CN105993944A - Method for propagating indigowoad leaf by using large cherry rootstock - Google Patents
Method for propagating indigowoad leaf by using large cherry rootstock Download PDFInfo
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- CN105993944A CN105993944A CN201610340734.3A CN201610340734A CN105993944A CN 105993944 A CN105993944 A CN 105993944A CN 201610340734 A CN201610340734 A CN 201610340734A CN 105993944 A CN105993944 A CN 105993944A
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- folium isatidis
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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- Developmental Biology & Embryology (AREA)
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Abstract
The invention discloses a method for propagating an indigowoad leaf by using a large cherry rootstock, and belongs to the technical field of plant propagation. The method comprises the following five steps: selection treatment of bud stems, selection of a culture medium, inoculation, hardening and transplanting. MS is selected as a basic culture medium, and then explants are inoculated on the corresponding culture medium, and then transferred to a rooting culture medium to be cultured after initial culture and the subsequent multiplication culture; rooted test tube plantlets are transplanted to a mixed medium and then transplanted to a field nursery after greenhouse culture and open-air culture. The survival rate of the transplanted indigowoad leaf seedlings is high and up to over 90%.
Description
Technical field
The present invention relates to the propagation method of a kind of large cherry stock Folium Isatidis, belong to plant propagation skill field.
Background technology
The blade of large cherry stock Folium Isatidis is roomy dark green, well developed root system, and vertical root is more, is difficult to lodging, and winter resistance is strong, high anti-root knot.Folium Isatidis is cooked stock grafting large cherry, and survival rate is high, " bound feet " phenomenon seldom occurs, and sapling up-growth is healthy and strong, the best fruiting period high yield, stable yields, be one of the excellent stock of large cherry.But Folium Isatidis cuttage is difficult to take root, mound layering emergence rate is low, can not meet far away the demand in market.
Summary of the invention
It is an object of the invention to meet the market demand instantly, it is provided that the Folium Isatidis propagation method that a kind of emergence rate is high.
For the above-mentioned technical problem solved, the invention provides the propagation method of large cherry stock Folium Isatidis, comprise the following steps:
The first step, bud stem process: adopt high-quality Folium Isatidis branch, cleaning surface with detergent after removing all blades, tap water rinses and within 25 minutes, is placed on workbench, with 70% ~ 75% ethanol sterilizing 1 ~ 2 minute, soaking 5 ~ 10 minutes with 0.1% mercuric chloride, 10 rear cutout stem with bud of aseptic water washing are standby again；
Second step, culture medium select: choosing MS minimal medium, sucrose concentration takes 1 ~ 5%, and agar 0.5 ~ 0.8%, pH value is 6.0；Culturing room's temperature 25 DEG C, light intensity 1500 ~ 2000Lx, illumination every day 10 ~ 15 hours；
3rd step inoculation: outer implant is seeded in the culture medium of MS+6 BA 0.3 ~ 0.7mg/L+NAA 0.1 ~ 0.5 mg/L and carries out initial incubation, after 3 ~ 5 days, the sprout that initial incubation is induced is proceeded to MS+6 BA 0.3 ~ 0.6mg/L+NAA
Breeding in the culture medium of 0.15 ~ 0.25 mg/L, enrichment culture, after 2 ~ 3 weeks, takes 1.5 ~ 2.0cm young sprout and transfers to 1/2MS+NAA 0.4 ~ 0.7
On the root media of mg/L+15 ~ 25g sucrose, cultivate about 25 days；
4th step seedling exercising: take out the test tube Seedling taken root, cleans the culture medium adhered to, in transfer to mixed-matrix, the Small plastic shed seedling exercising being placed in greenhouse, and shed temperature controls at 21 DEG C ~ 23 DEG C, relative humidity transplant Initial stage of culture be 90% and more than；
After 5th step hot-house culture 30 ~ 45 days, outdoor cultivation 4 ~ 9 days, transplant in nursery, land for growing field crops.
As preferably, with 70% ethanol sterilizing 90 seconds when bud stem processes, then soak 8 minutes with 0.1% mercuric chloride.
As preferably, sucrose concentration takes 2%, and agar 0.6%, pH value is 6.0；Culturing room's temperature 25 DEG C, light intensity 1800Lx, illumination every day 12 hours.
As preferably, during inoculation, outer implant is seeded in the culture medium of MS+6 BA 0.5mg/L+NAA 0.2mg/L and carries out initial incubation, proceed to the sprout that initial incubation is induced after 4 days breed in the culture medium of MS+6 BA 0.5mg/L+NAA 0.2 mg/L, after enrichment culture 15 days, take 1.8cm young sprout and transfer on the root media of 1/2MS+NAA 0.5 mg/L+18g sucrose.
As preferably, during seedling exercising, shed temperature controls at 22 DEG C, and relative humidity is 91% transplanting Initial stage of culture.
As preferably, during transplanting after hot-house culture 40 days, outdoor cultivation 8 days.
The beneficial effects of the present invention is: the present invention is greatly improved Folium Isatidis and takes root efficiency, and mound layering emergence rate is high, gained Folium Isatidis transplantation of seedlings, up to more than 90%, to meet market cultivation needs.
Detailed description of the invention
Below in conjunction with embodiment, the propagation method of the present invention is described further.
Embodiment one
The first step, bud stem process: adopt high-quality Folium Isatidis branch, cleaning surface with detergent after removing all blades, tap water rinses and within 25 minutes, is placed on workbench, with 70% ethanol sterilizing 1 minute, soaking 5 minutes with 0.1% mercuric chloride, 10 rear cutout stem with bud of aseptic water washing are standby again；
Second step, culture medium select: choosing MS minimal medium, sucrose concentration takes 1%, and agar 0.5%, pH value is 6.0.Culturing room's temperature 25 DEG C, light intensity 1500Lx, illumination every day 10 hours；
3rd step, inoculation: outer implant is seeded in the culture medium of MS+6 BA 0.3mg/L+NAA 0.1 mg/L and carries out initial incubation, proceed to the sprout that initial incubation is induced after 3 days breed in the culture medium of MS+6 BA 0.3mg/L+NAA 0.15 mg/L, after enrichment culture 2 weeks, take 1.5cm young sprout and transfer on the root media of 1/2MS+NAA 0.4mg/L+15g sucrose, cultivate about 25 days；
4th step, seedling exercising: take out the test tube Seedling taken root, clean the culture medium adhered to, in transfer to mixed-matrix, the Small plastic shed seedling exercising being placed in greenhouse, and shed temperature controls at 21 DEG C, and relative humidity is 90% transplanting Initial stage of culture；
5th step, transplanting: after hot-house culture 30 days, outdoor cultivation 4 days, transplant in nursery, land for growing field crops.
Embodiment two
The first step, bud stem process: adopt high-quality Folium Isatidis branch, cleaning surface with detergent after removing all blades, tap water rinses and within 25 minutes, is placed on workbench, with 75% ethanol sterilizing 2 minutes, soaking 10 minutes with 0.1% mercuric chloride, 10 rear cutout stem with bud of aseptic water washing are standby again；
Second step, culture medium select: choosing MS minimal medium, sucrose concentration takes 5%, and agar 0.8%, pH value is 6.0.Culturing room's temperature 25 DEG C, light intensity 2000Lx, illumination every day 15 hours；
3rd step, inoculation: outer implant is seeded in the culture medium of MS+6 BA 0.7mg/L+NAA 0.5 mg/L and carries out initial incubation, proceed to the sprout that initial incubation is induced after 5 days breed in the culture medium of MS+6 BA 0.6mg/L+NAA0.25 mg/L, after enrichment culture 3 weeks, take 2.0cm young sprout and transfer on the root media of 1/2MS+NAA0.7 mg/L+25g sucrose, cultivate about 25 days；
4th step, seedling exercising: take out the test tube Seedling taken root, clean the culture medium adhered to, in transfer to mixed-matrix, the Small plastic shed seedling exercising being placed in greenhouse, and shed temperature controls at 23 DEG C, and relative humidity is 98% transplanting Initial stage of culture；
5th step, transplanting: after hot-house culture 45 days, outdoor cultivation 9 days, transplant in nursery, land for growing field crops.
Embodiment three
The first step, bud stem process: adopt high-quality Folium Isatidis branch, cleaning surface with detergent after removing all blades, tap water rinses and within 25 minutes, is placed on workbench, with 70% ethanol sterilizing 90 seconds, soaking 8 minutes with 0.1% mercuric chloride, 10 rear cutout stem with bud of aseptic water washing are standby again；
Second step, culture medium select: choosing MS minimal medium, sucrose concentration takes 2%, and agar 0.6%, pH value is 6.0.Culturing room's temperature 25 DEG C, light intensity 1800Lx, illumination every day 12 hours；
3rd step, inoculation: outer implant is seeded in the culture medium of MS+6 BA 0.5mg/L+NAA 0.2mg/L and carries out initial incubation, proceed to the sprout that initial incubation is induced after 4 days breed in the culture medium of MS+6 BA 0.5mg/L+NAA 0.2 mg/L, after enrichment culture 15 days, take 1.8cm young sprout and transfer on the root media of 1/2MS+NAA 0.5 mg/L+18g sucrose, cultivate about 25 days
4th step, seedling exercising: take out the test tube Seedling taken root, clean the culture medium adhered to, in transfer to mixed-matrix, the Small plastic shed seedling exercising being placed in greenhouse, and shed temperature controls at 22 DEG C, and relative humidity is 91% transplanting Initial stage of culture；
5th step, transplanting: after hot-house culture 40 days, outdoor cultivation 8 days, transplant in nursery, land for growing field crops.
The survival rate of this method gained Folium Isatidis transplantation of seedlings is high, up to more than 90%.
Above the detailed description of the invention of the present invention is described, but the present invention is not limited to above description.For a person skilled in the art, any equal amendment to the technical program and replacement are all within the scope of the invention.Therefore, the impartial conversion made without departing from the spirit and scope of the invention and amendment, all should contain within the scope of the invention.
Claims (6)
1. the propagation method of a large cherry stock Folium Isatidis, it is characterised in that comprise the following steps:
The first step, bud stem process: adopt high-quality Folium Isatidis branch, cleaning surface with detergent after removing all blades, water rinses and within 25 minutes, is placed on workbench, with 70% ~ 75% ethanol sterilizing 1 ~ 2 minute, soaking 5 ~ 10 minutes with 0.1% mercuric chloride, 10 rear cutout stem with bud of aseptic water washing are standby again；
Second step, culture medium select: choosing MS minimal medium, sucrose concentration takes 1 ~ 5%, and agar 0.5 ~ 0.8%, pH value is 6.0；Culturing room's temperature 25 DEG C, light intensity 1500 ~ 2000Lx, illumination every day 10 ~ 15 hours；
3rd step, inoculation: outer implant is seeded in the culture medium of MS+6 BA 0.3 ~ 0.7mg/L+NAA 0.1 ~ 0.5 mg/L and carries out initial incubation, proceed to the sprout that initial incubation is induced after 3 ~ 5 days breed in the culture medium of MS+6 BA 0.3 ~ 0.6mg/L+NAA 0.15 ~ 0.25 mg/L, after enrichment culture 2 ~ 3 weeks, take 1.5 ~ 2.0cm young sprout and transfer on the root media of 1/2MS+NAA 0.4 ~ 0.7 mg/L+15 ~ 25g sucrose, cultivate about 25 days；
4th step, seedling exercising: take out the test tube Seedling taken root, clean the culture medium adhered to, in transfer to mixed-matrix, the Small plastic shed seedling exercising being placed in greenhouse, and shed temperature controls at 21 DEG C ~ 23 DEG C, relative humidity transplant Initial stage of culture be 90% and more than；
5th step, transplanting: after hot-house culture 30 ~ 45 days, outdoor cultivation 4 ~ 9 days, transplant in nursery, land for growing field crops.
The propagation method of large cherry stock Folium Isatidis the most according to claim 1, it is characterised in that: with 70% ethanol sterilizing 90 seconds when described bud stem processes, then soak 8 minutes with 0.1% mercuric chloride.
The propagation method of large cherry stock Folium Isatidis the most according to claim 2, it is characterised in that: described sucrose concentration takes 2%, and agar 0.6%, pH value is 6.0；Culturing room's temperature 25 DEG C, light intensity 1800Lx, illumination every day 12 hours.
The propagation method of large cherry stock Folium Isatidis the most according to claim 3, it is characterized in that: during described inoculation, outer implant is seeded in the culture medium of MS+6 BA 0.5mg/L+NAA 0.2mg/L and carries out initial incubation, proceed to the sprout that initial incubation is induced after 4 days breed in the culture medium of MS+6 BA 0.5mg/L+NAA 0.2 mg/L, after enrichment culture 15 days, take 1.8cm young sprout and transfer on the root media of 1/2MS+NAA 0.5 mg/L+18g sucrose.
5. according to the propagation method of the large cherry stock Folium Isatidis described in any one of Claims 1-4, it is characterised in that: during described seedling exercising, shed temperature controls at 22 DEG C, and relative humidity is 91% transplanting Initial stage of culture.
The propagation method of large cherry stock Folium Isatidis the most according to claim 5, it is characterised in that: during transplanting after hot-house culture 40 days, outdoor cultivation 8 days.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201610340734.3A CN105993944A (en) | 2016-05-23 | 2016-05-23 | Method for propagating indigowoad leaf by using large cherry rootstock |
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| CN201610340734.3A CN105993944A (en) | 2016-05-23 | 2016-05-23 | Method for propagating indigowoad leaf by using large cherry rootstock |
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| CN201610340734.3A Pending CN105993944A (en) | 2016-05-23 | 2016-05-23 | Method for propagating indigowoad leaf by using large cherry rootstock |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108235947A (en) * | 2016-12-23 | 2018-07-03 | 句容市绿润苗木有限公司 | The mixed-matrix propagation method of large cherry stock folium isatidis |
| CN108235946A (en) * | 2016-12-23 | 2018-07-03 | 句容市绿润苗木有限公司 | The organic substrate propagation method of large cherry stock folium isatidis |
| CN108235945A (en) * | 2016-12-23 | 2018-07-03 | 句容市绿润苗木有限公司 | The propagation method of large cherry stock folium isatidis |
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| JP2005095055A (en) * | 2003-09-24 | 2005-04-14 | Japan Science & Technology Agency | Method for inducing adventitious bud of sweet cherry fruit cultivated species by high-temperature treatment |
| CN104145814A (en) * | 2014-07-24 | 2014-11-19 | 四川农业大学 | Method for obtaining regeneration plants by stem tissue culture of cerasus cerasoides (var. cerasoides) |
| CN105325299A (en) * | 2015-11-23 | 2016-02-17 | 枣庄市农业科学研究院 | Tissue culture method and culture media for large cherry rootstock Gisela |
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2016
- 2016-05-23 CN CN201610340734.3A patent/CN105993944A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2005095055A (en) * | 2003-09-24 | 2005-04-14 | Japan Science & Technology Agency | Method for inducing adventitious bud of sweet cherry fruit cultivated species by high-temperature treatment |
| CN104145814A (en) * | 2014-07-24 | 2014-11-19 | 四川农业大学 | Method for obtaining regeneration plants by stem tissue culture of cerasus cerasoides (var. cerasoides) |
| CN105325299A (en) * | 2015-11-23 | 2016-02-17 | 枣庄市农业科学研究院 | Tissue culture method and culture media for large cherry rootstock Gisela |
Non-Patent Citations (2)
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108235947A (en) * | 2016-12-23 | 2018-07-03 | 句容市绿润苗木有限公司 | The mixed-matrix propagation method of large cherry stock folium isatidis |
| CN108235946A (en) * | 2016-12-23 | 2018-07-03 | 句容市绿润苗木有限公司 | The organic substrate propagation method of large cherry stock folium isatidis |
| CN108235945A (en) * | 2016-12-23 | 2018-07-03 | 句容市绿润苗木有限公司 | The propagation method of large cherry stock folium isatidis |
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Application publication date: 20161012 |