CN101233826B - Detoxifying fast breeding technique for edible lily - Google Patents
Detoxifying fast breeding technique for edible lily Download PDFInfo
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- CN101233826B CN101233826B CN2008100341814A CN200810034181A CN101233826B CN 101233826 B CN101233826 B CN 101233826B CN 2008100341814 A CN2008100341814 A CN 2008100341814A CN 200810034181 A CN200810034181 A CN 200810034181A CN 101233826 B CN101233826 B CN 101233826B
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Abstract
The invention relates to a virus elimination and rapid propagation technique for edible lily. The technique comprises the following processes in sequence: edible lily stem tip is taken to obtain virus elimination lily tissue culture seedling by starting and subculturing strong seedling; strong seedling induces regeneration lepisphere; regeneration lepisphere induces multiple shoot or adventitious bud; the multiple shoot or the adventitious bud induces regeneration lepisphere after subculturing of strong seedling and then the regeneration lepisphere induces the multiple shoot or the adventitious bud again. Batch expanding propagation is carried out through the cyclic alternating. The culture of tissue culture seedling of the virus elimination lily comprises the following steps: edible lilybulb is sand-cultured for 7-10 days; the outer scale is stripped and then the stripped lily bulb is cleaned and sterilized; growing points are peeled under an anatomical lens to get stem tip of 0.1-0.2mm as initial material of expanding propagation; the micro stem tip is inoculated on an initial medium of MS+6BA 0.5-2.5mg/L+NAA 0.1-0.5mg/L to be cultured; the tissue culture seedling of the virus elimination lily with non virus ratio of 100 percent through virus detection is obtained after 3-4 days of dark culture and then light culture. Industrial tissue culture and rapid propagation can be realized.
Description
Technical field
The present invention relates to a kind of detoxifying fast breeding technique of edible lily, especially a kind of at Yixing lily, have the high detoxification efficient and the quick group culturation rapid propagating technology of reproductive efficiency.
Background technology
The lily disease viral disease distributes wide, and harm is big, is one of main disease during edible lily production and fresh cut-flowers are produced, and is subjected to global attention.In recent years, along with China lily is introduced a fine variety increasing sharply of quantity and cultivated area, and nonstandard kind of ball be from numerous personal, and virus disease begins at each lily growing area of China popular, and the general incidence of disease is 40～50%, two generation the kind ball band poison rate more than 90%.Behind the lily infective virus, lifelong band poison is injured for a long time, destroys the normal physiological function of plant, mainly shows as growth potential decline, viral symptoms such as yellow, floral leaf; Propagate by insect amboceptors such as aphids, enlarged viral damaging range; Because virus harm descends edible lily output, the product qualitative change is bad, views and admires the commodity flower degradation of lily; Virus still is difficult to carry out directly effectively control with chemical agent or biologic product.Therefore, the countermeasure that prevents lily infective virus disease is to adopt the detoxification culture technique to obtain detoxic seedling, isolates breeding, produces the healthy ball of planting, and carry out strict virus at each production link and detect.
Lily is various in style, can be divided into officinal lily, edible lily and view and admire lily according to purposes, and this pre-induction lily carries out fast numerous technology and mainly concentrates in the selection to medium.
Number of patent application 200710068034.4 discloses a kind of detoxifying fast breeding method to ornamental type hybridization lily, inducing culture is MS+6BA 1.0～2.0mg/L+IBA 0.1～0.5mg/L+AC 1.0～3.0mg/L, and proliferated culture medium is MS+6BA 0.2～1.0mg/L+IBA 0.1～0.5mg/L.But the sign difference of viewing and admiring lily and edible lily is totally different, and the medium that therefore is used for fast breeding technique is also different because of kind.
Edible lily is well received by consumers as the health-care special vegetable, along with the raising of people's level of consumption, and production and consumption amount cumulative year after year.Yixing lily, lanzhou lily, Lilium longiflorum are to be three big edible lilies, wherein the highest with Yixing lily cultivated area maximum, output again, it has bitter and sweet flavour, bulb is rich in nutritions such as starch, protein, fat, mineral, and have that tonifying middle-Jiao and Qi, reason spleen are good for the stomach, multiple medicinal functions such as antitoxic heart-soothing and sedative, be called as tonic medicine lily, just enjoy the laudatory title of " ginseng in Taihu Lake " in history.Be usually used in making heat-clearing sweets etc. with white fungus, mung bean, the seed of jog's tears, lotus nut etc.Conventional lily propagation method is the clove breeding, and reproduction coefficient is low, and easily infected virus influences its economic worth.Because virus infections output descends, area under cultivation reaches the good species ball that several ten thousand mu kind ball need be replaced by detoxification to Huoshan county peasant in the lily planting process of Yixing.
Patent No. ZL03135142.5 discloses the method for quickly breeding of lanzhou lily in the edible lily, and induce and the fast breeding culture medium of its scale indefinite bud is MS+BA 2mg/L+NAA 0.2mg/L.But this invention mainly concentrates on fast numerous technology, and the present invention carries out numerously soon to the edible lily detoxifying tissue cultivating seedling, and reproduction coefficient reaches more than 15 times, has certain practical value.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of edible lily detoxifying fast breeding technology, breeds fast when obtaining tissue cultural seedlings of free, sets up the great detoxification clone of radix rapidly.
The present invention solves the problems of the technologies described above the technical scheme of being taked: a kind of edible lily detoxifying fast breeding technology, comprise in regular turn: get the edible lily stem apex through start, subculture strong sprout, obtain the detoxification tissue-cultured derived plant lily, be after testing virus-free after, by regeneration induction lepisphere in strong sprout, regeneration lepisphere sheet induced bundle is sprouted or indefinite bud, grow thickly bud or indefinite bud subculture, regeneration induction lepisphere again after strong sprout, regeneration lepisphere sheet induced bundle is again sprouted or indefinite bud, alternate cycles like this, carry out batch and expand numerously, wherein, described detoxification tissue-cultured derived plant lily incubation step comprises:
The first step is got edible lily kind ball warp and is crossed husky training 7～10 days, peels off outer scale, behind the cleaning and sterilizing, shells growing point under anatomical lens, cut open get 0.1～0.2mm size stem apex as expanding numerous starting material;
Second step, little stem apex is seeded in and starts medium MS+6BA 0.5～2.5mg/L+NAA0.1～last cultivation of 0.5mg/L, the dark cultivation changeed light cultivation acquisition detoxification tissue-cultured derived plant lily after 3～4 days, wherein, cultivation temperature is 24 ± 2 ℃, intensity of illumination 2000～3000Lx, light application time 12～14 hours/day.
The numerous starting material of expansion of the present invention is the little stem apex of edible lily, and little stem apex obtains the detoxification tissue-cultured derived plant lily through above-mentioned cultivation, and virus-free after testing back is expanded numerous.
On the such scheme basis, described subculture is with detoxification tissue-cultured derived plant lily subculture, strong sprout on subculture strong seedling culture base MS+6BA 0～2.5mg/L+NAA 0.1～0.5mg/L strong sprout, cultivation temperature is 24 ± 2 ℃, intensity of illumination 2000～3000Lx, light application time 12～14 hours/day.
Described regeneration induction lepisphere is with the Cheng Miao of described subculture after strong sprout regeneration induction lepisphere on regeneration induction lepisphere and root media 1/2MS+6BA 0～1.0mg/L+NAA 0～0.5mg/L, condition of culture is 24 ± 2 ℃ of temperature, intensity of illumination 2000～3000Lx, light application time 12～14 hours/day, growing can obtain detoxification edible lily group in 30～60 days and cultivate ball.
Described evoking adventive bud is detoxification edible lily group to be cultivated the ball scale plant in starting evoking adventive bud on the medium, and condition of culture is 24 ± 2 ℃ of temperature, intensity of illumination 2000～3000Lx, light application time 12～14 hours/day.
The present invention has studied scale evoking adventive bud, adventitious buds proliferation, blade stripping and slicing evoking adventive bud, scale and has induced four kinds of propagation modes of lepisphere, and compares, wherein,
(1) scale evoking adventive bud: the lily bud scale crosscut is become two sections, the inoculation of cardinal extremity base portion, most advanced and sophisticated otch is seeded in evoking adventive bud on the medium.
(2) the blade stripping and slicing is induced newly and sprouted: lily group training aseptic seedling blade is cut into the segment about 0.5cm, evoking adventive bud on medium, and the ability of comparison blade different parts generation indefinite bud, the result is the highest with the blade base adventitious bud induction frequency.
(3) adventitious buds proliferation: on different medium, induce adventitious buds proliferation.
(4) scale is induced lepisphere: get lily bud scale and directly induce lepisphere, the mode of directly breeding by lepisphere expands numerous.
Conclusion is: scale evoking adventive bud reproduction rate is the highest, and reproduction speed is the fastest.
On the basis of such scheme, the little stem apex of described edible lily obtains the detoxification tissue-cultured derived plant lily through startup, subculture after strong sprout, be after testing virus-free after, by strong sprout regeneration induction lepisphere, regeneration lepisphere sheet induced bundle sprout or indefinite bud, grow thickly bud or indefinite bud subculture, after strong sprout again regeneration induction lepisphere, regeneration lepisphere sheet again induced bundle sprout or indefinite bud, so alternate cycles is carried out batch and is expanded numerous.
The invention has the beneficial effects as follows:
The present invention has broken through lily detoxifying fast breeding technology in the past, choose the lily Shoot Tip Culture of 0.1～0.2mm size, survive, start propagation, breed into clone by little growing point cultivation, after testing, virus detects virus-free rate up to 100%, and realizes the industrialized tissue culture and rapid propagation of lily in high detoxification success rate.
Embodiment
Embodiment 1
A kind of edible lily detoxifying fast breeding technology, comprise in regular turn: get the little stem apex of edible lily and obtain the detoxification tissue-cultured derived plant lily after strong sprout through startup, subculture, be after testing virus-free after, by regeneration induction lepisphere in strong sprout, regeneration lepisphere sheet induced bundle is sprouted, the bud subculture of growing thickly, regeneration induction lepisphere, regeneration lepisphere sheet evoking adventive bud more again after strong sprout, alternate cycles like this, carry out batch and expand numerously, wherein, described detoxification tissue-cultured derived plant lily is cultivated and is comprised the steps:
The first step is got edible lily kind ball warp and is crossed husky training 7～10 days, peels off outer scale, behind the cleaning and sterilizing, shells growing point under anatomical lens, cuts the stem apex of 0.1～0.2mm size;
Second step, little stem apex is seeded in to start on the medium MS+6BA 1.0mg/L+NAA 0.1mg/L and cultivates, and secretly cultivates and changes the light cultivation after 3～4 days, and cultivation temperature is 24 ± 2 ℃, the intensity of illumination that light is cultivated is 2000～3000Lx, and light application time is 12～14 hours/day.Cultivate under these conditions, little Shoot Tip Culture survival rate reaches 58.9%, and gained tissue cultivating seedling virus detects virus-free rate and reaches 100%.
In subculture strong seedling culture base MS+6BA 0.8mg/L+NAA 0.1mg/L subculture, strong sprout, cultivation temperature is 24 ± 2 ℃, intensity of illumination 2000～3000Lx, light application time 12～14 hours/day.
With the Cheng Miao of described subculture after strong sprout regeneration induction lepisphere on regeneration induction lepisphere and root media 1/2MS+6BA 0mg/L+NAA 0.1mg/L, cultivation temperature is 24 ± 2 ℃, intensity of illumination 2000～3000Lx, light application time 12～14 hours/day, growing can obtain detoxification edible lily group in 45 days and cultivate ball.
The edible lily group is cultivated the ball scale plant in starting on the medium MS+6BA 0.8mg/L+NAA 0.1mg/L induced bundle and sprout, cultivation temperature is 24 ± 2 ℃, intensity of illumination 2000～3000Lx, light application time 12～14 hours/day.But the bud of growing thickly is through subculture Cheng Miaohou regeneration induction lepisphere again, and alternate cycles like this can expand numerously in enormous quantities, and it expands numerous coefficient and can reach more than 15 times.
Embodiment 2
A kind of edible lily detoxifying fast breeding technology, on the basis of embodiment 1, regeneration lepisphere sheet evoking adventive bud, comprise in regular turn: get the little stem apex of edible lily and obtain the detoxification tissue-cultured derived plant lily after strong sprout through startup, subculture, be after testing virus-free after, by regeneration induction lepisphere in strong sprout, regeneration lepisphere sheet evoking adventive bud, indefinite bud subculture, regeneration induction lepisphere, regeneration lepisphere sheet evoking adventive bud more again after strong sprout, alternate cycles like this, carry out batch and expand numerously, wherein, described detoxification tissue-cultured derived plant lily incubation step comprises:
The first step is got edible lily kind ball warp and is crossed husky training 7～10 days, peels off outer scale, behind the cleaning and sterilizing, shells growing point under anatomical lens, cut open get 0.1～0.2mm size stem apex as expanding numerous starting material;
Second step, little stem apex is seeded in to start on the medium MS+6BA 1.5mg/L+NAA 0.1mg/L and cultivates, the dark cultivation changeed light cultivation acquisition detoxification tissue-cultured derived plant lily after 3～4 days, wherein, cultivation temperature is 24 ± 2 ℃, intensity of illumination 2000～3000Lx, light application time 12～14 hours/day can get the detoxification tissue-cultured derived plant lily.
With detoxification tissue-cultured derived plant lily subculture, strong sprout on subculture strong seedling culture base MS+6BA 2.5mg/L+NAA0.1mg/L, cultivation temperature is 24 ± 2 ℃, intensity of illumination 2000～3000Lx, light application time 12～14 hours/day.
With the Cheng Miao of described subculture after strong sprout regeneration induction lepisphere on regeneration induction lepisphere and root media 1/2MS+6BA 0mg/L+NAA 0.5mg/L, condition of culture is 24 ± 2 ℃ of temperature, intensity of illumination 2000～3000Lx, light application time 12～14 hours/day, growing can obtain detoxification edible lily group in 35 days and cultivate ball.
Detoxification edible lily group is cultivated the ball scale plant in starting medium MS+6BA 0.5mg/L+NAA 0.5mg/L and go up evoking adventive bud, condition of culture is to be 24 ± 2 ℃ in the light cultivation temperature, intensity of illumination 2000～3000Lx, light application time 12～14 hours/day.But indefinite bud is through subculture Cheng Miaohou regeneration induction lepisphere again, and so alternate cycles can expand numerous in enormous quantities.
Claims (1)
1. the detoxifying fast breeding method of an edible lily, comprise in regular turn: get the edible lily stem apex and obtain the detoxification tissue-cultured derived plant lily through starting, subculture obtains detoxification Cheng Miao strong sprout, detects to after virus-free, and the Cheng Miao that subculture was obtained after strong sprout is used for the regeneration induction lepisphere, regeneration lepisphere sheet induced bundle is sprouted or indefinite bud, bud or the indefinite bud subculture regeneration induction lepisphere again after strong sprout of growing thickly, regeneration lepisphere sheet induced bundle is again sprouted or indefinite bud, so alternate cycles, carrying out batch expands numerous
It is characterized in that:
Described edible lily stem apex comprises through starting acquisition detoxification tissue-cultured derived plant lily incubation step:
The first step is got edible lily kind ball warp and is crossed husky training 7～10 days, peels off outer scale, behind the cleaning and sterilizing, shells growing point under anatomical lens, cut open get 0.1～0.2mm size stem apex as expanding numerous starting material;
Second step, stem apex is seeded in and starts medium MS+6BA 0.5～2.5mg/L+NAA0.1～last cultivation of 0.5mg/L, secretly cultivates and changes light cultivation acquisition detoxification tissue-cultured derived plant lily, 24 ± 2 ℃ of cultivation temperature after 3～4 days, intensity of illumination 2000～3000Lx that light is cultivated, light application time 12～14 hours/day;
Described subculture strong sprout: be with detoxification tissue-cultured derived plant lily subculture strong sprout on subculture strong seedling culture base MS+6BA0.8～2.5mg/L+NAA 0.1～0.5mg/L, cultivation temperature is 24 ± 2 ℃, intensity of illumination 2000～3000Lx, light application time 12～14 hours/day obtains detoxification Cheng Miao; With described one-tenth seedling regeneration induction lepisphere, detoxification Cheng Miao is the regeneration induction lepisphere on regeneration induction lepisphere and root media 1/2MS+NAA 0.1～0.5mg/L, condition of culture is 24 ± 2 ℃ of temperature, intensity of illumination 2000～3000Lx, light application time 12～14 hours/day, 30～60 days acquisition detoxification edible lily groups of growing are cultivated ball;
Described regeneration lepisphere sheet induced bundle is sprouted or indefinite bud, be the edible lily group of detoxification to be cultivated the ball scale plant in starting on the medium induced bundle and sprout or indefinite bud, 24 ± 2 ℃ of cultivation temperature, intensity of illumination 2000～3000Lx, light application time 12～14 hours/day.
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| CN102771391B (en) * | 2012-07-17 | 2013-09-11 | 安徽霍山鹏飞现代农业科技有限公司 | Forcing culture technique of virus-free lily by industrial tissue culture and low-temperature bulb treatment |
| CN103733994B (en) * | 2013-12-18 | 2016-01-06 | 浙江大学 | The method in a kind of oriental hybrid lily plantlet in vitro strong sprout |
| CN109673519B (en) * | 2019-03-08 | 2020-12-15 | 长江师范学院 | Method for virus-free rapid propagation of lily bulbels |
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| CN1903017A (en) * | 2005-07-29 | 2007-01-31 | 潘利军 | Method for separating bulbs of lily tissue culture |
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| CN1903017A (en) * | 2005-07-29 | 2007-01-31 | 潘利军 | Method for separating bulbs of lily tissue culture |
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