CN107135952A - A kind of method for promoting tissue culture seedlings of bananas to take root with strengthening root - Google Patents
A kind of method for promoting tissue culture seedlings of bananas to take root with strengthening root Download PDFInfo
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- CN107135952A CN107135952A CN201710571144.6A CN201710571144A CN107135952A CN 107135952 A CN107135952 A CN 107135952A CN 201710571144 A CN201710571144 A CN 201710571144A CN 107135952 A CN107135952 A CN 107135952A
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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
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- 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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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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Abstract
A kind of steps such as the inoculation co-cultured the invention provides method for promoting tissue culture seedlings of bananas to take root with strengthening root, culture, the preparation of banana root media, India's pyriform spore and tissue culture seedlings of bananas including India's pyriform spore.Taken root by using the promotion tissue culture seedlings of bananas that colonizes of India's pyriform spore, this method is simple to operate, shortens the tissue culture seedling rooting cycle, can remarkably promote tissue culture seedlings of bananas and take root and have certain strengthening root effect.Workable, effect of the invention is notable.
Description
Technical field
The present invention relates to Banana Tissue culture field, more particularly to a kind of side for promoting tissue culture seedlings of bananas to take root with strengthening root
Method.
Background technology
Banana is second-biggest-in-the-world fruit, while being also the fourth-largest cereal crops.China is the important original producton location of banana, is planted
Area is planted up to 6,200,000 mu, more than 1,000 ten thousand tons are produced per year, is second largest banana production state in the world.Banana industry has turned into China of China
The important Major agro-industry in southern area.With tradition inhale bud division propagation mode compared with, tissue-cultured seedling have stabilization characteristics of genetics,
The features such as growing way uniformity, breeding are quickly, resistance against diseases is strong, therefore select tissue-cultured seedling in banana production more.Tissue-cultured seedling root system
Number number and fibrous root it is plentiful degree be weigh tissue-cultured seedling quality major criterion, while strengthening seedling and rooting be also determine transplant survival
The key link of rate.
The problem of few number of taking root and weaker root system often being faced in tissue culture seedlings of bananas factorial praluction.India's pyriform spore is earliest
Separate and obtain in the rhizosphere soil of two plants of shrubs in India Thar Desert, be that a kind of class mycorhiza that can carry out pure culture is true
Bacterium, it is typically characterised by its chlamydospore and resembled a pear in shape, and hence obtain one's name India's pyriform spore.Research finds India's pyriform spore to many plantations
The formation of thing root system, grow and number of taking root has obvious facilitation, it is regrettably had not yet to see so far in banana especially
The report applied in tissue culture seedlings of bananas production.If promoting taking root and strengthening root for tissue culture seedlings of bananas using the bacterium, improve root system and live
Power, undoubtedly can be of great advantage to the sound development of tissue culture seedlings of bananas factorial praluction and banana industry.
The content of the invention
, can be by culture it is an object of the invention to provide a kind of method for promoting tissue culture seedlings of bananas to take root with strengthening root
India's pyriform spore fungus block is added in base and promotes tissue culture seedling rooting, increase tissue culture seedling rooting number and rugosity.
To realize the above, the present invention is adopted the following technical scheme that：
A kind of method for promoting tissue culture seedlings of bananas to take root with strengthening root, culture, banana culture of rootage including India's pyriform spore
The steps such as the inoculation that preparation, India's pyriform spore and the tissue culture seedlings of bananas of base are co-cultured.Rush is colonized by using India's pyriform spore
Enter tissue culture seedlings of bananas to take root, this method is simple to operate, shorten the tissue culture seedling rooting cycle, tissue culture seedlings of bananas life can be remarkably promoted
Root simultaneously has certain strengthening root effect.Workable, effect of the invention is notable.
Specifically include following steps：
（1）The culture of India's pyriform spore
Potato is cleaned into peeling to weigh 200 g, tiny bulk or strip is cut into, plus 800ml distilled water enters pot and boiled to horse
Bell potato is well-done, 4 layers of filtered through gauze, weighs 20 g glucose and 20 g agar are separately added into filtrate, distilled water is settled to 1 L simultaneously
Dissolving is boiled, triangular flask mesohigh steam sterilizing is dispensed into（121 DEG C, 20 min）, it is slightly cool after pour into sealing in culture dish plate and treat
With being configured to PDA solid mediums；
India's pyriform spore bacterial strain is activated, a diameter of 5 mm of colony edge fungus block is taken with card punch, access PDA solid mediums are put down
In the middle of plate, 28 DEG C of inversion dark culturings are standby after 7 days；
（2）The preparation of tissue culture seedlings of bananas root media
The formula of root media is the g/L agar of MS+0.3 mg/L NAA+30 g/L sucrose+6, and pH 5.8 boils packing extremely
In tissue culture bottle, every bottle of 30 ml, high pressure steam sterilization（121 DEG C, 20 min）It is stand-by；
（3）India's pyriform spore is co-cultured with tissue culture seedlings of bananas
The tissue culture seedlings of bananas of normal succeeding preservation culture is cut into individual plant seedling, life is transferred to after retaining top vane excision fibrous root
In root culture medium, every bottle connects 2 plants, takes PDA to be inverted 28 DEG C of dark culturings India's pyriform spore of 7 days, colony edge is taken with card punch
A diameter of 5 mm fungus block, accesses the culture medium center of tissue culture seedlings of bananas, apart from plant 1cm or so, each tissue culture bottle connects
Plant India's pyriform spore fungus block, 25 ± 2 DEG C of cultivation temperature, 2000 Lx, the h/d of light application time 12.
Compared with conventional tissue culture seedlings of bananas rooting method, the present invention has advantages below：
（1）India's pyriform spore of the present invention, can manually pure culture, training method is simple, it is easy to prepare with scale.
（2）India's pyriform spore fungus block is inoculated with the medium, and the mode co-cultured with banana is simple to operate, and effect is notable.
（3）The present invention can shorten tissue culture seedlings of bananas and take root the cycle, and batch production tissue culture seedlings of bananas produces general culture of rootage
Cycle is 4-5 weeks or so, and cycle time can be taken root to 3 weeks by adding India's pyriform spore.
（4）Counted and root system rugosity using being taken root invention significantly improves banana, effectively strengthen root activity.
（5）The present invention is applied to a variety of banana germplasm, simply using a kind of collective media with regard to the good hestening rooting of energy,
The production link of effectively save special culture media configuration.
Brief description of the drawings
Fig. 1 is 1 culture of rootage of embodiment Sanming City wild banana root growth situation map of 30 days；
Fig. 2 is 2 culture of rootage of embodiment, 30 days ' Tian Bao any of several broadleaf plants ' root growth situation map；
Fig. 3 is 3 culture of rootage of embodiment No. 1 root growth situation map of Foochow dwarf banana of 30 days；
Fig. 4 is 4 culture of rootage of embodiment Foochow bajiao banana root system growing state figure of 30 days；
Note：-P.indicaFor without India's pyriform spore control；+P.indicaFor inoculation India pyriform spore fungus block.
Embodiment
With reference to specific embodiment, with In Fujian Province characteristic germplasm wild banana（Sanming City wild banana）With cultivation any of several broadleaf plants（' my god
Precious any of several broadleaf plants ', Foochow dwarf banana No. 1, Foochow bajiao banana）For material, technical scheme is described further, but the present invention is not
It is only limitted to this.
Embodiment 1：
1st, the culture of India's pyriform spore
Potato is cleaned into peeling to weigh 200 g, tiny bulk or strip is cut into, plus 800ml distilled water enters pot and boils 20-30
Min is well-done to potato, 4 layers of filtered through gauze, weighs 20 g glucose and 20 g agar are separately added into filtrate, distilled water constant volume
To 1 L and dissolving is boiled, be dispensed into triangular flask mesohigh steam sterilizing（121 DEG C, 20 min）, it is slightly cool after pour into culture dish plate
Sealing is stand-by, is configured to PDA solid mediums.
India's pyriform spore bacterial strain is activated, a diameter of 5 mm of colony edge fungus block is taken with card punch, PDA solid cultures are accessed
In the middle of base flat board, 28 DEG C of inversion dark culturings are standby after 7 days.
2nd, the preparation of tissue culture seedlings of bananas root media
The formula of root media is the g/L agar of MS+0.3 mg/L NAA+30 g/L sucrose+6, and pH 5.8 boils packing extremely
In tissue culture bottle, every bottle of 30 ml or so, high pressure steam sterilization（121 DEG C, 20 min）It is stand-by.
3rd, India's pyriform spore and tissue culture seedlings of bananas are co-cultured
Sanming City wild banana tissue-cultured seedling of normal succeeding preservation culture is cut into individual plant seedling, turned after retaining top vane excision fibrous root
It is connected in root media, every bottle connects 2 plants, takes PDA to be inverted 28 DEG C of dark culturings India's pyriform spore of 7 days, bacterium is taken with card punch
Fall the fungus block that edge diameter is 5 mm, the culture medium center of tissue culture seedlings of bananas is accessed, apart from plant 1cm or so, each group
Train bottle and be inoculated with India's pyriform spore fungus block.25 ± 2 DEG C of cultivation temperature, 2000 Lx, the h/d of light application time 12.
It is inoculated with after India's pyriform spore every Sanming City wild banana tissue-cultured seedling plant height of measurement in 10 days, the number of blade and number of taking root,
Photographed to record after 30 days and see Fig. 1, data concrete outcome is shown in Table 1, table 2.India's pyriform spore is inoculated with it can be seen from table 1, table 2
Sanming City wild banana tissue-cultured seedling plant height and the number of blade and control group difference be not obvious；And averagely take root and count apparently higher than control group,
Especially after inoculation India pyriform spore 10 days and 20 days difference it is extremely notable, averagely taking root for counting when being inoculated with 30 days is still significantly higher than
Control group, efficiently solves the problem of Sanming City wild banana takes root weak and difficult.
Influence of the India's pyriform spore of table 1 to Sanming City wild banana tissue-cultured seedling plant height, the number of blade
Influence of the India's pyriform spore of table 2 to Sanming City wild banana tissue culture seedling rooting number
Embodiment 2：
1st, the culture of India's pyriform spore
Potato is cleaned into peeling to weigh 200 g, tiny bulk or strip is cut into, plus 800 ml distilled water enter pot and boil 20-
30 min are well-done to potato, 4 layers of filtered through gauze, weigh 20 g glucose and 20 g agar are separately added into filtrate, distilled water
It is settled to 1 L and boils dissolving, is dispensed into triangular flask mesohigh steam sterilizing（121 DEG C, 2 0min）, it is slightly cool after pour into culture dish
Seal stand-by in plate, be configured to PDA solid mediums.
India's pyriform spore bacterial strain is activated, a diameter of 5 mm of colony edge fungus block is taken with card punch, PDA solid cultures are accessed
In the middle of base flat board, 28 DEG C of inversion dark culturings are standby after 7 days.
2nd, the preparation of tissue culture seedlings of bananas root media
The formula of root media is the g/L agar of MS+0.3 mg/L NAA+30 g/L sucrose+6, and pH5.8 boils packing to group
Train in bottle, every bottle of 30 ml or so, high pressure steam sterilization（121 DEG C, 20 min）It is stand-by.
3rd, India's pyriform spore and tissue culture seedlings of bananas are co-cultured
' Tian Bao any of several broadleaf plants ' tissue-cultured seedling of normal succeeding preservation culture is cut into individual plant seedling, transferred after retaining top vane excision fibrous root
Into root media, every bottle connects 2 plants, takes PDA to be inverted 28 DEG C of dark culturings India's pyriform spore of 7 days, bacterium colony is taken with card punch
Edge diameter is 5mm fungus block, the culture medium center of tissue culture seedlings of bananas is accessed, apart from cm of plant 1 or so, each tissue culture
Bottle one India's pyriform spore fungus block of inoculation.25 ± 2 DEG C of cultivation temperature, 2000 Lx, the h/d of light application time 12.
' Tian Bao any of several broadleaf plants ' tissue-cultured seedling plant height, number of blade, take root number and root were measured once after inoculation India pyriform spore every 10 days
It is long, photographed to record after 30 days and see Fig. 2, data concrete outcome is shown in Table 3, table 4.As can be seen that inoculation India pyriform from table 3, table 4
Tian Bao any of several broadleaf plants tissue-cultured seedling plant height average value of spore is slightly above control group；20 days and 30 days after inoculation, number of blade average value was significantly high
In control group, average root length was significantly better than control at 20 days；10 days after inoculation, it is vaccinated with Tian Bao any of several broadleaf plants group of India's pyriform spore
Training seedling average pole of taking root is significantly higher than control group, and root system is more vigorous, and fibrous root number and root length are significantly better than control, and root activity is more
By force.
Influence of the India's pyriform spore of table 3 to ' Tian Bao any of several broadleaf plants ' tissue-cultured seedling plant height, the number of blade
The India's pyriform spore of table 4 is to ' Tian Bao any of several broadleaf plants ' tissue culture seedling rooting number, the influence of root length
Embodiment 3：
1st, the culture of India's pyriform spore
Potato is cleaned into peeling to weigh 200 g, tiny bulk or strip is cut into, plus 800ml distilled water enters pot and boils 20-30
Min is well-done to potato, 4 layers of filtered through gauze, weighs 20 g glucose and 20 g agar are separately added into filtrate, distilled water constant volume
To 1 L and dissolving is boiled, be dispensed into triangular flask mesohigh steam sterilizing（121 DEG C, 20 min）, it is slightly cool after pour into culture dish plate
Sealing is stand-by, is configured to PDA solid mediums.
India's pyriform spore bacterial strain is activated, a diameter of 5 mm of colony edge fungus block is taken with card punch, PDA solid cultures are accessed
In the middle of base flat board, 28 DEG C of inversion dark culturings are standby after 7 days.
2nd, the preparation of tissue culture seedlings of bananas root media
The formula of root media is the g/L agar of MS+0.3 mg/L NAA+30 g/L sucrose+6, and pH 5.8 boils packing extremely
In tissue culture bottle, every bottle of 30 ml or so, high pressure steam sterilization（121 DEG C, 20 min）It is stand-by.
3rd, India's pyriform spore and tissue culture seedlings of bananas are co-cultured
No. 1 tissue-cultured seedling of Foochow dwarf banana of normal succeeding preservation culture is cut into individual plant seedling, retained after top vane excision fibrous root
It is transferred in root media, every bottle connects 2 plants, takes PDA to be inverted 28 DEG C of dark culturings India's pyriform spore of 7 days, taken with card punch
A diameter of 5 mm of colony edge fungus block, accesses the culture medium center of tissue culture seedlings of bananas, apart from cm of plant 1 or so, each
Tissue culture bottle is inoculated with India's pyriform spore fungus block.25 ± 2 DEG C of cultivation temperature, 2000 Lx, the h/d of light application time 12.
It is inoculated with after India's pyriform spore every No. 1 tissue-cultured seedling plant height of 10 days Foochow dwarf bananas of measurement, the number of blade, number of taking root,
Photographed to record after 30 days and see Fig. 3, data concrete outcome is shown in Table 5, table 6.As can be seen that inoculation India pyriform spore from table 5, table 6
No. 1 tissue-cultured seedling plant height of Foochow dwarf banana be significantly higher than within 20 days control after inoculation, be above on the number of blade and several average values of taking root
Control group, root system is more vigorous plentiful and more sturdy.
Influence of the India's pyriform spore of table 5 to No. 1 tissue-cultured seedling plant height of Foochow dwarf banana, the number of blade
Influence of the India's pyriform spore of table 6 to No. 1 tissue culture seedling rooting number of Foochow dwarf banana
Embodiment 4：
1st, the culture of India's pyriform spore
Potato is cleaned into peeling to weigh 200 g, tiny bulk or strip is cut into, plus 800ml distilled water enters pot and boils 20-30
Min is well-done to potato, 4 layers of filtered through gauze, weighs 20 g glucose and 20 g agar are separately added into filtrate, distilled water constant volume
To 1 L and dissolving is boiled, be dispensed into triangular flask mesohigh steam sterilizing（121 DEG C, 20min）, it is slightly cool after pour into culture dish plate
Sealing is stand-by, is configured to PDA solid mediums.
India's pyriform spore bacterial strain is activated, a diameter of 5 mm of colony edge fungus block is taken with card punch, PDA solid cultures are accessed
In the middle of base flat board, 28 DEG C are inverted dark culturing 7 days, obtain India's pyriform spore.
2nd, the preparation of tissue culture seedlings of bananas root media
The formula of root media is the g/L agar of MS+0.3 mg/L NAA+30 g/L sucrose+6, and pH 5.8 boils packing extremely
In tissue culture bottle, every bottle of 30 ml or so, high pressure steam sterilization（121 DEG C, 20 min）It is stand-by.
3rd, India's pyriform spore and tissue culture seedlings of bananas are co-cultured
Foochow bajiao banana tissue-cultured seedling of normal succeeding preservation culture is cut into individual plant seedling, transferred after retaining top vane excision fibrous root
Into root media, every bottle connects 2 plants, takes PDA to be inverted 28 DEG C of dark culturings India's pyriform spore of 7 days, bacterium colony is taken with card punch
Edge diameter is 5 mm fungus block, the culture medium center of tissue culture seedlings of bananas is accessed, apart from cm of plant 1 or so, each tissue culture
Bottle one India's pyriform spore fungus block of inoculation.25 ± 2 DEG C of cultivation temperature, 2000 Lx, the h/d of light application time 12.
It is inoculated with after India's pyriform spore every No. 1 tissue-cultured seedling plant height of 10 days Foochow dwarf bananas of measurement, the number of blade, number of taking root,
Photographed to record after 30 days and see Fig. 4, data concrete outcome is shown in Table 7, table 8.India's pyriform spore is inoculated with it can be seen from table 7, table 8
In Foochow bajiao banana tissue-cultured seedling plant height, blade with control group difference less, take root and count upper extremely notable with control group difference, about
For 2 times of control, root system is more vigorous and more sturdy, and growth-promoting effect is obvious.
Influence of the India's pyriform spore of table 7 to Foochow bajiao banana tissue-cultured seedling plant height, the number of blade
Influence of the India's pyriform spore of table 8 to Foochow bajiao banana tissue culture seedling rooting number
As can be seen from the above embodiments：Promote the effect of plant establishment using India's pyriform spore, be seeded to culture of rootage
Co-cultured in base with tissue culture seedlings of bananas, take root number and the root system rugosity of banana seedlings can be remarkably promoted within 30 days after inoculation, and by reality
Test and draw, can reach conventional tissue culture transplantation of seedlings requirement within 3 weeks or so in inoculation India pyriform spore, shorten tissue culture seedlings of bananas and take root
In the cycle, it is worth with larger production application.In this experiment, using same root media, in access India pyriform spore
The growth of 4 kinds of banana germplasm tissue-cultured seedling root systems more than promoting afterwards.In addition, the banana seedlings obtained using the present invention have India's pears
Shape spore is colonized, and can promote absorption of the banana seedlings to soil media, growth and the anti-adversity ability of banana seedlings is improved, with high
Economic value.
Above content is to combine the further description that specific preferred embodiment is made to the present invention, it is impossible to assert this
The specific implementation of invention is confined to these explanations, for general technical staff of the technical field of the invention, not
On the premise of departing from present inventive concept, some simple deduction or replace can also be made, the present invention should be all considered as belonging to and be carried
The protection domain that claims of friendship are determined.
Claims (1)
1. a kind of method for promoting tissue culture seedlings of bananas to take root with strengthening root, it is characterised in that：Comprise the following specific steps that：
（1）The culture of India's pyriform spore
Potato is cleaned into peeling to weigh 200 g, tiny bulk or strip is cut into, plus 800ml distilled water enters pot and boiled to horse
Bell potato is well-done, 4 layers of filtered through gauze, weighs 20 g glucose and 20 g agar are separately added into filtrate, distilled water is settled to 1 L simultaneously
Boil dissolving, be dispensed into triangular flask mesohigh steam sterilizing, it is slightly cool after pour into culture dish plate and seal stand-by, be configured to PDA and consolidate
Body culture medium；
India's pyriform spore bacterial strain is activated, a diameter of 5 mm of colony edge fungus block is taken with card punch, access PDA solid mediums are put down
In the middle of plate, 28 DEG C of inversion dark culturings are standby after 7 days；
（2）The preparation of tissue culture seedlings of bananas root media
The formula of root media is the g/L agar of MS+0.3 mg/L NAA+30 g/L sucrose+6, and pH 5.8 boils packing extremely
In tissue culture bottle, every bottle of 30 ml, high pressure steam sterilization is stand-by；
（3）India's pyriform spore is co-cultured with tissue culture seedlings of bananas
The tissue culture seedlings of bananas of normal succeeding preservation culture is cut into individual plant seedling, life is transferred to after retaining top vane excision fibrous root
In root culture medium, every bottle connects 2 plants, takes PDA to be inverted 28 DEG C of dark culturings India's pyriform spore of 7 days, colony edge is taken with card punch
A diameter of 5 mm fungus block, accesses the culture medium center of tissue culture seedlings of bananas, apart from plant 1cm, each tissue culture bottle inoculation one
Individual India's pyriform spore fungus block, 25 ± 2 DEG C of cultivation temperature, 2000 Lx, the h/d of light application time 12.
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| CN107778090A (en) * | 2017-11-30 | 2018-03-09 | 福建农林大学 | A kind of preparation for promoting stock root growth |
| CN109122095A (en) * | 2018-08-10 | 2019-01-04 | 福建农林大学 | A method of it cultivating high-quality passionflower seedling and improves plant resistance |
| CN110122304A (en) * | 2019-06-14 | 2019-08-16 | 福建农林大学 | A kind of method of banana water planting hardening |
| CN110923187A (en) * | 2019-10-31 | 2020-03-27 | 长江大学 | Agricultural waste product culture medium suitable for growth of piriformospora indica |
| CN112438169A (en) * | 2019-08-30 | 2021-03-05 | 贵州省果树科学研究所 | Planting method for improving nutrient content of pitaya |
| CN116138164A (en) * | 2022-07-06 | 2023-05-23 | 三明市农业科学研究院 | Gerbera strong seedling cultivation method |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107778090A (en) * | 2017-11-30 | 2018-03-09 | 福建农林大学 | A kind of preparation for promoting stock root growth |
| CN109122095A (en) * | 2018-08-10 | 2019-01-04 | 福建农林大学 | A method of it cultivating high-quality passionflower seedling and improves plant resistance |
| CN110122304A (en) * | 2019-06-14 | 2019-08-16 | 福建农林大学 | A kind of method of banana water planting hardening |
| CN112438169A (en) * | 2019-08-30 | 2021-03-05 | 贵州省果树科学研究所 | Planting method for improving nutrient content of pitaya |
| CN110923187A (en) * | 2019-10-31 | 2020-03-27 | 长江大学 | Agricultural waste product culture medium suitable for growth of piriformospora indica |
| CN116138164A (en) * | 2022-07-06 | 2023-05-23 | 三明市农业科学研究院 | Gerbera strong seedling cultivation method |
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