CN106172011B - A kind of dwarf banana high temperature light culture tissue culture and rapid propagation method - Google Patents
A kind of dwarf banana high temperature light culture tissue culture and rapid propagation method Download PDFInfo
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- CN106172011B CN106172011B CN201610711045.9A CN201610711045A CN106172011B CN 106172011 B CN106172011 B CN 106172011B CN 201610711045 A CN201610711045 A CN 201610711045A CN 106172011 B CN106172011 B CN 106172011B
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- high temperature
- temperature light
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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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- 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
Abstract
The present invention relates to field of plant tissue culture technique, a kind of particularly dwarf banana high temperature light culture tissue culture and rapid propagation method.A kind of dwarf banana high temperature light culture tissue culture and rapid propagation method, includes the following steps：Explant preparation, squamous subculture, culture of rootage.The method subculture bud of the present invention is sprouted, and early, differentiation is early, differentiation is fast, and breeding coefficient is high, and effective bud rate is high；Rooted seedling planting percent is high, and kind seedling leaf and false stem good toughness, root system is not easily to fall off, and temporary planting survival rate is high.Using high temperature light culture technology, subculture cycle shortens 10~15d, has and reduces energy consumption, improves space availability ratio, improves production efficiency, reduces production cost advantage.
Description
Technical field
The present invention relates to field of plant tissue culture technique, a kind of particularly dwarf banana high temperature light culture tissue-culturing rapid propagation side
Method.
Background technology
Dwarf banana (Musa ABB Pisang Awak) also known as Annan any of several broadleaf plants, Saigon any of several broadleaf plants and milk any of several broadleaf plants etc., it is (referred to as fragrant with fragrant tooth any of several broadleaf plants
Any of several broadleaf plants) (Musa AAA Cavendish) belong to the two big types of China staple food any of several broadleaf plants.Mainstream kind " bronze 1 currently on the market
Number " due to fruit thin skin, pulp color is yellowish or yellow, and meat is slided, sweet, is had micro-perfume and is liked by consumers in general.Closely
In the past few years dwarf banana not only rises year by year in south China consumption figure, and northerly market also begins to open, and dwarf banana sales volume is gradual
Increase.With the expansion of dwarf banana industry, the dwarf banana seedling of high-quality health is badly in need of merchandized handling, the seedling tissue-culturing rapid propagation skill of dwarf banana
Art just seems especially important.In the dwarf banana tissue-culturing rapid propagation of early stage, the squamous subculture stage there are breeding coefficient it is low, " Buds of The Pineapple " it is more,
Steep the problems such as bud ratio is big, simple bud rate is high, period length (reaching more than 40d)；The culture of rootage stage there are rooting rate is low, radical is few,
The problems such as root is long, root is easy to fall off, the production of dwarf banana tissue culture is caused to occur, and variation risk is big, pollutes high, low production efficiency and production
The drawbacks such as of high cost.In production generally breeding coefficient is adjusted by changing the phytohormone concentration in culture medium.Improve plant
Hormone concentration can improve breeding coefficient, but can increase the risk of variation simultaneously；And the concentration for reducing plant hormone can reduce change
Different rate, but breeding coefficient also decreases, it cannot be guaranteed that the high-quality dwarf banana seedling of rapid, high volume breeding health.
At present, there is the influence that scholar has studied different temperatures and different illumination grow tissue culture seedlings of bananas, research shows that warm
Degree is proliferated tissue culture seedlings of bananas and takes root and have pole significant impact, but related is reported to what the illumination of dwarf banana tissue-cultured seedling and temperature regulates and controls
Road is seldom.
Research team where inventor compares by practice for many years and multiple experiment, by adjusting " bronze 1 "
The temperature condition and illumination condition of dwarf banana seedling tissue-culturing rapid propagation, exploration obtain a kind of dwarf banana high temperature light culture tissue culture and rapid propagation method, should
Method maintains low aberration rate while higher breeding coefficient is obtained, and ensure that the quality of seedling, improves production efficiency, into
One step reduces production cost.
Invention content
The object of the present invention is to provide a kind of dwarf banana high temperature light culture tissue culture and rapid propagation method, this method is obtaining higher breeding
Low aberration rate is maintained while coefficient, the quality of seedling is ensure that, improves production efficiency, further reduced and be produced into
This.
Technical solution provided by the present invention is：
A kind of dwarf banana high temperature light culture tissue culture and rapid propagation method, includes the following steps：
(1) explant preparation and Primary culture：It acquires the excellent 50~100cm high of elite stand of dwarf banana and inhales bud, rinsed through tap water
Totally, extra leaves and stems are cut off, appearance is cleaned 2~3 times with alcohol, peels leaf sheath off on superclean bench and be whittled into growing point
Centered on diameter 3cm, high 5cm cylinder scapus, then it is 0.3~0.5cm to separate the length of side from cylinder scapus center
The small sprout tuber of blockage, as explant is inoculated into 25~30d of culture in primary culture medium, obtains clump bud；
(2) squamous subculture；The clump bud of activated culture is inoculated on subculture medium, is cultivated under the conditions of high temperature light culture
25~30d；
(3) culture of rootage：13~16d of culture in root media is first placed under the conditions of high temperature light culture；It is put again
15~25d is cultivated under the conditions of natural scattered light, obtains tissue-cultured seedling of taking root；
(4) hardening：When under growth root tissue-cultured seedling grows supreme 3~5cm, has 3~4 expansion leaves, it is moved out of culturing room
Go out, be placed on the shady and cool place in outdoor with natural scattered light condition, 7~10d of hardening obtains rooted seedling；
(5) it is planted：Rooted seedling is taken out, washes away root culture medium, is planted in greenhouse in sand bed, drenching sufficient root water, and hide
The moon is periodically sprayed water, and relative air humidity is 75~85%, and temperature is 27~35 DEG C.
Preferably, step (2), step (3) high temperature light culture condition of culture are：32~35 DEG C of temperature, illumination are<
10Lux。
Preferably, the primary culture medium and subculture medium of step (1) and step (2) are MS culture mediums+6-BA
2.0~3.0mg/L+IBA0.2mg/L.
Preferably, root media is MS culture medium+NAA 1.0mg/L+IBA 0.2mg/L, pH in step (3)
5.8。
Preferably, there is matrix in step (5) in sand bed, the matrix is by pig manure：Soil：Peat soil is by volume
3:85:12 compositions.
Preferably, the preparation method of the matrix is：Pig manure, soil and peat soil, mixing is measured to be spread according to seedling growth greenhouse
Seedbed is set as, covers the river sand of 2cm thickness above, disinfection is applied with 800 times of carbendazim leaching.
Compared with prior art, the present invention has the advantages that：
(1) cylinder scapus is not required to using disinfection in method of the invention, directly separates explant, improves work effect
Rate, while avoid injury of the antiseptic solutions such as mercuric chloride to explant.
(2) method subculture bud of the invention is sprouted, and early, differentiation is early, differentiation is fast, and breeding coefficient is high, and effective bud rate is high.
(3) the rooted seedling planting percent that method of the invention obtains is high, and kind seedling leaf and false stem good toughness are not easily broken, and
It is high to be planted survival rate.
(4) method subculture cycle of the invention shortens 10~15d, has and reduces energy consumption, improves space availability ratio, improves life
It produces efficiency, reduce production cost advantage.
Specific embodiment
With reference to concrete example, the present invention is described in further detail.
Embodiment 1：
A kind of dwarf banana high temperature light culture tissue culture and rapid propagation method, includes the following steps：
(1) explant preparation and Primary culture：It acquires the excellent elite stand 50cm high of dwarf banana and inhales bud, rinsed well through tap water,
Extra leaves and stems are cut off, appearance is cleaned 2 times with alcohol, peels leaf sheath off on superclean bench and be whittled into centered on growing point
The cylinder scapus of diameter 3cm, high 5cm, then the blockage that the length of side is 0.3cm is separated from cylinder scapus center, it is as outer
The small sprout tuber of implant, is inoculated into primary culture medium and cultivates 25d, obtain clump bud；Primary culture medium is MS culture mediums+6-BA
2.0mg/L+IBA0.2mg/L；
(2) squamous subculture；The clump bud of activated culture is inoculated on subculture medium, is cultivated under the conditions of high temperature light culture
25d；High temperature light culture condition of culture is：32 DEG C of temperature, illumination are<10Lux；Primary culture medium and subculture medium are MS trainings
Support base+6-BA 2.0mg/L+IBA0.2mg/L；
(3) culture of rootage：It is first placed in root media under the conditions of high temperature light culture and cultivates 13d；It places it in again certainly
15d so is cultivated under the conditions of scattering light, obtains tissue-cultured seedling of taking root；High temperature light culture condition of culture is：32 DEG C of temperature, illumination are<
10Lux；Root media is MS culture medium+NAA 1.0mg/L+IBA 0.2mg/L, pH 5.8；
(4) hardening：When under growth root tissue-cultured seedling grows supreme 3cm, has 3 expansion leaves, it out of culturing room is removed, is placed on
The shady and cool place in outdoor with natural scattered light condition, hardening 7d obtain rooted seedling；
(5) it is planted：Rooted seedling is taken out, washes away root culture medium, is planted in greenhouse in sand bed, drenching sufficient root water, and hide
The moon is periodically sprayed water, relative air humidity 75%, and temperature is 27 DEG C；There is matrix in sand bed, the matrix is by pig manure：Soil：
Peat soil is 3 by volume:85:12 compositions；The preparation method of the matrix is：Measurement pig manure, soil and peat soil, mixing,
Seedbed is laid to according to seedling growth greenhouse, covers the river sand of 2cm thickness above, disinfection is applied with 800 times of carbendazim leaching.
Embodiment 2：
A kind of dwarf banana high temperature light culture tissue culture and rapid propagation method, includes the following steps：
(1) explant preparation and Primary culture：It acquires the excellent elite stand 100cm high of dwarf banana and inhales bud, rinsed well through tap water,
Extra leaves and stems are cut off, appearance is cleaned 3 times with alcohol, peels leaf sheath off on superclean bench and be whittled into centered on growing point
The cylinder scapus of diameter 3cm, high 5cm, then the blockage that the length of side is 0.5cm is separated from cylinder scapus center, it is as outer
The small sprout tuber of implant, is inoculated into primary culture medium and cultivates 30d, obtain clump bud；Primary culture medium is MS culture mediums+6-BA
3.0mg/L+IBA0.2mg/L；
(2) squamous subculture；The clump bud of activated culture is inoculated on subculture medium, is cultivated under the conditions of high temperature light culture
30d；High temperature light culture condition of culture is：35 DEG C of temperature, illumination are<10Lux；Primary culture medium and subculture medium are MS trainings
Support base+6-BA3.0mg/L+IBA0.2mg/L；
(3) culture of rootage：It is first placed in root media under the conditions of high temperature light culture and cultivates 16d；It places it in again certainly
25d so is cultivated under the conditions of scattering light, obtains tissue-cultured seedling of taking root；High temperature light culture condition of culture is：35 DEG C of temperature, illumination are<
10Lux；Root media is MS culture medium+NAA 1.0mg/L+IBA 0.2mg/L, pH 5.8；
(4) hardening：When under growth root tissue-cultured seedling grows supreme 5cm, has 4 expansion leaves, it out of culturing room is removed, is placed on
The shady and cool place in outdoor with natural scattered light condition, hardening 10d obtain rooted seedling；
(5) it is planted：Rooted seedling is taken out, washes away root culture medium, is planted in greenhouse in sand bed, drenching sufficient root water, and hide
The moon is periodically sprayed water, relative air humidity 85%, and temperature is 35 DEG C；There is matrix in sand bed, the matrix is by pig manure：Soil：
Peat soil is 3 by volume:85:12 compositions；The preparation method of the matrix is：Measurement pig manure, soil and peat soil, mixing,
Seedbed is laid to according to seedling growth greenhouse, covers the river sand of 2cm thickness above, disinfection is applied with 800 times of carbendazim leaching.
Embodiment 3：
A kind of dwarf banana high temperature light culture tissue culture and rapid propagation method, includes the following steps：
(1) explant preparation and Primary culture：It acquires the excellent elite stand 80cm high of dwarf banana and inhales bud, rinsed well through tap water,
Extra leaves and stems are cut off, appearance is cleaned 3 times with alcohol, peels leaf sheath off on superclean bench and be whittled into centered on growing point
The cylinder scapus of diameter 3cm, high 5cm, then the blockage that the length of side is 0.4cm is separated from cylinder scapus center, it is as outer
The small sprout tuber of implant, is inoculated into primary culture medium and cultivates 28d, obtain clump bud；Primary culture medium is MS culture mediums+6-BA
2.5mg/L+IBA0.2mg/L；
(2) squamous subculture；The clump bud of activated culture is inoculated on subculture medium, is cultivated under the conditions of high temperature light culture
28d；High temperature light culture condition of culture is：33 DEG C of temperature, illumination are<10Lux；Primary culture medium and subculture medium are MS trainings
Support base+6-BA 2.2mg/L+IBA0.2mg/L；
(3) culture of rootage：It is first placed in root media under the conditions of high temperature light culture and cultivates 15d；It places it in again certainly
20d so is cultivated under the conditions of scattering light, obtains tissue-cultured seedling of taking root；High temperature light culture condition of culture is：33 DEG C of temperature, illumination are<
10Lux；Root media is MS culture medium+NAA 1.0mg/L+IBA 0.2mg/L, pH 5.8；
(4) hardening：When under growth root tissue-cultured seedling grows supreme 4cm, has 3 expansion leaves, it out of culturing room is removed, is placed on
The shady and cool place in outdoor with natural scattered light condition, hardening 8d obtain rooted seedling；
(5) it is planted：Rooted seedling is taken out, washes away root culture medium, is planted in greenhouse in sand bed, drenching sufficient root water, and hide
The moon is periodically sprayed water, relative air humidity 80%, and temperature is 30 DEG C；There is matrix in sand bed, the matrix is by pig manure：Soil：
Peat soil is 3 by volume:85:12 compositions；The preparation method of the matrix is：Measurement pig manure, soil and peat soil, mixing,
Seedbed is laid to according to seedling growth greenhouse, covers the river sand of 2cm thickness above, disinfection is applied with 800 times of carbendazim leaching.
Embodiment 4：The implementation result of the method for the present invention
Condition of culture：3 temperature gradients and 2 lighting gradients, respectively high temperature (32-35 DEG C), medium temperature (28-30 are set
DEG C), room temperature (24-26 DEG C)；Light (2000Lux, illumination 12h/d), it is dark (<10Lux hides upper all curtain daytime, does not turn on light i.e.
Can).6 different illumination and treatment of different temperature combination are set altogether, respectively：High temperature light culture (method of the invention), high temperature
Optical culture (control 1), medium temperature light culture (control 2), medium temperature optical culture (control 3), room temperature light culture (control 4) and normal-temperature light training
It supports (control 5).
It should be noted that in the processing of control 1-5, the condition of culture in step (2), step (3) is different, other steps
Operation it is identical with embodiment 1-3.
Influence of the method for 4.1 present invention to subculture cycle and breeding coefficient
Influence of the method for 1 present invention of table to squamous subculture breeding coefficient
Note：There are significant difference (P between the different alphabetical expression processing of same column<0.05), similarly hereinafter.
As shown in Table 1, high temperature light culture processing breeding coefficient is being inoculated with 12d up to 2.0 or so (1.98), and to impinging upon inoculation
Just reach 2.0 or so (being respectively 2.01,2.01,1.91,2.02,1.97) during 25d, the processing of high temperature light culture is faster than control treatment
13d illustrates that high temperature light culture can promote the bud to sprout, break up that early, differentiation is fast.The differentiation of subculture bud is fast, and bud is grown in a short time
Time it is just more, bud is just relatively strong, can shorten subculture cycle, it is more than 40d that early stage dwarf banana tissue culture, which produces subculture cycle, and sharp
With high temperature light culture technology, subculture cycle shorten to 25~30d, saves the squamous subculture time of 10~15d.Squamous subculture
After 25d, the breeding coefficient of the embodiment of the present invention 1-3 reaches as high as 2.62, is all higher than other 5 control treatments, illustrates this hair
The method for tissue culture of bright dwarf banana can improve breeding coefficient.
Influence of the method for 4.2 present invention to subculture bud quality
The method of the present invention of table 2 is to subculture bud into the influence of bud rate and growing state
As shown in Table 2, the effective bud rates of the embodiment of the present invention 1-3 are maximum, are 96.74%, illustrate the dwarf banana of the present invention
Method for tissue culture can improve effective bud rate.
The blade of subculture bud expansion and the root grown increase the risk polluted during switching, increase workload, reduce work effect
Rate.The embodiment of the present invention 1-3 buds expansion number of blade is 0, is more conducive to subculture, reduces pollution risk during switching, improves
Working efficiency.
Foaming bud is to weigh the important indicator that squamous subculture bud is abnormal and makes a variation, and foaming bud is more, illustrates subculture bud
More abnormal, the risk of variation is bigger, and bud foaming is also unfavorable for further transferring.The embodiment of the present invention 1-3 foaming bud rate is most
Small, subculture bud is high-quality.
Influence of the method for 4.3 present invention to rooted seedling and temporary planting
Influence of the method for 3 present invention of table to rooted seedling planting percent and growing state
As shown in table 3, the planting percent highest of taking root of method culture of rootage of the invention, reaches 100% planting percent, shows
It writes higher than control treatment, rooted seedling height of seedling, average radical and the expansion number of blade per young plant can reach production requirement.
Influence of the method for 4 present invention of table to sand culture shoot survival percent
By seedling temporary planting in preprepared sand bed, after being planted 3 weeks, survival rate is investigated.As shown in table 4,
It is 100% that the method for the present invention, which obtains processing shoot survival percent, higher than other control groups.
Influence of the method for 4.4 present invention to energy consumption and space utilization situation
The culturing rack for being 1.5m × 0.5m with individual layer length and width in production, each 4 layers of shelf.The method of the present invention is after being commissioned to train
Placement can be laminated in the stage of supporting, culture bottle, and every frame can put 1008 bottles, and culture bottle cannot stack during optical culture, and every frame can only
432 bottles are put, therefore, method of the invention improves 133.33% compared with optical culture mode space availability ratio.Per layer frame during optical culture
The fluorescent tube of 2 40W of son installation, a culturing rack each squamous subculture period (unified to be calculated by 25d) equivalent 96.00 degree of electricity consumption,
The method of the present invention can then save this part energy consumption, while also enhance production security.Therefore, subculture light culture pair
Energy consumption is reduced, space availability ratio is improved, improves tissue culture production efficiency, reducing production cost and improve benefit and have remarkable result.
In conclusion the method subculture bud of the present invention is sprouted, early, differentiation is early, differentiation is fast, and breeding coefficient is high, effective bud rate
It is high；Rooted seedling planting percent is high, and kind seedling leaf and false stem good toughness, root system is not easily to fall off, and temporary planting survival rate is high.It is secretly trained using high temperature
The technology of supporting, subculture cycle shorten 10~15d, have and reduce energy consumption, improve space availability ratio, improve production efficiency, reduce life
Produce cost advantages.
The description of the aforementioned specific exemplary embodiment to the present invention is in order to illustrate and illustration purpose.These descriptions
It is not wishing to limit the invention to disclosed precise forms, and it will be apparent that according to the above instruction, can much be changed
And variation.The purpose of selecting and describing the exemplary embodiment is that explain that the specific principle of the present invention and its reality should
With so that those skilled in the art can realize and utilize the present invention a variety of different exemplary implementations and
Various chooses and changes.The scope of the present invention is intended to be limited by claims and its equivalents.
Claims (3)
1. a kind of dwarf banana high temperature light culture tissue culture and rapid propagation method, which is characterized in that include the following steps：
(1) explant preparation and Primary culture：It acquires the excellent 50~100cm high of elite stand of dwarf banana and inhales bud, rinsed well through tap water,
Extra leaves and stems are cut off, appearance is cleaned 2~3 times with alcohol, peels leaf sheath off on superclean bench and be whittled into using growing point in
The diameter 3cm of the heart, the cylinder scapus of high 5cm, then separate the small side that the length of side is 0.3~0.5cm from cylinder scapus center
The small sprout tuber of block, as explant is inoculated into 25~30d of culture in primary culture medium, obtains clump bud；
(2) squamous subculture；The clump bud of activated culture is inoculated on subculture medium, under the conditions of high temperature light culture cultivate 25~
30d；
(3) culture of rootage：13~16d of culture in root media is first placed under the conditions of high temperature light culture；It places it in again certainly
15~25d so is cultivated under the conditions of scattering light, obtains tissue-cultured seedling of taking root；
(4) hardening：When under growth root tissue-cultured seedling grows supreme 3~5cm, has 3~4 expansion leaves, it out of culturing room is removed, is put
In the shady and cool place in outdoor with natural scattered light condition, 7~10d of hardening obtains rooted seedling；
(5) it is planted：Rooted seedling is taken out, washes away root culture medium, is planted in sand bed, drenching sufficient root water in greenhouse, and shade, determines
Phase sprays water, and relative air humidity is 75~85%, and temperature is 27~35 DEG C；
Step (2), step (3) high temperature light culture condition of culture are：32~35 DEG C of temperature, illumination are<10Lux；
The primary culture medium and subculture medium of step (1) and step (2) are MS culture mediums+6-BA2.0~3.0mg/L+
IBA0.2mg/L；
Root media is MS culture medium+NAA 1.0mg/L+IBA 0.2mg/L, pH5.8 in step (3).
2. dwarf banana high temperature light culture tissue culture and rapid propagation method according to claim 1, which is characterized in that sand bed in step (5)
In have matrix, the matrix is by pig manure：Soil：Peat soil is 3 by volume:85:12 compositions.
3. dwarf banana high temperature light culture tissue culture and rapid propagation method according to claim 2, which is characterized in that described in step (5)
The preparation method of matrix is：It measures pig manure, soil and peat soil, mixing and seedbed is laid to according to seedling growth greenhouse, it is thick to cover 2cm above
River sand, apply disinfection with the leaching of 800 times of carbendazim.
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| CN106688887B (en) * | 2016-12-08 | 2019-08-20 | 广东省农业科学院果树研究所 | The tissue culture and rapid propagation method of miscellaneous No. 1 dwarf banana of powder |
| CN106922537A (en) * | 2017-04-21 | 2017-07-07 | 中国热带农业科学院海口实验站 | A kind of cultural method and its culture medium of kylin fruit seedling |
| CN113142055B (en) * | 2021-04-29 | 2022-10-28 | 广西壮族自治区农业科学院 | In-vitro proliferation preservation method for banana germplasm resources |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102726302A (en) * | 2012-07-18 | 2012-10-17 | 广西植物组培苗有限公司 | Dark culture method for banana tissue culture |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102726302A (en) * | 2012-07-18 | 2012-10-17 | 广西植物组培苗有限公司 | Dark culture method for banana tissue culture |
Non-Patent Citations (1)
| Title |
|---|
| 温度对香蕉组培瓶苗根系生长的影响;黎兆安;《广西农业科学》;19971231;第232页第2.1节至233页至第234页第2.4节 * |
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