CN103314846A - Adventitious bud induction method for amygdalus pedunculata pall explants - Google Patents
Adventitious bud induction method for amygdalus pedunculata pall explants Download PDFInfo
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- CN103314846A CN103314846A CN2013100942732A CN201310094273A CN103314846A CN 103314846 A CN103314846 A CN 103314846A CN 2013100942732 A CN2013100942732 A CN 2013100942732A CN 201310094273 A CN201310094273 A CN 201310094273A CN 103314846 A CN103314846 A CN 103314846A
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Abstract
The invention discloses a method for increasing adventitious bud induction number of explants by using potassium ion. On the basis of the conventional tissue culture of amygdalus pedunculata pall, by adding potassium ion, the adventitious bud induction number of each explant of amygdalus pedunculata pall is increased substantially. The method of the invention is simple and practicable, high in efficiency, and usable for rapid artificial breeding of amygdalus pedunculata pall.
Description
Technical field
The present invention relates to a kind of Amygdalus pedunculata explant adventitious bud inducing method, belong to field of plant tissue culture technique.
Background technology
Amygdalus pedunculata (
Amygdalus pedunculataPall.) be the rose family (Rosaceae) peach belong to (
Amygdalus) the almond subgenus (
Amygdalus) machaka, have another name called wild cherry, handle almond, Nanking cherry, be the distinctive Amygdalus wild species of China.The good characteristics such as Amygdalus pedunculata has wide accommodation, drought resisting, barren-resistant, weather-proof, the ability of fixing the sand is strong, cultivating existing ecological benefits at Arid desert region has again certain economic benefit.The Amygdalus pedunculata kernel contains crude fat 531g/kg, crude protein 255.9g/kg, and iron 39.88mg/kg, zinc 42.16mg/kg, calcium 0.224mg/kg is of high nutritive value.Contain in the fruit to cause and rush down composition brush-cherry seed glucoside (Amygdaluside, 3-rhamnoglucosylkaempferol), can be widely used in medical treatment, health care aspect, tracheitis, hypertension, neurasthenia, pneumonia, diabetes are had certain curative effect, also can be used to auxiliary for treating cancer; Amygdalus pedunculata benevolence also can be made into sedative and anodyne; Be suitable for processing health products, its oil content can be made industrial raw material up to 45.19%～52.00%, is a kind of bioenergy plant of great exploitation potential for its.But owing to naturally and artificially destroying, the distribution area of Amygdalus pedunculata reduces increasingly, has been listed in the secondary endangered plants.Need development Amygdalus pedunculata tissue rapid propagation technology in the production badly, and technique also is in the exploratory stage at present.
The modes of reproduction of Amygdalus pedunculata is still take sowing, cuttage as main at present.Breeding cycle of growing directly from seeds is long, and technical difficulty is little, but because Amygdalus pedunculata is cross pollination, the seedling interindividual variation is large, is difficult to the economic character that keeps good.And long-term cottage propagation easily causes the accumulation of fungi and virus, causes deterioration of variety, and perfect tissue culture technique can become the important means of Amygdalus pedunculata rearing new variety and Mechanism of Physiological and Biochemical research.The Amygdalus pedunculata tissue is cultivated not only can Fast-propagation Amygdalus pedunculata good seed, is used for the production of large tracts of land artificial cultivation, meets the need of market, and also sets up good basis for next step screens choiceness.
Summary of the invention
The object of the present invention is to provide a kind of method of utilizing potassium ion to improve adventitious bud inducing quantity on the Amygdalus pedunculata explant.
To achieve these goals, the present invention takes following technical solution:
A kind of Amygdalus pedunculata explant adventitious bud inducing method comprises cultivation and the indefinite spore induction of Amygdalus pedunculata of Amygdalus pedunculata seed seedling, adds the formation of soluble potassium salt evoking adventive bud in Amygdalus pedunculata adventitious bud induction culture base.
OnStating the adventitious bud induction culture base is to add soluble potassium salt, 6-benzyl aminoadenine and methyl α-naphthyl acetate in the MS minimal medium, and wherein potassium concentration is 1000～1600mg/L, and 6-benzyl aminoadenine concentration is 2.5～15 μ M, and concentration of NAA is 0～5 μ M.Preferred condition is: potassium concentration is 1000～1400mg/L, and 6-benzyl aminoadenine concentration is 5～10 μ M, and concentration of NAA is 0.5～2.5 μ M.Described soluble potassium salt is selected from potassium nitrate, potassium chloride, potassium dihydrogen phosphate.
The present invention choose Amygdalus pedunculata [
Amygdalus pedunculataPall.] seed is initial material, under aseptic condition, sprout and generate the seed seedling, explant take the joint (axillalry bud) of seed seedling as adventitious bud inducing, by improving the potassium ion consumption of medium, can significantly improve the indefinite spore induction quantity of Amygdalus pedunculata, and can not increase the brownization degree of indefinite bud.It is simple to operate, is easy to improve the stripped indefinite spore induction efficient of Amygdalus pedunculata, for the Fast-propagation that manually carries out on a large scale Amygdalus pedunculata provides important technical foundation.
Description of drawings
Fig. 1 is the 6-BA that adds 5.0 μ M in the MS minimal medium, the NAA of 5.0 μ M;
Fig. 2 adds 2200mg/L K in the MS minimal medium
+(with 6.04g/L KNO
3Form add), the 6-BA of 5 μ M, the NAA of 2.0 μ M;
Fig. 3 is the 6-BA that adds 10 μ M in the MS minimal medium, the NAA of 2.5 μ M;
Fig. 4 adds 1200mg/L K in the MS minimal medium
+(with 3.29g/L KNO
3Form add), the 6-BA of 10 μ M, the NAA of 2.5 μ M;
Fig. 5 adds 1200mg/L K in the MS minimal medium
+The 6-BA of (form with 0.29g/L KCl adds), 10 μ M, the NAA of 2.5 μ M;
Fig. 6 adds 2200mg/L K in the MS minimal medium
+(with 7.68g/L KH
2PO
4Form add), the 6-BA of 10 μ M, the NAA of 2.5 μ M.
Embodiment
Specifically, the present invention utilizes the method for adventitious bud inducing quantity on the potassium ion raising Amygdalus pedunculata explant to comprise the following steps:
(1) conventional Aseptic seedling culture is adopted in the cultivation of Amygdalus pedunculata seed seedling
Amygdalus pedunculata [
Amygdalus pedunculataPall.] seed is rinsed well through running water, seed is immersed mass fraction in superclean bench and be 60s in 70% the ethanolic solution, then places the mercuric chloride (HgCl of concentration 0.1%
2) soak 8min in the solution, with aseptic water washing 3 times.Blot with the moisture of the filter paper that passes through sterilization with the surface of the seed, then plant in the MS solid culture medium and cultivate, the intensity of illumination between culture period is 4000Lx, and the photoperiod is 16h, and temperature is 25 ± 0.5 ℃.
(2) the indefinite spore induction of Amygdalus pedunculata
Get the Amygdalus pedunculata seed seedling that grows to 5cm, cut the axillalry bud of Amygdalus pedunculata with the scalpel through sterilization, then the axillalry bud of Amygdalus pedunculata is inoculated in the adventitious bud induction culture base and cultivates, intensity of illumination between culture period is 4000Lx, photoperiod is 16h, temperature is 25 ± 0.5 ℃, and incubation time is 35 days.
Consisting of of described adventitious bud induction culture base: add soluble potassium salt, 6-benzyl aminoadenine and methyl α-naphthyl acetate in the MS minimal medium, wherein potassium concentration is 1000～1600mg/L, 6-benzyl aminoadenine (6-BA) concentration is 2.5～15 μ M, and methyl α-naphthyl acetate (NAA) concentration is 0～5 μ M.Preferred mode is that potassium concentration is 1000～1400mg/L, and 6-benzyl aminoadenine (6-BA) concentration is 5～10 μ M, and methyl α-naphthyl acetate (NAA) concentration is 0.5～2.5 μ M.
The inventor studies show that, in the Amygdalus pedunculata tissue culture procedures, indefinite spore induction is a restrictive factor.The applicant is in screening Amygdalus pedunculata tissue is cultivated, additive capacity to the potassium ion in the medium in the Induction Process of indefinite bud carries out deep research, the potassium ion consumption of finding suitable raising medium can improve the indefinite spore induction quantity of Amygdalus pedunculata significantly, and can not increase the vitrifying of indefinite bud and the degree of brownization.
The MS minimal medium forms: the NH of 1.65g/L
4NO
3, 1.9g/L KNO
3, 0.17g/L KH
2PO
4, 0.37g/L MgSO
47H
2The CaCl of O, 0.44g/L
22H
2The KI of O, 0.83mg/L, the H of 6.2mg/L
3BO
3, 22.3mg/L MnSO
44H
2The ZnSO of O, 8.6mg/L
47H
2The Na of O, 0.25mg/L
2MoO
42H
2The CuSO of O, 0.025mg/L
45H
2The CoCl of O, 0.025mg/L
26H
2The Na of O, 37.3mg/L
2The FeSO of EDTA, 27.8mg/L
47H
2O, inositol 100mg/L, glycine 2mg/L, thiamine hydrochloride 0.1mg/L, puridoxine hydrochloride 0.5mg/L, nicotinic acid 0.5mg/L, sucrose 30g/L.
It below is the specific embodiment that the inventor provides.
Embodiment 1
(1) cultivation of Amygdalus pedunculata seed seedling
The Amygdalus pedunculata seed is rinsed well through running water, seed is immersed mass fraction in superclean bench and be 60s in 70% the ethanolic solution, then places the mercuric chloride (HgCl of concentration 0.1%
2) soak 8min in the solution, use again aseptic water washing 3 times.Blot with the filter paper of the bacterium of the going out moisture with the surface of the seed, then plant in the MS solid culture medium and cultivate.Intensity of illumination between culture period is 4000Lx, and the photoperiod is 16h, and temperature is 25 ± 0.5 ℃.
(2) the indefinite spore induction of Amygdalus pedunculata
Get the Amygdalus pedunculata seed seedling that grows to 5 cm, cut the joint (axillalry bud) of Amygdalus pedunculata with the scalpel through sterilization, then will save (being the adventitious bud induction culture base) cultivation in the MS medium of NAA that (axillalry bud) be inoculated into the 6-BA that contains 5.0 μ M and 5.0 μ M.Intensity of illumination is 4000 Lx, and the photoperiod is 16 h, and temperature is 25 ± 0.5 ℃, incubation time 35 days, and after cultivation is finished, 1.6 of the quantity average out to (Fig. 1) of indefinite bud on the axillalry bud.
Embodiment 2
Similar to Example 1, different is that the adventitious bud induction culture base is to add the 6-BA of 15 μ M and the NAA of 2.0 μ M in the MS minimal medium.After cultivation is finished, 2.2 of the quantity average out to of indefinite bud on the axillalry bud.
Embodiment 3
Similar to Example 1, different is that the adventitious bud induction culture base is the 6-BA that adds 2.5 μ M in the MS minimal medium; After cultivation is finished, 0.8 of the quantity average out to of indefinite bud on the axillalry bud.
Embodiment 4
Similar to Example 1, different is that the adventitious bud induction culture base is the 6-BA that adds 10 μ M in the MS minimal medium; After cultivation is finished, 1.2 of the quantity average out to of indefinite bud on the axillalry bud.
Embodiment 5
Similar to Example 1, different is that the adventitious bud induction culture base is to add the 6-BA of 10 μ M and the NAA of 2.5 μ M in the MS minimal medium; After cultivation is finished, 2.3 of the quantity average out to (Fig. 3) of indefinite bud on the axillalry bud.
Embodiment 6
Similar to Example 1, different is that the adventitious bud induction culture base is KNO in the MS minimal medium
3Content (be equivalent to 500 mg/L K to 1.37g/L
+Dosage), the 6-BA of 10 μ M and the NAA of 2.5 μ M.After cultivation is finished, 1.5 of the quantity average out to of indefinite bud on the axillalry bud.
Embodiment 7
Similar to Example 1, different is that the adventitious bud induction culture base is to have added 1200mg/L K in the MS minimal medium
+(with 3.29g/L KNO
3Form add), the 6-BA of 10 μ M; After cultivation is finished, 1.8 of the quantity average out to of indefinite bud on the axillalry bud.
Embodiment 8
Similar to Example 1, different is that the adventitious bud induction culture base is the K that has added variable concentrations in the MS minimal medium
+, K
+With KNO
3Form add (seeing Table 1), the 6-BA of 10 μ M and the NAA of 2.5 μ M.After cultivation was finished, the par of indefinite bud saw Table 1 on the axillalry bud.
As seen from Table 1, K
+Concentration in 800～1600mg/L scope, can evoking adventive bud the increase of quantity, K
+Concentration is preferably in 1000～1400mg/L(Fig. 4), greater than 2200mg/L, brownization appears.
Embodiment 9
Similar to Example 1, that different is K
+It is the form adding with KCl.After cultivation was finished, the par of indefinite bud saw Table 2 on the axillalry bud.
As seen from Table 2, K
+Concentration in 800～1600mg/L scope, can evoking adventive bud the increase of quantity, K
+Concentration is preferably in 1000～1600mg/L(Fig. 5), greater than 2200mg/L, brownization appears.
Embodiment 10
Similar to Example 1, that different is K
+With KH
2PO
4Form add.After cultivation was finished, the par of indefinite bud saw Table 3 on the axillalry bud.
As seen from Table 3, K
+Concentration in 800～1600mg/L scope, can evoking adventive bud the increase of quantity, K
+Concentration is preferably in 1000～1600mg/L, greater than 2200mg/L, brownization (Fig. 6) occur.
Embodiment 11
Similar to Example 1, different is to add 2200mg/L K in the MS minimal medium
+(with 6.04g/L KNO
3Form add), the 6-BA of 5 μ M, the NAA of 2.0 μ M.After cultivation is finished, 1.2 of the quantity average out to of indefinite bud on the axillalry bud, brownization of indefinite bud serious (Fig. 2).
Claims (4)
1. Amygdalus pedunculata explant adventitious bud inducing method comprises it is characterized in that cultivation and the indefinite spore induction of Amygdalus pedunculata of Amygdalus pedunculata seed seedling: add the formation of soluble potassium salt evoking adventive bud in Amygdalus pedunculata adventitious bud induction culture base.
2. Amygdalus pedunculata explant adventitious bud inducing method according to claim 1, it is characterized in that: described adventitious bud induction culture base is to add soluble potassium salt, 6-benzyl aminoadenine and methyl α-naphthyl acetate in the MS minimal medium, wherein potassium concentration is 1000～1600mg/L, 6-benzyl aminoadenine concentration is 2.5～15 μ M, and concentration of NAA is 0～5 μ M.
3. Amygdalus pedunculata explant adventitious bud inducing method according to claim 2, it is characterized in that: potassium concentration is 1000～1400mg/L, and 6-benzyl aminoadenine concentration is 5～10 μ M, and concentration of NAA is 0.5～2.5 μ M.
4. according to claim 1 to 3 one of any described Amygdalus pedunculata explant adventitious bud inducing methods, it is characterized in that: described soluble potassium salt is selected from potassium nitrate, potassium chloride, potassium dihydrogen phosphate.
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|---|---|---|---|
| CN201310094273.2A CN103314846B (en) | 2013-03-22 | 2013-03-22 | Adventitious bud induction method for amygdalus pedunculata pall explants |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103907536A (en) * | 2014-04-16 | 2014-07-09 | 西北大学 | Method for shortening rooting time of amygdalus pedunculata pall test-tube plantlets |
| CN110063260A (en) * | 2019-05-31 | 2019-07-30 | 内蒙古农业大学 | A kind of Amygdalus pedunculata rapid propagation method |
| CN118177078A (en) * | 2024-03-21 | 2024-06-14 | 中国林业科学研究院经济林研究所 | Culture system and method for amygdalus pedunculata tissue culture |
| CN118177078B (en) * | 2024-03-21 | 2024-10-22 | 中国林业科学研究院经济林研究所 | Culture system and method for amygdalus pedunculata tissue culture |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114631534A (en) * | 2022-04-02 | 2022-06-17 | 安徽农业大学 | Exogenous mixture for improving yield of wheat infected by flowering period, application method and application thereof |
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| JPH0654632A (en) * | 1992-08-06 | 1994-03-01 | Hashimoto Corp | Plant belonging to new cultivar of purunus amygdalus and method for breeding thereof |
| CN101461319A (en) * | 2009-01-15 | 2009-06-24 | 西北农林科技大学 | Rapid seedling growing method for longstalck peaches |
-
2013
- 2013-03-22 CN CN201310094273.2A patent/CN103314846B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0654632A (en) * | 1992-08-06 | 1994-03-01 | Hashimoto Corp | Plant belonging to new cultivar of purunus amygdalus and method for breeding thereof |
| CN101461319A (en) * | 2009-01-15 | 2009-06-24 | 西北农林科技大学 | Rapid seedling growing method for longstalck peaches |
Non-Patent Citations (2)
| Title |
|---|
| BARBARA NOWAK ET.AL: "The effect of total inorganic nitrogen and the balance between its ionic forms on adventitious bud formation and callus growth of ‘We˛gierka Zwykła’ plum(Prunus domestica L.)", 《ACTA PHYSIOL PLANT》, vol. 29, 31 May 2007 (2007-05-31), pages 483 - 2 * |
| 吴恩岐等: "沙地植物柄扁桃的组织培养与植株再生", 《植物生理学通讯》, vol. 42, no. 6, 21 December 2006 (2006-12-21), pages 1132 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103907536A (en) * | 2014-04-16 | 2014-07-09 | 西北大学 | Method for shortening rooting time of amygdalus pedunculata pall test-tube plantlets |
| CN110063260A (en) * | 2019-05-31 | 2019-07-30 | 内蒙古农业大学 | A kind of Amygdalus pedunculata rapid propagation method |
| CN118177078A (en) * | 2024-03-21 | 2024-06-14 | 中国林业科学研究院经济林研究所 | Culture system and method for amygdalus pedunculata tissue culture |
| CN118177078B (en) * | 2024-03-21 | 2024-10-22 | 中国林业科学研究院经济林研究所 | Culture system and method for amygdalus pedunculata tissue culture |
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