CN104541665A - 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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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, particularly relate to a kind of method breaking northern aconitum seeds dormancy.
Background technology
Ranunculaceae aconitum plant is divided into rhizome of Chinese monkshood subgenus (Subgen Aconitum), Root of Nakedstamen Monkshood subgenus (Subgen Paraconitum) and the flat subgenus of ox (Subgen Gymnaconitum).Wherein, rhizome of Chinese monkshood subgenus is divided into again rhizome of Chinese monkshood group and many fruit rhizome of Chinese monkshood groups, and rhizome of Chinese monkshood group is divided into again rhizome of Chinese monkshood system of Xingan, aconitum brachypodum system, brown purple monkshood root system, Baoshan rhizome of Chinese monkshood system, aconitum soongoricum Stapf system, rock rhizome of Chinese monkshood system, Aconitum rotunifolium Kar. Et kir system, aobvious post rhizome of Chinese monkshood system, aconitum tanguticumStapf system, twining monkshood root system.Aconitum plant is many containing alkaloids such as aconitines, and the block root of most kind has severe toxicity.About have in aconitum plant 36 kinds can hyoscine.But rhizome of Chinese monkshood morphological feature not of the same race, habit of growth, medical value all have difference.Such as:
The rhizome of Chinese monkshood (Aconitum carmichaeli Debx.) belongs to rhizome of Chinese monkshood subgenus twining monkshood root system, rhizome of Chinese monkshood main product Sichuan, Shaanxi.Yunnan, Guizhou, Hebei, Hunan, Hubei, Jiangxi, Gansu etc. are economized cultivation.Give birth in grass slope, mountain region or shrubbery.Long 3 ~ 3.2 millimeters of aconitum seeds, prismatic, only at two horizontal film wings of close life.In 9 ~ October of florescence, really October phase is to November." Chinese Pharmacopoeia 2010 " specifies that its dried root is for " monkshood ".Have dispel rheumatism, antalgic.Have a pain and narcotic analgesic for wind-cold-dampness arthralgia, arthralgia, trusted subordinate's crymodynia, cold hernia.
Root of Nakedstamen Monkshood (Aconitum gymnandrum Maxim.) belongs to Root of Nakedstamen Monkshood subgenus, is distributed in Tibet, western Sichuan, Qinghai, SOUTH OF GANSU.Be born in mountain region Cao Po, meadow, limit, field or river bank sandy ground.Seed falls ovoid, the horizontal narrow wing of close life.6 ~ August of florescence.There is analgesic effect of dispeling the wind, for wines used as antirheumatic, pain relieving, desinsection etc.
Radix coniti coreani (Aconitum coreanum (H.L é v.) Rapaics) belongs to rhizome of Chinese monkshood subgenus aconitum brachypodum system, is distributed in China, Korea, Far-east Area of Russia, is born in grass slope, mountain region or sparse woods.Seed is about ellipse, tool three vertical ribs, 8 ~ September of florescence.Control the diseases such as migraine and general headache, numbness and ache of cold dampness, facial paralysis, external application can control mange.Block root infusion can used as pesticides, prevents and treats the stem rust of wheat.
The north rhizome of Chinese monkshood (Aconitum kusnezoffii Rchb) belongs to rhizome of Chinese monkshood subgenus twining monkshood root system, is mainly distributed in Shanxi, Hebei, the Inner Mongol, Liaoning, Jilin and Heilungkiang, is born on hillside or grassy marshland in China.The seed of the north rhizome of Chinese monkshood is about 2.5 millimeters, and flat ellipse is spherical, along the narrow wing of rib tool, only to look unfamiliar horizontal film wing one.7 ~ September of florescence." Chinese Pharmacopoeia 2010 " specifies that its dried root is for " radix aconiti agrestis ", have dispel rheumatism, the function of antalgic, for wind-cold-dampness arthralgia, arthralgia, trusted subordinate's crymodynia, cold hernia is had a pain and narcotic analgesic.At present, radix aconiti agrestis has shown good application foundation in cardiac stimulant, analgesia, anti-inflammatory, immunity moderation etc." Cinobufagin " that take radix aconiti agrestis as primary raw material is compared with current conventional antalgesic " Sauteralgyl ", evident in efficacy in treatment cancer pain at late stage etc., and without additive, is widely used.The demand of radix aconiti agrestis resource is improved day by day.
But due to ecotope heavy damage and artificial excavating, northern rhizome of Chinese monkshood wild resource is tending towards exhausted.Because yellow radix aconiti agrestis, the dried root that splits the rhizome of Chinese monkshood, the melon leaf rhizome of Chinese monkshood, the circular cone sequence rhizome of Chinese monkshood, Aconitum Sungpanense Hand-mazz and the too white rhizome of Chinese monkshood are also used when radix aconiti agrestis by the place of production and customary difference more, cause radix aconiti agrestis base source, market product chaotic, the validity of Chinese medicine, safety and stability are difficult to control.Therefore, carry out northern rhizome of Chinese monkshood introduction and acclimatization, make wild change man kind, the in situ conservation and the sustainable use that realize northern rhizome of Chinese monkshood resource seem particularly important.There is embryo after-ripening and physiology after-ripening in north aconitum seeds, although the form of i.e. seed is reached maturity, embryo not yet completes growth, embryo small volume, structure imperfection, must again after growing and making embryo maturation after a while, and seed just possesses the condition of sprouting.This characteristic of north aconitum seeds makes the northern rhizome of Chinese monkshood occur in the process of seminal propagation, and germination rate is low, slow, irregular phenomenon of germinateing of germinateing.The problems such as its resting stage is long, germination rate is low become the Main Bottleneck of northern rhizome of Chinese monkshood large-scale production, have had a strong impact on the seminal propagation effect of the northern rhizome of Chinese monkshood.
At present, although the artificial planting north rhizome of Chinese monkshood can adopt block root propagation or cut the vegetative manner such as bud breeding, vegetative propagation is utilized to there is many defects, as (1) adopts block root propagation, its reproduction coefficient is lower, and the wild product of a strain only can breed 3 ~ 5 strain cultivation product; (2) after employing block root carries out many generation breedings, the aspects such as seedling will occur degradation phenomena, and concrete manifestation resistance is poor, disease is many, yield poorly; (3) block root propagation is adopted can to cause destructive injury to wild resource equally.
Therefore, break northern aconitum seeds dormancy, improve its germination rate, will the key technology of for a change northern rhizome of Chinese monkshood cultivation mode be become, the development of northern rhizome of Chinese monkshood industry will be promoted simultaneously.
Summary of the invention
In view of this, the technical problem to be solved in the present invention is to provide a kind of method broken northern aconitum seeds dormancy fast, improve northern germination rate of monkshood seed.
The method breaking northern aconitum seeds dormancy provided by the invention comprises:
After the aconitum seeds sterilization of north, take concentration as the gibberellin process of 300mg/L ~ 400mg/L;
Seed after gibberellin process is carried out after high and low temperature alternative cultivation in 0 DEG C ~ 5 DEG C cultivations;
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; The temperature of high-temperature cultivation is 19 DEG C ~ 21 DEG C, and the time is 11h ~ 13h.
The germination rate of north aconitum seeds is low is the key problem causing current northern aconitum seeds breeding bottleneck.Ubiquity embryo after-ripening phenomenon in aconitum plant seed, but existing result of study shows: and aconitum plant not of the same race is also different to the response of vernalization condition, shows as: not of the same race is different to kindred plant hormone optimum concentration; Not of the same race different to the response of different plant hormones; Not of the same race also different to the response of inducing temperature.The present invention proves by experiment, and after northern aconitum seeds sterilization, after gibberellin immersion and Fluctuation temperature culture, seed germination rate can reach more than 78%, and the resting stage of seed is shortened to about 40d.
As preferably, the temperature of Low-temperature culture is 15 DEG C, and the time is 12h; The temperature of high-temperature cultivation is 20 DEG C, and the time is 12h; In 3 DEG C of cultivations after high and low temperature alternative cultivation.
As preferably, the air humidity of Fluctuation temperature culture is 65% ~ 70%.
The dormancy time temperature influence of north aconitum seeds is comparatively large, and the present invention proves by experiment, adopts Temp change method provided by the invention to cultivate, can significantly improve the germination rate of seed.
As preferably, seed is placed in Fluctuation temperature culture on two-layer moistening filter paper.
As preferably, the method breaking northern aconitum seeds dormancy provided by the invention is carried out under dark condition.
As preferably, after alternate culture, the embryo rate of northern aconitum seeds is not less than 80%.
Embryo rate refers to that embryo is long and accounts for the long percentage of endosperm.Research practice shows, embryo rate reaches more than 80%, and now embryo has differentiated cotyledon, plumule, plumular axis, radicle, completion morphology after-ripening substantially.But now seed can't be sprouted, must carry out physiology after-ripening under cryogenic, between physiological afterripening period morphologically there is not any change in embryo, and just idiosome increases.Only have and can sprout after northern aconitum seeds completes form after-ripening and physiology after-ripening, and there is strict succession and irreversibility between form after-ripening and physiology after-ripening, therefore, only have when northern aconitum seeds is after Fluctuation temperature culture to embryo rate is not less than 80%, carrying out Low-temperature culture makes it enter physiology after-ripening, the percentage of seedgermination that guarantee is final.
As preferably, the time that high and low temperature alternative is cultivated is 18d ~ 22d.
Preferably, the time that high and low temperature alternative is cultivated is 20d.
As preferably, 0 DEG C ~ 5 DEG C times of cultivating are 17d ~ 25d.
Preferably, 0 DEG C ~ 5 DEG C times of cultivating are 20d.
As preferably, the concentration of gibberellin is 300mg/L.
As preferably, the time of gibberellin process is 24h.
Gibberellin is a Plant Hormone, can stimulate the growth of leaf and bud.Prior art shows, the optimum concentration of gibberellin to aconitum plant not of the same race is inconsistent.But current, and have no gibberellin for breaking the report of northern aconitum seeds dormancy.And, nutrition due to seed germination places one's entire reliance upon endosperm, and when gibberellin excessive concentration, although embryo may be made to grow up rapidly under the stimulation of gibberellin, but the nutriment of endosperm storage may be made to be hydrolyzed in a large number, can not be grown by embryonic development in time and utilize, cause endosperm to liquefy, finally cause seed decay, death.The present invention confirms by experiment, when gibberellin concentration is 300mg/L, is conducive to the sprouting of seed most, and rotten kind of rate is lower.
As preferably, sterilization adopts mass fraction to be the aqueous sodium hypochlorite solution of 1%.
As preferably, the time of sterilization is 5 ~ 10min.
As preferably, between sterilization and gibberellin process, also comprise the step of distilled water flushing; The number of times rinsed is 3 ~ 5 times.
It is because carry caused by a large amount of bacteriums or fungi in its epidermis that seed goes rotten in incubation.The present invention adopt mass fraction be 1% aqueous sodium hypochlorite solution sterilization processing is carried out to seed after, then seed to be cultivated.Experiment confirms, adopt method provided by the invention to carry out sterilization, rotten kind of rate of northern aconitum seeds is no more than 10%.
The method breaking northern aconitum seeds dormancy provided by the invention comprises: after northern aconitum seeds sterilization, take concentration as the gibberellin process of 300mg/L ~ 400mg/L; Seed after gibberellin process is carried out after high and low temperature alternative cultivation in 0 DEG C ~ 5 DEG C cultivations; 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; The temperature of high-temperature cultivation is 19 DEG C ~ 21 DEG C, and the time is 11h ~ 13h.The present invention is in conjunction with gibberellin and intermittent warming north aconitum seeds; northern aconitum seeds dormancy can be broken fast; by under northern aconitum seeds conventional art condition, at least the resting stage of 90d foreshortens to about 40d; germination rate brings up to 78.65%, and processing method of the present invention is easy, efficiently; northern aconitum seeds dormancy can be removed within a short period of time; make it sprout, for northern rhizome of Chinese monkshood large-scale production sowing pre-treatment and indoor seed testing provide technical measures fast and efficiently, have broad application prospects.
Embodiment
The invention provides a kind of method breaking northern aconitum seeds dormancy, those skilled in the art can use for reference present disclosure, and suitable improving technique parameter realizes.Special needs to be pointed out is, all similar replacements and change apparent to those skilled in the art, they are all deemed to be included in the present invention.Method of the present invention and application are described by preferred embodiment, related personnel obviously can not depart from content of the present invention, spirit and scope methods and applications as herein described are changed or suitably change with combination, realize and apply the technology of the present invention.
The raw material that the present invention adopts is all common commercially available product, all can be buied by market.
Wherein, 1, northern aconitum seeds: gather Qu Chaihe town, Panshi City, Jilin Province for the examination northern rhizome of Chinese monkshood (Aconitum kusnezoffii Reichb.) seed in October, 2014, identified by Gao Yun Wang Yingping researcher.After the seed natural air drying gathered, the impurity such as removing carpopodium, shell, mildew seed, store shady and cool dry place for subsequent use.
2, chemical reagent: gibberellin GA3 is purchased from Yuan Ye bio tech ltd, Shanghai, and clorox is purchased from Tianjin good fortune chemical reagent factory in morning.
3, instrument and equipment: slide measure (0 ~ 150mm), digital display temperature control refrigerator (BCD-251WBSV), illumination box able to programme (MGC-250BP-2)
Below in conjunction with embodiment, set forth the present invention further:
Embodiment 1 ~ 2
Get northern aconitum seeds, removing impurity, with liquor natrii hypochloritis's sterilization 5 ~ 10min of 1%, again with distilled water flushing 3 ~ 5 times, seed after sterilization is soaked 24h respectively in the Gibberellins solution of different quality concentration, then select in the same size, the seed of full grains is placed in the culture dish being lined with two layers of filter paper, 100 seeds are put in each culture dish, each process repetition 3 times, the seed of each process is carried out treatment of different temperature, and the embryo of each process seed is long and endosperm is long, calculating embryo rate, germination rate, rotten kind of rate to adopt slide measure to measure.Embryo rate is determined as measured value when to cultivate 20 days under this condition; Germination rate is cultivation 40 days measured values, and all rotten kind of rate minutes are cultivation 40 days.Embodiment arranges as shown in table 1:
Table 1 embodiment is arranged
Comparative example 1 ~ 19
Get northern aconitum seeds, removing impurity, with liquor natrii hypochloritis's sterilization 5 ~ 10min of 1%, again with distilled water flushing 3 ~ 5 times, seed after sterilization is soaked 24h respectively in the Gibberellins solution of different quality concentration, then select in the same size, the seed of full grains is placed in the culture dish being lined with two layers of filter paper, 100 seeds are put in each culture dish, each process repetition 3 times, the seed of each process is carried out treatment of different temperature, and the embryo of each process seed is long and endosperm is long, calculating embryo rate, germination rate, rotten kind of rate to adopt slide measure to measure.Comparative example 1 ~ 13: embryo rate is determined as measured value when to cultivate 20 days under this condition; Germination rate is cultivation 40 days measured values, and all rotten kind of rate minutes are cultivation 40 days.Comparative example arranges as shown in table 2:
Table 2 comparative example is arranged
Embodiment 3
Utilize the statistical data of SAS8.0 software to embodiment 1 ~ 2 and comparative example 1 ~ 19 to carry out one-way analysis of variance, different disposal group difference significance adopts Fisher's LSD method to analyze.Computing formula is:
Embryo rate (%)=embryo length/endosperm long × 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
Statistics is as shown in table 3:
Table 3 result is added up
Result shows: northern aconitum seeds exists deep-sleep, and under normal condition, its resting stage is long, and germination rate is low, and through mass concentration 300mgL
-1~ 400mgL
-1gibberellin process, embryonic development speed is significantly higher than contrast, and gibberellin optimization process concentration is 300mgL
-1better in conjunction with 15 DEG C (12h), 20 DEG C of (12h) intermittent warming effects, embryo rate can reach more than 80% at cultivation 20d, and after cultivate through 3 DEG C of cryogenic thermostats, germination rate is significantly higher than other process and control group, visible, the method can break northern aconitum seeds dormancy at short notice, promotes its seed germination.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (10)
1. break a method for northern aconitum seeds dormancy, it is characterized in that, comprising:
After the aconitum seeds sterilization of north, take concentration as the gibberellin process of 300mg/L ~ 400mg/L;
Seed after gibberellin process is carried out after high and low temperature alternative cultivation in 0 DEG C ~ 5 DEG C cultivations;
In described 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; The temperature of high-temperature cultivation is 19 DEG C ~ 21 DEG C times is 11h ~ 13h.
2. method according to claim 1, is characterized in that, the temperature of described Low-temperature culture is 15 DEG C, and the time is 12h; The temperature of described high-temperature cultivation is 20 DEG C, and the time is 12h; In 3 DEG C of cultivations after described high and low temperature alternative cultivation.
3. method according to claim 1, is characterized in that, after described high and low temperature alternative is cultivated, the embryo rate of northern aconitum seeds is not less than 80%.
4. method according to claim 1, is characterized in that, the time that described high and low temperature alternative is cultivated is 18d ~ 22d.
5. method according to claim 1, is characterized in that, described 0 DEG C ~ 5 DEG C times of cultivating are 17d ~ 25d.
6. method according to claim 1, is characterized in that, the concentration of described gibberellin is 300mg/L.
7. method according to claim 1, is characterized in that, the time of described gibberellin process is 24h.
8. method according to claim 1, is characterized in that, described sterilization adopts mass fraction to be the aqueous sodium hypochlorite solution of 1%.
9. method according to claim 1, is characterized in that, the time of described sterilization is 5 ~ 10min.
10. method according to claim 1, is characterized in that, between described sterilization and gibberellin process, also comprises the step of distilled water flushing; The number of times of described flushing is 3 ~ 5 times.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111657084A (en) * | 2020-06-28 | 2020-09-15 | 国药种业有限公司 | Seedling raising method for aconitum sinomontanum seeds |
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| CN101731094A (en) * | 2009-12-30 | 2010-06-16 | 雅安三九中药材科技产业化有限公司 | Method for breeding aconite |
| CN102090179A (en) * | 2009-12-15 | 2011-06-15 | 四川省中医药科学院 | Method for processing aconitum seeds |
| CN103766040A (en) * | 2014-01-17 | 2014-05-07 | 甘肃农业大学 | Method for improving seed germination rate of wild aconitum gymnandrum maxim |
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Patent Citations (4)
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| CN102090179A (en) * | 2009-12-15 | 2011-06-15 | 四川省中医药科学院 | Method for processing aconitum seeds |
| CN101715665A (en) * | 2009-12-30 | 2010-06-02 | 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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Non-Patent Citations (1)
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| CN111657084A (en) * | 2020-06-28 | 2020-09-15 | 国药种业有限公司 | Seedling raising method for aconitum sinomontanum seeds |
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