CN106613969A - Method for batch production of mat grass through one-step culture - Google Patents
Method for batch production of mat grass through one-step culture Download PDFInfo
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- CN106613969A CN106613969A CN201611113343.4A CN201611113343A CN106613969A CN 106613969 A CN106613969 A CN 106613969A CN 201611113343 A CN201611113343 A CN 201611113343A CN 106613969 A CN106613969 A CN 106613969A
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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
The invention discloses a method for batch production of mat grass through one-step culture. The method comprises the following steps of (1) acquiring basic seedlings: selecting tillering nodes of find mat grass plants as explants, sterilizing the explants, culturing the sterilized explants on 1/2MS+6-BA 1.2 to 4.5 mg/L of solid agar culture, and inducing the tillering nodes; (2) carrying out batch production: inoculating the tillering nodes acquired through the step (1) onto a solid agar culture with 1/2MS+6-BA 1.2 to 4.5 mg/L + NAA 0.02 to 1.0 mg/L + IAA 0.02 to 1.0mg/L + paclobutrazol 0.5 to 5.0mg/L for propagating and rooting; (3) carrying out field production: before taking seedlings out of a test-tube for transplanting, opening a test-tube cap for hardening the seedlings, taking out for cleaning the culture medium on the root part, and then directly transplanting the seedlings into a paddy field. The method is excellent in culture effect, high in propagation factor, good in stability and low in production cost, and is a fast propagation method for acquiring a large number of test-tube plantlets within the shortest time.
Description
Technical field
The invention belongs to the tissue culture technology field of seat grass, and in particular to the method for seat grass forming seedling through one step culture mass production.
Background technology
Seat grass (Juncus effusus L.) is a kind of important industrial crops, with soft and high resilience, and is difficult
Fade, also moisture absorption, deodorization, insect prevention, mothproof and with characteristics such as natural hay-scenteds, make seat grass be well suited for weaving tatami, straw mat,
Straw mattress, straw fan and straw basket etc., its product has ventilation, moisture absorption, refrigerant effect.Because seat grass can adjust dry and wet, institute with itself
So that appropriate drying can be kept with product summer made by seat grass, the paresthesia of skin of people is made comfortably, Winter protection performance is good
It is good, therefore seat straw product has become family's articles for daily use important in people's daily life, at the same can also as interior when
Still ornament, deep to be favored by consumers in general, and its knitting has the knitting such as those plastics, chemical fibre, bamboo incomparable
Characteristic and advantage.Seat grass roots stem can make medicinal, have diuresis, it is sleeping, the effects such as stop blooding, control nocturnal fretfulness in infants, therefore Japanese likes
It is to use Lin's straw product most also to have its deeper reason Japan as interior decoration and sleep with seat straw product
Country, at least existing history of more than 500 years.With the continuous improvement of our people's living standard and lifting for back to nature heat
Rise, with natural Lin grass as the green knitting of raw material, people will be caused more and more to pay close attention to.
Seat grass is perennial root herbaceous plant, and by division propagation, one is that year by year division propagation causes to conventional breeding
Quality Down, is not suitable for processing.The present invention is by method for tissue culture, the advantage of seat grass of purificating and rejuvenating, and natural plant height is attended in creation
The best approach of effect seedling, sets up the cultivating system of a set of efficient low-consume amount, is converted into achievement in research with most fast speed existing
Grow directly from seeds force of labor.
The content of the invention
It is an object of the invention to provide a kind of seat grass method for quickly breeding of efficient low-consume amount, the method culture effect is good,
Growth coefficient is high, and good stability, low production cost are a kind of quick breeding sides that a large amount of test tube seedlings are obtained within the most short time
Method.
The purpose of the present invention is achieved in the following ways：
A kind of method of seat grass forming seedling through one step culture mass production, comprises the following steps：
(1) basic seedling is obtained：The tillering node for selecting excellent seat grass plant is explant, in 1/2MS+6- after explant sterilization
Cultivate on the Solid agar culture of BA1.2～4.5mg/L, induce tiller bud；
(2) mass production：The tiller bud that step (1) is obtained, is connected on 1/2MS+6-BA1.2～4.5mg/L+
Increased on the Solid agar culture of NAA0.02～1.0mg/L+IAA0.02～1.0mg/L+ 0.5～5.0mg/L of paclobutrazol
Grow and take root；
(3) field production：Before seedling bottle outlet is transplanted, bottle cap hardening is opened, cleaning root culture medium is taken out, subsequently by seedling
Directly transplant to paddy field.
The whole strain of seat grass is dug out from land for growing field crops in step (1), root is cleaned up, placed in the water of cleaning and cultivate 2 weeks, will
Tillering node is divided into individual plant, then root and base portion old leaf are removed, and it is high that tillering node retains 1.8-2.2cm.
When tillering node is sterilized in step (1), cleaned up with washing powder first, with aseptic washing 5 on aseptic operating platform
～7 times, infiltrated -1 minute 30 seconds with 70% alcohol, place into 0.1% mercuric chloride solution and sterilize 12～18 minutes, sterilized water punching
Wash 5～6 times, it is standby.
During the clump bud squamous subculture that step (1) is induced, it is divided into 4-6 strains/clump, then proceeds to step (1) identical culture medium
In, every 30d subcultures once.
Step (2) is to scale off the Multiple Buds that step (1) is obtained, and is divided into 4-6 strains/clump, and every bottle of 7-10 clump is bred
With take root.
During step (2) squamous subculture, it is divided into 4-6 strains/clump, every bottle of 7-10 clump, in proceeding to step (2) same medium.
The cultivation temperature of step (1) and (2) is 24 DEG C ± 2, in 1000～2000lx low light environments, daily 10～12 hours
Illumination cultivation.
Step (3) is that the root leaf for obtaining step (2) is various, and 5-6cm high seedling bottle outlet is transplanted, and first opens bottle cap refining
Seedling 5-7d, further takes out cleaning root culture medium, subsequently seedling is divided into 4-6 strains/Cong Yizhi paddy fields；Field production water level keeps
In 1-2cm depths.
The Solid agar culture that step (1) is preferably used is：1/2MS+6-BA3.5mg/L；Or use culture medium：1/
2MS+6-BA4.0mg/L。
The Solid agar culture that step (2) is preferably used is：1/2MS+6-BA2.5mg/L+NAA0.6mg/L+
IAA0.5mg/L+ paclobutrazol 3.0mg/L；Or use culture medium：1/2MS+6-BA3.0mg/L+NAA0.6mg/L+
IAA0.4mg/L+ paclobutrazol 4.0mg/L.
The present invention compared with prior art, the beneficial effects of the present invention is：
1st, in place of in view of the shortcomings of the prior art, the inventive method improves seat grass good characteristic, after its clone
In generation, without callus phase, can keep the high consistency with maternal good characteristic.
2nd, the present invention needs to design cultural method according to production, first with a large amount of tiller buds of one-step inducing, suppresses the formation of root,
Proliferation times are improved, accelerates the breeding of basic seedling, proliferation times reach 7-10；Per bottle obtains more than 70 plants of effective seedling, carries significantly
High culture efficiency, has saved production cost.
3rd, the mass production stage is with one-step inducing tiller bud, adventitious root, breeds and is reduced to a step and completes with taking root, i.e., and " one
Step seedling " method, forming whole plant only needs 30d or so, has saved the rootage duration of 30d or so.
4th, directly land for growing field crops is transplanted from bottle, eliminates the greenhouse hardening stage, and 4-6 strains/Cong Jinhang is transplanted, seedling-slowing stage
Short, transplanting survival rate is up to more than 95%.
5th, the present invention selects to carry out division propagation for control without the tillering node of tissue culture propagation；Both transplanting conditions are consistent,
Topdress in good time once；Transplant 7 months or so, seedling is emerald green compared to control stem color for the plant of tissue culture test tube seedling, fresh grass diameter
Relatively thick, grass blade is longer, and top grass is long.
6th, calculated according to the method for forefathers' cost budgeting, the technology of the present invention produces 1,000,000 plants of seat grass test tube seedlings, and per bottle effectively
Seedling of taking root is more than 70 plants, it is only necessary to which the basic seedling of about 0.5 ten thousand bottles of production, about 1.5 ten thousand bottles of seedlings of taking root add up to 20,000 bottles.The present invention is not only
Sterile working step is reduced, and has saved culturing room space, simplify health seedling production routine, reduced and produce into
This, improves production efficiency.
Description of the drawings
Fig. 1 is the growing state of seat grass effectively seedling seedling；
Fig. 2 is the growing state of seat grass effectively seedling root；
Fig. 3 is growing state of the seat grass tissue culture test tube seedling in land for growing field crops；
Fig. 4 is growing state of the seat grass axillary seedling in land for growing field crops.
Specific embodiment
It is intended to further illustrate the present invention with reference to embodiments, and the unrestricted present invention.
Embodiment 1, tissue-culturing quick-propagation seat grass
First, culture medium is prepared and sterilized
Just it is used for basic seedling breeding for subculture medium：1/2MS+6-BA3.5mg/L；
Forming seedling through one step culture mass production culture medium：1/2MS+6-BA2.5mg/L+NAA0.6mg/L+IAA0.5mg/L+ multiple-effect
Azoles 3.0mg/L.
At 121 DEG C of above culture medium, sterilize 20 minutes.
2nd, basic seedling breeding
Seat grass (Juncus effusus L.) tillering node is taken, the whole strain of seat grass is dug out from land for growing field crops, root is cleaned up,
Place in the water of cleaning and cultivate 2 weeks or so, then tillering node is divided into individual plant, then root, base portion old leaf and top are removed, tillering node
Retain 2cm or so height；During sterilization, cleaned up with washing powder first, with aseptic washing 5～7 times on aseptic operating platform, used
70% alcohol infiltrates -1 minute 30 seconds, places into 0.1% mercuric chloride solution and sterilizes 12～18 minutes, aseptic water washing 5～6
It is secondary, it is standby.Tillering node is proceeded to equipped with plastic culture bottle in Initial culture base (80 milliliters of capacity), each blake bottle explant
Number is 10 or so, is cultivated under following condition of culture：24 DEG C or so, daily illumination in 10 hours, intensity of illumination is
1000～2000lx or so；Tillering node is inoculated with 90.Culture 3 weeks or so, from tillering node base portion tiller bud is initially formed, and continues to train
Support 2 weeks, the tiller bud of 100% tillering node base portion formation more than 5, up to 10.During squamous subculture, clump bud is divided into 4-6
Strain/clump, in each subculture, each tiller bud breeds 7-10 times every 30d subcultures once.This step needs the formation for suppressing root,
In case affecting the breeding of basic seedling.According to production scale, sufficient basic seedling is bred, then carry out mass production.
3rd, mass production
90 strains taken in above-mentioned 90 strains carry out successive propagation, and concrete grammar is as follows：By dividing that step (1) is obtained
Tiller bud clump is pressed the specification of 4-6 strains/clump and separates, and inoculation is cultivated in the medium, the same step of culture medium (2), and condition of culture is same
Step (2)；After culture 30d or so, proliferation times are 7-10, and height of seedling 5-6cm, synchronous rooting rate is up to 100%, per bottle of acquisition root
More than 70 plants of the various effective seedling of leaf.Effectively the growing state of seedling seedling is as shown in figure 1, growing state such as Fig. 2 institutes of effective seedling root
Show.
4th, test tube transplantation of seedlings：
Step (3) is that the root leaf for obtaining step (2) is various, and 5-6cm high seedling bottle outlet is transplanted, and first opens bottle cap refining
Seedling 5-7d, further takes out cleaning root culture medium, subsequently seedling is divided into 4-6 strains/clump or so and moves to paddy field, selects numerous without tissue culture
The seat grass suckering plant grown carries out division propagation to compare, and water level is held in 1-2cm depths；Both transplanting conditions are consistent with method.
Transplanting time is on October 22nd, 2015, and 10d or so restoration ecosystems, 30d or so grows young leaves, 60d or so 100%
Grow young leaves.The growing state for transplanting 8 months seat grass test tube seedlings is as shown in Figure 3；Transplant growing state such as Fig. 4 of control in 8 months
It is shown.
Seat grass tissue culture propagation provided by the present invention, tillering node is explant, is drawn materials easily, and quantity is big, loses
Pass stability high, the advantage such as well-grown.During Initial culture, without callus phase, directly from tillering node base
Portion forms tiller bud, not only fast growth, and breeding coefficient is big.In the mass production stage, taken root using synchronous propagation,
The method of " forming seedling through one step culture ", simplifies health seedling production routine.The technology of the present invention produces 1,000,000 plants of seats grass test tube seedlings, per bottle
Seedling of effectively taking root is more than 70 plants, it is only necessary to which the basic seedling of about 0.5 ten thousand bottles of production, about 1.5 ten thousand bottles of seedlings of taking root add up to 20,000 bottles.The present invention
Sterile working step is not only reduced, and has saved culturing room space, simplify health seedling production routine, reduce production
Cost, improves production efficiency.
Embodiment 2, tissue-culturing quick-propagation seat grass
First, culture medium is prepared and sterilized
Just for subculture medium：1/2MS+6-BA4.0mg/L；
Forming seedling through one step culture mass production culture medium：1/2MS+6-BA3.0mg/L+NAA0.6mg/L+IAA0.4mg/L+ multiple-effect
Azoles 4.0mg/L.
Seat grass (Juncus effusus L.) tillering node is taken, basic operation method is consistent with embodiment 1；Effect and enforcement
Example 1 is more or less the same.
Claims (10)
1. the method for a kind of seat grass forming seedling through one step culture mass production, it is characterised in that comprise the following steps：
(1) basic seedling is obtained：The tillering node for selecting excellent seat grass plant is explant, in 1/2MS+6- after explant sterilization
Cultivate on the Solid agar culture of BA1.2～4.5mg/L, induce tiller bud；
(2) mass production：By step (1) obtain tiller bud, be connected on 1/2MS+6-BA1.2～4.5mg/L+NAA0.02～
Bred and taken root on the Solid agar culture of 1.0mg/L+IAA0.02～1.0mg/L+ 0.5～5.0mg/L of paclobutrazol；
(3) field production：Before seedling bottle outlet is transplanted, bottle cap hardening is opened, take out cleaning root culture medium, it is subsequently that seedling is direct
Transplant to paddy field.
2. the method for seat according to claim 1 grass forming seedling through one step culture mass production, it is characterised in that will in step (1)
The whole strain of seat grass is dug out from land for growing field crops, and root is cleaned up, and is placed in the water of cleaning and is cultivated 2 weeks, and tillering node is divided into individual plant, then will
Root and base portion old leaf are removed, and it is high that tillering node retains 1.8-2.2cm.
3. the method for seat according to claim 1 grass forming seedling through one step culture mass production, it is characterised in that in step (1) point
When tiller section is sterilized, cleaned up with washing powder first, with aseptic washing 5～7 times on aseptic operating platform, infiltrated with 70% alcohol
- 1 minute 30 seconds, place into 0.1% mercuric chloride solution and sterilize 12～18 minutes, aseptic water washing 5～6 times is standby.
4. the method for the careless forming seedling through one step culture mass production of seat according to claim 1 or 2 or 3, it is characterised in that step
(1) during the clump bud squamous subculture for inducing, be divided into 4-6 strains/clump, then proceed in step (1) identical culture medium, every 30d after
In generation, is once.
5. the method for seat according to claim 1 grass forming seedling through one step culture mass production, it is characterised in that step (2) be by
The Multiple Buds that step (1) is obtained are scaled off, and are divided into 4-6 strains/clump, and every bottle of 7-10 clump is bred and taken root.
6. the method for seat grass forming seedling through one step culture mass production according to claim 1 or 5, it is characterised in that step (2) after
During culture, it is divided into 4-6 strains/clump, every bottle of 7-10 clump, in proceeding to step (2) same medium.
7. the method for seat according to claim 1 grass forming seedling through one step culture mass production, it is characterised in that step (1) and (2)
Cultivation temperature be 24 DEG C ± 2, in 1000～2000lx low light environments, daily 10～12 hours illumination cultivations.
8. the method for seat according to claim 1 grass forming seedling through one step culture mass production, it is characterised in that step (3) be by
The root leaf that step (2) is obtained is various, and 5-6cm high seedling bottle outlet is transplanted, and first opens bottle cap hardening 5-7d, further takes out cleaning root
Portion's culture medium, is subsequently divided into 4-6 strains/Cong Yizhi paddy fields by seedling；Field production water level is held in 1-2cm depths.
9. the method for seat according to claim 1 grass forming seedling through one step culture mass production, it is characterised in that step (1) is used
Solid agar culture be：1/2MS+6-BA3.5mg/L；Or use culture medium：1/2MS+6-BA4.0mg/L.
10. the method for seat according to claim 1 grass forming seedling through one step culture mass production, it is characterised in that step (2) is used
Solid agar culture be：1/2MS+6-BA2.5mg/L+NAA0.6mg/L+IAA0.5mg/L+ paclobutrazol 3.0mg/L；Or
Using culture medium：1/2MS+6-BA3.0mg/L+NAA0.6mg/L+IAA0.4mg/L+ paclobutrazol 4.0mg/L.
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| CN201611113343.4A CN106613969B (en) | 2016-12-07 | 2016-12-07 | A kind of method of seat grass forming seedling through one step culture mass production |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107646570A (en) * | 2017-11-10 | 2018-02-02 | 来安县义坤农业发展有限公司 | A kind of implantation methods of Lin's grass |
| CN109169132A (en) * | 2018-09-27 | 2019-01-11 | 岭南生态文旅股份有限公司 | A kind of Hipeastrum vittalum (L Her.) Herb.- Amaryllisvittata Ait tissue-cultured seedling crop field efficient cultivation method |
| CN113197042A (en) * | 2021-04-25 | 2021-08-03 | 福建生物工程职业技术学院 | Breeding method of polygonatum cyrtonema multi-bud tillering variety |
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| CN101569288A (en) * | 2009-06-08 | 2009-11-04 | 浙江大学 | Method for culturing in-vitro tissues of rush mat |
| CN104642108A (en) * | 2015-02-03 | 2015-05-27 | 中国科学院亚热带农业生态研究所 | Method suitable for tissue culture mass production of multiple plants |
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2016
- 2016-12-07 CN CN201611113343.4A patent/CN106613969B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101569288A (en) * | 2009-06-08 | 2009-11-04 | 浙江大学 | Method for culturing in-vitro tissues of rush mat |
| CN104642108A (en) * | 2015-02-03 | 2015-05-27 | 中国科学院亚热带农业生态研究所 | Method suitable for tissue culture mass production of multiple plants |
Non-Patent Citations (2)
| Title |
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| L. XU ET.AL.,: "Development of an efficient tissue culture protocol for callus formation and plant regeneration of wetland species Juncus effusus L.", 《IN VITRO CELL.DEV.BIOL.—PLANT》 * |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107646570A (en) * | 2017-11-10 | 2018-02-02 | 来安县义坤农业发展有限公司 | A kind of implantation methods of Lin's grass |
| CN109169132A (en) * | 2018-09-27 | 2019-01-11 | 岭南生态文旅股份有限公司 | A kind of Hipeastrum vittalum (L Her.) Herb.- Amaryllisvittata Ait tissue-cultured seedling crop field efficient cultivation method |
| CN113197042A (en) * | 2021-04-25 | 2021-08-03 | 福建生物工程职业技术学院 | Breeding method of polygonatum cyrtonema multi-bud tillering variety |
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