...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/008
Inventor 张明生吴彦秋吕享叶睿华李小兰
Owner GUIZHOU UNIV
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