CN105028196A - Quick tissue culture seedling breeding method of decaisne fargesii - Google Patents
Quick tissue culture seedling breeding method of decaisne fargesii Download PDFInfo
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- CN105028196A CN105028196A CN201510361990.6A CN201510361990A CN105028196A CN 105028196 A CN105028196 A CN 105028196A CN 201510361990 A CN201510361990 A CN 201510361990A CN 105028196 A CN105028196 A CN 105028196A
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Abstract
The invention discloses a quick tissue culture seedling breeding method of decaisne fargesii. The method comprises the following steps of seed selection, seed treatment, medium treatment, inoculation, root induction and transplantation, and comprises the following concrete steps of (a) seed selection: cleaning and airing seeds, and stripping testa; (b) seed treatment: treating through warm water, soaking in disinfectant, and rinsing through aquae sterilisata; (c) medium treatment: sprouting a medium, breeding the medium, rooting the medium, adjusting a PH value, and sterilizing; (d) inoculating; (e) root inducting; (f) transplantation: hardening-seedling, transplanting into the medium, and watering sufficiently. According to the quick tissue culture seedling breeding method of the decaisne fargesii, aseptic seedlings germinated from the seeds of the decaisne fargesii are adopted as explants, aseptic seedling stems induce multiple shoots so as to improve the coefficient of propagation, the medium is selected, the hormone combination is adjusted, the bud grows strongly and highly, the growth is quick, more clumpy buds can be produced, the root system is developed, the regeneration-seedling survival rate is high, and seedlings are provided to be planted in batch.
Description
[technical field]
The present invention relates to the technical field of breeding and seedling, particularly a kind of technical field of tissue culturing fast seedling-cultivating method of cat dung melon.
[background technology]
Cat dung melon, has another name called cat dung melon, terrible finger, cat dung cylinder.The cock intestines again in Zhaotong region of Yunnan Province, Lardizabalaceae Mao Shiguashu defoliation bush plant, up to 5 meters.Among the remote mountains in the towering Qinling Mountains, the Bashan Mountain, grow a kind of wild bush, people are its cat dung melon.After its fruit maturation, in bluish violet, many juices, the dung that shape is drawn the spitting image of cat, so be commonly called as cat dung melon, also known as Decaisnea fargesii.
Cat dung melon is a kind of wild economic plant having very much Development volue.Be rich in rubber in its pericarp, its content is about 10% ~ 20% of dry weight.And the chemical constitution of this rubber is identical with general natural rubber, admixes with butadiene-styrene rubber and use, can general rubber product be made.Cat dung melon is a kind of wild rubber plant having very much value.Root and fruit can be used as medicine, and its taste is sweet, pungent, and property is put down, and has removing heat from the lung to relieve cough, dispels rheumatism, clearing heat and detoxicating effect, cure mainly the diseases such as pulmonary tuberculosis, cough, rheumatic arthralgia, pruritus vulvae, hernia and Tumor suppression.In river, decocting liquid washout private parts, the anus of western root among the people, have good antipruritic effect, and fruit concentrated decoction is used for chapped skin, takes orally and control hernia.
Cat dung melon is a kind of nutritious wild fruit.Carbohydrate, protein, fat, pectin, vitamin and mineral nutrition is rich in its pulp.Taste is sweet in honey, can for eating and process sugaring, wine brewing, food, pectin, jelly, jam etc. raw.Its seed oil content is 18% ~ 22%.Oil yield is close with cottonseed, oily edible, and its taste is fragrant beautiful, being rich in nutrition, also can soap system, makes iundustrial oil.
Utilize traditional method sowing cultivation cat dung melon cycle length, length consuming time, the demand in market can not be met.Current introduces a fine variety just in active research stage, not yet large-scale promotion to the nursery of cat dung melon, and technical scheme is also little.
[summary of the invention]
Object of the present invention solves the problems of the prior art exactly, a kind of tissue culturing fast seedling-cultivating method of cat dung melon is proposed, Selective agar medium, adjustment hormone combination, bud robust growth and high, growth is soon, produce more clump bud, root system is very flourishing, and regrowth survival rate is high, for plantation provides seedling in a large number.
For achieving the above object, the present invention proposes a kind of tissue culturing fast seedling-cultivating method of cat dung melon, comprise initial point selection, seed treatment, medium treatment, inoculation, root induction and transplanting, concrete steps are as follows:
A () initial point selection: the ripe and fruit of natural cracking on cat dung melon plant of gathering, extrudes seed, dries after cleaning, peel off exosper;
B () seed treatment: the seed after peeling is put into warm water, carries out warm water process, use medicining liquid dipping after warm water process, then uses rinsed with sterile water 6 ~ 8 times;
(c) medium treatment: induction seed germination medium is that 1/2MS medium adds gibberellin 1.0mg/L, proliferated culture medium is that 1/2MS medium adds methyl α-naphthyl acetate NAA0.2mg/L, basic element of cell division 6-BA1.0mg/L, active carbon 0.2g/L, root media is that 1/2MS medium adds methyl α-naphthyl acetate NAA0.1mg/L, active carbon 0.2g/L, adjustment pH value 5.8 ~ 6.0, after medium preparing and packaging, carry out sterilization treatment;
D () is inoculated: be inoculated on Plates for germination media by the seed after process, in the induction seed germination stage, first light culture in culturing room, cultivate proceed to light again after seedling is formed under, as aseptic height of seedling 3 ~ 4cm, aseptic seedling is divided into the top stem section containing the multiple internode of terminal bud and bottom stem section, proliferated culture medium is transferred to respectively by primary long polar orientation, select optimal explant material, the induction of indefinite bud is carried out in proliferated culture medium, indefinite bud after propagation, expansion is carried out numerous in the proliferated culture medium that 20% ~ 40% access is fresh, remaining indefinite bud access root media,
E () root induction: start Zhan Yeshi at indefinite bud, is divided into it single seedling to proceed to root induction in root media in time, treats single seedling rooting and form complete regeneration plant;
F () transplants: regeneration plant needs to carry out hardening, and hardening is carried out for 5 ~ 7 days before field planting, accepts scattered light, to leak informaton cooling, reduce irrigation amount, transplant after hardening, wash away the medium of base portion, be transplanted in ready medium, after transplanting, water sufficient water, then use plastic film covering moisture-heat preservation, matrix with moistening, not ponding be advisable, take off film after one week, suitably shade and ventilation, spray weekly the nutrient solution of the MS macroelement once diluting 10 times.
As preferably, in described step (b), the temperature of warm water process is 35 ~ 45 DEG C, and the time is 20 ~ 30 hours; Thimerosal is 0.1% mercury chloride, and soak time is 3 ~ 5 minutes.
As preferably, in described step (c), sterilization treatment Conditions Temperature is 120 DEG C, pressure is 1.1kg/cm
2, the time sterilizing 18 ~ 25min of sterilization treatment.
As preferably, in described step (d), cultivate under the light of seedling, illuminance is 2000lx, and light application time is 10 ~ 12h/d, and temperature is 22 ~ 26 DEG C.
As preferably, in described step (f), transplant medium and select humus soil and river sand blending agent, the ratio of humus soil and river sand is 1:1
As preferably, in described step (f), transplant rear film and keep humidity about 90%, temperature controls at 25 ~ 27 DEG C.
Beneficial effect of the present invention:
The present invention is with the aseptic seedling of the seed germination of cat dung melon for explant, and aseptic seedling top sturdy stem section is the best explant that induced bundle is sprouted, and the basic element of cell division and growth hormone play an important role to the induction of Multiple Buds and growth.Cat dung melon aseptic seedling stem sections induced bundle is adopted to sprout and can improve reproduction coefficient, Selective agar medium, adjustment hormone combination, optimum adventitious buds proliferation, the differentiation of optimum bud and root, bud robust growth and high, growth is fast, and produce more clump bud, root system is very flourishing, regrowth survival rate is high, for plantation provides seedling in a large number.
Feature of the present invention and advantage will be described in detail by embodiment.
[embodiment]
The tissue culturing fast seedling-cultivating method of a kind of cat dung of the present invention melon, comprise initial point selection, seed treatment, medium treatment, inoculation, root induction and transplanting, concrete steps are as follows:
A () initial point selection: the ripe and fruit of natural cracking on cat dung melon plant of gathering, extrudes seed, dries after cleaning, peel off exosper;
B () seed treatment: the seed after peeling is put into warm water, carries out warm water process, use medicining liquid dipping after warm water process, then uses rinsed with sterile water 6 ~ 8 times;
(c) medium treatment: induction seed germination medium is that 1/2MS medium adds gibberellin 1.0mg/L, proliferated culture medium is that 1/2MS medium adds methyl α-naphthyl acetate NAA0.2mg/L, basic element of cell division 6-BA1.0mg/L, active carbon 0.2g/L, root media is that 1/2MS medium adds methyl α-naphthyl acetate NAA0.1mg/L, active carbon 0.2g/L, adjustment pH value 5.8 ~ 6.0, after medium preparing and packaging, carry out sterilization treatment;
D () is inoculated: be inoculated on Plates for germination media by the seed after process, in the induction seed germination stage, first light culture in culturing room, cultivate proceed to light again after seedling is formed under, as aseptic height of seedling 3 ~ 4cm, aseptic seedling is divided into the top stem section containing the multiple internode of terminal bud and bottom stem section, proliferated culture medium is transferred to respectively by primary long polar orientation, select optimal explant material, the induction of indefinite bud is carried out in proliferated culture medium, indefinite bud after propagation, expansion is carried out numerous in the proliferated culture medium that 20% ~ 40% access is fresh, remaining indefinite bud access root media,
E () root induction: start Zhan Yeshi at indefinite bud, is divided into it single seedling to proceed to root induction in root media in time, treats single seedling rooting and form complete regeneration plant;
F () transplants: regeneration plant needs to carry out hardening, and hardening is carried out for 5 ~ 7 days before field planting, accepts scattered light, to leak informaton cooling, reduce irrigation amount, transplant after hardening, wash away the medium of base portion, be transplanted in ready medium, after transplanting, water sufficient water, then use plastic film covering moisture-heat preservation, matrix with moistening, not ponding be advisable, take off film after one week, suitably shade and ventilation, spray weekly the nutrient solution of the MS macroelement once diluting 10 times.
Embodiment:
The first step, initial point selection: the ripe and fruit of natural cracking on cat dung melon plant of gathering, extrude seed, dry after cleaning, peel off exosper;
Second step, seed treatment: the seed after peeling is put into warm water, carry out warm water process, and temperature is 40 DEG C, and the time is 25 hours, uses medicining liquid dipping after warm water process, and thimerosal is 0.1% mercury chloride, and soak time is 4 minutes, then use rinsed with sterile water 8 times;
3rd step, medium treatment: induction seed germination medium is that 1/2MS medium adds gibberellin 1.0mg/L, proliferated culture medium is that 1/2MS medium adds methyl α-naphthyl acetate NAA0.2mg/L, basic element of cell division 6-BA1.0mg/L, active carbon 0.2g/L, root media is that 1/2MS medium adds methyl α-naphthyl acetate NAA0.1mg/L, active carbon 0.2g/L, adjustment pH value 5.8, after medium preparing and packaging, carry out sterilization treatment, sterilization treatment Conditions Temperature is 120 DEG C, pressure is 1.1kg/cm
2, the time sterilizing 20min of sterilization treatment;
4th step, inoculation: the seed after process is inoculated on Plates for germination media, in the induction seed germination stage, first light culture in culturing room, cultivate proceed to light again after seedling is formed under, illuminance is 2000lx, light application time is 12h/d, temperature is 25 DEG C, as aseptic height of seedling 4cm, aseptic seedling is divided into the top stem section containing the multiple internode of terminal bud and bottom stem section, proliferated culture medium is transferred to respectively by primary long polar orientation, select optimal explant material, the induction of indefinite bud is carried out in proliferated culture medium, indefinite bud after propagation, expansion is carried out numerous in the proliferated culture medium that 40% access is fresh, remaining indefinite bud access root media,
5th step, root induction: start Zhan Yeshi at indefinite bud, be divided into it single seedling to proceed to root induction in root media in time, treat single seedling rooting and form complete regeneration plant;
6th step, transplant: regeneration plant needs to carry out hardening, hardening is carried out in field planting for first 7 days, accept scattered light, to leak informaton cooling, reduce irrigation amount, transplant after hardening, wash away the medium of base portion, be transplanted in ready medium, transplant medium and select humus soil and river sand blending agent, the ratio of humus soil and river sand is 1:1, sufficient water is watered after transplanting, then plastic film covering moisture-heat preservation is used, transplant rear film and keep humidity about 90%, temperature controls at 25 DEG C, matrix is with moistening, ponding is not advisable, film is taken off after one week, suitably shade and ventilation, spray weekly the nutrient solution of the MS macroelement once diluting 10 times.The transplanting medium adaptability of cat dung melon regrowth is relatively wider, but with loose ventilative, the matrix that the content of organic matter is high is good.
Transplant after 1 month and observe, Plant Leaf look dark green, and robust growth grows fine, and the survival rate of regrowth is to 96.8%.
The present invention, with the aseptic seedling of the seed germination of cat dung melon for explant, aseptic seedling top sturdy stem section is the best explant that induced bundle is sprouted, and the basic element of cell division and growth hormone play an important role to the induction of Multiple Buds and growth.Cat dung melon aseptic seedling stem sections induced bundle is adopted to sprout and can improve reproduction coefficient, Selective agar medium, adjustment hormone combination, optimum adventitious buds proliferation, the differentiation of optimum bud and root, bud robust growth and high, growth is fast, and produce more clump bud, root system is very flourishing, regrowth survival rate is high, for plantation provides seedling in a large number.
Above-described embodiment is to explanation of the present invention, is not limitation of the invention, anyly all belongs to protection scope of the present invention to the scheme after simple transformation of the present invention.
Claims (6)
1. a tissue culturing fast seedling-cultivating method for cat dung melon, comprise initial point selection, seed treatment, medium treatment, inoculation, root induction and transplanting, concrete steps are as follows:
A () initial point selection: the ripe and fruit of natural cracking on cat dung melon plant of gathering, extrudes seed, dries after cleaning, peel off exosper;
B () seed treatment: the seed after peeling is put into warm water, carries out warm water process, use medicining liquid dipping after warm water process, then uses rinsed with sterile water 6 ~ 8 times;
(c) medium treatment: induction seed germination medium is that 1/2MS medium adds gibberellin 1.0mg/L, proliferated culture medium is that 1/2MS medium adds methyl α-naphthyl acetate NAA0.2mg/L, basic element of cell division 6-BA1.0mg/L, active carbon 0.2g/L, root media is that 1/2MS medium adds methyl α-naphthyl acetate NAA0.1mg/L, active carbon 0.2g/L, adjustment pH value 5.8 ~ 6.0, after medium preparing and packaging, carry out sterilization treatment;
D () is inoculated: be inoculated on Plates for germination media by the seed after process, in the induction seed germination stage, first light culture in culturing room, cultivate proceed to light again after seedling is formed under, as aseptic height of seedling 3 ~ 4cm, aseptic seedling is divided into the top stem section containing the multiple internode of terminal bud and bottom stem section, proliferated culture medium is transferred to respectively by primary long polar orientation, select optimal explant material, the induction of indefinite bud is carried out in proliferated culture medium, indefinite bud after propagation, expansion is carried out numerous in the proliferated culture medium that 20% ~ 40% access is fresh, remaining indefinite bud access root media,
E () root induction: start Zhan Yeshi at indefinite bud, is divided into it single seedling to proceed to root induction in root media in time, treats single seedling rooting and form complete regeneration plant;
F () transplants: regeneration plant needs to carry out hardening, and hardening is carried out for 5 ~ 7 days before field planting, accepts scattered light, to leak informaton cooling, reduce irrigation amount, transplant after hardening, wash away the medium of base portion, be transplanted in ready medium, after transplanting, water sufficient water, then use plastic film covering moisture-heat preservation, matrix with moistening, not ponding be advisable, take off film after one week, suitably shade and ventilation, spray weekly the nutrient solution of the MS macroelement once diluting 10 times.
2. the tissue culturing fast seedling-cultivating method of a kind of cat dung melon as claimed in claim 1, is characterized in that: in described step (b), and the temperature of warm water process is 35 ~ 45 DEG C, and the time is 20 ~ 30 hours; Thimerosal is 0.1% mercury chloride, and soak time is 3 ~ 5 minutes.
3. the tissue culturing fast seedling-cultivating method of a kind of cat dung melon as claimed in claim 1, it is characterized in that: in described step (c), sterilization treatment Conditions Temperature is 120 DEG C, pressure is 1.1kg/cm
2, the time sterilizing 18 ~ 25min of sterilization treatment.
4. the tissue culturing fast seedling-cultivating method of a kind of cat dung melon as claimed in claim 1, it is characterized in that: in described step (d), cultivate under the light of seedling, illuminance is 2000lx, and light application time is 10 ~ 12h/d, and temperature is 22 ~ 26 DEG C.
5. the tissue culturing fast seedling-cultivating method of a kind of cat dung melon as claimed in claim 1, is characterized in that: in described step (f), and transplant medium and select humus soil and river sand blending agent, the ratio of humus soil and river sand is 1:1.
6. the tissue culturing fast seedling-cultivating method of a kind of cat dung melon as claimed in claim 1, is characterized in that: in described step (f), and transplant rear film and keep humidity about 90%, temperature controls at 25 ~ 27 DEG C.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106171975A (en) * | 2016-07-07 | 2016-12-07 | 三峡大学 | The in-vitro culture method of Radix decaisneae fargesii |
| CN108184672A (en) * | 2018-03-07 | 2018-06-22 | 贵州省山地资源研究所 | A kind of Stauntonia latifolia callus from stem segment method for inducing and cultivating |
| CN115413444A (en) * | 2022-10-12 | 2022-12-02 | 安徽博帕特智能科技发展有限公司 | Plant seed screening method |
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| CN102090327A (en) * | 2009-12-15 | 2011-06-15 | 湖南农业大学 | Method for quickly breeding Akebia trifoliata Koidz fruit seedling in test tube |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106171975A (en) * | 2016-07-07 | 2016-12-07 | 三峡大学 | The in-vitro culture method of Radix decaisneae fargesii |
| CN106171975B (en) * | 2016-07-07 | 2019-03-12 | 三峡大学 | The in-vitro culture method of Decaisnea fargesii |
| CN108184672A (en) * | 2018-03-07 | 2018-06-22 | 贵州省山地资源研究所 | A kind of Stauntonia latifolia callus from stem segment method for inducing and cultivating |
| CN115413444A (en) * | 2022-10-12 | 2022-12-02 | 安徽博帕特智能科技发展有限公司 | Plant seed screening method |
| CN115413444B (en) * | 2022-10-12 | 2024-01-05 | 合肥创农生物科技有限公司 | Screening method of plant seeds |
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