CN105638462A

CN105638462A - Method for domesticating and transplanting taxus chinensis var mairei tissue culture seedling

Info

Publication number: CN105638462A
Application number: CN201511022864.4A
Authority: CN
Inventors: 李炎林; 熊兴耀; 李莉
Original assignee: Hunan Agricultural University
Current assignee: Hunan Agricultural University
Priority date: 2015-12-30
Filing date: 2015-12-30
Publication date: 2016-06-08
Anticipated expiration: 2035-12-30
Also published as: CN105638462B

Abstract

The invention discloses a method for domesticating and transplanting a taxus chinensis var mairei tissue culture seedling. The method comprises the following steps of (1) selecting the tissue culture seedling by means of selecting the taxus chinensis var mairei tissue culture seedling with a well-grown root system and more than one true leaf; (2) preliminarily domesticating the tissue culture seedling by means of putting a tissue culture container with the tissue culture seedling into a greenhouse for 3 to 5 days; (3) preparing a transplanting container and a culture substrate by means of selecting a seedling culturing box, and filling vermiculite, turf or nonwoven-fabric wrapped fresh coconut chaff in holes of the seeding culturing box; (4) transplanting and hardening seedling by means of planting the tissue culture seedling in the middle of the culture substrate obtained in the step (3), watering sufficiently, closing a seedling culturing box cover, putting the box into the greenhouse with 50 percent of sunshield screen, culturing at an environmental temperature of not more than 35 DEG C, and opening the seedling culturing box cover and performing a conventional seedling management mode after two weeks, wherein the survival rate of transplanted tissue culture seedlings reaches 100 percent at most. The method disclosed by the invention has the advantages that the disadvantages in the prior art are overcome, and the effects of simplicity in operation, low cost, high efficiency, stability and reusability of the seedling culturing box are achieved.

Description

A kind of rooting culture method of Taxus mairei tissue cultured seedling

Technical field

The invention belongs to Ramulus et folium taxi cuspidatae Fast-propagation field, be specifically related to a kind of rooting culture method of Taxus mairei tissue cultured seedling.

Background technology

Taxus mairei (Taxuschinensisvar.mairei) is gymnosperm, belongs to taxaceae (Taxaceae) Taxus (Taxus) plant, aiphyllium or shrub, dioecian plant. Taxus mairei is China's Relict Plant in the distinctive Tertiary Period, integrate view and admire, the important resource plant of medicinal and material. Since 20 century 70s, since from yewtree, extraction separation obtains natural anti-cancer Product Taxol, Ramulus et folium taxi cuspidatae is as a kind of important medicinal plants, and its medical value is more and more approved. Ramulus et folium taxi cuspidatae is except having antitumor pharmacological action, also have as antiinflammatory, antibacterial, protect the liver, pain relieving, convulsion, blood sugar lowering, antioxidation, stain resistance, many-sided drug effect (Cao Jiwu such as bring down a fever, Deng. southern china Ramulus et folium taxi cuspidatae is studied. China Forestry Publishing House, 2014,2 3). Taxus mairei taxol in bark content up to 0.049% 0.069%, be one that in this genus, content of taxol is the highest (eat civilization and iron army. Taxus mairei ecological study. China Forestry Publishing House, 2012,1). Taxus mairei natural propagation ability is relatively low, and natural resources lack, and interests are become in addition, and the denudation phenomenon of Taxus mairei remains incessant after repeated prohibition, and natural Ramulus et folium taxi cuspidatae woods number is fewer and feweri, and it has been in Critical Condition, is considered first-grade state protection plant.

The breeding of Ramulus et folium taxi cuspidatae mainly has cottage propagation and seminal propagation to be main. Taxus mairei is shallow root system plant, and main root is undeveloped, side root horizontal development, and expanding surface is relatively wide, easily lodges. And cottage propagation root system of plant is without significant main root, by the plant of seminal propagation, there is obvious main root, well developed root system, grow strong. Therefore the seminal propagation of Taxus mairei has stronger advantage, but Chinese yew genus plants seed includes wax coat, phellem layer, stone cell layer etc. from inside to outside because of seed coat structure, the structure of this hard and compact is unfavorable for the sprouting of seed and suppresses the after-ripening with embryo that sheds of seed germination material, cause the long dormancy of seed, generally could germinate through two summers in spring one under field conditions (factors). The succession that obvious Taxus mairei plant relies on seminal propagation to expand its colony under Natural habitat, explore more opportunity for survival be extremely slowly (eat civilization and iron army. Taxus mairei ecological study. China Forestry Publishing House, 2012,1).Therefore adopting seed to breed in vitro with slight epispastic breeding Ramulus et folium taxi cuspidatae is one of its important modes of reproduction, and the tissue cultured seedling of current Ramulus et folium taxi cuspidatae obtains and has been achieved for certain progress, but the research of training tissue culture seedling rooting culture also has room for promotion.

Affect factor that Ramulus et folium taxi cuspidatae micro-propagation seedling seedling exercising survives except kind factor, also relevant with factors such as the management of moisture, illumination managements in the quality of tissue cultured seedling, the substrate of seedling exercising, temperature and seedling exercising process. When the adventitious root of the tissue cultured seedling of Ramulus et folium taxi cuspidatae grows to 1.5 cm, take out from bottle, after processing 20 minutes with the carbendazim solution of 1.2g/L, when constant temperature and humidity, three time-of-weeks are divided to gradually reduce cultivation temperature and cultivate humidity, transplant in the domestication substrate of lichen, perlite and river sand domestication survival rate respectively 91%, 26% and 73% (Cheng Guangyou and Wang Mingqi. the slight epispastic of Ramulus et folium taxi cuspidatae. Journal of northeast Forestry university, 2000.28 (2): 13 16). Adopt the nutritive cube mixed-matrix of fluvial sand+peat (substrate be) transplant the tissue cultured seedling of Taxus mairei temperature be 25 DEG C, humidity be more than 75% transplant about 30, survival rate of plant is 90.1%, but the culture medium (Tang Daofang in nutritive cube do not handed in detail in article, Deng. the tissue culture of Taxus mairei and plant regeneration research, Agriculture in Hunan science, 2008, (2): 21 23). Taxus mairei stem section is induced high about 2cm stalwartness seedling and proceeds to DCR+NAA0.1mg/L culture medium root induction, within about 45 days, root length is had to go out, when root length is about 0.5cm, transplant to peat fine sand Mixed culture substrate, cultivate about 30 days under temperature 25 DEG C, more than 65% humidity, be gradually increased intensity of illumination, survival rate of plant is up to 57.3% (Zhang Shengxi, Deng. Taxus mairei Jing Duan tissue culture. Chinese countryside well-to-do level science and technology .2010,6:35 36,52.). Take root after Taxus chinensis var. mairei Callus seedling differentiation tissue cultured seedling on the mixed-matrix of river sand+peat planting percent up to more than 95% (leaf adds the Wang Yumei that sues for peace. Taxus mairei tissue cultivating and seedling is tested. Fujian Forestry science and technology, 2008,35 (3): 102 105).

As can be seen here, the rooting culture survival rate of the significant impact Ramulus et folium taxi cuspidatae tissue cultured seedling such as Ramulus et folium taxi cuspidatae variety classes, different outer implant origin, the different control taming seedling exercising substrate, different rooting culture temperature, humidity and planting percent. So far, report but without a kind of rooting culture method of quick, easy embryo germination in vitro Taxus mairei Tissue vaccine.

Summary of the invention

It is an object of the invention to provide a kind of rooting culture method of Taxus mairei tissue cultured seedling, the method is quick, easy, transplants seedling exercising survival rate high, it is possible to achieve the large-scale production of Taxus mairei seedling.

It is an object of the invention to be accomplished by.

A kind of rooting culture method of Taxus mairei tissue cultured seedling, comprises the following steps:

(1) selection of tissue cultured seedling: select root system development good, the Taxus mairei tissue cultured seedling of more than 1 true leaf;

(2) tissue cultured seedling is tentatively tamed: being put into tissue cultured seedling by group culture container in the greenhouse of 50% sunshade net 35 days, indoor temperature is less than 35 DEG C;

(3) preparation of transplanting container and cultivation matrix: select seedling-raising box, filling Vermiculitum, the peat composed of rotten mosses or with the fresh coconut palm bran of non-woven wraps toward seedling-raising box hole in;

(4) transplanting and seedling exercising: in the middle of the cultivation matrix in tissue cultured seedling implantation step (3), water foot moisture until the substrate of seedling-raising box hole tray has water to overflow, close nursery lid, put in the warmhouse booth with 50% sunshade net and cultivate, ambient temperature, less than 35 DEG C, is opened seedling-raising box lid and is entered conventional seeling industry management mode after 2 weeks.

The substrate of described step (3) acclimatization and transplants is with the fresh coconut palm bran powder of non-woven wraps or is directly Vermiculitum, substrate is inserted in seedling-raising box hole, flattens.

Described step (4) is by the middle of the cultivation matrix in tissue cultured seedling implantation step (3), and disposable the watering after enough of moisture need not rewater.

Seedling-raising box lid height >=5 centimetre in described method; The height (not including the height of lid) of seedling-raising box >=6 centimetres; Diameter >=3 centimetre of supporting hole tray, the hole tray degree of depth is highly consistent with seedling-raising box. The specification of hole number and seedlings nursing plate is unrestricted.

The invention have the advantages that

(1) present invention provides the complete operation flow process of a set of rooting culture based on Taxus mairei embryo germination in vitro seedling first;

(2) present invention explores the substrate being suitable for Taxus mairei training tissue culture seedling by lot of experiments, including Vermiculitum, the peat composed of rotten mosses and by fresh three kinds of coconut palm bran powder of non-woven wraps, and supporting seedling-raising box device;

(3) compared with existing Ramulus et folium taxi cuspidatae training tissue culture seedling technology, seedling exercising survival rate is once transplanted high, it is not necessary to the work of loaded down with trivial details temperature and water management aspect;

(4) compared with prior art, the seedling exercising time of the present invention is short, convenient transplanting after seedling exercising, and the seedling of applicable Internet of Things pattern is sold.

(5) present invention can realize the large-scale production of Taxus mairei seedling.

Accompanying drawing explanation

Fig. 1 is the partial routine that the employing present invention obtains the rooting culture method of Taxus mairei tissue cultured seedling;

The tissue cultured seedling that wherein figure A Taxus mairei embryo germination in vitro is formed;

B Taxus mairei prepares to transplant after tentatively taming;

C D Taxus mairei tissue cultured seedling Vermiculitum seedling-raising box system acclimatization and transplants 21 days, rooting culture survival rate 100%;

E F Taxus mairei tissue cultured seedling non-woven wraps fresh coconut palm bran seedling-raising box system acclimatization and transplants 21 days, rooting culture survival rate 100%;

G H Taxus mairei tissue cultured seedling peat composed of rotten mosses seedling-raising box system acclimatization and transplants 21 days, rooting culture survival rate 91.67%;

I J Taxus mairei tissue cultured seedling perlite seedling-raising box system acclimatization and transplants 21 days, rooting culture survival rate 66.66%; Taxus mairei blade color is partially yellow at later stages;

K L Taxus mairei tissue cultured seedling garden mould seedling-raising box system acclimatization and transplants 21 days, rooting culture survival rate 0%;

M N Taxus mairei tissue cultured seedling river sand seedling-raising box system acclimatization and transplants 21 days, rooting culture survival rate 33.33%; Taxus mairei blade color is partially yellow at later stages.

Detailed description of the invention

Following example are further intended to the present invention is described, without limiting the present invention.

Preferred embodiment 1

(2) tissue cultured seedling is tentatively tamed: be placed in greenhouse 35 days by group culture container band tissue cultured seedling;

(3) preparation of transplanting container and cultivation matrix: select seedling-raising box, fills Vermiculitum in seedling-raising box hole;

(4) transplanting and seedling exercising: in the middle of the cultivation matrix in tissue cultured seedling implantation step (3), water sufficient moisture, close nursery lid, put in the warmhouse booth with 50% sunshade net and cultivate, ambient temperature is less than 35 DEG C, can opening seedling-raising box lid after 2 weeks and enter conventional seeling industry management mode, tissue cultured seedling transplanting survival rate is 100%.

Preferred embodiment 2

(3) preparation of transplanting container and cultivation matrix: select seedling-raising box, fills the peat composed of rotten mosses in seedling-raising box hole;

(4) transplanting and seedling exercising: in the middle of the cultivation matrix in tissue cultured seedling implantation step (3), water sufficient moisture, close nursery lid, put in the warmhouse booth with 50% sunshade net and cultivate, ambient temperature is less than 35 DEG C, can opening seedling-raising box lid after 2 weeks and enter conventional seeling industry management mode, tissue cultured seedling transplanting survival rate is 91.67%.

Preferred embodiment 3

(3) preparation of transplanting container and cultivation matrix: select seedling-raising box, fills the fresh coconut palm bran powder of non-woven wraps in seedling-raising box hole;

Claims

1. the rooting culture method of a Taxus mairei tissue cultured seedling, it is characterised in that comprise the following steps:

(2) tissue cultured seedling is tentatively tamed: put into tissue cultured seedling by group culture container in the greenhouse of 50% sunshade net 35 days;

2. method according to claim 1, it is characterised in that

3. method according to claim 1, it is characterised in that

4. method according to claim 1, it is characterised in that seedling-raising box lid height >=5 centimetre; Height >=6 centimetre of seedling-raising box; Diameter >=3 centimetre of supporting hole tray.