CN110384044B - Cultivation method of virus-free seed stems of taros - Google Patents
Cultivation method of virus-free seed stems of taros Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
- A01G24/12—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material containing soil minerals
- A01G24/15—Calcined rock, e.g. perlite, vermiculite or clay aggregates
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/20—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
- A01G24/28—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material containing peat, moss or sphagnum
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G31/00—Soilless cultivation, e.g. hydroponics
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/20—Reduction of greenhouse gas [GHG] emissions in agriculture, e.g. CO2
- Y02P60/21—Dinitrogen oxide [N2O], e.g. using aquaponics, hydroponics or efficiency measures
Abstract
The invention discloses a cultivation method of a virus-free seed stem of taro, which relates to the field of cultivation of virus-free seedlings of taro, and specifically comprises the following steps: preparing a culture medium by using 6-BA, NAA, agar and sucrose, and placing the culture medium in a test tube; selecting a healthy bulb with a regular shape for accelerating germination, cutting a stem tip, then inoculating the stem tip into a stem tip growth culture medium for induction culture to form a detoxified test-tube plantlet, and inoculating the test-tube plantlet onto the bulb induction culture medium to obtain a detoxified seed bulb; transplanting the detoxified seed stems into a matrix of grass carbon and vermiculite to harden the seedlings, and observing and recording the growth condition of the taros during the culture period. The method provided by the invention has the advantages that the seed stem is cultured in the culture medium by using the virus-free test-tube seedling to obtain the virus-free seed stem, then the obtained virus-free seed stem is transplanted into the matrix of turf and vermiculite to harden the seedling, the induction of the virus-free seed stem of the taro can effectively increase the planting survival rate, the serious differentiation condition in the test-tube seedling planting process is reduced, the seed stem is high in yield, the storage and the transportation are easy, and the guarantee is provided for the later-stage taro planting production.
Description
Technical Field
The invention relates to the technical field of cultivation of virus-free seedlings of taros, in particular to a cultivation method of virus-free seed stems of taros.
Background
According to long-term research by scientists, the cause of crop quality deterioration is mainly infection by plant viruses. In order to obtain high yield, virus-free seedlings, i.e. virus-free seedlings, must be cultured in production, which is the concept of virus-free seedlings. Agricultural scientists in China use biotechnology measures to obtain virus-free seedling materials from virus-infected plants. For example, the apical meristem of potato plants has little or no virus, and apical meristem culture techniques are often used to obtain disease-free plants, i.e., virus-free potatoes. Potato blocks grown by the plant are used as stock seeds to propagate more virus-free seed potatoes for large-area popularization and use in production; the virus-free seedling technology is also applied to agricultural production of flowers, fruit trees, konjak and the like, so that high-quality non-toxic flower bulbs, virus-free fruit tree seedlings and the like can be produced.
At present, the taro virus-free seedlings are obtained by culturing the stem tip meristem of the taro, but the survival rate of the virus-free seedlings is low, the transportation is inconvenient, cluster seedlings are easy to appear after the seedlings are planted, the seed stem yield is low, and the popularization of the taro virus-free seedlings is seriously influenced.
The invention patent of patent application publication No. CN208081437U discloses a tissue culture and rapid propagation technology of red bud taro virus-free seedlings, which is characterized in that a strong and disease-free bulb is selected, sterilized and disinfected, and a stem tip with the size of 0.2mm is selected and inoculated into an induction medium for differentiation culture; after culturing into cluster buds, cutting off the top ends of the cluster buds, and putting the cluster buds into a proliferation culture medium for rapid propagation; when the cluster bud leaves are unfolded, putting the cluster bud leaves into a rooting culture medium for strong seedling rooting culture, and then transferring the cluster bud leaves into a hardening seedling. The method adopts twice detoxification technique in propagation, picks stem tip of test-tube plantlet, and improves detoxification effect. The formula is optimized in the preparation of the culture medium, and the growth of test-tube plantlets is facilitated. The method overcomes the defects of large seed consumption and high cost of the traditional method for using the corms as the propagation material, can artificially control the culture growth condition, is not influenced by natural conditions, has small material taking amount, is economic in culture material, short in growth period and large in propagation coefficient, reduces the production cost, maintains the variety property, improves the yield and the quality, and can realize industrial seedling culture and industrial production.
However, the technical scheme still has more defects in actual application, such as difficulty in storage and inconvenience in transportation of the obtained detoxified seedlings, the phenomena of mass death of the detoxified seedlings and difficulty in survival in the later period in the transportation process, and the obtained detoxified seedlings are easy to differentiate after being planted and can not ensure the production of the planted taro in the later period.
Disclosure of Invention
In order to overcome the defects of the prior art, the embodiment of the invention provides a cultivation method of the virus-free taro seed stems, wherein the virus-free test tube plantlets are cultivated by using a culture medium of MS +1.0mg/L6-BA +0.5mg/LNAA +7g/L agar +8% sucrose to obtain the test tube seed stems, and then the obtained test tube seed stems are transplanted into a matrix of grass carbon + vermiculite to be cultivated again.
In order to achieve the purpose, the invention provides the following technical scheme: a cultivation method of a virus-free seed stem of taro comprises the following steps:
s1, preparing a stem tip culture medium and a seed stem induction culture medium, adding 6-BA, NAA, agar and sucrose with certain mass into a basic culture medium MS, mixing, and carrying out autoclaving for later use;
s2, selecting strong and disease and insect-free corms, cleaning the surfaces of the corms, accelerating germination, cutting stem tips of taro single buds with the length of about 0.2-0.3mm under a microscope when the buds grow to 1.5-2cm, and then inoculating the stem tips into a culture medium for induction culture to form virus-free test-tube plantlets;
s3, inoculating the test-tube plantlet obtained in the S2 to a seed stem induction culture medium for culture to obtain a virus-free seed stem;
and S4, mixing the grass peat and the vermiculite according to a certain proportion to obtain a seed stem transplanting matrix, transplanting the detoxified seed stems obtained in the step S3 into the grass peat and vermiculite matrix for hardening seedlings, and observing and recording the growth condition of the taro seed stems during the culture period.
In a preferred embodiment, in step S1, the stem tip induction medium is MS +2mg/L6-BA +0.5mg/LNAA +7g/L agar +3% sucrose, and the pH is adjusted to 5.7.
In a preferred embodiment, in step S1, the seed stem induction medium is MS +0.8mg/L6-BA +0.4mg/LNAA +6.8g/L agar +6% sucrose, and the pH is adjusted to 5.7.
In a preferred embodiment, in step S1, the seed stem induction medium is MS +1.0mg/L6-BA +0.5mg/LNAA +7g/L agar +8% sucrose, and the pH is adjusted to 5.7.
In a preferred embodiment, in step S1, the seed stem induction medium is MS +1.2mg/L6-BA +0.6mg/LNAA +7.2g/L agar +10% sucrose, and the pH is adjusted to 5.7.
In a preferred embodiment, in the step S3, the culture temperature of the seed stems is 20-30 ℃.
In a preferred embodiment, in step S3, the illumination intensity of the seed stem induction culture is 4000lx, and the illumination time is 13-15h/d.
In a preferred embodiment, in the step S4, the ratio of grass peat to vermiculite is 1-3.
The invention has the technical effects and advantages that:
according to the method, the seed stems are cultured by using the culture medium of MS +1.0mg/L6-BA +0.5mg/LNAA +7g/L agar +8% of cane sugar to obtain the virus-free seed stems, then the obtained virus-free seed stems are transplanted into the matrix of turf + vermiculite to carry out seedling hardening, the survival rate of planting can be effectively increased, differentiation is reduced, the seed stems are high in yield, the virus-free seed stems are easy to store and transport, the virus-free seed stems are not differentiated after being planted, and the guarantee is provided for later-stage seed taro production.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1:
the invention provides a cultivation method of a virus-free seed stem of taro, which comprises the following steps:
s1, preparing a stem tip culture medium and a seed stem induction culture medium, adding 6-BA, NAA, agar and sucrose with certain mass into a basic culture medium MS, wherein the stem tip culture medium is MS +2mg/L6-BA +0.5mg/LNAA +7g/L agar +3% sucrose, adjusting the pH to be 5.7, the seed stem induction culture medium is MS +0.8mg/L6-BA +0.4mg/LNAA +6.8g/L agar +6% sucrose, mixing to obtain a culture medium, adjusting the pH to be 5.7, and carrying out autoclaving on the obtained culture medium for later use;
s2, selecting strong and disease and insect-free corms, cleaning the surfaces of the corms, accelerating germination, cutting stem tips of taro with the length of about 0.2-0.3mm from a single bud under a microscope when the buds grow to 1.5cm, and then inoculating the stem tips into a culture medium to be induced and cultured into virus-free seedlings;
s3, inoculating the virus-free seedlings to a seed stem induction culture medium for culture, wherein the culture temperature is 20 ℃, the illumination intensity of seed stem induction culture is 4000lx, and the illumination time is 13h/d to obtain virus-free seed stems;
s4, mixing the grass peat and the vermiculite according to a certain ratio to obtain a transplanting matrix, transplanting the detoxified seed stems obtained in the step S3 into the grass peat and vermiculite matrix for hardening seedlings, wherein the ratio of the grass peat to the vermiculite is 2:1, observing and recording the growth condition of the taro seed stems during the culture period.
Example 2:
the invention provides a cultivation method of a virus-free seed stem of taro, which comprises the following steps:
s1, preparing a stem tip culture medium and a bulb induction culture medium, adding 6-BA, NAA, agar and sucrose with certain mass into a basic culture medium MS, wherein the stem tip culture medium is MS +2mg/L6-BA +0.5mg/LNAA +7g/L agar +3% sucrose, adjusting the pH to be 5.7, the seed stem induction culture medium is MS +1.0mg/L6-BA +0.5mg/LNAA +7g/L agar +8% sucrose, mixing to obtain a culture medium, adjusting the pH to be 5.7, and carrying out autoclaving on the obtained culture medium for later use;
s2, selecting strong and disease and insect-free corms, washing the surfaces of the corms, accelerating germination, cutting stem tips of taro single buds with the length of about 0.2-0.3mm under a microscope when the buds grow to 1.7cm, and then inoculating the stem tips into a culture medium to be induced and cultured into test-tube plantlets;
s3, inoculating the S2 test-tube plantlet to a seed stem induction culture medium, wherein the seed stem culture temperature is 25 ℃, the illumination intensity is 4000lx, and the illumination time is 14h/d to obtain a virus-free seed stem;
s4, mixing the grass peat and the vermiculite according to a certain ratio to obtain a seed stem transplanting matrix, transplanting the detoxified seed stems obtained in the step S3 into the grass peat and vermiculite matrix for hardening seedlings, wherein the ratio of the grass peat to the vermiculite is 2:1, observing and recording the growth condition of the taro seed stems during the culture period.
Example 3:
the invention provides a cultivation method of a virus-free seed stem of taro, which comprises the following steps:
s1, preparing a stem tip culture medium and a corm induction culture medium, adding 6-BA, NAA, agar and sucrose with certain mass into a basic culture medium MS, wherein the stem tip culture medium is specifically MS +2mg/L6-BA +0.5mg/LNAA +7g/L agar +3% sucrose, adjusting the pH to 5.7, the seed stem induction culture medium is specifically MS +1.2mg/L6-BA +0.6mg/LNAA +7.2g/L agar +10% sucrose, adjusting the pH to 5.7, mixing to obtain a culture medium, and carrying out high-pressure sterilization on the obtained culture medium for later use;
s2, selecting strong and disease and pest-free corms, cleaning the surfaces of the corms, accelerating germination, cutting stem tips of taros with single buds of 0.2-0.3mm in length under a microscope when the buds grow to 2cm, and then inoculating the stem tips into a culture medium to induce and culture the stem tips into virus-free seedlings;
s3, inoculating the virus-free seedlings obtained in the S2 onto a seed stem induction culture medium, wherein the culture temperature is 30 ℃, the illumination intensity of the induction culture is 4000lx, and the illumination time is 15h/d to obtain virus-free seed stems;
s4, mixing the grass peat and the vermiculite according to a certain ratio to obtain a seed stem transplanting matrix, transplanting the detoxified seed stems obtained in the step S3 into the grass peat and vermiculite matrix for hardening seedlings, wherein the ratio of the grass peat to the vermiculite is 2:1, during the culture period, the growth of seed stems is observed and recorded.
Example 4:
the invention provides a cultivation method of a virus-free seed stem of taro, which comprises the following steps:
s1, preparing a stem tip culture medium and a seed stem induction culture medium, adding 6-BA, NAA, agar and sucrose with certain mass into a basic culture medium MS, wherein the stem tip culture medium is MS +2mg/L6-BA +0.5mg/LNAA +7g/L agar +3% sucrose, adjusting the pH to be 5.7, the seed stem induction culture medium is MS +1.0mg/L6-BA +0.5mg/LNAA +7g/L agar +8% sucrose, mixing to obtain a culture medium, adjusting the pH to be 5.7, and performing high-pressure sterilization on the obtained culture medium for later use;
s2, selecting strong and disease and insect-free corms, cleaning the surfaces of the corms, accelerating germination, cutting stem tips of taro with the length of about 0.2-0.3mm from a single bud under a microscope when the buds grow to 1.7cm, and then inoculating the stem tips into a culture medium to be induced and cultured into virus-free seedlings;
s3, inoculating the virus-free seedlings obtained in the S2 onto a seed stem induction culture medium, wherein the culture temperature is 25 ℃, the illumination intensity of the induction culture is 4000lx, and the illumination time is 14h/d to obtain virus-free seed stems;
s4, mixing the grass peat and the vermiculite according to a certain proportion to obtain a seed stem transplanting matrix, transplanting the detoxified seed stems obtained in the step S3 into the grass peat and vermiculite matrix for hardening, wherein the proportion of the grass peat to the vermiculite is 1, and observing and recording the growth condition of the taro seed stems during culture.
Example 5:
the invention provides a cultivation method of a virus-free seed stem of taro, which comprises the following steps:
s1, preparing a stem tip culture medium and a seed stem induction culture medium, adding 6-BA, NAA, agar and sucrose with certain mass into a basic culture medium MS, wherein the stem tip culture medium is MS +2mg/L6-BA +0.5mg/LNAA +7g/L agar +3% sucrose, adjusting the pH to be 5.7, the seed stem induction culture medium is MS +1.0mg/L6-BA +0.5mg/LNAA +7g/L agar +8% sucrose, mixing to obtain a culture medium, adjusting the pH to be 5.7, and performing high-pressure sterilization on the obtained culture medium for later use;
s2, selecting strong and disease and insect-free corms, cleaning the surfaces of the corms, accelerating germination, cutting stem tips of taro single buds with the length of about 0.2-0.3mm under a microscope when the buds grow to 1.7cm, and then inoculating the stem tips into a culture medium in a test tube to be induced and cultured into virus-free seedlings;
s3, inoculating the virus-free seedlings obtained in the S2 onto a seed stem induction culture medium, wherein the culture temperature is 25 ℃, the illumination intensity of the induction culture is 4000lx, and the illumination time is 14h/d to obtain virus-free seed stems;
and S4, mixing the grass peat and the vermiculite according to a certain ratio to obtain a transplanting matrix, transplanting the test tube seed stems obtained in the step S3 into the grass peat + vermiculite matrix for re-cultivation, wherein the ratio of the grass peat to the vermiculite is 3.
Example 6:
the growth data of 30 colocasia esculenta virus-free seed stems cultured in the above examples 1-5 during the culture period were randomly extracted, and the following table was obtained:
as can be seen from the above table, the ratio of raw materials in the culture medium and the transplanting matrix in the embodiment 2 is moderate, and the temperature and the illumination in the culture process are both proper, so that the culture of the virus-free seed stems of the taros is facilitated, the rooting rate and the survival rate of the obtained virus-free seedlings are greatly improved, and the differentiation after planting is less.
And finally: the above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that are within the spirit and principle of the present invention are intended to be included in the scope of the present invention.
Claims (1)
1. A cultivation method of a taro virus-free seed stem is characterized in that: the specific cultivation method comprises the following steps:
s1, preparing a stem tip culture medium and a seed stem induction culture medium, adding 6-BA, NAA, agar and sucrose with certain mass into a basic culture medium MS, wherein the stem tip culture medium is specifically MS +2mg/L6-BA +0.5mg/LNAA +7g/L agar +3% sucrose, adjusting the pH to 5.7, the seed stem induction culture medium is specifically MS +1.0mg/L6-BA +0.5mg/LNAA +7g/L agar +8% sucrose, mixing to obtain a culture medium, adjusting the pH to 5.7, and carrying out autoclaving on the obtained culture medium for later use;
s2, selecting strong and disease and pest-free corms, cleaning the surfaces of the corms, accelerating germination, cutting stem tips of taros with single buds of 0.2-0.3mm in length under a microscope when the buds grow to 1.7cm, and then inoculating the stem tips into a stem tip culture medium to induce and culture the stem tips into test-tube plantlets;
s3, inoculating the S2 test-tube plantlet to a seed stem induction culture medium, wherein the seed stem culture temperature is 25 ℃, the illumination intensity is 4000Lx, and the illumination time is 14h/d, so as to obtain a virus-free seed stem;
s4, mixing the grass peat and the vermiculite according to a certain ratio to obtain a seed stem transplanting matrix, transplanting the detoxified seed stems obtained in the step S3 into the grass peat and vermiculite matrix for hardening seedlings, wherein the ratio of the grass peat to the vermiculite is 2:1, during the culture period, the growth condition of the taro seed stems is observed and recorded.
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| CN113455365B (en) * | 2021-08-02 | 2022-05-31 | 江西省超级水稻研究发展中心(江西省农科院海南水稻育种中心) | Method for preserving potted plant of common wild rice seed stems |
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| CN1073344C (en) * | 1997-10-07 | 2001-10-24 | 南京农业大学 | Binglang taro seed bulb rapid propagation method |
| CN102217550A (en) * | 2011-06-09 | 2011-10-19 | 马宗新 | Fast-propagation technology and composition of culture medium for virus-free plantlets of red bud taros |
| CN103070071A (en) * | 2013-01-22 | 2013-05-01 | 武汉市蔬菜科学研究所 | Method for fast propagation of test tube taro by detoxification |
| USPP27733P2 (en) * | 2015-02-10 | 2017-02-28 | Brian's Botanicals | Colocasia plant named ‘Painted Black Gecko’ |
| CN111919747A (en) * | 2020-07-27 | 2020-11-13 | 衡阳市蔬菜研究所 | Method for improving inductivity and rooting rate based on detoxified areca taro culture |
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| EP2927323A2 (en) * | 2011-04-11 | 2015-10-07 | Targeted Growth, Inc. | Identification and the use of krp mutants in plants |
| CN106234229A (en) * | 2016-09-30 | 2016-12-21 | 广西壮族自治区农业科学院 | A kind of method that Fructus Colocasiae Esculentae tissue cultured seedling is heeled in |
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