CN108811584A - A method of promoting Paris polyphylla seed fast germination - Google Patents
A method of promoting Paris polyphylla seed fast germination Download PDFInfo
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- CN108811584A CN108811584A CN201810666071.3A CN201810666071A CN108811584A CN 108811584 A CN108811584 A CN 108811584A CN 201810666071 A CN201810666071 A CN 201810666071A CN 108811584 A CN108811584 A CN 108811584A
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- seed
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
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Abstract
The invention belongs to field of biotechnology, and in particular to a method of promote Paris polyphylla seed fast germination.Specially：Remove kind of a skin；Using sodium tungstate solution, 10~14h is impregnated；Reuse 6~10h of hydrogen peroxide dipping；It reuses potassium nitrate solution and Gibberellins solution joint impregnates 36~48h；Reuse 6~10h of hydrogen peroxide dipping；It reuses potassium nitrate solution and Gibberellins solution joint impregnates 36~48h；25~30s is impregnated with alcohol again；It reuses mercuric chloride solution and continues 6~10min of immersion, clean, gnotobasis is dried, and is seeded on 1/2MS culture medium, vaccination area >=0.09cm2/.The present invention has general applicability to paris plant seed；And germination is early, germination percentage is high：Rudiment is begun to when 30 days, average germination percentage is up to 80% or more within 60 days.The present invention takes full advantage of the physiological property of Paris polyphylla seed, easy to operate, the cost of raw material is cheap, extends in practical application.
Description
Technical field
The invention belongs to field of biotechnology, and in particular to a method of promote Paris polyphylla seed fast germination.
Background technique
Paris plant is liliaceous plant, and subordinate mainly has Yunnan Rhizoma Paridis, paris polyphylla etc., and rhizome can be used as medicine,
As important Chinese medicine, it has also become Gongxuening capsule, Wound plaster of yannan Baiyao, Yunnan Baiyao spray, particle for relieving heat and cough,
The main raw material(s) of a variety of Chinese patent drugs and new drug such as antiviral granule.Since Paris polyphylla is huge medicinal and economic value, so that city
Head's phase, supply falls short of demand, and wild resource accelerates atrophy, therefore has expedited the emergence of family's kind mode of Paris polyphylla, main vegetative propagation containing stem tuber with
The two ways such as seed seedling-raising sexual propagation.But Paris polyphylla seed has both two kinds of characteristics of morphological dormacy and physiological dormancy simultaneously, needs
It could be sprouted by one summer of two winters, and the planting percent after sprouting is relatively low, cause seed seedling-raising breeding cycle length, risk big.Cause
This family plants Paris polyphylla and still breeds mostly by radical bud or stem tuber, causes the opposite reduction of commodity amount in the market, it is difficult to meet market need
It asks.Therefore the dormancy period for shortening Paris polyphylla seed, promotes its fast-germination, and efficiently, quickly producing sterile seedling is to alleviate Paris polyphylla
The effective approach of resource pressure.
Forefathers, which have been directed to Paris polyphylla seed and sprout the problems such as difficult, have been done correlative study and has applied for a patent, such as Patent No.
201410502503.9 application for a patent for invention discloses " a kind of to make four months long of paris polyphylla seed, eight months places come into leaves
Reason method ", low temperature places or so two weeks after which mainly dries in the shade mature seed, and 20 DEG C of crust are peelled off mid-October
Left and right is placed 3 months, is gone out to root long, and low-temperature cold store is placed 15 days, is sown in 2800 meters of height above sea level of big field, then 2 lis of lid later
The thick pine needle of rice.But the technical treatment time is long, and is limited by natural conditions larger.For another example Patent No.
201510163234.2 application for a patent for invention discloses " a kind of suspension culture method of Paris polyphylla nucellar cell ", this technology is main
Be using plus the B5 fluid nutrient medium of hormone suspension culture is carried out to the nucellar cell of Paris polyphylla, to obtain the weight containing effective ingredient
Building cell culture.More complex on this technical operation, practicability in operation is also less strong.
Summary of the invention
The object of the present invention is to provide a kind of methods for promoting Paris polyphylla seed fast germination.
For achieving the above object, the technical scheme adopted by the invention is that：One kind promoting Paris polyphylla seed fast germination
Method, include the following steps：
(1) pretreatment of seed：Remove kind of a skin；
(2) it impregnates for the first time：The sodium tungstate solution for the use of concentration being 0.05~0.3%, room temperature, impregnate the seed 10~
14h takes out seed；
(3) it impregnates for second：The hydrogen peroxide solution for the use of concentration being 1~3%, impregnates the 6~10h of seed by 3~5 DEG C,
Take out seed；
(4) third time is impregnated：The potassium nitrate solution and concentration for the use of concentration being 18~20mmol/L are 450~550mg/L
Gibberellins solution, at room temperature, joint impregnates the 36~48h of seed；
(5) the 4th immersions：The hydrogen peroxide for the use of concentration being 1~3%, impregnates the 6~10h of seed by 3~5 DEG C, takes out
Seed；
(6) the 5th immersions：The potassium nitrate solution and concentration for the use of concentration being 18~20mmol/L are 450~550mg/L
Gibberellins solution, room temperature, joint impregnate 36~48h of seed, take out seed, dry in the shade, then with concentration be 70% alcohol immersion
25~30s of seed immediately cleans seed；
(7) the 6th immersions：Using the mercuric chloride solution that concentration is 0.05~0.3% continue to impregnate the seed 6~
10min takes out seed, cleans, gnotobasis is dried, and is seeded on 1/2MS culture medium, vaccination area >=0.09cm2/.
Preferably, the sodium tungstate solution concentration that the first time impregnates is 0.1%, and the soaking time is 12h.
Preferably, described second hydrogen peroxide concentration impregnated is 2%, and the soaking time is 8h.
Preferably, the potassium nitrate solution concentration that the third time is impregnated is 19mmol/L, and gibberellin concentration is 500mg/L,
The soaking time is 42h.
Preferably, the hydrogen peroxide concentration of the 4th immersion is 2%, and the soaking time is 8h.
Preferably, the potassium nitrate solution concentration of the 5th immersion is 19mmol/L, and gibberellin concentration is 500mg/L,
The soaking time is 42h.
Preferably, the mercuric chloride solution concentration of the 6th immersion is 0.1%, and the soaking time is 8min.
8. the method according to claim 1 for promoting Paris polyphylla seed fast germination, it is characterised in that：The first time
The room temperature of immersion is 20 DEG C；Described second temperature impregnated is 4 DEG C.
9. the method according to claim 1 for promoting Paris polyphylla seed fast germination, it is characterised in that：The third time
The room temperature of immersion is 20 DEG C；The temperature of 4th immersion is 4 DEG C.
10. the method according to claim 1 for promoting Paris polyphylla seed fast germination, it is characterised in that：Described 5th
The room temperature of secondary immersion is 20 DEG C.
The invention has the advantages that：
1, the Paris polyphylla seed handled using technical solution of the present invention has general applicability, needle to paris plant seed
To a variety of Paris polyphylla seeds in belonging to, there is excellent rush rudiment effect.
2, the Paris polyphylla seed that the present invention is handled, germination is early, and germination percentage is high：Rudiment, 60 balances are begun to when 30 days
Equal germination percentage can reach 80% or more, and 90 days germination percentages are up to 95% or more.
3, the present invention takes full advantage of the physiological property of Paris polyphylla seed, easy to operate, the cost of raw material is cheap, extends to
In practical application.
Detailed description of the invention
Fig. 1 is the Paris polyphylla seed base-root just prominent schematic diagram that breaks in the seed coat；
Fig. 2 is that Paris polyphylla seed base-root is long to portable status diagram；
Fig. 3 is that the Paris polyphylla seed after transplanting grows up to seedling schematic diagram.
Specific embodiment
Concrete operation step of the invention is as follows：
1, the pretreatment of seed：Paris polyphylla seed is taken, external red kind skin is removed in rubbing, and aseptic deionized water is cleaned, natural
It dries in the shade；
2, it impregnates for the first time：The sodium tungstate solution for the use of concentration being 0.05~0.3%, impregnates the seed to dry in the shade at 20 DEG C
10~14h takes out seed；
3, it impregnates for second：The hydrogen peroxide solution for the use of concentration being 1~3% continues to impregnate 6~10h of seed at 4 DEG C,
Take out seed；
4, third time is impregnated：The potassium nitrate solution and concentration for the use of concentration being 18~20mmol/L are 450~550mg/L's
Gibberellins solution, joint impregnates 36~48h of seed at 20 DEG C；
5, the 4th immersion：The hydrogen peroxide for the use of concentration being 1~3% continues to impregnate 6~10h of seed at 4 DEG C, take out
Seed；
6, the 5th immersion：The potassium nitrate solution and concentration for the use of concentration being 18~20mmol/L are 450~550mg/L's
Gibberellins solution, joint impregnates 36~48h of seed at 20 DEG C, takes out seed, dries in the shade at room temperature, then the wine for being 70% with concentration
Essence impregnates seed 30s, immediately cleans seed with sterile water；
7, the 6th immersion：Continue to impregnate 6~10min of seed using the mercuric chloride solution that concentration is 0.05~0.3%, take out
Seed is placed in superclean bench and is dried, is then seeded on 1/2MS culture medium with sterile water wash seed 5~6 times, inoculation
Area >=0.09cm2/.Carry out seed germination culture.To seed base-root it is prominent break in the seed coat and grow to 2~3cm, cotyledon formed,
When endosperm has been absorbed, seed is chosen, is transplanted in the cylindrical culture bottle of new addition WPM (inoculation radius >=1cm).
It is emphasized that immersion sequence of the invention and time, the medical fluid impregnated each time and concentration are all through excessive
Amount test and finally confirm after proving repeatedly, cancel certain primary soaking concentration for impregnating, exchanging immersion sequence, change medical fluid,
All it is unable to reach effect of the invention；
Below with reference to specific embodiment, further explaination is done to the present invention.Because length limits, only choose most representative
Part test as embodiment, the test in verification process is far above several following.
Embodiment one：It impregnates number and the time is preferred
1, choose current year fresh mature, uniform in size, state is suitable, the identical Yunnan Rhizoma Paridis seed 1900 of storage time
, be divided into 19 groups, every group 100, seed weight in each group and between group equal difference it is not significant.Seed is carried out using the above method
Rudiment.Wherein, soaking time is "/", and expression does not carry out the secondary immersion；Concentration of sodium tungstate is 0.1%, and hydrogen peroxide concentration is
2%, nitric acid potassium concn is 19mmol/L, and gibberellin concentration is 500mg/L, and mercuric chloride concentration is 0.1%.Each group it is specific
Operating parameter is as shown in table 1.
The other specific preparation parameter of 1 each group of table
2, above-mentioned each group was counted respectively in 30 days, 45 days, 60 days, 90 days, 180 days average germination percentages.Wherein, 180 days
The seed not germinateed also is counted as germination failure；180 days germination percentages are counted as scheme failure lower than 90%.Concrete outcome is as shown in table 2.
The other germination of 2 each group of table
As can be seen from the above table：When omitting certain primary immersion, germination percentage is significantly lower than each group normally handled.Use this hair
The Paris polyphylla seed of bright technical solution processing, 90 days average germination percentages are 90% or more, and the mode of selection group 2, and germination percentage is most
It is high.When improving the soaking time of certain medical fluids, germination percentage is declined, this may be long soaking time, and Paris polyphylla seed exists
The germination later period is easy rotten reason.
3, swashed using the endogenous of Paris polyphylla seed before Elisa kit (being purchased from bronze object Bioisystech Co., Ltd) test processes
Plain GA, IAA and ABA content；And when counting germination percentage, i.e., each group is tested in 30 days, 45 days, 60 days, 90 days and 180 days respectively
The Endogenous Hormone Contents in Vitro state of the Paris polyphylla seed slightly dried after processing.
The results are shown in Table 3.
The other Endogenous Hormone Contents in Vitro of 3 each group of table
As can be seen from the above table, the Paris polyphylla seed (group 1~13) handled using technical solution of the present invention, each endogenous hormones
Variation is：Gibberellin (GA) content obviously increases, and effectively breaks seed dormancy；Auxin (IAA) content also obviously increases, side
Help promotion seed growth；Abscisic acid (ABA) content is substantially reduced, and further helps to break seed dormancy.Wherein, using group 2
Technical solution, in seed the variation of each endogenous hormones most beneficial for seed rudiment and growth, this result also with step 2
Germination test result is consistent.
Embodiment two：Immersion sequence is preferred
1, choose current year fresh mature, uniform in size, state is suitable, the identical Yunnan Rhizoma Paridis seed 1100 of storage time
, be divided into 11 groups, every group 100, seed weight in each group and between group equal difference it is not significant.Use the above method, adjustment leaching
Bubble sequence, carries out seed germination.Concentration of sodium tungstate is 0.1%, and hydrogen peroxide concentration is 2%, and nitric acid potassium concn is
19mmol/L, gibberellin concentration are 500mg/L, and mercuric chloride concentration is 0.1%.
Wherein, X time is represented with digital X to impregnate, such as：Immersion sequence be：12345, it indicates sequentially to carry out first time leaching
Bubble impregnates for second until the 5th immersion；For another example immersion sequence is：13452, indicate immersion sequence
For：It impregnates for the first time, third time is impregnated, is impregnated for the 4th time, is impregnated for the 5th time, impregnating for second.
The specific immersion sequence of each group is as shown in table 4.
The other immersion sequence of 4 each group of table
2, above-mentioned each group was counted respectively in 30 days, 45 days, 60 days, 90 days, 180 days average germination percentages.Wherein, 180 days
The seed not germinateed also is counted as germination failure；180 days germination percentages are counted as scheme failure lower than 90%.Concrete outcome is as shown in table 5.
The other germination of 5 each group of table
3, by the method for embodiment one, endogenous hormones GA, ZR, IAA and ABA content of Paris polyphylla seed before measurement is handled；And
When counting germination percentage, i.e., 30 days, 45 days, 60 days, 90 days and 180 days, Paris polyphylla seed is endogenous after test each group is handled respectively
Hormone-content.The results are shown in Table 6.
The other Endogenous Hormone Contents in Vitro of 6 each group of table
Embodiment three：The concentration for impregnating medical fluid is preferred
1, choose current year fresh mature, uniform in size, state is suitable, the identical Yunnan Rhizoma Paridis seed 1300 of storage time
, be divided into 13 groups, every group 100, seed weight in each group and between group equal difference it is not significant.Seed is carried out using the above method
Rudiment.Wherein, the concentration of each group immersion medical fluid is as shown in table 7.
The concentration of each group of table 7 immersion medical fluid
2, above-mentioned each group was counted respectively in 30 days, 45 days, 60 days, 90 days, 180 days average germination percentages.Wherein, 180 days
The seed not germinateed also is counted as germination failure；180 days germination percentages are counted as scheme failure lower than 90%.Concrete outcome is as shown in table 8.
The other germination of 8 each group of table
3, by the method for embodiment one, endogenous hormones GA, ZR, IAA and ABA content of Paris polyphylla seed before measurement is handled；And
When counting germination percentage, i.e., 30 days, 45 days, 60 days, 90 days and 180 days, Paris polyphylla seed is endogenous after test each group is handled respectively
Hormone-content.The results are shown in Table 9.
The other Endogenous Hormone Contents in Vitro of 9 each group of table
Claims (10)
1. a kind of method for promoting Paris polyphylla seed fast germination, it is characterised in that：Include the following steps：
(1) pretreatment of seed：Remove kind of a skin；
(2) it impregnates for the first time：The sodium tungstate solution for the use of concentration being 0.05~0.3%, room temperature impregnate the 10~14h of seed,
Take out seed；
(3) it impregnates for second：The hydrogen peroxide solution for the use of concentration being 1~3%, impregnates the 6~10h of seed by 3~5 DEG C, takes out
Seed；
(4) third time is impregnated：The potassium nitrate solution and concentration for the use of concentration being 18~20mmol/L are the red of 450~550mg/L
Mycin solution, at room temperature, joint impregnate the 36~48h of seed；
(5) the 4th immersions：The hydrogen peroxide for the use of concentration being 1~3% 3~5 DEG C, impregnates the 6~10h of seed, takes out kind
Son；
(6) the 5th immersions：The potassium nitrate solution and concentration for the use of concentration being 18~20mmol/L are the red of 450~550mg/L
Mycin solution, room temperature, joint impregnate 36~48h of seed, take out seed, dry in the shade, then the alcohol for being 70% with concentration impregnates seed
25~30s immediately cleans seed；
(7) the 6th immersions：Continue to impregnate the 6~10min of seed using the mercuric chloride solution that concentration is 0.05~0.3%, take
Seed out is cleaned, and gnotobasis is dried, and is seeded on 1/2MS culture medium, vaccination area >=0.09cm2/.
2. the method according to claim 1 for promoting Paris polyphylla seed fast germination, it is characterised in that：The first time impregnates
Sodium tungstate solution concentration be 0.1%, the soaking time be 12h.
3. the method according to claim 1 for promoting Paris polyphylla seed fast germination, it is characterised in that：Second of immersion
Hydrogen peroxide concentration be 2%, the soaking time be 8h.
4. the method according to claim 1 for promoting Paris polyphylla seed fast germination, it is characterised in that：The third time is impregnated
Potassium nitrate solution concentration be 19mmol/L, gibberellin concentration be 500mg/L, the soaking time be 42h.
5. the method according to claim 1 for promoting Paris polyphylla seed fast germination, it is characterised in that：4th immersion
Hydrogen peroxide concentration be 2%, the soaking time be 8h.
6. the method according to claim 1 for promoting Paris polyphylla seed fast germination, it is characterised in that：5th immersion
Potassium nitrate solution concentration be 19mmol/L, gibberellin concentration be 500mg/L, the soaking time be 42h.
7. the method according to claim 1 for promoting Paris polyphylla seed fast germination, it is characterised in that：6th immersion
Mercuric chloride solution concentration be 0.1%, the soaking time be 8min.
8. the method according to claim 1 for promoting Paris polyphylla seed fast germination, it is characterised in that：The first time impregnates
Room temperature be 20 DEG C；Described second temperature impregnated is 4 DEG C.
9. the method according to claim 1 for promoting Paris polyphylla seed fast germination, it is characterised in that：The third time is impregnated
Room temperature be 20 DEG C；The temperature of 4th immersion is 4 DEG C.
10. the method according to claim 1 for promoting Paris polyphylla seed fast germination, it is characterised in that：5th leaching
The room temperature of bubble is 20 DEG C.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1210663A (en) * | 1998-09-08 | 1999-03-17 | 中国科学院昆明植物研究所 | Seed sprouting and quick propagating technology of Yunnan Rhizoma Paridis |
| JP2003081858A (en) * | 2001-09-13 | 2003-03-19 | Nippon Yakuyo Shokuhin Kenkyusho:Kk | Prophylactic and therapeutic agent for diabetes mellitus and health food |
| CN102487627A (en) * | 2011-12-09 | 2012-06-13 | 四川农业大学 | Method for effectively breaking dormancy of Paris polyphylla seeds |
| CN103621557A (en) * | 2013-12-20 | 2014-03-12 | 贵州大学 | Special treatment agent for polyphylla seed germination |
| CN106233871A (en) * | 2016-07-29 | 2016-12-21 | 柳州创宇科技有限公司 | A kind of processing method improving Rhizoma Paridis seed germination rate |
| CN107810958A (en) * | 2017-10-19 | 2018-03-20 | 中国农业大学 | A kind of method for promoting Yunnan Paris polyphylla seed to sprout |
-
2018
- 2018-06-26 CN CN201810666071.3A patent/CN108811584A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1210663A (en) * | 1998-09-08 | 1999-03-17 | 中国科学院昆明植物研究所 | Seed sprouting and quick propagating technology of Yunnan Rhizoma Paridis |
| JP2003081858A (en) * | 2001-09-13 | 2003-03-19 | Nippon Yakuyo Shokuhin Kenkyusho:Kk | Prophylactic and therapeutic agent for diabetes mellitus and health food |
| CN102487627A (en) * | 2011-12-09 | 2012-06-13 | 四川农业大学 | Method for effectively breaking dormancy of Paris polyphylla seeds |
| CN103621557A (en) * | 2013-12-20 | 2014-03-12 | 贵州大学 | Special treatment agent for polyphylla seed germination |
| CN106233871A (en) * | 2016-07-29 | 2016-12-21 | 柳州创宇科技有限公司 | A kind of processing method improving Rhizoma Paridis seed germination rate |
| CN107810958A (en) * | 2017-10-19 | 2018-03-20 | 中国农业大学 | A kind of method for promoting Yunnan Paris polyphylla seed to sprout |
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