CN104686348A - Tissue culture rapid propagation technique of moringa oleifera Lam. - Google Patents
Tissue culture rapid propagation technique of moringa oleifera Lam. Download PDFInfo
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- CN104686348A CN104686348A CN201510086066.1A CN201510086066A CN104686348A CN 104686348 A CN104686348 A CN 104686348A CN 201510086066 A CN201510086066 A CN 201510086066A CN 104686348 A CN104686348 A CN 104686348A
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Abstract
The invention discloses a tissue culture rapid propagation technique of moringa oleifera Lam. and relates to a seedling growing method for obtaining strain property stable moringa oleifera Lam. seedlings from the moringa oleifera Lam. by use of a vegetative rapid propagation technique. According to the method, the axillary buds of the moringa oleifera Lam. are taken as the explants, and the rapid propagation of the moringa oleifera Lam. seedlings is realized by virtue of the processes of bud induction, bud proliferation, rooting culture, acclimatization and transplanting and the like; the tissue culture rapid propagation technique has important practical significance on acceleration of the introduction and the artificial cultivation of the moringa oleifera Lam. and culturing of high-quality moringa oleifera Lam. seedlings.
Description
Technical field
The present invention relates to the method for Plant Tissue Breeding in agricultural biotechnologies, specifically, relate to a kind of Moringa group culturation rapid propagating technology.
Background technology
Moringa (
moringa oleiferalam
.) be Moringaceae Moringa plant, be branch list platymiscium.Moringa originates in the Himalayas of north India, nineteen sixty Moringa introduce Southwestern China area as greening-tree from India, current Main Cultivation in Fujian, Yunnan, Guangxi, Guangdong, the ground such as Hainan.Moringa nutrition is super abundant, available in all varieties to mineral elements such as calcium, iron, potassium from vitamin A, B, C race, protein, therefore has the title of vegetables tree, and the Moringa in seedling stage can be cooked in whole strain.The fruit pod of fresh prematurity or maturation, immature embryo, leaf, flower, the equal edible of root, but the Moringa consumer group mainly concentrates on India at present, and therefore the edibility of Moringa needs to develop.Moringa is with luxuriant foliage and spreading branches in leafy profusion, and biomass is large, is important feed resource, and improving quality of agricultural product has positive role.Moringa embryo is rich in grease, without the need to heating before squeezing, is a kind of pure natural plant oil, can be used as the raw material of the industrial products such as perfume, senior lubricant, senior pigment, senior natural soap.Be rich in electrically charged albumen in Moringa embryo, therefore its powder can be used as water scavengine agent, the particle suspended in the muddy water that not only can flocculate can also precipitate pathogenic microorganisms, and has no side effect.The powder of a Moringa embryo can purify 1 liter of muddy water, and Moringa embryo is purified waste water simple to operate, and cost is low, and prediction Moringa is infinite in the application aspect potentiality that purify water.Moringa is usually used in the treatments such as diabetes, hypertension, cardiovascular disease, rheumatism, scurvy, is a kind of medicinal plant with multiple pharmacological function.
Moringa is multiple use woody plant, and the human lives such as human food, health care, medicine, fuel, dyestuff, environmental protection, Water warfare and healthy closely bound up industry, industry can play life will be acted on.Moringa has wide adaptability, cultivates extensive biological nature, has good market and commercial promise in China's development Moringa large-scale planting and product development.At present in Guangdong, Fujian, the ground such as Yunnan introduces a fine variety Moringa, and the product of exploitation also engenders.Moringa seedling is bred mainly through seminal propagation, cottage propagation, cannot the demand of large-scale planting seedling in the short time.Therefore, in order to alleviate market Moringa seedling present situation in short supply, the Moringa group culturation rapid propagating technology setting up complete set is necessary very much.At present, complete Moringa group culturation rapid propagating technology there is not yet report.
Summary of the invention
The object of the present invention is to provide out a kind of Moringa group culturation rapid propagating technology, the present invention includes with Moringa axillalry bud be explant, by processes such as bud inducement, Shoot propagation, culture of rootage, hardening and transplantings, achieve the Fast-propagation of Moringa seedling, thus achieve object of the present invention.
A kind of Moringa group culturation rapid propagating technology of the present invention, comprises the following steps:
(1) Acquire and process of explant: the raw then full branch of internal organs bud selecting Moringa fine individual plant, in washing powder water soaking 10 ~ 30min after defoliation, tap water 1 ~ 2h after outwash, be cut into the stem section that long 1 ~ 3cm is with axillalry bud, first use after 75% ethanol disinfection 5 ~ 30s with aseptic washing 3 ~ 5 times, again with 0.1% mercuric chloride solution sterilization 10 ~ 25min, with for subsequent use after aseptic water washing 4 ~ 6 times.
(2) bud inducement: the stem segment with axillary bud that the process of step (1) obtains is inoculated on bud inducement medium, first full light culture 3 ~ 5 days under 25 ~ 28 DEG C of conditions, then be placed in illumination every day 10 ~ 12 hours, intensity of illumination is 1500 ~ 2000lx, until induced synthesis Multiple Buds.
(3) Shoot propagation: the Multiple Buds of step (2) is proceeded on proliferated culture medium and carries out Multiplying culture, first full light culture 1 ~ 5 day under 25 ~ 28 DEG C of conditions after inoculation, then illumination every day is placed in 12 ~ 16 hours, intensity of illumination is 2000 ~ 3000lx, being placed in cultivation temperature is cultivate under the condition of 25 ~ 28 DEG C, and switching in 20 ~ 25 days once.
(4) culture of rootage: Multiple Buds step (1) or (2) being highly about 3 ~ 5cm is cut to be inoculated on root media and carried out culture of rootage, first full light culture 1 ~ 3 day under 25 ~ 28 DEG C of conditions after inoculation, then illumination every day is placed in 14 ~ 16 hours, intensity of illumination is 3000 ~ 5000lx, and cultivation temperature is be cultured under the condition of 25 ~ 28 DEG C to take root.
(5) acclimatization and transplants: the rooting tube plantlet decap of height about 8 ~ 10cm was placed in natural lighting lower refining seedling after 5 ~ 7 days, test-tube plantlet is taken out from blake bottle, wash root medium off, to plant in the matrix be mixed into by peat soil and yellow sand mud and to be colonizated in large Tanaka.
Bud inducement medium described in above-mentioned steps (2) is: MS+6-BA2 ~ 8mg/L+ sucrose 15 ~ 30g/L+ agar 3.5 ~ 6.0g/L, pH is 5.4 ~ 5.8.
Proliferated culture medium described in above-mentioned steps (3) is: MS+6-BA0.5 ~ 2.0mg/L+NAA0.1 ~ 1.0mg/L+ sucrose 15 ~ 30g/L+ agar 3.5 ~ 6.0g/L, pH is 5.4 ~ 5.8.
Root media described in above-mentioned steps (4) is: 1/2MS+IAA0.3 ~ 1.0mg/L+ sucrose 15 ~ 30g/L+ agar 3.5 ~ 6.0g/L, pH is 5.4 ~ 5.8.
Advantage of the present invention is: domestic at present not yet have report about Moringa group culturation rapid propagating technology, and the present invention has simple, easy, economic feature.The Moringa test-tube seedling transplanting survival rate be bred as by the present invention, to more than 90%, effectively can solve the high-quality Moringa seedling supply problem in Moringa large-scale planting.
Embodiment
Following examples further illustrate of the present invention, is not limitation of the present invention.
Embodiment 1:
(1) Acquire and process of explant: the raw then full branch of internal organs bud selecting Moringa fine individual plant, in washing powder water soaking 10min after defoliation, tap water 1h after outwash, be cut into the stem section that long 1 ~ 3cm is with axillalry bud, first use after 75% alcohol disinfecting 5s with aseptic washing 3 times, again with 0.1% mercuric chloride solution sterilization 10min, with for subsequent use after aseptic water washing 5 times.
(2) bud inducement: the stem segment with axillary bud that the process of step (1) obtains is inoculated on bud inducement medium, first full light culture 3 days under 28 DEG C of conditions, then be placed in illumination every day 10 hours, intensity of illumination is that 1500 ~ 2000lx cultivates and can form Multiple Buds in 18 days, and inductivity reaches 87.4%.Described bud inducement medium is: MS+6-BA4mg/L+ sucrose 25g/L+ agar 4g/L, pH is 5.8.
(3) Shoot propagation: the Multiple Buds of step (2) is proceeded on proliferated culture medium and carries out Multiplying culture, first full light culture 1 day under 28 DEG C of conditions after inoculation, then illumination every day is placed in 12 hours, intensity of illumination is 2000lx, being placed in cultivation temperature is cultivate under the condition of 28 DEG C, once, growth coefficient reaches 6.72 in switching in 20 days.Described proliferated culture medium is: MS+6-BA1.5mg/L+NAA0.5mg/L+ sucrose 20g/L+ agar 3.5g/L, pH is 5.8.
(4) culture of rootage: Multiple Buds step (1) or (2) being highly about 3 ~ 5cm is cut to be inoculated on root media and carried out culture of rootage, first full light culture 1 day under 28 DEG C of conditions after inoculation, then illumination every day is placed in 14 hours, intensity of illumination is 3000lx, cultivation temperature is cultivate under the condition of 28 DEG C namely to take root for 21 days, and rooting rate reaches 90.3%.Described root media is: 1/2MS+IAA0.5mg/L+ sucrose 15g/L+ agar 3.5g/L, pH is 5.8.
(5) acclimatization and transplants: the rooting tube plantlet decap of height about 8 ~ 10cm was placed in natural lighting lower refining seedling after 5 days, test-tube plantlet is taken out from blake bottle, wash root medium off, planting in the matrix be mixed into by peat soil and yellow sand mud and to be colonizated in large Tanaka, transplanting survival rate is 89.6%.
Embodiment 2:
(1) Acquire and process of explant: the raw then full branch of internal organs bud selecting Moringa fine individual plant, in washing powder water soaking 15min after defoliation, tap water 3h after outwash, be cut into the stem section that long 1 ~ 3cm is with axillalry bud, first use after 75% alcohol disinfecting 10s with aseptic washing 3 times, again with 0.1% mercuric chloride solution sterilization 15min, with for subsequent use after aseptic water washing 5 times.
(2) bud inducement: the stem segment with axillary bud that the process of step (1) obtains is inoculated on bud inducement medium, first full light culture 3 days under 28 DEG C of conditions, then be placed in illumination every day 11 hours, intensity of illumination is that 1500 ~ 2000lx cultivates and can form Multiple Buds in 21 days, and inductivity reaches 94.1%.Described bud inducement medium is: MS+6-BA7mg/L+ sucrose 30g/L+ agar 5g/L, pH is 5.8.
(3) Shoot propagation: the Multiple Buds of step (2) is proceeded on proliferated culture medium and carries out Multiplying culture, first full light culture 3 days under 28 DEG C of conditions after inoculation, then illumination every day is placed in 13 hours, intensity of illumination is 2500lx, being placed in cultivation temperature is cultivate under the condition of 28 DEG C, once, growth coefficient reaches 7.02 in switching in 23 days.Described proliferated culture medium is: MS+6-BA1.0mg/L+NAA0.2mg/L+ sucrose 25g/L+ agar 4g/L, pH is 5.8.
(4) culture of rootage: Multiple Buds step (1) or (2) being highly about 3 ~ 5cm is cut to be inoculated on root media and carried out culture of rootage, first full light culture 3 days under 28 DEG C of conditions after inoculation, then illumination every day is placed in 14 hours, intensity of illumination is 3500lx, cultivation temperature is cultivate under the condition of 28 DEG C namely to take root for 21 days, and rooting rate reaches 93.7%.Described root media is: 1/2MS+IAA0.8mg/L+ sucrose 15g/L+ agar 3.5g/L, pH is 5.8.
(5) acclimatization and transplants: the rooting tube plantlet decap of height about 8 ~ 10cm was placed in natural lighting lower refining seedling after 5 days, test-tube plantlet is taken out from blake bottle, wash root medium off, planting in the matrix be mixed into by peat soil and yellow sand mud and to be colonizated in large Tanaka, transplanting survival rate is 90.8%.
Claims (4)
1. a Moringa group culturation rapid propagating technology, is characterized in that comprising the following steps:
(1) Acquire and process of explant: the raw then full branch of internal organs bud selecting Moringa fine individual plant; in washing powder water soaking 10 ~ 30min after defoliation; tap water 1 ~ 2h after outwash; be cut into the stem section that long 1 ~ 3cm is with axillalry bud; first use after 75% ethanol disinfection 5 ~ 30s with aseptic washing 3 ~ 5 times; again with 0.1% mercuric chloride solution sterilization 10 ~ 25min, with for subsequent use after aseptic water washing 4 ~ 6 times;
(2) bud inducement: the stem segment with axillary bud that the process of step (1) obtains is inoculated on bud inducement medium; first full light culture 3 ~ 5 days under 25 ~ 28 DEG C of conditions; then be placed in illumination every day 10 ~ 12 hours, intensity of illumination is 1500 ~ 2000lx, until induced synthesis Multiple Buds;
(3) Shoot propagation: the Multiple Buds of step (2) is proceeded on proliferated culture medium and carries out Multiplying culture; first full light culture 1 ~ 5 day under 25 ~ 28 DEG C of conditions after inoculation; then illumination every day is placed in 12 ~ 16 hours; intensity of illumination is 2000 ~ 3000lx; being placed in cultivation temperature is cultivate under the condition of 25 ~ 28 DEG C, and switching in 20 ~ 25 days once;
(4) culture of rootage: Multiple Buds step (1) or (2) being highly about 3 ~ 5cm is cut to be inoculated on root media and carried out culture of rootage; first full light culture 1 ~ 3 day under 25 ~ 28 DEG C of conditions after inoculation; then illumination every day is placed in 14 ~ 16 hours; intensity of illumination is 3000 ~ 5000lx, and cultivation temperature is be cultured under the condition of 25 ~ 28 DEG C to take root;
(5) acclimatization and transplants: the rooting tube plantlet decap of height about 8 ~ 10cm was placed in natural lighting lower refining seedling after 5 ~ 7 days, test-tube plantlet is taken out from blake bottle, wash root medium off, to plant in the matrix be mixed into by peat soil and yellow sand mud and to be colonizated in large Tanaka.
2. a kind of Moringa group culturation rapid propagating technology according to claim 1, is characterized in that the bud inducement medium described in step (2) is: MS+6-BA2 ~ 8mg/L+ sucrose 15 ~ 30g/L+ agar 3.5 ~ 6.0g/L, pH is 5.4 ~ 5.8.
3. a kind of Moringa group culturation rapid propagating technology according to claim 1, it is characterized in that the proliferated culture medium described in step (3) is: MS+6-BA0.5 ~ 2.0mg/L+NAA0.1 ~ 1.0mg/L+ sucrose 15 ~ 30g/L+ agar 3.5 ~ 6.0g/L, pH is 5.4 ~ 5.8.
4. a kind of Moringa group culturation rapid propagating technology according to claim 1, is characterized in that the root media described in step (4) is: 1/2MS+IAA0.3 ~ 1.0mg/L+ sucrose 15 ~ 30g/L+ agar 3.5 ~ 6.0g/L, pH is 5.4 ~ 5.8.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105961197A (en) * | 2016-05-09 | 2016-09-28 | 华南农业大学 | High efficiency regeneration method of Moringa oleifera |
| CN107094618A (en) * | 2017-03-23 | 2017-08-29 | 上海杉植物科技有限公司 | A kind of method for tissue culture of Moringa tissue-cultured seedling |
| CN109042217A (en) * | 2018-09-19 | 2018-12-21 | 福建农林大学 | A kind of Moringa tissue-cultured seedling rapidly and efficiently method for transplanting |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102150619A (en) * | 2011-03-09 | 2011-08-17 | 云南省农业科学院花卉研究所 | Moringa embryo callus induction and plant regeneration method |
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- 2015-02-24 CN CN201510086066.1A patent/CN104686348A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102150619A (en) * | 2011-03-09 | 2011-08-17 | 云南省农业科学院花卉研究所 | Moringa embryo callus induction and plant regeneration method |
Non-Patent Citations (1)
| Title |
|---|
| 张婧: ""辣木组织培养及有效成分分析"", 《中国优秀硕士学位论文全文数据库 基础科学辑》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105961197A (en) * | 2016-05-09 | 2016-09-28 | 华南农业大学 | High efficiency regeneration method of Moringa oleifera |
| CN107094618A (en) * | 2017-03-23 | 2017-08-29 | 上海杉植物科技有限公司 | A kind of method for tissue culture of Moringa tissue-cultured seedling |
| CN109042217A (en) * | 2018-09-19 | 2018-12-21 | 福建农林大学 | A kind of Moringa tissue-cultured seedling rapidly and efficiently method for transplanting |
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