CN1903018A - Induction bulb forming method for lily tissue culture - Google Patents
Induction bulb forming method for lily tissue culture Download PDFInfo
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- CN1903018A CN1903018A CN 200510046957 CN200510046957A CN1903018A CN 1903018 A CN1903018 A CN 1903018A CN 200510046957 CN200510046957 CN 200510046957 CN 200510046957 A CN200510046957 A CN 200510046957A CN 1903018 A CN1903018 A CN 1903018A
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- lily
- tissue culture
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Abstract
A tissue culture method for inducing bulb of lily features that the tissue-cultured seedlings of lily 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 lily tissue culture is induced the method for balling-up.
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.Now all lily tissue culture finally all are to carry out transfer with the lily seedling of taking root, and this method training tissue culture seedling difficulty, transfer survival rate are lower.
Summary of the invention
The object of the present invention is to provide a kind of lily tissue culture to induce the method for balling-up; Lily seedling stalwartness, transfer survival rate height, shortening form the time of commodity ball after the balling-up.
For achieving the above object, the technical solution used in the present invention is:
The lily tissue culture is induced the method for balling-up, cultivate inducing the numerous tissue-cultured derived plant lily of expansion to be inoculated in the group training medium (as: MS medium), sugar and/or concentration of sucrose are 60～120g/l in the medium, and hormone BA concentration is 0.1～0.5mg/l, and NAA concentration is 0.01～0.2mg/l.
Described cultivation is in 2000～3000LX, the illumination of 12～14 hours every days, cultivates under 20～26 ℃ the condition; The preferred concentration of sugar and/or sucrose is 75～105g/l in the medium, is preferably 90g/l; The preferable concentration of BA is 0.2～0.3mg/l, is preferably 0.2mg/l; The preferable concentration of NAA is 0.05～0.15mg/l, is preferably 0.1mg/l.
The present invention has following advantage:
1. the present invention continues the seedling of taking root of lily to cultivate balling-up, carries out transfer with the tissue cultivating seedling after the balling-up, tissue cultivating seedling stalwartness during transfer, and survival rate can reach more than 95%.
2. the present invention adopts improved MS medium, and tissue-cultured derived plant lily balling-up speed is fast, and diameter is big, and growing way is good.
Embodiment
Embodiment 1 lily tissue culture is induced balling-up:
1) induces the numerous tissue-cultured derived plant lily of expansion: when lily bud length 5～10cm, get 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; Indefinite bud is repeated to change in the above-mentioned medium, and per 25～35 days (present embodiment is 30 days) transfers once, forms the seedling of the about 2～3cm of many height;
2) tissue cultivating seedling balling-up: the tissue-cultured derived plant lily (seedling) that will expand in numerous is split into individual plant, be inoculated in the following medium: MS+BA 0.1～0.5+NAA 0.01～0.2+ sucrose 60～120g/l, 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, cultivation through 55～65 days (present embodiment is 60 days), tissue cultivating seedling just grows up to the bulblet of lilium about the about 1.3cm of diameter, carry out transfer then, survival rate can reach more than 96%;
3) observe its different hormone concentration and media and different sucrose inducing action effect to bulblet of lilium:
The tissue-cultured derived plant lily that expands in numerous is split into individual plant, is inoculated in the improved MS medium of following experiment, medium is: MS+BA+NAA+ sucrose or sugar; After 60 days cultivation, its growing way situation such as following table 1-4.
A. when BA and one timing (BA 0.2mg/l+NAA 0.1mg/l) of NAA concentration, sugared content the inducing of variable concentrations to lily bulb.(following sugar or cane sugar content are meant the total content of sugar in the medium or sucrose)
Table 1
| Sugar concentration (g/l) | 45 | 60 | 75 | 90 | 105 | 120 |
| Bulb diameter size (mm) | 3.3 | 8.5 | 13.3 | 13.9 | 11.2 | 7.3 |
As can be seen from the above table, when BA and NAA concentration and other conditions were constant, along with the concentration of sugar increases, bulb diameter also constantly increased, when sugared concentration 90g/l, the maximum average out to 13.9mm of bulb diameter, be higher than 90g/l after, bulb diameter begins to descend gradually, so the best sugared concentration of inducing balling-up is 90g/l, induce the diameter maximum, growing way is normal, and the transfer survival rate is the highest.
B. when BA and one timing (BA 0.2mg/l+NAA 0.1mg/l) of NAA concentration, the cane sugar content of variable concentrations is induced lily bulb.
Table 2
| Sucrose concentration (g/l) | 45 | 60 | 75 | 90 | 105 | 120 |
| Bulb diameter size (mm) | 3.3 | 8.5 | 13.3 | 13.9 | 11.2 | 7.3 |
As can be seen from the above table: when BA and NAA concentration and other conditions were constant, along with the concentration of sugar increases, bulb diameter also constantly increased, when sucrose concentration 90g/l, the maximum average out to 13.9mm of bulb diameter, be higher than 90g/l after, bulb diameter begins to descend gradually.
C. when sucrose concentration 90g/l, BA are 0.2mg/l, NAA content the inducing of variable concentrations to lily bulb.
Table 3
| NAA concentration (mg/) | 0.01 | 0.05 | 0.1 | 0.15 | 0.2 | 0.25 |
| Bulb diameter size (mm) | 5.3 | 11.9 | 13.8 | 12.0 | 9.4 | 6.1 |
As can be seen from the above table: when BA and sugared concentration and other conditions were constant, NAA concentration is bigger, the best then genus NAA 0.1mg/l of lepisphere diameter between 0.05～0.15 the time.
D. when sucrose concentration 90g/l, NAA are 0.1mg/l, BA content the inducing of variable concentrations to lily bulb.
Table 4
| BA concentration (mg/) | 0.1 | 0.15 | 0.2 | 0.3 | 0.4 | 0.5 | 0.6 |
| Bulb diameter size (mm) | 9.8 | 11.3 | 13.9 | 10.9 | 8.4 | 6.3 | 3.2 |
As can be seen from the above table: when NAA and sugared concentration and other conditions were constant, BA concentration is bigger, the best then genus BA0.2mg/l of lepisphere diameter between 0.15～0.3 the time.
From table 1-4 as can be seen, inducing in the process of balling-up in the lily tissue culture, mainly is that the influence of sucrose or sugared concentration and hormone concentration and media is bigger.Its best lily induces the balling-up medium to be: MS+BA 0.2mg/l+NAA 0.1mg/l+ sucrose 90g/l, and average diameter reaches 13.9mm, and growing way is normal.
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 the concentration for sugar, BA and NAA of outstanding inducing action in the present invention, and need not strict the qualification for each component consumption in the improved MS medium, can adopt shown in the improved MS table 5 composed as follows in of the present invention group of training, 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 5
| 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 lily tissue culture is induced the method for balling-up, it is characterized in that: will induce the numerous tissue-cultured derived plant lily of expansion to be inoculated in the group training medium and cultivate, sugar and/or concentration of sucrose are 60～120g/l in the medium, hormone BA concentration is 0.1～0.5mg/l, NAA concentration is 0.01～0.2mg/l, and tissue cultivating seedling grows up to bulblet of lilium can carry out transfer.
2. induce the method for balling-up according to the described lily tissue culture of claim 1, 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 55～65 days cultivation, tissue cultivating seedling grows up to bulblet of lilium can carry out transfer.
3. induce the method for balling-up according to the described lily tissue culture of claim 1, it is characterized in that: sugar and/or concentration of sucrose are 75～105g/l in the described medium, and BA concentration is 0.2～0.3mg/l, and NAA concentration is 0.05～0.15mg/l.
4. induce the method for balling-up according to the described lily tissue culture of claim 1, it is characterized in that: sugar and/or concentration of sucrose are 90g/l in the described medium, and BA concentration is 0.2mg/l, and NAA concentration is 0.1mg/l.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200510046957 CN1903018A (en) | 2005-07-29 | 2005-07-29 | Induction bulb forming method for lily tissue culture |
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|---|---|---|---|
| CN 200510046957 CN1903018A (en) | 2005-07-29 | 2005-07-29 | Induction bulb forming method for lily tissue culture |
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|---|---|---|---|
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Cited By (3)
| 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 |
| CN102771391A (en) * | 2012-07-17 | 2012-11-14 | 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 |
| CN103238610A (en) * | 2013-04-23 | 2013-08-14 | 内蒙古农业大学 | Chemical composition for promoting gladiolus corm to quickly grow as well as preparation method and application of chemical composition |
-
2005
- 2005-07-29 CN CN 200510046957 patent/CN1903018A/en active Pending
Cited By (4)
| 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 |
| CN102771391A (en) * | 2012-07-17 | 2012-11-14 | 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 |
| CN103238610A (en) * | 2013-04-23 | 2013-08-14 | 内蒙古农业大学 | Chemical composition for promoting gladiolus corm to quickly grow as well as preparation method and application of chemical composition |
| CN103238610B (en) * | 2013-04-23 | 2015-03-25 | 内蒙古农业大学 | Chemical composition for promoting gladiolus corm to quickly grow as well as preparation method and application of chemical composition |
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Open date: 20070131 |