CN109463281A - A kind of black wooden Tissue Cultured Acacia seedling standardized production technique - Google Patents
A kind of black wooden Tissue Cultured Acacia seedling standardized production technique Download PDFInfo
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- CN109463281A CN109463281A CN201811523806.3A CN201811523806A CN109463281A CN 109463281 A CN109463281 A CN 109463281A CN 201811523806 A CN201811523806 A CN 201811523806A CN 109463281 A CN109463281 A CN 109463281A
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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
- A01H4/001—Culture apparatus for tissue culture
Abstract
A kind of black wooden Tissue Cultured Acacia seedling standardized production technique, including the production of proliferation seedling and rooted seedling production, proliferation seedling production content are related to inoculum density, Multiplying culture period switching cycle and the switching cycle for producing nursery stock period；Standing time and hardening greenhouse standing time when rooted seedling production content includes inoculum density, nursery stock cutting-height, nursery stock cutting degree, rooted seedling culture, the following steps are included: after a) obtaining the black wooden Tissue Cultured Acacia seedling, the Multiplying culture for first carrying out 15-30 days obtains proliferation seedling；B) transfer proliferation seedling is inoculated with；C) proliferation seedling culture 25-40 days of inoculation；D) simple bud in cutting proliferation seedling higher than 2.0cm is as rooted seedling；E) rooted seedling, f are inoculated with) rooted seedling culture 4-24 days；G) hardening, time are 6-26 days.Standardized production of the present invention realizes the increase of nursery stock yield, improves seedling quality, improves production efficiency, reduces production cost, pushes the black wooden yearning between lovers choiceness large-scale production, realizes the black wooden yearning between lovers Developing Clonal Forestry high effect culture.
Description
Technical field
The invention belongs to black wooden Tissue Cultured Acacia seedling standardized production technology fields.
Technical background
Black wood yearning between lovers (Acacia melanoxylon) is Mimosaceae Acacia tree species, is yearning between lovers class highest megaphanerophyte
One of, originate in the ground such as Australia, the new several sub- and Indonesia of Papua.Black wood yearning between lovers wood grain is beautiful, and material is good, has
Nitrogen-fixing root nodule, litter are abundant, and performance of improving the soil is good, introduce the 1990s behind China in Guangdong, Fujian, Guangxi, Hainan etc.
It successfully plants on ground.
In this century, our unit is by Clone Selection And Reproduction, and it is more to overcome branch, the unconspicuous problem of trunk, black wood yearning between lovers
Have become the multipurpose tree species such as South China, China fast growing, precious material and ecological public good.According to incompletely statistics, 2015
It rises, black wood yearning between lovers clonal planting area more than the speed of 10,000 mus/year to develop.To meet the black wooden yearning between lovers Clonal Plantations
The needs of large-scale development, in this century, the black wooden Tissue Cultured Acacia research is carried out on domestic more ground in succession, but most tissue culture factories exist
The problems such as subculture cycle is long, and proliferation rate, rooting rate difference are big, leads to tissue-cultured seedling low output, at high cost, could not realize industry substantially
Metaplasia produces.My unit acquires 10 choiceness and carries out group training research, wherein 7 clones on the basis of Seedling selection
Higher production efficiency is obtained, large-scale production has been entered.Find that the black wooden yearning between lovers tissue cultures Plays metaplasia produces work in research
Skill improves seedling quality and plays an important role to nursery stock yield is increased.The black wooden Tissue Cultured Acacia seedling production effect can be improved using the present invention
Rate pushes the black wooden Tissue Cultured Acacia seedling large-scale production process, lays the foundation for the black wooden yearning between lovers Developing Clonal Forestry.Both at home and abroad to black wood
Tissue Cultured Acacia is concentrated mainly in the basic research such as culture medium prescription, is had not been reported to production technology Standardization Research.
Summary of the invention
In order to solve the above problem, the purpose of the present invention is to provide a kind of black wooden Tissue Cultured Acacia seedling standardized production technique,
It realizes that the black wooden Tissue Cultured Acacia seedling standardized production can increase nursery stock yield, improves seedling quality, improve production efficiency, reduce production
Cost pushes the black wooden yearning between lovers choiceness large-scale production process, realizes the black wooden yearning between lovers Developing Clonal Forestry high effect culture.
In order to achieve the above object, the invention adopts the following technical scheme:
A kind of black wooden Tissue Cultured Acacia seedling standardized production technique, comprising the following steps: after a) obtaining the black wooden Tissue Cultured Acacia seedling,
The Multiplying culture for first carrying out 15-30 days obtains proliferation seedling；B) transfer proliferation seedling is inoculated with；C) the proliferation seedling culture 25- being inoculated with
40 days；D) simple bud in cutting proliferation seedling higher than 2.0cm is as rooted seedling；E) it is inoculated with rooted seedling；F) rooted seedling culture 4-24 days；
G) hardening, time are 6-26 days；
Further, the more preferably black wooden Tissue Cultured Acacia seedling standardized production technique obtains the black wooden Tissue Cultured Acacia in step a)
Miao Hou first carries out 15-20 days Multiplying cultures；Proliferation seedling culture 35-40 days be inoculated in step c)；Rooted seedling is trained in step f)
It supports 4-8 days；Hardening in step g), time are 22-26 days；
Proliferation seedling inoculum density is to connect with 200mL cylinder jam jar (base diameter 5.5cm, high 9.5cm) in step b)
Kind container, culture medium thickness 1.5cm, 8-12 clumps of nursery stocks of every bottle of inoculation, every Cong Miaomu are 4-6 plants；
The culture medium of proliferation seedling culture is added in every liter of minimal medium using MS as minimal medium in step c)
6BA0.4-0.6mg, white sugar 25-30g, agar 5-6g, pH value are adjusted to 5-6, culture medium dispense after in advance in 1.0-1.2kg/
cm2Sterilize 18min under high pressure, 121 DEG C of high temperature；
The quantity for the rooted seedling cut in step d) is 25-50 plants, requires not destroy Miao Cong when cutting rooted seedling, not damage
Seedling cuts the visual seedling growth situation of the Miao Cong left, continues Multiplying culture；
Rooted seedling inoculum density is to connect with 200mL cylinder jam jar (base diameter 5.5cm, high 9.5cm) in step e)
Kind of container, culture medium thickness 1.5cm, every bottle 10-25 plants of inoculation nursery stock；
The culture medium of rooted seedling culture is added in every liter of minimal medium using 1/2MS as minimal medium in step f)
IBA1.0-1.5mg, white sugar 25-30g, agar 5-6g, pH value are adjusted to 5-6, culture medium dispense after in advance in 1.0-1.2kg/
cm2Sterilize 18min under high pressure, 121 DEG C of high temperature；
The cultivation temperature of rooted seedling is 23-27 DEG C in step f), and daytime adds 1500-2000Lx artificial lighting 10-11h；
Hardening temperature is 25-35 DEG C, illumination 7000-10000Lx in step g), and seeding room is daylight glass greenhouse, is had outside
70% sunshade net facility and cooling installation,.
The black wooden yearning between lovers nursery stock obtained by the production technology, rooting rate reach 85%, average height of seedling 2.5cm, seedling
The wooden well developed root system, blade are unfolded, stalk is dark green.
Bring technical effect of the present invention are as follows: the increasing of nursery stock yield may be implemented in black wood Tissue Cultured Acacia seedling standardized production
Add, improve seedling quality, improve production efficiency, reduce production cost, push the black wooden yearning between lovers choiceness large-scale production into
Journey realizes the black wooden yearning between lovers Developing Clonal Forestry high effect culture, the black wooden yearning between lovers nursery stock obtained by standardized production, well developed root system,
Blade is unfolded, stalk is dark green, can be directly used for nursery transplanting.
Specific embodiment
Below with reference to embodiment, the present invention is described in further detail, it should be understood that these embodiments are merely to illustrate this
It invents rather than limits the scope of the invention, after the present invention has been read, those skilled in the art are to of the invention
It is as defined in the appended claims that the modification of various equivalent forms falls within the application.
Embodiment 1
A) after obtaining the black wooden Tissue Cultured Acacia seedling, 15 days Multiplying cultures are first carried out, obtain proliferation seedling；
B) transfer proliferation seedling is inoculated with: with 200mL cylinder jam jar (base diameter 5.5cm, high 9.5cm) for inoculation
Container, culture medium thickness 1.5cm are inoculated with 20 bottles, and 8 clumps of nursery stocks of every bottle of inoculation, every Cong Miaomu is 4-6 plants, the monthly proliferation of nursery stock
Rate is 3.35%；
C) after being proliferated seedling culture 40 days, enter the nursery stock production stage later, from high to low in proliferation seedling when nursery stock production
Select nursery stock cutting simple bud of the height of seedling greater than 2.0cm as rooted seedling in clump, the quantity for cutting rooted seedling is 45-50 plants, cutting
It requires not destroy Miao Cong when rooted seedling, does not damage seedling；
D) it is inoculated with rooted seedling: with 200mL cylinder jam jar (base diameter 5.5cm, high 9.5cm) for inoculation container, training
Support base thickness 1.5cm, every bottle of 20 plants of inoculation nursery stock；
E) cultivate rooted seedling: the nursery stock of culture of rootage is placed 4 days in culturing room, and it is 23 DEG C that temperature, which is arranged, in culturing room, daytime
Additional 1500Lx artificial lighting 11h；
F) after 4 days, a small amount of nursery stock base portion exposes the tip of a root of white, and transfer rooted seedling enters heliogreenhouse and refined at this time
Seedling, hardening temperature are 25 DEG C, illumination 10000Lx, and the time is 26 days, and the rooting rate for obtaining nursery stock is 98.3%, and height of seedling exists
2.71 ± 0.08cm is taken root number 7.0 ± 0.6.
Embodiment 2
A) after obtaining the black wooden Tissue Cultured Acacia seedling, 20 days Multiplying cultures are first carried out, obtain proliferation seedling；
B) transfer proliferation seedling is inoculated with: with 200mL cylinder jam jar (base diameter 5.5cm, high 9.5cm) for inoculation
Container, culture medium thickness 1.5cm are inoculated with 20 bottles, and 9 clumps of nursery stocks of every bottle of inoculation, every Cong Miaomu is 4-6 plants, the monthly proliferation of nursery stock
Rate is 3.41%；
C) after being proliferated seedling culture 35 days, enter the nursery stock production stage later, from high to low in proliferation seedling when nursery stock production
Select nursery stock cutting simple bud of the height of seedling greater than 2.0cm as rooted seedling in clump, the quantity for cutting rooted seedling is 45-50 plants, cutting
It requires not destroy Miao Cong when rooted seedling, does not damage seedling；
D) it is inoculated with rooted seedling: with 200mL cylinder jam jar (base diameter 5.5cm, high 9.5cm) for inoculation container, training
Support base thickness 1.5cm, every bottle of 15 plants of inoculation nursery stock；
E) cultivate rooted seedling: the nursery stock of culture of rootage is placed 8 days in culturing room, and it is 24 DEG C that temperature, which is arranged, in culturing room, daytime
Additional 2000Lx artificial lighting 10h；
F) after 8 days, part nursery stock base portion exposes the tip of a root of white, and transfer rooted seedling enters heliogreenhouse and refined at this time
Seedling, hardening temperature are 30 DEG C, illumination 8000Lx, and the time is 22 days, and the rooting rate for obtaining nursery stock is 91.3%, and height of seedling is 2.89
± 0.09cm is taken root number 6.2 ± 0.4.
Embodiment 3
A) after obtaining the black wooden Tissue Cultured Acacia seedling, 25 days Multiplying cultures are first carried out, obtain proliferation seedling；
B) transfer proliferation seedling is inoculated with: with 200mL cylinder jam jar (base diameter 5.5cm, high 9.5cm) for inoculation
Container, culture medium thickness 1.5cm are inoculated with 20 bottles, and 11 clumps of nursery stocks of every bottle of inoculation, every Cong Miaomu is 4-6 plants, the monthly proliferation of nursery stock
Rate is 2.59%；
C) after being proliferated seedling culture 30 days, enter the nursery stock production stage later, from high to low in proliferation seedling when nursery stock production
Select nursery stock cutting simple bud of the height of seedling greater than 2.0cm as rooted seedling in clump, the quantity for cutting rooted seedling is 30-35 plants, cutting
It requires not destroy Miao Cong when rooted seedling, does not damage seedling；
D) it is inoculated with rooted seedling: with 200mL cylinder jam jar (base diameter 5.5cm, high 9.5cm) for inoculation container, training
Support base thickness 1.5cm, every bottle of 25 plants of inoculation nursery stock；
E) cultivate rooted seedling: the nursery stock of culture of rootage is placed 12 days in culturing room, and it is 26 DEG C that temperature, which is arranged, in culturing room, daytime
Additional 1800Lx artificial lighting 11h；
F) after 12 days, nursery stock base portion grows the root of white, shifts rooted seedling at this time and enters heliogreenhouse progress hardening, hardening
Temperature is 30 DEG C, illumination 9000Lx, and the time is 18 days, and the rooting rate for obtaining nursery stock is 84.4%, height of seedling 2.92 ±
0.10cm is taken root number 4.8 ± 0.3.
Embodiment 4
A) after obtaining the black wooden Tissue Cultured Acacia seedling, 30 days Multiplying cultures are first carried out, obtain proliferation seedling；
B) transfer proliferation seedling is inoculated with: with 200mL cylinder jam jar (base diameter 5.5cm, high 9.5cm) for inoculation
Container, culture medium thickness 1.5cm are inoculated with 20 bottles, and 12 clumps of nursery stocks of every bottle of inoculation, every Cong Miaomu is 4-6 plants, the monthly proliferation of nursery stock
Rate is 2.50%；
C) after being proliferated seedling culture 25 days, enter the nursery stock production stage later, from high to low in proliferation seedling when nursery stock production
Select nursery stock cutting simple bud of the height of seedling greater than 2.0cm as rooted seedling in clump, the quantity for cutting rooted seedling is 25-30 plants, cutting
It requires not destroy Miao Cong when rooted seedling, does not damage seedling；
D) it is inoculated with rooted seedling: with 200mL cylinder jam jar (base diameter 5.5cm, high 9.5cm) for inoculation container, training
Support base thickness 1.5cm, every bottle of 10 plants of inoculation nursery stock；
E) cultivate rooted seedling: the nursery stock of culture of rootage is placed 24 days in culturing room, and it is 27 DEG C that temperature, which is arranged, in culturing room, daytime
Additional 2000Lx artificial lighting 10h；
F) after 24 days, nursery stock base portion grows the root of white, shifts rooted seedling at this time and enters heliogreenhouse progress hardening, hardening
Temperature is 35 DEG C, illumination 7000Lx, and the time is 6 days, and the rooting rate for obtaining nursery stock is 63.2%, height of seedling 2.13 ±
0.06cm is taken root number 1.6 ± 0.2.
It is basic culture medium that the culture medium of seedling culture is proliferated in above embodiments with MS, is added in every liter of minimal medium
6BA0.4-0.6mg, white sugar 25-30g, agar 5-6g, pH value are adjusted to 5-6, culture medium dispense after in advance in 1.0-1.2kg/
cm2Sterilize 18min under high pressure, 120-123 DEG C of high temperature.
And rooted seedling culture medium is basic culture medium with 1/2MS, and 6BA0.4-0.6mg, white is added in every liter of minimal medium
Sugared 25-30g, agar 5-6g, pH value are adjusted to 5-6, culture medium dispense after in advance in 1.0-1.2kg/cm2High pressure, 120-123
Sterilize 18min under DEG C high temperature.
Below with reference to the optimal black wooden Tissue Cultured Acacia seedling standardized production technique of data exhibiting:
1) the black wooden yearning between lovers transfer period
Consistent black wooden 60 bottles of seedling of the Tissue Cultured Acacia of selection growth, the Multiplying culture period is respectively 15,20,25,30 days, culture
It is counted after 60 days, i.e., cultivation cycle is that 15 days corotation connect 4 times, i.e. cultivation cycle is that 20 days corotation connect 3 times, i.e., cultivation cycle is
25,30 days corotation connect 2 times, as a result to cultivate 15 days switching moon proliferation rate highests, cultivate 20 days switching proliferation rates and take second place.Therefore black
The tissue-cultured seedling Multiplying culture period of the wooden yearning between lovers is preferably 15-20 days.
Table 1 is proliferated influence of the seedling switching cycle to seedling growth
2) the black wooden yearning between lovers is proliferated seedling inoculum density
The consistent black wooden Tissue Cultured Acacia of selection growth is proliferated seedling, is 4-5 plants/clump and 5-6 plants/clump the cutting of proliferation seedling, often
Bottle culture medium is respectively connected to 8,9,10,11,12 clumps, and 20 bottles of vaccinization, totally 1000 clumps, 9 clumps of proliferation is as a result inoculated with every bottle
Rate highest, the quantity greater than 2.0cm nursery stock are most；8 clumps of proliferation rates are inoculated with every bottle and occupy the greater than the quantity of 2.0cm nursery stock
2 (tables 2), proliferation seedling cutting is that 4-5 plants/clump is identical with 5-6 plants/clump result.Therefore proliferation seedling cutting be 4-6 plants/clump, every bottle
It is inoculated with 8-9 clumps of production efficiency highests.
Influence of 2 inoculum density of table to the black wooden Tissue Cultured Acacia growth
3) switching cycle of production phase
The consistent black wooden Tissue Cultured Acacia of selection growth is proliferated 60 bottles of seedling, cultivate 25 respectively in culturing room after inoculation, 30,35,
Rooted seedling is produced after 40 days, most rooted seedlings are as a result obtained for 40 days with culture, and culture is taken second place for 35 days, surplus after cutting rooted seedling
Remaining Miao Cong continues Multiplying culture, and next phase can continue to produce rooted seedling.Therefore production phase best switching cycle is 35-
40 days.
Influence of the 3 rooted seedling switching cycle of table to yield
4) rooted seedling inoculum density
10,15,20,25 plants of the rooted seedling point of cutting is seeded in respectively in root media and is cultivated 20 days, as a result to connect
20 plants of kind obtain highest rooting rates, and height of seedling and biomass of individual tree are to be inoculated with 10 plants/bottle as highest.
Therefore comprehensively consider from production cost and nursery stock rooting rate, every bottle of rooted seedling is to be inoculated with preferably 20 plants.
Influence of the 4 rooted seedling inoculum density of table to seedling growth
5) the rooted seedling hardening time
Inoculated rooted seedling is unified incubation time 30 days, places different time in culturing room and heliogreenhouse respectively,
First culturing room place 4,8,12,16,20,24 days, after accordingly move in heliogreenhouse hardening culture 26,22,18,14,10,6
It, as a result rooting rate, number of taking root are placed 4-8 days with elder generation in culturing room, after heliogreenhouse place 22-26 days it is preferable.
The influence of 5 different time culture of rootage of table and hardening culture to nursery stock
It above are only part specific embodiment of the invention, but design concept of the invention is not limited to this, all utilizations
This design makes a non-material change to the present invention, and should all belong to behavior that violates the scope of protection of the present invention.As long as without departing from
The content of technical solution of the present invention, it is to the above embodiments according to the technical essence of the invention any type of simply to repair
Change, equivalent variations and remodeling, still falls within the protection scope of technical solution of the present invention.
Claims (9)
1. a kind of black wooden Tissue Cultured Acacia seedling standardized production technique, which comprises the following steps: a) obtain the black wooden yearning between lovers
After tissue-cultured seedling, 15-30 days Multiplying cultures are first carried out, obtain proliferation seedling；B) transfer proliferation seedling is inoculated with；C) proliferation being inoculated with
Seedling culture 25-40 days；D) simple bud in cutting proliferation seedling is as rooted seedling；E) it is inoculated with rooted seedling；F) rooted seedling culture 4-24 days；
G) hardening, time are 6-26 days.
2. the black wooden Tissue Cultured Acacia seedling standardized production technique according to claim 1, which is characterized in that obtained in step a)
After black wood Tissue Cultured Acacia seedling, 15-20 days Multiplying cultures are first carried out；The proliferation seedling being inoculated in step c) culture 35-40 days；Step
F) rooted seedling culture 4-8 days in；Hardening in step g), time are 22-26 days.
3. the black wooden Tissue Cultured Acacia seedling standardized production technique according to claim 1, which is characterized in that be proliferated in step b)
Seedling inoculum density is 8-12 clumps of nursery stocks of every bottle of inoculation, and every Cong Miaomu is 4-6 plants.
4. the black wooden Tissue Cultured Acacia seedling standardized production technique according to claim 1 or 2 or 3, which is characterized in that step c)
6BA0.4-0.6mg, white sugar 25- is added using MS as minimal medium in the culture medium of middle proliferation seedling culture in every liter of minimal medium
30g, agar 5-6g, pH value are adjusted to 5-6, culture medium dispense after in advance in 1.0-1.2 kg/cm2It sterilizes under high pressure, high temperature.
5. the black wooden Tissue Cultured Acacia seedling standardized production technique according to claim 1, which is characterized in that cut in step d)
Rooted seedling quantity be 25-50 plants.
6. the black wooden Tissue Cultured Acacia seedling standardized production technique according to claim 1, which is characterized in that take root in step e)
Seedling inoculum density is every bottle 10-25 plants of inoculation nursery stock.
7. the black wooden Tissue Cultured Acacia seedling standardized production technique according to claim 4, which is characterized in that take root in step f)
IBA1.0-1.5mg, white sugar 25- is added using 1/2MS as minimal medium in the culture medium of seedling culture in every liter of minimal medium
30g, agar 5-6g, pH value are adjusted to 5-6, culture medium dispense after in advance in 1.0-1.2 kg/cm2It sterilizes under high pressure, high temperature.
8. the black wooden Tissue Cultured Acacia seedling standardized production technique according to claim 1 or 2, which is characterized in that in step f)
The cultivation temperature of rooted seedling is 23-27 DEG C, and daytime adds 1500-2000Lx artificial lighting 10-11h.
9. the black wooden Tissue Cultured Acacia seedling standardized production technique according to claim 1 or 2, which is characterized in that in step g)
Hardening temperature is 25-35 DEG C, illumination 7000-10000 Lx.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110731267A (en) * | 2019-10-24 | 2020-01-31 | 中国林业科学研究院热带林业研究所 | Domestication process of acacia melanoxylon hard rooting clone tissue culture seedlings |
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| CN110731267A (en) * | 2019-10-24 | 2020-01-31 | 中国林业科学研究院热带林业研究所 | Domestication process of acacia melanoxylon hard rooting clone tissue culture seedlings |
| CN110731267B (en) * | 2019-10-24 | 2021-06-18 | 中国林业科学研究院热带林业研究所 | Domestication process of acacia melanoxylon hard rooting clone tissue culture seedlings |
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