CN1903019A - Induction buds sprouting method for lily tissue culture seedling - Google Patents
Induction buds sprouting method for lily tissue culture seedling Download PDFInfo
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- CN1903019A CN1903019A CN 200510046958 CN200510046958A CN1903019A CN 1903019 A CN1903019 A CN 1903019A CN 200510046958 CN200510046958 CN 200510046958 CN 200510046958 A CN200510046958 A CN 200510046958A CN 1903019 A CN1903019 A CN 1903019A
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- lily
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Abstract
A tissue culture method for inducing buds of lily features that the tissue on the stem tip of sterilized lily plant are cultured in the culture medium containing sugar and/or cane sugar, hormone BA and NAA proportionally to grow bulbs.
Description
Technical field
The present invention relates to Plant Tissue Breeding, specifically the method for lily tissue culture induction buds sprouting.
Background technology
Plant Tissue Breeding is meant under the condition that exsomatizes, and with the part of plant corpus, as a bit of stem, a fritter leaf even a cell, is seeded on the medium, makes them form the method for a whole plant again.The step of tissue culture have the making of medium and inoculation material sterilization, inoculation, plant induction, take root and transplant.
Wherein Chang Yong medium is the MS medium, and its prescription is:
| Composition | Working concentration (mg/L) | |
| Macroelement | Potassium nitrate KNO 3 | 1900 |
| Ammonium nitrate NH 4NO 3 | 1650 | |
| Potassium dihydrogen phosphate KH 2PO 4 | 170 | |
| Magnesium sulfate MgSO 4·7H 2O | 370 | |
| Calcium chloride CaCl 2·2H 2O | 440 | |
| Trace element | Potassium iodide KI | 0.83 |
| Boric acid H 3BO 3 | 6.2 | |
| Manganese sulphate MnSO 4·4H 2O | 22.3 | |
| Zinc sulphate ZnSO 4·7H 2O | 8.6 | |
| Sodium molybdate Na 2MoO 4·2H 2O | 0.25 | |
| Copper sulphate CuSO 4·5H 2O | 0.025 | |
| Cobalt chloride CoCl 2 | 0.025 | |
| Molysite | Disodium ethylene diamine tetraacetate Na 2·EDTA | 37.3 |
| Ferrous sulfate FeSO 2·7H2O | 27.8 | |
| Organic principle | Inositol | 100 |
| Glycine | 2 | |
| Thiamine hydrochloride VB1 | 0.1 | |
| Puridoxine hydrochloride VB6 | 0.5 | |
| Nicotinic acid VB5 or VPP | 0.5 | |
| Sucrose sucrose | 30g/L | |
| Agar agar | 7g/L |
Lily flower is by the asexual process breeding, and virus is transmitted accumulation by generation, and harm is on the rise.Separate the growing point of flower plant 0.1-0.5 millimeter size, what cultivation obtained is virus-free seedling basically; After the virus removal, the plant strain growth gesture is strong, and it is big that flower becomes, lovely luster, and anti-adversity ability improves, and produces flower quantity and rises.This technology will be widely used.Breeding expansion numerous in the present lily tissue culture all is to transfer to the cutting and separating of grow thickly bud or callus to carry out successive transfer culture on the new medium, accelerates; But existing tissue-cultured derived plant lily is induced and is all adopted above-mentioned common MS medium in the differentiation, wherein hormone BA concentration is 1.0mg/l, growth hormone NAA concentration is 0.1mg/l, the inductivity of its bud is generally 76%, inductivity is lower, and the numerous coefficient of expansion of its differentiation back bud is not higher than 4 usually yet, and it is not high to expand numerous coefficient.
Summary of the invention
The method that the object of the present invention is to provide a kind of inductivity height, expands the high lily tissue culture induction buds sprouting of numerous coefficient.
For achieving the above object, the technical solution used in the present invention is:
The method of lily tissue culture induction buds sprouting, with the lily plant through behind the sterilization, get stem-tip tissue be inoculated in group training medium (as; The MS medium) cultivate in, hormone BA concentration is 0.4～1.5mg/l in the medium, and NAA concentration is 0.1～0.5mg/l, and stem-tip tissue directly differentiation sprouts.
Described cultivation is in 2000～3000LX, the illumination of 12～14 hours every days, cultivates under 20～26 ℃ the condition, through 25～40 days cultivation; BA concentration is preferably 0.4～0.6mg/l in the medium, is preferably 0.5mg/l; NAA concentration is preferably 0.3～0.5mg/l, is preferably 0.5mg/l.
Advantage of the present invention is: adopt improved MS medium, tissue-cultured derived plant lily induces differentiation speed fast, has shortened the time of lily formation commodity balls; Inductivity height (can reach 92.7%), and numerous coefficient of expansion of its differentiation back bud is up to 6.2, the bud growing way is good, helps lily carried out next step successive transfer culture operation.
Embodiment
The method of embodiment 1 lily tissue culture induction buds sprouting:
1) when lily bud length 5～10cm, gets its shoot apical meristem, be inoculated in the MS medium, be added into BA (6-benzylaminopurine) 0.5mg/l, NAA (methyl) 0.5mg/l and sucrose 30g/l and induce; Under the illumination condition of 2000～3000LX (present embodiment is 2000LX) 12～14 hours every days (present embodiment is 14 hours), under 20～26 ℃ the temperature condition, after 30～40 days (present embodiment is 30 days), directly induce the indefinite bud of many densifications, if the prolongation incubation time then directly forms the bud clump; To repeat to change over to after the indefinite bud cutting and separating in the above-mentioned medium, per 25～35 days (present embodiment is 30 days) transfers once, can breed in a large number, forms the seedling of the about 2～3cm of many height; The numerous coefficient of average expansion can reach 6.2.
2) observe the inducing action effect of its different hormone concentration and media to the lily bud:
The lily plant through behind the sterilization, is got stem-tip tissue and is inoculated in the improved MS medium of following experiment, and medium is: MS+BA+NAA; After 30 days cultivation, its growing way situation such as following table 1-2.
A. when BA was 0.5mg/l, the NAA content of variable concentrations was induced differentiation lily bud.
Table 1
| NAA concentration (mg/) | 0.1 | 0.3 | 0.4 | 0.45 | 0.5 |
| The quantity of bud | Few | Generally | More | Many | At most |
As can be seen from the above table: when BA concentration and other conditions are constant, all right, the best then genus NAA 0.5mg/l of quantity of differentiation of NAA concentration lily bud between 0.3～0.5 time; NAA concentration is big more, and the quantity of bud differentiation is many more.
B. when NAA was 0.5mg/l, the NAA content of variable concentrations was to the influence of differentiation lily bud.
Table 2
| BA concentration (mg/) | 0.4 | 0.45 | 0.5 | 0.6 | 1.0 |
| The quantity of bud | Generally | More | At most | More | Few |
As can be seen from the above table: when NAA concentration and other conditions are constant, all right, the best then genus BA 0.5mg/l of quantity of differentiation of BA concentration lily bud between 0.4～0.6 time; In the time of between 0.4～0.5, BA concentration is big more, and the quantity of bud differentiation is many more, and BA concentration surpasses at 0.5 o'clock, and BA concentration is big more, and the quantity of bud differentiation is few more.
From table 1-2 as can be seen, in the process of lily tissue culture induction buds sprouting, the influence of major hormone concentration proportioning is bigger.Its best lily induction buds sprouting medium is: MS+BA 0.5mg/l+NAA 0.5mg/l, inductivity are up to 92.7%, and the numerous coefficient of expansion of differentiation back bud is up to 6.2, and the bud growing way is good.
Because conventional MS medium and WPM medium all can be used in the lily tissue culture procedures, and the consumption of each component differs bigger in their composition, and play outstanding inducing action in the present invention be BA and NAA, and need not strict the qualification for each component consumption in the improved MS medium, can adopt in the tissue culture of the present invention shown in the improved MS table 3 composed as follows, wherein also should be added with potassium nitrate (KNO
3) 1000～1900mg/L, potassium dihydrogen phosphate (KH
2PO
4) 170-340mg/L, potassium iodide (KI) 0.75～0.83mg/L and cobalt chloride (CoCl
2) 0.025～0.5mg/L, or be added with Ca (NO
3)
24H
2O 350-556mg/L and K
2SO
4990～1450mg/L.
Table 3
| Composition | Working concentration (mg/L) | |
| Macroelement | Ammonium nitrate NH 4NO 3 | 400～1650 |
| Magnesium sulfate MgSO 4·7H 2O | 370 | |
| Calcium chloride CaCl 2·2H 2O | 96～440 | |
| Trace element | Boric acid H 3BO 3 | 6.2 |
| Manganese sulphate MnSO 4·4H 2O | 22.3～22.5 | |
| Zinc sulphate ZnSO 4·7H 2O | 8.6 | |
| Sodium molybdate Na 2MoO 4·2H 2O | 0.25 | |
| Copper sulphate CuSO 4·5H 2O | 0.025～0.25 | |
| Molysite | Disodium ethylene diamine tetraacetate Na 2·EDTA | 37.3 |
| Ferrous sulfate FeSO 2·7H2O | 37.3～37.5 | |
| Organic principle | Inositol | 100 |
| Glycine | 2 | |
| Thiamine hydrochloride VB1 | 0.1～1.0 | |
| Puridoxine hydrochloride VB6 | 0.5 | |
| Nicotinic acid VB5 or VPP | 0.5 | |
| Sucrose sucrose | 30g/L | |
| Agar agar | 7g/L |
Claims (4)
1. the method for lily tissue culture induction buds sprouting, it is characterized in that: the lily plant through behind the sterilization, is got stem-tip tissue and is inoculated in the group training medium and cultivates, and hormone BA concentration is 0.4～1.5mg/l in the medium, NAA concentration is 0.1～0.5mg/l, and stem-tip tissue directly differentiation sprouts.
2. according to the method for the described lily tissue culture of claim 1 induction buds sprouting, it is characterized in that: described cultivation is in 2000～3000LX, the illumination of 12～14 hours every days, cultivate under 20～26 ℃ the condition, through 25～40 days cultivation, stem-tip tissue directly differentiation sprouted.
3. according to the method for the described lily tissue culture of claim 1 induction buds sprouting, it is characterized in that: BA concentration is 0.4～0.6mg/l in the described medium, and NAA concentration is 0.3～0.5mg/l.
4. according to the method for the described lily tissue culture of claim 1 induction buds sprouting, it is characterized in that: BA concentration is 0.5mg/l in the described medium, and NAA concentration is 0.5mg/l.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200510046958 CN1903019A (en) | 2005-07-29 | 2005-07-29 | Induction buds sprouting method for lily tissue culture seedling |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200510046958 CN1903019A (en) | 2005-07-29 | 2005-07-29 | Induction buds sprouting method for lily tissue culture seedling |
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| Publication Number | Publication Date |
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| CN1903019A true CN1903019A (en) | 2007-01-31 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN 200510046958 Pending CN1903019A (en) | 2005-07-29 | 2005-07-29 | Induction buds sprouting method for lily tissue culture seedling |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102030575A (en) * | 2010-11-15 | 2011-04-27 | 青海大学 | Oriental lily hybrid bulb propagation and cutting-flower cultivation nutrient fertilizer |
-
2005
- 2005-07-29 CN CN 200510046958 patent/CN1903019A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102030575A (en) * | 2010-11-15 | 2011-04-27 | 青海大学 | Oriental lily hybrid bulb propagation and cutting-flower cultivation nutrient fertilizer |
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Open date: 20070131 |