CN103299905B - Strawberry subculture method - Google Patents
Strawberry subculture method Download PDFInfo
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- CN103299905B CN103299905B CN201310212115.2A CN201310212115A CN103299905B CN 103299905 B CN103299905 B CN 103299905B CN 201310212115 A CN201310212115 A CN 201310212115A CN 103299905 B CN103299905 B CN 103299905B
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Abstract
The invention discloses a strawberry subculture method. The method comprises the following steps: inoculating a strawberry explant subjected to embryogeny callus group induction culture onto a strawberry subculture medium, culturing for 5 days in a low-temperature artificial climate dark box of 2-3 DEG C at first, and then, carrying out dark culturing for 25-30 days under the conditions that the room temperature is 25+/-1 DEG C in the day and is 20+/-2 DEG C at night. Low temperature-room temperature culture conditions are adopted to benefit the somatic embryo maturation, the maturing rate of somatic embryos can be greatly improved, the differentiation rate of upper and lower hypocotyl embryoids is averagely increased by above 25%, and the matched callus can no longer be increased and turn light or get brown, so that a problem that a conventional subculture medium easily causes genotype obstacles to influence normal culture or cause yellow leaves and plant malformation is solved.
Description
Technical field
The present invention relates to a kind of cultural method, especially relate to a kind of strawberry subculture method, belong to phytobiology technical field.
Background technology
Strawberry obtains large-area popularizing planting with its outstanding economic worth and ecological functions in China.And in the face of need of production and existing working condition, strawberry all awaits further improvement in output, quality and resistance etc.Biotechnology being combined with conventional breeding methods and carry out New Strawberry Variety cultivation, is the effective way improving breeding strawberry efficiency.
Traditional strawberry subculture method is: the formula of the medium chosen is unfavorable for the squamous subculture of plant, component agar has the inherent shortcoming that cannot overcome in Plant Tissue Breeding, have impact on the whole structure of medium, and usually choose normal temperature or higher temperature due to condition of culture temperature, cause embryo subculture not easily ripe, extend and cultivate the time limit, reduce efficiency.
Summary of the invention
For solving the deficiencies in the prior art, the object of the present invention is to provide a kind of strawberry subculture method that effectively can improve strawberry reproduction speed.
For achieving the above object, the present invention is achieved by the following technical solutions:
A kind of strawberry subculture method, it is characterized in that, step is as follows: be inoculated in strawberry subculture medium by the strawberry explant after embryo generation callus group Fiber differentiation, first carry out cultivating in the 2-3 DEG C of low temperature artificial climate camera bellows of 5 days, then light culture 25-30 days under room temperature 25 ± 1 DEG C, the night condition of 20 ± 2 DEG C by day.
Further, described strawberry subculture medium comprises 1/2 improvement SH+2-4mg/L2, and 4-D+0.2-0.5mg/L6-BA+30g/L sucrose+3 ~ 4g/L phytagel, pH value is 5.6-5.8.
And 1/2 described improvement SH comprises grand nutrition element, micronutrient element and organic reagent.
Wherein, described grand nutrition element comprises:
Ammonium sulfate 450mg/L;
Potassium nitrate 2900 mg/L;
Epsom salt 200 mg/L;
Anhydrous potassium dihydrogenphosphate 450 mg/L;
Iron edta sodium salt 1.4 mg/L.
Described micronutrient element comprises:
Manganese sulfate monohydrate 12 mg/L;
Zinc sulphate 1.2 mg/L;
Boric acid 4.0 mg/L;
Potassium iodide 0.8 mg/L;
Sodium molybdate 0.2 mg/L;
Copper sulphate 0.1 mg/L;
Cobalt chloride 0.1 mg/L.
And described organic reagent comprises:
Thiamine hydrochloride 6 mg/L;
Nicotinic acid 2.2 mg/L;
Puridoxine hydrochloride 2.5 mg/L.
The invention has the beneficial effects as follows: the present invention is owing to have employed the condition of culture of low temperature-normal temperature, be conducive to the maturation of somatic embryo, the maturing rate of somatic embryo can be increased substantially, upper and lower plumular axis embryoid bodies differentiate rate is made on average to improve more than 25%, join callus also no longer increase shoal or occur brown, overcome traditional subculture medium and easily occur genotype barrier, just can not grow cultivation, or there is yellow leaf, the problem of plant deformityization.
Embodiment
Below in conjunction with specific embodiment, concrete introduction is carried out to the present invention.
embodiment 1
300 strawberry explants after embryo generation callus group Fiber differentiation are inoculated in strawberry subculture medium, first carry out cultivating in 2 DEG C of low temperature artificial climate camera bellows of 5 days, then light culture 25 days under room temperature 25 ± 1 DEG C, the night condition of 20 ± 2 DEG C by day, wherein, the formula of described strawberry subculture medium is: 1/2 improvement SH+2mg/L2,4-D+0.2mg/L6-BA+30g/L sucrose+4g/L phytagel, pH value is 5.6.
embodiment 2
300 strawberry explants after embryo generation callus group Fiber differentiation are inoculated in strawberry subculture medium, first carry out cultivating in 3 DEG C of low temperature artificial climate camera bellows of 5 days, then light culture 28 days under room temperature 25 ± 1 DEG C, the night condition of 20 ± 2 DEG C by day, wherein, the formula of described strawberry subculture medium is: 1/2 improvement SH+3mg/L2,4-D+0.3mg/L6-BA+30g/L sucrose+4g/L phytagel, pH value is 5.6.
embodiment 3
300 strawberry explants after embryo generation callus group Fiber differentiation are inoculated in strawberry subculture medium, first carry out cultivating in 2 DEG C of low temperature artificial climate camera bellows of 5 days, then light culture 30 days under room temperature 25 ± 1 DEG C, the night condition of 20 ± 2 DEG C by day, wherein, the formula of described strawberry subculture medium is: 1/2 improvement SH+4mg/L2,4-D+0.5mg/L6-BA+30g/L sucrose+3g/L phytagel, pH value is 5.7.
embodiment 4
300 strawberry explants after embryo generation callus group Fiber differentiation are inoculated in strawberry subculture medium, first carry out cultivating in 3 DEG C of low temperature artificial climate camera bellows of 5 days, then light culture 28 days under room temperature 25 ± 1 DEG C, the night condition of 20 ± 2 DEG C by day, wherein, the formula of described strawberry subculture medium is: 1/2 improvement SH+2.5mg/L2,4-D+0.4mg/L6-BA+30g/L sucrose+3.2g/L phytagel, pH value is 5.8.
embodiment 5
300 strawberry explants after embryo generation callus group Fiber differentiation are inoculated in strawberry subculture medium, first carry out cultivating in 2 DEG C of low temperature artificial climate camera bellows of 5 days, then light culture 30 days under room temperature 25 ± 1 DEG C, the night condition of 20 ± 2 DEG C by day, wherein, the formula of described strawberry subculture medium is: 1/2 improvement SH+3.4mg/L2,4-D+0.4mg/L6-BA+30g/L sucrose+3.5g/L phytagel, pH value is 5.8.
In above-described embodiment 1-5,1/2 improvement SH medium comprises grand nutrition element, micronutrient element and organic reagent.Wherein, described grand nutrition element comprises: ammonium sulfate 450mg/L; Potassium nitrate 2900 mg/L; Epsom salt 200 mg/L; Anhydrous potassium dihydrogenphosphate 450 mg/L; Iron edta sodium salt 1.4 mg/L.Described micronutrient element comprises: manganese sulfate monohydrate 12 mg/L; Zinc sulphate 1.2 mg/L; Boric acid 4.0 mg/L; Potassium iodide 0.8 mg/L; Sodium molybdate 0.2 mg/L; Copper sulphate 0.1 mg/L; Cobalt chloride 0.1 mg/L.And described organic reagent comprises: thiamine hydrochloride 6 mg/L; Nicotinic acid 2.2 mg/L; Puridoxine hydrochloride 2.5 mg/L.
Cultivation results is: after embryo subculture is cultivated and terminated, under above-mentioned condition of culture, owing to have employed the condition of culture of low temperature-normal temperature, be conducive to the maturation of somatic embryo, the maturing rate of somatic embryo can be increased substantially, upper and lower plumular axis embryoid bodies differentiate rate is made on average to improve more than 25%, join callus also no longer increase shoal or occur brown, overcome traditional subculture medium and easily occur genotype barrier, just can not grow cultivation, or there is yellow leaf, the problem of plant deformityization.
Therefore can drawing the supplementary measures such as the composition by adjusting condition of culture and improvement minimal medium, effectively overcoming the genotype barrier problem in strawberry squamous subculture, increasing substantially the conclusion of somatic embryo occurrence frequency and regeneration frequencies.
Be illustrated according to above-described embodiment, should be appreciated that above-described embodiment does not limit the present invention in any form, all employings are equal to replacement or the technical scheme that obtains of equivalent transformation mode, all drop within protection scope of the present invention.
Claims (1)
1. a strawberry subculture method, it is characterized in that, step is as follows: be inoculated in strawberry subculture medium by the strawberry explant after embryo generation induction of callus, first carry out cultivating in the 2-3 DEG C of low temperature artificial climate camera bellows of 5 days, then light culture 25-30 days under room temperature 25 ± 1 DEG C, the night condition of 20 ± 2 DEG C by day, described strawberry subculture medium is by 1/2 improvement SH+2-4mg/L2,4-D+0.2-0.5mg/L6-BA+30g/L sucrose+3 ~ 4g/L phytagel forms, and pH value is 5.6-5.8;
And 1/2 described improvement SH is made up of grand nutrition element, micronutrient element and organic reagent;
Described grand nutrition element is composed of the following components:
Ammonium sulfate 450mg/L,
Potassium nitrate 2900 mg/L,
Epsom salt 200 mg/L,
Anhydrous potassium dihydrogenphosphate 450 mg/L,
Iron edta sodium salt 1.4 mg/L;
Described micronutrient element is composed of the following components:
Manganese sulfate monohydrate 12 mg/L,
Zinc sulphate 1.2 mg/L,
Boric acid 4.0 mg/L,
Potassium iodide 0.8 mg/L,
Sodium molybdate 0.2 mg/L,
Copper sulphate 0.1 mg/L,
Cobalt chloride 0.1 mg/L;
Described organic reagent is composed of the following components:
Thiamine hydrochloride 6 mg/L,
Nicotinic acid 2.2 mg/L,
Puridoxine hydrochloride 2.5 mg/L.
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| CN105010123B (en) * | 2014-04-28 | 2017-06-09 | 上海市农业科学院 | The method and culture medium of strawberry distant hybrid are obtained by rescue isolated culture |
| CN105409777A (en) * | 2015-12-16 | 2016-03-23 | 青岛百瑞吉生物工程有限公司 | Growth and differentiation culture medium for somatic embryo of pear tree |
| CN105961201A (en) * | 2016-05-16 | 2016-09-28 | 玉溪市六合农业科技发展有限公司 | Tissue culture method for strawberry transplant seedlings |
Citations (2)
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| CN101049090A (en) * | 2007-05-22 | 2007-10-10 | 南京农业大学 | Method for carrying out taking off poison and quick breeding by using strawberry anther |
| CN102668987A (en) * | 2012-05-31 | 2012-09-19 | 句容市白兔镇云兔草莓专业合作社 | Multiplying culture media for strawberry |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101049090A (en) * | 2007-05-22 | 2007-10-10 | 南京农业大学 | Method for carrying out taking off poison and quick breeding by using strawberry anther |
| CN102668987A (en) * | 2012-05-31 | 2012-09-19 | 句容市白兔镇云兔草莓专业合作社 | Multiplying culture media for strawberry |
Non-Patent Citations (6)
| Title |
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| APLICACION DE TECNICAS BIOTECNOLOGICAS EN FRUTALES, UNA VIA VALIOSA PARA EL RESCATE Y LA CONSERVACION DE ESTAS ESPECIES;Argelys Kessel;《Cultivos Tropicales》;20081231;第29卷(第3期);27-37 * |
| Dayuan Wang等.Somatic Embryogenesis and Plant Regeneration from Immature Embryos of Strawberry.《HrotScience》.1984,第19卷(第1期),71-72. * |
| 李卫东等.草莓花药愈伤组织类型与状态调控研究.《河北农业大学学报》.2004,第27卷(第2期),59-63. * |
| 李青等.草莓人工种子培育的研究.《果树科学》.1995,(第12期),133-135. * |
| 胡忠等.影响宁夏枸杞愈伤组织体细胞胚发生的因素.《汕头大学学报》.2006,第21卷(第4期),57-63. * |
| 草莓花药愈伤组织继代培养和不定芽发生研究;吴伟民等;《江苏农业研究》;19991231;第20卷(第4期);19-23 * |
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