CN104541665B - Method for breaking dormancy of aconitum kusnezoffii reichb seeds - Google Patents
Method for breaking dormancy of aconitum kusnezoffii reichb seeds Download PDFInfo
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- CN104541665B CN104541665B CN201510033569.2A CN201510033569A CN104541665B CN 104541665 B CN104541665 B CN 104541665B CN 201510033569 A CN201510033569 A CN 201510033569A CN 104541665 B CN104541665 B CN 104541665B
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Abstract
The invention relates to the technical field of traditional Chinese medicine cultivation, and in particular relates to a method for breaking the dormancy of aconitum kusnezoffii reichb seeds. The method comprises the steps of sterilizing the aconitum kusnezoffii reichb seeds, and then treating the seeds with 300-400mg/L gibberellin; and carrying out high and low temperature alternative cultivation on the seeds treated with the gibberellin, and then culturing at the temperature of 0-5 DEG C, wherein in the process of the high and low temperature alternative cultivation, the temperature of the low temperature cultivation is controlled to 14-16 DEG C, the time of the low temperature cultivation is controlled to 11-13 hours, the temperature of the high temperature cultivation is controlled to 19-21 DEG C, and the time of the high temperature cultivation is controlled to 11-13 hours. According to the method, the aconitum kusnezoffii reichb seeds are treated by combining the gibberellin and variable temperature, so that the dormancy of the aconitum kusnezoffii reichb seeds can be rapidly broken, of the dormancy period, being at least 90 days, of the aconitum kusnezoffii reichb seeds under traditional technical conditions is shortened to about 40 days, and the germination rate is increased to 78.65%.
Description
Technical field
The present invention relates to technical field of Chinese herbal medicine cultivation, more particularly to one kind breaks the seed dormant method of Aconitum kusnezoffii Reichb.
Background technology
Ranunculaceae aconitum plant is divided into Aconitum carmichjaelii Debx. subgenus (Subgen Aconitum), Radix aconiti gymnandri subgenus (Subgen
) and Herba Aconiti Ochranthi subgenus (Subgen Gymnaconitum) Paraconitum.Wherein, Aconitum carmichjaelii Debx. subgenus is divided into Aconitum carmichjaelii Debx. group and many fruit crows again
Head group, Aconitum carmichjaelii Debx. group are divided into Aconitum carmichjaelii Debx. system of Xingan, aconitum brachypodum system, brown purple monkshood root system, Baoshan Aconitum carmichjaelii Debx. system, Aconitum soongaricum system, rock again
Aconitum carmichjaelii Debx. system, Aconitum rotunifolium Kar. Et kir. system, aobvious post Aconitum carmichjaelii Debx. system, aconitum tanguticumStapf system, Radix aconiti volubiliss system.Containing biologies such as aconitines more than aconitum plant
Alkali, the tuber of most species have severe toxicity.36 kinds are there are about in aconitum plant can hyoscine.But Aconitum carmichjaelii Debx. morphological characteristic not of the same race,
Habit, medical value all have difference.For example：
Aconitum carmichjaelii Debx. (Aconitum carmichaeli Debx.) category Aconitum carmichjaelii Debx. subgenus Radix aconiti volubiliss system, Aconitum carmichjaelii Debx. main product Sichuan, Shaanxi.
Yunnan, Guizhou, Hebei, Hunan, Hubei, Jiangxi, Gansu etc. save cultivation.In raw mountain region grass slope or shrubbery.Aconitum seeds length 3～
3.2 millimeters, prismatic, only in the horizontal film wing of the close life in two faces.In 9～October of florescence, fruiting period October is to November.《Chinese Pharmacopoeia 2010》Rule
Fixed its dried root is " Radix Aconiti ".With expelling wind and removing dampness, antalgic.For anemofrigid-damp arthralgia, arthralgia, trusted subordinate's crymodynia, cold
Hernia is had a pain and narcotic analgesic.
Radix aconiti gymnandri (Aconitum gymnandrum Maxim.) belongs to Radix aconiti gymnandri subgenus, is distributed in Tibet, Si Chuanxi
Portion, Qinghai, SOUTH OF GANSU.It is born in mountain region Cao Po, TIANBIANCAO ground or river bank sandy ground.Seed fall ovoid, the horizontal narrow wing of close life.Florescence
6～August.Effect with dispelling wind and eliminating pain, for wind-damp dispelling, pain relieving, parasite killing etc..
Radix Aconiti Coreani (Aconitum coreanum (H.L é v.) Rapaics) category Aconitum carmichjaelii Debx. subgenus aconitum brachypodum system, is distributed in
China, Korea, Far-east Area of Russia, are born in mountain region grass slope or sparse woods.Seed is about ellipse, has three vertical ribs, florescence 8
～September.The diseases such as migraine and general headache, arthralgia due to cold-dampnes, facial hemiparalysiss are controlled, external can control scabies.Tuber infusion can used as pesticides, prevent and treat it is little
Straw rust.
Aconitum kusnezoffii Reichb (Aconitum kusnezoffii Rchb) category Aconitum carmichjaelii Debx. subgenus Radix aconiti volubiliss system, is mainly distributed in China
Shanxi, Hebei, the Inner Mongol, Liaoning, Jilin and Heilungkiang, are born on hillside or grassy marshland.The seed of Aconitum kusnezoffii Reichb is about 2.5 millimeters,
Flat ellipse is spherical, has narrow wing along rib, only looks unfamiliar horizontal film wing one.7～September of florescence.《Chinese Pharmacopoeia 2010》Specify its dried pieces
Root is " Radix Aconiti Kusnezoffii ", the function with expelling wind and removing dampness, antalgic, and for anemofrigid-damp arthralgia, arthralgia, trusted subordinate's crymodynia, colic of cold type are made
Pain and narcotic analgesic.At present, Radix Aconiti Kusnezoffii has shown good application foundation at aspects such as heart tonifying, analgesia, antiinflammatory, regulation immunity.
" cinobufagin " with Radix Aconiti Kusnezoffii as primary raw material is treating cancer pain at late stage compared with current conventional analgesic " dolantin "
Etc. aspect it is evident in efficacy, and without additive, be widely used.The demand of Radix Aconiti Kusnezoffii resource is increasingly improved.
But, due to ecological environment heavy damage and artificial excavation, Aconitum kusnezoffii Reichb wild resource tends to exhausted.Because of the place of production and
It is customary different also by drying that is Dendrobium denneanum Kerr. black, splitting more Aconitum carmichjaelii Debx., Radix aconiti hemsleyani, circular cone sequence Aconitum carmichjaelii Debx., Aconitum Sungpanense Hand-mazz and too white Aconitum carmichjaelii Debx.
Tuber is used when Radix Aconiti Kusnezoffii, causes market Radix Aconiti Kusnezoffii base source product chaotic, and the effectiveness of Chinese medicine, safety and stability are difficult to control to.Therefore,
Aconitum kusnezoffii Reichb introduction and acclimatization is carried out, wild change man kind is made, the in situ conservation and sustainable use for realizing Aconitum kusnezoffii Reichb resource seems particularly
It is important.There is embryo after-ripening and physiology after-ripening in Aconitum kusnezoffii Reichb seed, although having reached maturity in the form of seed, embryo is not yet complete
Into development, embryo small volume, structure imperfection, it is necessary to which, again after a period of time development makes embryo maturation, seed just possesses
The condition of sprouting.This characteristic of Aconitum kusnezoffii Reichb seed makes Aconitum kusnezoffii Reichb occur during seminal propagation that germination rate is low, germination is slow,
Germinate irregular phenomenon.Its rest period length, the low problem of germination percentage become the Main Bottleneck of Aconitum kusnezoffii Reichb large-scale production, seriously
Have impact on the seminal propagation effect of Aconitum kusnezoffii Reichb.
At present, although the vegetative manners such as bud breeding, profit can be bred or be cut to artificial growth Aconitum kusnezoffii Reichb using tuber
There is many defects with asexual propagation, such as (1) is bred using tuber, its breeding coefficient is relatively low, one plant of wild product be only capable of breeding 3～
5 plants of cultivation product；(2) after many generation breedings are carried out using tuber, seedling will appear from degradation phenomena, and concrete manifestation resistance is poor, disease
The low aspect of many, yield；(3) destructive injury equally can be caused using tuber breeding to wild resource.
Therefore, break Aconitum kusnezoffii Reichb seed dormancy, improve its germination rate, the crucial skill for changing Aconitum kusnezoffii Reichb cultivation mode will be become
Art, while the development of Aconitum kusnezoffii Reichb industry will be promoted.
The content of the invention
In view of this, the technical problem to be solved in the present invention is to provide one kind quickly break Aconitum kusnezoffii Reichb seed dormancy, carry
The method of high Aconitum kusnezoffii Reichb percentage of seedgermination.
The seed dormant method of Aconitum kusnezoffii Reichb of breaking that the present invention is provided includes：
After the sterilization of Aconitum kusnezoffii Reichb seed, with concentration as 300mg/L～gibberellin treatment of 400mg/L；
Seed after gibberellin treatment is carried out high and low temperature alternative culture to cultivate after 0 DEG C～5 DEG C；
In high and low temperature alternative incubation, the temperature of Low- temperature culture is 14 DEG C～16 DEG C, and the time is 11h～13h；High temperature is trained
Foster temperature is 19 DEG C～21 DEG C, and the time is 11h～13h.
The low key problem for being to cause current Aconitum kusnezoffii Reichb seminal propagation bottleneck of the germination rate of Aconitum kusnezoffii Reichb seed.Aconitum plant
Generally existing embryo after-ripening phenomenon in seed, but existing result of study shows：Aconitum plant not of the same race is to accelerating germination condition
Response is also different, shows as：Different inter-species are different to kindred plant hormone optimum concentration；Different inter-species are to different phytohormone
Response is different；Response of the different inter-species to inducing temperature is also different.The present invention is experimentally confirmed, after the sterilization of Aconitum kusnezoffii Reichb seed,
Jing after gibberellins immersion and Fluctuation temperature culture, seed germination rate is up to more than 78%, and it is left that the rest period of seed is shorten to 40d
It is right.
Preferably, the temperature of Low- temperature culture is 15 DEG C, the time is 12h；The temperature of high-temperature cultivation is 20 DEG C, and the time is
12h；High and low temperature alternative culture is cultivated after 3 DEG C.
Preferably, the air humidity of Fluctuation temperature culture is 65%～70%.
The dormancy time temperature influence of Aconitum kusnezoffii Reichb seed is larger, and the present invention is experimentally confirmed, using present invention offer
Temp change method culture, the germination percentage of seed can be significantly improved.
Preferably, seed is placed in Fluctuation temperature culture on the filter paper of two-layer moistening.
Preferably, the seed dormant method of Aconitum kusnezoffii Reichb of breaking that the present invention is provided is carried out under dark condition.
Preferably, after alternate culture, the embryo rate of Aconitum kusnezoffii Reichb seed is not less than 80%.
Embryo rate refers to that embryo length accounts for the percentage ratio of endosperm length.Research practice shows that, up to more than 80%, now embryo is differentiated for embryo rate
Go out cotyledon, plumule, plumular axis, radicle, be substantially finished form after-ripening.But, now seed can't be sprouted, it is necessary in low temperature bar
Physiology after-ripening is carried out under part, embryo does not morphologically occur any change between physiological afterripening period, simply idiosome increase.Only when northern crow
Head seed can be sprouted after completing form after-ripening and physiology after-ripening, and exists between form after-ripening and physiology after-ripening strict
Succession and irreversibility, therefore, only when Aconitum kusnezoffii Reichb seed is Jing after Fluctuation temperature culture is not less than 80% to embryo rate, carry out low temperature training
Support and make it into physiology after-ripening, just can guarantee that final percentage of seedgermination.
Preferably, the time of high and low temperature alternative culture is 18d～22d.
Preferably, the time of high and low temperature alternative culture is 20d.
Preferably, the time of 0 DEG C～5 DEG C cultures is 17d～25d.
Preferably, the time of 0 DEG C～5 DEG C cultures is 20d.
Preferably, the concentration of gibberellins is 300mg/L.
Preferably, the time of gibberellin treatment is 24h.
Gibberellins are a Plant Hormones, can stimulate the growth of leaf and bud.The prior art indicate that, gibberellins are not to of the same race
The optimum concentration of aconitum plant is inconsistent.But it is current, and gibberellins are had no for breaking the seed dormant report of Aconitum kusnezoffii Reichb
Road.Also, as the nutrition that seed is sprouted places one's entire reliance upon endosperm, and work as gibberellins excessive concentration, although embryo may be made
Rapidly grow up under the stimulation of gibberellins, the nutrient substance of endosperm storage may but hydrolyzed in a large number, it is impossible in time by embryo
Growth promoter is utilized, and is caused endosperm to liquefy, is ultimately resulted in seed decay, death.The present invention experiments prove that, when gibberellins it is dense
Spend for 300mg/L when, most beneficial for the sprouting of seed, and rotten kind of rate is relatively low.
Preferably, the aqueous sodium hypochlorite solution for adopting mass fraction for 1% that sterilizes.
Preferably, the time of sterilization is 5～10min.
Preferably, sterilization and gibberellin treatment between, also including distilled water flushing the step of；The number of times of flushing is 3～5
It is secondary.
Seed is gone rotten caused by carrying substantial amounts of antibacterial or funguses in being because its epidermis in incubation.The present invention is adopted
Seed is carried out after sterilization processing with the aqueous sodium hypochlorite solution that mass fraction is 1%, then seed is cultivated.Experiment card
Real, the method provided using the present invention is sterilized, and rotten kind of rate of Aconitum kusnezoffii Reichb seed is less than 10%.
The seed dormant method of Aconitum kusnezoffii Reichb of breaking that the present invention is provided includes：After the sterilization of Aconitum kusnezoffii Reichb seed, with concentration it is
The gibberellin treatment of 300mg/L～400mg/L；Seed after gibberellin treatment is carried out into high and low temperature alternative culture after 0 DEG C～5
DEG C culture；In high and low temperature alternative incubation, the temperature of Low- temperature culture is 14 DEG C～16 DEG C, and the time is 11h～13h；High temperature is trained
Foster temperature is 19 DEG C～21 DEG C, and the time is 11h～13h.The present invention combines gibberellins and intermittent warming Aconitum kusnezoffii Reichb seed, can
Quickly break Aconitum kusnezoffii Reichb seed dormancy, by under the conditions of Aconitum kusnezoffii Reichb seed conventional art at least the rest period of 90d to foreshorten to 40d left
The right side, germination percentage bring up to 78.65%, and processing method of the present invention is easy, efficiently, can release Aconitum kusnezoffii Reichb seed within a short period of time and stop
Sleep so as to sprout, pre-treatment is sowed for Aconitum kusnezoffii Reichb large-scale production and indoor seed testing provides fast and efficiently technology
Measure, has broad application prospects.
Specific embodiment
The invention provides one kind breaks the seed dormant method of Aconitum kusnezoffii Reichb, those skilled in the art can be used for reference in herein
Hold, be suitably modified technological parameter realization.Specifically, all similar replacements and change are to those skilled in the art
For be it will be apparent that they are considered as being included in the present invention.The method of the present invention and application have passed through preferably enforcement
Example is described, related personnel substantially can in without departing from present invention, spirit and scope to method described herein and
Using be modified or suitably change and combine, realize and apply the technology of the present invention.
The raw material that the present invention is adopted is all common commercially available product, all can be buied by market.
Wherein, 1, Aconitum kusnezoffii Reichb seed：For trying Aconitum kusnezoffii Reichb (Aconitum kusnezoffii Reichb.) seed in 2014
October in year collection Jilin Province Panshi City Qu Chaihe towns, are identified by Gao Yun Wang Yingping researcher.Adopt
After the seed natural air drying of collection, the impurity such as carpopodium, shell, mildew seed are removed, store standby at shady and cool drying.
2nd, chemical reagent：Gibberellins GA3 is purchased from Shanghai Yuan Ye bio tech ltd, and sodium hypochlorite is purchased from Tianjin good fortune
Morning chemical reagent factory.
3rd, instrument and equipment：Slide gauge (0～150mm), digital display temperature control refrigerator (BCD-251WBSV), programmable optical is according to training
Foster case (MGC-250BP-2)
With reference to embodiment, the present invention is expanded on further：
Embodiment 1～2
Take Aconitum kusnezoffii Reichb seed, remove impurity, with 1% liquor natrii hypochloritises 5～10min of sterilization, then with distilled water
Rinse 3～5 times, the seed after sterilization is soaked into 24h respectively in the Gibberellins solution of different quality concentration, size is then selected
Unanimously, the seed of full grains is placed in and is lined with the culture dish of two layers of filter paper, puts 100 seeds, at each in each culture dish
Reason is repeated 3 times, and the seed of each process is carried out treatment of different temperature, determines each embryo length and embryo for processing seed using slide gauge
Breast is long, calculates embryo rate, germination rate, rotten kind of rate.Measured value when embryo rate is cultivated 20 days under the conditions of being determined as this；Germination rate is culture 40
Its measured value, all rotten kind of rate minutes are culture 40 days.Embodiment arranges as shown in table 1：
1 embodiment of table is arranged
Comparative example 1～19
Take Aconitum kusnezoffii Reichb seed, remove impurity, with 1% liquor natrii hypochloritises 5～10min of sterilization, then with distilled water
Rinse 3～5 times, the seed after sterilization is soaked into 24h respectively in the Gibberellins solution of different quality concentration, size is then selected
Unanimously, the seed of full grains is placed in and is lined with the culture dish of two layers of filter paper, puts 100 seeds, at each in each culture dish
Reason is repeated 3 times, and the seed of each process is carried out treatment of different temperature, determines each embryo length and embryo for processing seed using slide gauge
Breast is long, calculates embryo rate, germination rate, rotten kind of rate.Comparative example 1～13：Measured value when embryo rate is cultivated 20 days under the conditions of being determined as this；Sprout
The rate of sending out is 40 days measured values of culture, and all rotten kind of rate minutes are culture 40 days.Comparative example arranges as shown in table 2：
2 comparative example of table is arranged
Embodiment 3
One factor analysis of variance is carried out to the statistical data of embodiment 1～2 and comparative example 1～19 using SAS8.0 softwares,
Different disposal group difference significance is analyzed using Fisher's LSD methods.Computing formula is：
Embryo rate (%)=embryo length/endosperm length × 100%
Subnumber × 100% is planted experimentally in rotten kind of rate (%)=seed number/confession of going rotten
Subnumber × 100% is planted experimentally in germination rate (%)=germination seed number/confession
Statistical result is as shown in table 3：
3 result of table is counted
As a result show：There is deep-sleep in Aconitum kusnezoffii Reichb seed, under normal condition, its rest period is long, and germination percentage is low, and Jing matter
Amount concentration 300mgL-1～400mgL-1Gibberellin treatment, embryonic development speed are significantly higher than control, and gibberellins optimization process is dense
Spend for 300mgL-1, better with reference to 15 DEG C (12h), 20 DEG C of (12h) intermittent warmings, embryo rate can reach 80% in culture 20d
More than, and after 3 DEG C of cryogenic thermostat cultures of Jing, germination rate be significantly higher than other process and matched group, it is seen then that the method can be in short-term
It is interior to break Aconitum kusnezoffii Reichb seed dormancy, promote its seed to sprout.
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (7)
1. one kind breaks the seed dormant method of Aconitum kusnezoffii Reichb, it is characterised in that include：
After the sterilization of Aconitum kusnezoffii Reichb seed, with concentration as 300mg/L～gibberellin treatment of 400mg/L；
The time of the gibberellin treatment is 24h；
Seed after gibberellin treatment is carried out high and low temperature alternative culture to cultivate after 0 DEG C～5 DEG C；
In the high and low temperature alternative incubation, the temperature of Low- temperature culture is 14 DEG C～16 DEG C, and the time is 11h～13h；High temperature is trained
It is 11h～13h 19 DEG C～21 DEG C times that foster temperature is；
The time of the high and low temperature alternative culture is 18d～22d；
The time of described 0 DEG C～5 DEG C cultures is 17d～25d.
2. method according to claim 1, it is characterised in that the temperature of the Low- temperature culture is 15 DEG C, and the time is 12h；
The temperature of the high-temperature cultivation is 20 DEG C, and the time is 12h；The high and low temperature alternative culture is cultivated after 3 DEG C.
3. method according to claim 1, it is characterised in that after the high and low temperature alternative culture, the embryo of Aconitum kusnezoffii Reichb seed
Rate is not less than 80%.
4. method according to claim 1, it is characterised in that the concentration of the gibberellins is 300mg/L.
5. method according to claim 1, it is characterised in that the sterilization adopt mass fraction for 1% sodium hypochlorite
Aqueous solution.
6. method according to claim 1, it is characterised in that the time of the sterilization is 5～10min.
7. method according to claim 1, it is characterised in that between the sterilization and gibberellin treatment, also including distillation
The step of water is rinsed；The number of times of the flushing is 3～5 times.
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| CN101715665B (en) * | 2009-12-30 | 2012-08-22 | 雅安三九中药材科技产业化有限公司 | Method for improving germination rate of monkshood seed |
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