CN107094618A - A kind of method for tissue culture of Moringa tissue-cultured seedling - Google Patents
A kind of method for tissue culture of Moringa tissue-cultured seedling Download PDFInfo
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- CN107094618A CN107094618A CN201710177055.3A CN201710177055A CN107094618A CN 107094618 A CN107094618 A CN 107094618A CN 201710177055 A CN201710177055 A CN 201710177055A CN 107094618 A CN107094618 A CN 107094618A
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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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
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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
- A01H4/001—Culture apparatus for tissue culture
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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
A kind of method for tissue culture of Moringa tissue-cultured seedling, including：(1) experiment material must be selected；(2) specific experiment step：(2.1) acquisition of aseptic explant：Blade is cut off, retains terminal bud lobus cardiacus, petiole stays 0.1 0.2cm, cuts into 0.5 0.8cm stem section, 12 axillary buds of every section of band；(2.3) Moringa squamous subculture；(2.4) after cultivating 21 25 days, plant longitudinal growth plant height is sprouted in more than 4.5cm, plant lateral bud, and statistics multiplied ratio averagely reaches 1:6；(2.5) after being cultivated 18 days in the culture mediums of LM 8, plant height averagely reaches 4cm, and rooting rate reaches 98%, and root length is more than 2cm, and root bar number is more than 3；(2.6) tissue-cultured seedling is tamed：Moringa tissue-cultured seedling of taking root is moved in greenhouse, and unscrewing bottle cap hardening can transplant to sand bed, survival rate to more than 95% for 23 days.The present invention solves the phenomenon of raw, the fragile frangibility of plant jaundice in Moringa tissue-cultured seedling tissue culture procedures, and realizes that expanding a numerous, step of taking root completes, and saves cost more than 40% in bottle.
Description
Technical field
The present invention relates to a kind of tissue culture method, and in particular to a kind of method for tissue culture of Moringa tissue-cultured seedling, belongs to biological
Technical field.
Background technology
Moringa (Moringa oleifera Lam.), also known as drumstick tree, milk tree, foreign Ailanthus altissima, horseradish, sound of vomiting beans etc. are chanted in a loud voice,
It is fallen leaves woody vegetables and the oilseed plant of Moringaceae (Moringaceae) Moringa (adans), Yin Qigen has acid,
Therefore the Moringa ¨ that thus gains the name.Moringa originates in the Himalayas southern foot of north India, now plants extensively in Asia, Africa and U.S.
More than the 30 of continent tropical, subtropical countries and area.
Moringa whole body is all precious, and its root, stem, leaf, flower, fruit, seed etc. are available, and contains abundant nutrients
Matter, is widely used in the multiple fields such as agricultural, industry, animal husbandry, medicine, beauty, is that one kind collects delicious, nutrition and health care in one
The rare dish of body, is described as " tree of miracle ".Moringa grows soon, and biomass is big.Moringa disease resistance is stronger, to bacterium,
The pathogenic organisms such as fungi have stronger resistivity.
Existing investigative technique：
It is the culture of propagation and substep progress of taking root mostly in the existing report on Moringa tissue cultures.Wu Xinqin
Deng《Hormon is with the influence for comparing the growth of Moringa tissue-cultured seedling》MS+6-BA0.02mg/L+NAA 0.02- are used in report
This formula of 0.5mg/L+3% white sugar carries out the Multiplying culture of Moringa, and proliferation times can reach 5.6-8.1, in 1/2MS+
Highest of being taken root on the culture medium of NAA0.2mg/L+ sucrose 2% only 79%；
Luo Yunxia etc. exists《Study on tissue culture of Moringa Oleifera Lam is studied》The stem section obtained in research by seed is studied, in MS
Shoot proliferation in+6-BA 0.6mg/L+NAA 0.1mg/L+ sucrose 30%+ agar 6g/L culture mediums, proliferation times are up to 5.8；；
Rooting induction can obtain the rooted seedling of stalwartness on 1/2MS+IBA 0.1mg/L+ sucrose 15%+ agar 6g/L culture mediums, raw
Root rate is up to 98%；Nursery survival rate is transplanted up to 80%.
Zhu's tail silver is waited《The tissue cultures of Moringa and quick study on reproduction》It is middle to propose,；It is to change to breed most suitable culture medium
Good MS+6-BA0.3mg/L+KT0.2mg/L+NAA0.1mg/L, average coefficient of proliferation reaches 3.46；；MS+ is improved 1/2
The rooted seedling of stalwartness is obtained on NAA0.1mg/L+IBA0.2mg/L+ sucrose 25g/L+ carragheen 6.5g/L culture mediums, rooting rate reaches
More than 90%, and plant and root system grow fine.
Li Guoyun using giving birth to Moringa seedling level pressure, having several lateral buds to be propagating materials then, in improvement MS+BA0.3-
Shoot proliferation on 1.0mg/L+NAA0.1-0.2mg/L+ sucrose 20g/L+ carragheens 9g/L, proliferation times more than 3.0 are taken root and lured
Lead and obtained on 1/3 improvement MS+NAA0.2-0.5mg/L+IBA0.2-0.5mg/L+ sucrose 10/L+ carragheens 10g/L culture medium
Healthy and strong rooted seedling, rooting induction rate more than 90%, transplant nursery succeed (《Moringa tissue culture technical research is total
Knot》).
Xiang Suqiong is in research《Moringa tissue cultures are induced with tetraploid plant》Work in, using sajina as material
, it is most preferably foster base in MS+BA 0.6mg/L+NAA 0.1mg/L solid medium, growth coefficient can reach 6.03, life
Root culture medium is most suitable with MS+IBA 0.2mg/L, and rooting rate is 90%, and rooted seedling is best in quality.
《The research of plant regeneration system is set up using Moringa stem section》In, the selection subculture medium such as Wang Hongfeng is MS+
6BA0.4rng/L+NAA0.2mg/L+ carragheens 5g/L+ sugar 30g/L, growth coefficient can reach 3.0；Root media is 1/
2MS+IBA0.4mg/L+NAA.2mgmg/L+ carragheen 7g/L+ sugar 20g/L, rooting rate is 100%.
Existing patent report：
The Patent data on Moringa tissue culture has at present,《A kind of Moringa group culturation rapid propagating technology》、《A kind of Moringa tissue cultures
In seed disinfection method》With《A kind of method of Moringa tissue-cultured seedling strengthening seedling and rooting》.
Studies have reported that weak point：
In the research report and Patent data delivered, the tissue culture mode of Moringa is propagation and substep of taking root is carried out
, Moringa is as woody vegetables, and its rudiment power is strong, and factorial praluction complex steps are carried out using traditional group training research method,
Increase cost of labor；Meanwhile, Moringa tissue culture seeding stem segment quality is crisp, frangibility, and waste is easily caused in process of production.
The content of the invention
It is an object of the present invention to provide a kind of method for tissue culture of Moringa tissue-cultured seedling, to overcome prior art to be deposited
Disadvantages mentioned above and deficiency.
The technical problem solved required for of the invention, can be achieved through the following technical solutions：
This experiment is directed to a kind of Subculture of Moringa, solve in a kind of Moringa tissue-culturing rapid propagation tissue-cultured seedling jaundice and
The problem of fragile frangibility.By optimized production process flow, Multiplying culture and training of taking root in Moringa tissue culture procedures are realized
Support it is synchronous carry out, shorten cultivation cycle and saved cost of labor and surpass 40%, improve the income of Moringa factorial praluction.
The technical problem solved required for of the invention, can be achieved through the following technical solutions：
A kind of quick breeding by group culture method of Moringa, it is characterised in that：Comprise the following steps：
(1) experiment material must be selected：Select that growing way is neat, plant height be 3.5-4.5cm, subculture cycle be 21-25 days,
Leaf color is used as culture materials in the Moringa tissue-cultured seedling of green or peak green；
(2) specific experiment step：
(2.1) cutting of material：Blade is cut off, retains terminal bud lobus cardiacus, petiole stays 0.1-0.2cm, cuts into 0.5-0.8cm
Stem section, 1-2 axillary bud of every section of band；
(2.2) Moringa squamous subculture；Terminal bud and stem section separate inoculation, are equably inoculated into subculture medium 8 (LM-8)
In；
Wherein, improvement MS (KNO in step (2.2) subculture medium 8 (LM-8)30.95-1.43g/L, NH4NO3
0.81-1.24g/L, MgSO4·7H2O 0.19-0.28g/L, KH2PO40.09-0.13g/L, CaCl2·2H2O 0.22-
0.44g/L, Na2EDTA·2H2O 0.002-0.04g/L,FeSO4·7H2O 0.015-0.03g/L,MnSO4·H2O 8.45-
16.9mg/L, H3BO33.1-6.2mg/L, ZnSO4·7H2O 4.3-8.6mg/L, KI 0.42-0.83mg/L, Na2MoO4·
2H2O 0.13-0.25mg/L, CoCl2·6H2O 0.01-0.025mg/L, CuSO4·5H2O 0.01-0.025mg/L, inositol 5-
10mg/L, nicotinic acid 0.025-0.05mg/L, thiamine hydrochloride 0.005-0.01mg/L, puridoxine hydrochloride 0.025-0.05mg/L,
Additional 6-BA0.0005-0.001mg/L, IBA 0.1mg/L, white granulated sugar 30g/L+ agar 5.5g/L, pH value are adjusted to 6.1.
(2.3) in step (2.2), it is that light application time is 12h/d, intensity of illumination that Moringa squamous subculture, which obtains condition of culture,
2000-3000lux, cultivation temperature is 26 ± 2 DEG C；
(2.4) after culture 21-25d, plant longitudinal growth plant height is sprouted in more than 4.5cm, plant lateral bud, statistics propagation
Ratio averagely reaches 1:6.01.
(2.5) after being cultivated 18 days in LM-8 culture mediums, plant height averagely reaches 4cm, and rooting rate reaches 98%, and root is long
More than 2cm, root bar number is more than 3；
(2.6) tissue-cultured seedling is tamed：Moringa tissue-cultured seedling of taking root is moved in greenhouse, unscrew bottle cap hardening can transplant within 2-3 days to
Sand bed, survival rate to more than 95%.
Beneficial effects of the present invention：
Nutritional ingredient needed for the present invention is directed to Moringa squamous subculture incubation is adjusted, and makes Moringa tissue-cultured seedling wooden
Change degree is improved, and solves the phenomenon of plant jaundice and frangibility during Moringa tissue-cultured seedling；
This experiment improves the tissue culture production technology of Moringa tissue-cultured seedling, using the hand for expanding numerous propagation and progress simultaneously of taking root
Section, realizes that one walks out of seedling, saves cost of labor, improves the economic well-being of workers and staff of production.
The present invention is cultivated, Moringa tissue-cultured seedling is in stem section lignifying journey using Moringa as test material using one kind formula
Degree is improved, and is difficult to be crushed, growth coefficient can reach 6, while rooting rate reaches more than 98%, is transplanted to greenhouse hardening, is survived
Rate realizes the factorial praluction of Moringa more than 95%, successfully.
Nutritional ingredient needed for the present invention is directed to Moringa tissue culture procedures is adjusted, and solves Moringa tissue-cultured seedling tissue
The phenomenon of raw, the fragile frangibility of plant jaundice in incubation, and realize that expanding a numerous, step of taking root completes, and shortens peppery in bottle
In the cycle of wooden tissue culture factorial praluction, cost is saved more than 40%.
Embodiment
Below in conjunction with specific embodiment, make progressive explanation to the present invention.It should be understood that following examples are merely to illustrate this hair
It is bright not for limit the scope of the present invention.
Embodiment 1
A kind of quick breeding by group culture method of Moringa, comprises the following steps：
(1) experiment material must be selected：Select that growing way is neat, plant height be 3.5-4.5cm, subculture cycle be 21-25 days,
Leaf color is used as culture materials in the Moringa tissue-cultured seedling of green or peak green；
(2) specific experiment step：
(2.1) cutting of material：Blade is cut off, retains terminal bud lobus cardiacus, petiole stays 0.1-0.2cm, cuts into 0.5-0.8cm
Stem section, 1-2 axillary bud of every section of band；
(2.2) Moringa squamous subculture；Terminal bud and stem section separate inoculation, are equably inoculated into subculture medium 8 (LM-8)
In；
Wherein, improvement MS (KNO in step (2.2)30.95-1.43g/L, NH4NO30.81-1.24g/L, MgSO4·
7H2O 0.19-0.28g/L, KH2PO40.09-0.13g/L, CaCl2·2H2O 0.22-0.44g/L, Na2EDTA·2H2O
0.002-0.04g/L,FeSO4·7H2O 0.015-0.03g/L,MnSO4·H2O 8.45-16.9mg/L, H3BO3 3.1-
6.2mg/L, ZnSO4·7H2O 4.3-8.6mg/L, KI 0.42-0.83mg/L, Na2MoO4·2H2O 0.13-0.25mg/L,
CoCl2·6H2O 0.01-0.025mg/L, CuSO4·5H2O 0.01-0.025mg/L, inositol 5-10mg/L, nicotinic acid 0.025-
0.05mg/L, thiamine hydrochloride 0.005-0.01mg/L, puridoxine hydrochloride 0.025-0.05mg/L, additional 6-BA0.0005-
0.001mg/L, IBA 0.1mg/L, white granulated sugar 30g/L+ agar 5.5g/L, pH value are adjusted to 6.1.
Wherein, in step (2.2), it is that light application time is 12h/d, intensity of illumination that Moringa squamous subculture, which obtains condition of culture,
2000-3000lux, cultivation temperature is 26 ± 2 DEG C；
Growth performance in each subculture medium is as described in Table 1：
Growth performance in each subculture medium of table 1
Table 1 test result indicates that：The presence of agar and carragheen influences the color and regularity of plant, with carragheen phase
Than the tissue-cultured seedling growth in agar is more neat, although leaf color is in slightly yellow, but does not influence propagation；Contain in Moringa autologous tissue
Natural growth regulatory substance, has there is a small amount of Callus formation in blank cultures, and 6-BA concentration exists
During 0.1mg/L, foam-like calli induction was both inhibited, block callus is also few.Carried out according to LM-4 formula
Operation, growth coefficient can reach 6.01.
Embodiment 2
A kind of quick breeding by group culture method of Moringa, comprises the following steps：
(1) experiment material must be selected：Select that growing way is neat, plant height be 3.5-4.5cm, subculture cycle be 21-25 days,
Leaf color is used as culture materials in the Moringa tissue-cultured seedling of green or peak green；
(2) specific experiment step：
(2.1) cutting of material：Blade is cut off, retains terminal bud lobus cardiacus, petiole stays 0.1-0.2cm, cuts into 0.5-0.8cm
Stem section, 1-2 axillary bud of every section of band；
(2.2) Moringa squamous subculture；Terminal bud and stem section separate inoculation, are equably inoculated into subculture medium 8 (LM-8)
In；
Wherein, improvement MS (KNO in step (2.2) subculture medium 8 (LM-8)30.95-1.43g/L, NH4NO3
0.81-1.24g/L, MgSO4·7H2O 0.19-0.28g/L, KH2PO40.09-0.13g/L, CaCl2·2H2O 0.22-
0.44g/L, Na2EDTA·2H2O 0.002-0.04g/L,FeSO4·7H2O 0.015-0.03g/L,MnSO4·H2O 8.45-
16.9mg/L, H3BO33.1-6.2mg/L, ZnSO4·7H2O 4.3-8.6mg/L, KI 0.42-0.83mg/L, Na2MoO4·
2H2O 0.13-0.25mg/L, CoCl2·6H2O 0.01-0.025mg/L, CuSO4·5H2O 0.01-0.025mg/L, inositol 5-
10mg/L, nicotinic acid 0.025-0.05mg/L, thiamine hydrochloride 0.005-0.01mg/L, puridoxine hydrochloride 0.025-0.05mg/L,
Additional 6-BA0.0005-0.001mg/L, IBA0.1mg/L, white granulated sugar 30g/L+ agar 5.5g/L, pH value are adjusted to 6.1.
Wherein, in step (2.2), it is that light application time is 12h/d, intensity of illumination that Moringa squamous subculture, which obtains condition of culture,
2000-3000lux, cultivation temperature is 26 ± 2 DEG C；
Growth performance in each subculture medium is as described in Table 2：
Growth performance in each subculture medium of table 2
In table 2, Moringa take root operation when, in LM-8 culture mediums obtain tissue-cultured seedling degree of lignification it is relatively high, wash seedling shifting
Lose small during cultivation, therefore squamous subculture is carried out again with the rooted seedling in culture medium, after 30 days, stem with bud is same
It can take root, compared with LM-4 proliferated culture mediums, tissue-cultured seedling is without vitrification phenomenon, while can obtain rooted seedling again.This is in work
A procedure is just reduced in factory's metaplasia production, the production cycle shortens, and corresponding operation is manually also reduced.
(2.3) after culture 28-30d, the rooted seedling of acquisition moves on to greenhouse, and unscrewing bottle cap hardening can transplant for 2-3 days, survive
Rate reaches more than 95%.
The embodiment to the present invention is illustrated above, but the present invention is not limited thereto, without departing from
Spirit of the invention, the present invention can also have various change.
Claims (3)
1. a kind of method for tissue culture of Moringa tissue-cultured seedling, it is characterised in that：Comprise the following steps：
(1) experiment material must be selected：Select that growing way is neat, plant height is that 3.5-4.5cm, subculture cycle are 21-25 days, leaf color
Culture materials are used as in the Moringa tissue-cultured seedling of green or peak green；
(2) specific experiment step：
(2.1) cutting of material：Blade is cut off, retains terminal bud lobus cardiacus, petiole stays 0.1-0.2cm, cuts into 0.5-0.8cm stem
Section, 1-2 axillary bud of every section of band；
(2.2) Moringa squamous subculture；Terminal bud and stem section separate inoculation, are equably inoculated into subculture medium 8 (LM-8)；
(2.3) after culture 21-25d, plant longitudinal growth plant height is sprouted in more than 4.5cm, plant lateral bud, counts multiplied ratio
Averagely reach 1:6.01；
(2.4) after being cultivated 18 days in LM-8 culture mediums, plant height averagely reaches 4cm, and rooting rate reaches 98%, and root length is more than
2cm, root bar number is more than 3；
(2.5) tissue-cultured seedling is tamed：Moringa tissue-cultured seedling of taking root is moved in greenhouse, and unscrewing bottle cap hardening can transplant to sand bed for 2-3 days,
Survival rate is to more than 95%.
2. a kind of method for tissue culture of Moringa tissue-cultured seedling according to claim 1, it is characterised in that：Step (2.2) after
For improvement MS (KNO in culture medium 8 (LM-8)30.95-1.43g/L, NH4NO30.81-1.24g/L, MgSO4·7H2O
0.19-0.28g/L, KH2PO40.09-0.13g/L, CaCl2·2H2O 0.22-0.44g/L, Na2EDTA·2H2O 0.002-
0.04g/L,FeSO4·7H2O 0.015-0.03g/L,MnSO4·H2O 8.45-16.9mg/L, H3BO33.1-6.2mg/L,
ZnSO4·7H2O 4.3-8.6mg/L, KI 0.42-0.83mg/L, Na2MoO4·2H2O 0.13-0.25mg/L, CoCl2·
6H2O 0.01-0.025mg/L, CuSO4·5H2O 0.01-0.025mg/L, inositol 5-10mg/L, nicotinic acid 0.025-0.05mg/
L, thiamine hydrochloride 0.005-0.01mg/L, puridoxine hydrochloride 0.025-0.05mg/L, additional 6-BA 0.0005-0.001mg/
L, IBA 0.1mg/L, white granulated sugar 30g/L+ agar 5.5g/L, pH value are adjusted to 6.1.
3. a kind of method for tissue culture of Moringa tissue-cultured seedling according to claim 1, it is characterised in that：In step (2.2),
The condition of culture of Moringa tissue cultures is that light application time is 12h/d, and intensity of illumination 2000-3000lux, cultivation temperature is 26 ± 2
℃。
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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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| CN104686348A (en) * | 2015-02-24 | 2015-06-10 | 陈桂容 | Tissue culture rapid propagation technique of moringa oleifera Lam. |
| CN105248280A (en) * | 2015-10-30 | 2016-01-20 | 王晓翔 | One-step seedling culturing tissue culture and rapid propagation method of pennisetum hydridum |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| 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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Application publication date: 20170829 |