CN109463279B - Rapid propagation method of curcuma wenyujin sterile seedlings - Google Patents

Abstract

The invention provides a rapid propagation technology of Curcuma wenyujin Y.H.Chen et C.Ling vaccine, which comprises the following steps: firstly, obtaining an explant; secondly, sterilizing the explants; inducing tissue culture seedling of cluster bud; induction of root tip callus; inducing the Curcuma wenyujin Y.H.Chen et C.Ling vaccine; sixthly, propagation of the curcuma wenyujin sterile seedlings. The callus is induced by the root tip meristem of the tissue culture seedling, and the sterile seedling is obtained by redifferentiation, so that the problem that the sterile meristem is difficult to obtain directly from the explant is effectively solved, the obtaining of the sterile seedling is also ensured, and the problem that the tissue culture seedling generated by the explant induction cannot be continuously propagated due to the carrying of endophyte is solved; the technology of the invention has the advantages that the sterile seedlings are continuously propagated and cultured in MS, 2.0 mg/L6-BA and 0.5mg/L NAA culture medium, the synchronous realization of propagation and rooting is realized, the process of rooting and seedling strengthening is omitted, and the tissue culture cost is saved.

Description

Rapid propagation method of curcuma wenyujin sterile seedlings

Technical Field

The invention relates to the technical field of common turmeric propagation, in particular to a rapid propagation method of common turmeric sterile seedlings.

Background

The common turmeric is perennial herb of curcuma of zingiberaceae, has the values of medical health care, eating, beauty treatment and garden appreciation, is an important economic crop, is a Wenzhou genuine medicinal material, and is one of eight Zhejiang flavors. The part of the common turmeric which is used as the traditional Chinese medicine is a tuber, and the common turmeric can form the common turmeric, the common turmeric and the common turmeric, is used for relieving qi stasis, relieving pain, resisting inflammation and the like, and the volatile oil containing sesquiterpene alcohol and sesquiterpene compounds extracted from the common turmeric root is frequently used clinically, wherein the components can effectively treat tumors and reduce the stroke of thrombus, so the common turmeric and the common turmeric are very effective medicines and have very wide development prospects. Meanwhile, the common turmeric root-tuber is also used for reducing the degree of platelet aggregation, protecting the liver, achieving good treatment on cardiovascular diseases and playing a certain role in the aspect of acute renal failure, and has a very wide medicinal range.

Since the seeds of Curcuma are propagated for a long time, while maintaining the excellent properties of female parent, the plants are susceptible to bacterial wilt caused by Pseudomonas solanacearum during production, vascular bundles of overground stem and underground stem of Zingiberaceae plant damaged by the bacteria are blocked and become rotten, the leaves are light yellow, and the plants die. Bacterial wilt often occurs in tropical, subtropical and temperate regions, and the bacterial wilt causes plant withering and death because the bacterial wilt grows in soil and contacts with roots and stems of plants, and if roots and stems of the plants have wounds, the bacterial wilt can invade, so that the vascular system can be blocked by heteropolysaccharide generated by the bacterial wilt, and normal water delivery can not be carried out. Investigation shows that ginger plant seed blocks often have bacterial wilt, the field incidence rate is not low, generally 10% -20%, the serious disease field is as high as 30% -40%, and even some field blocks can be harvested absolutely.

The traditional production mode of the curcuma wenyujin for long-term vegetative propagation is far from meeting the requirements of the market on the quality and the yield of the curcuma wenyujin and can not adapt to the modern production, so that the modern biological culture method, such as a tissue culture technology, is applied to improve the yield and the quality of the curcuma wenyujin so as to quickly and efficiently obtain the sterile seedlings. At present, the bacteria-removing and rapid propagation of the common turmeric is not reported in detail, so the experiment aims to provide a theoretical basis for utilizing the bacteria-removing seedlings of the common turmeric to rapidly produce the common turmeric, improve the quality of the common turmeric, improve the yield and implement the GAP of the common turmeric through a rapid propagation system of the common turmeric bacteria-removing seedlings, and simultaneously provide an effective way for industrially producing the seedlings and solving the problem of economic reduction caused by seed reservation of common turmeric farmers for long-term interference.

Disclosure of Invention

Solves the technical problem

Aiming at the defects of the prior art, the invention provides a rapid propagation method of curcuma wenyujin sterile seedlings.

Technical scheme

In order to achieve the purpose, the invention is realized by the following technical scheme: a rapid propagation method of Curcuma wenyujin Y.H.Chen et C.Ling vaccine comprises the following steps:

firstly, obtaining an explant: cutting off buds with the length of 0.5-1.0 cm from tuberous roots, placing the buds under running water for washing to remove surface dirt, carefully peeling off leaf sheaths, and cutting the buds into small segments with stem tips of about 0.5 cm;

② explant sterilization: moving the small sections with the stem tips to an ultra-clean workbench, sterilizing the small sections with 75% alcohol for 5-10 seconds, washing the small sections with sterile water for 3-4 times, placing the small sections in the sterile water, starting an ozone sterilization lamp and an ultraviolet sterilization lamp for one hour, draining the small sections, and placing the small sections in a sterile bottle;

③ inducing the tissue culture seedling of the cluster buds: inoculating the obtained explant temperature curcuma aromatica buds to an MS culture medium, adding 1.0-4.0mg/L of 6-BA and 0.1-0.5mg/L of NAA into the MS culture medium, and culturing for 30 days;

induction of root tip callus: continuously culturing the obtained Curcuma wenyujin tissue culture seedling on MS +2.0 mg/L6-BA +0.5mg/L NAA culture medium for 30 days to obtain a Curcuma wenyujin complete seedling with a thick root system, wherein the Curcuma wenyujin complete seedling has two thick and thin roots, selecting a meristem at the top of the thick root system, cutting the meristem by using a scalpel, inoculating the meristem on MS +0.5 mg/L6-BA +5 mg/L2, 4-D culture medium, and culturing for 30 days to obtain a callus;

induction of Curcuma wenyujin Y.H.Chen et C.Ling vaccine: inoculating the obtained Curcuma wenyujin callus on MS +3.0 mg/L6-BA +0.1 mg/L2, 4-D culture medium, and allowing Curcuma wenyujin plantlet to grow two weeks later to obtain Curcuma wenyujin sterile plantlet;

sixthly, propagation of the curcuma wenyujin degerming vaccine: the sterile seedlings are subjected to propagation culture on MS, 2.0 mg/L6-BA and 0.5mg/L NAA culture medium.

Preferably, 3.0mg/L of 6-BA and 0.5mg/L of NAA are added into the MS culture medium in the step (c).

Advantageous effects

The invention provides a rapid propagation method of curcuma wenyujin sterile seedlings, which has the following beneficial effects:

the technology adopts the meristem at the root tip of the tissue culture seedling to induce and generate callus, and obtains the sterile seedling through redifferentiation, thereby effectively solving the problem that the sterile meristem is difficult to obtain directly from the explant, ensuring the obtaining of the sterile seedling, and solving the problem that the tissue culture seedling generated through the induction of the explant can not be continuously propagated due to carrying endophyte;

the technology of the invention has the advantages that the sterile seedlings are continuously propagated and cultured in MS, 2.0 mg/L6-BA and 0.5mg/L NAA culture medium, the synchronous realization of propagation and rooting is realized, the process of rooting and seedling strengthening is omitted, and the tissue culture cost is saved.

Drawings

FIG. 1 is a schematic view of a cluster bud of Curcuma wenyujin according to the present invention;

FIG. 2 is a schematic view of the thick and thin roots of the whole seedling of Curcuma wenyujin Y.H.Chen et C.Ling of the present invention;

FIG. 3 is a schematic diagram of Curcuma wenyujin Y.H.Chen et C.Ling tip callus and its redifferentiation.

Detailed Description

The embodiment of the invention provides a rapid propagation method of Curcuma wenyujin Y.H.Chen et C.Ling vaccine, which comprises the following steps:

firstly, obtaining an explant: cutting off buds with the length of 0.5-1.0 cm from tuberous roots, placing the buds under running water for washing to remove surface dirt, carefully peeling off leaf sheaths, and cutting the buds into small sections with stem tips of about 0.5cm, which is beneficial to the starting induction of the curcuma wenyujin;

② explant sterilization: moving the small sections with the stem tips to an ultra-clean workbench, sterilizing the small sections with 75% alcohol for 5-10 seconds, washing the small sections with sterile water for 3-4 times, placing the small sections in the sterile water, starting an ozone sterilization lamp and an ultraviolet sterilization lamp for one hour, draining the small sections, and placing the small sections in a sterile bottle;

③ inducing the tissue culture seedling of the cluster buds: inoculating the obtained explant temperature curcuma aromatica buds to an MS culture medium, adding 1.0-4.0mg/L of 6-BA and 0.1-0.5mg/L of NAA into the MS culture medium, and culturing for 30 days;

preferably, 3.0mg/L of 6-BA and 0.5mg/L of NAA are added into the MS culture medium in the third step, and the formula has the highest induction rate, the best effect, strong buds, good growth vigor and details shown in the specification

Table 1 and figure 1.

Table 1: the induction effect of cluster buds is influenced by different hormone concentrations

Induction of root tip callus: culturing the obtained Curcuma wenyujin tissue culture seedling on MS +2.0 mg/L6-BA +0.5mg/L NAA culture medium for 30 days to obtain Curcuma wenyujin whole seedling with coarse root system, as shown in figure 2. The integral Curcuma wenyujin seedling has two kinds of thick and thin roots, a meristem at the top of a thick root system (namely a small white point at the top of the thick root system) is selected, cut by a scalpel and inoculated on a culture medium of MS +0.5 mg/L6-BA +5 mg/L2, 4-D, and the culture time is 30 days to obtain a callus; see figure 3 for details. The meristem at the top of the thick root system is selected, the size of the meristem is large, the color difference with the surrounding color is obvious, the meristem is beneficial to obtaining, and the meristem has no vascular bundle, so that the complete bacteria removal effect is beneficial to obtaining.

Induction of Curcuma wenyujin Y.H.Chen et C.Ling vaccine: inoculating the obtained Curcuma wenyujin callus on MS +3.0 mg/L6-BA +0.1 mg/L2, 4-D culture medium, and allowing Curcuma wenyujin plantlet to grow two weeks later to obtain Curcuma wenyujin sterile plantlet; see figure 3 for details.

Sixthly, propagation of the curcuma wenyujin degerming vaccine: the sterile seedlings are subjected to propagation culture on MS +2.0 mg/L6-BA +0.5mg/L NAA culture medium, the proliferation rate is more than 5 times, and large-scale production can be carried out, and details are shown in Table 2.

Table 2: test of proliferation rate of propagation culture of germ-free seedlings

Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.

Claims (1)

1. A rapid propagation method of Curcuma wenyujin Y.H.Chen et C.Ling vaccine is characterized by comprising the following steps:

firstly, obtaining an explant: cutting off buds with the length of 0.5-1.0 cm from tuberous roots, placing the buds under running water for washing to remove surface dirt, carefully peeling off leaf sheaths, and cutting the buds into small segments with stem tips of about 0.5 cm;

② explant sterilization: moving the small sections with the stem tips to an ultra-clean workbench, sterilizing the small sections with 75% alcohol for 5-10 seconds, washing the small sections with sterile water for 3-4 times, placing the small sections in the sterile water, starting an ozone sterilization lamp and an ultraviolet sterilization lamp for one hour, draining the small sections, and placing the small sections in a sterile bottle;

③ inducing the tissue culture seedling of the cluster buds: inoculating the obtained explant temperature curcuma aromatica buds to an MS culture medium, adding 3.0mg/L of 6-BA and 0.5mg/L of NAA into the MS culture medium, and culturing for 30 days;

induction of root tip callus: continuously culturing the obtained Curcuma wenyujin tissue culture seedling on MS +2.0 mg/L6-BA +0.5mg/L NAA culture medium for 30 days to obtain a Curcuma wenyujin complete seedling with a thick root system, wherein the Curcuma wenyujin complete seedling has two thick and thin roots, selecting a meristem at the top of the thick root system, cutting the meristem by using a scalpel, inoculating the meristem on MS +0.5 mg/L6-BA +5 mg/L2, 4-D culture medium, and culturing for 30 days to obtain a callus;

induction of Curcuma wenyujin Y.H.Chen et C.Ling vaccine: inoculating the obtained Curcuma wenyujin callus on MS +3.0 mg/L6-BA +0.1 mg/L2, 4-D culture medium, and allowing Curcuma wenyujin plantlet to grow two weeks later to obtain Curcuma wenyujin sterile plantlet;

sixthly, propagation of the curcuma wenyujin degerming vaccine: the sterile seedlings are subjected to propagation culture on MS, 2.0 mg/L6-BA and 0.5mg/L NAA culture medium.

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