CN101796899A - Method for sprouting seeds of polyphylla Smith var.ynynanensis (Franch.) Hand-Mazz - Google Patents
Method for sprouting seeds of polyphylla Smith var.ynynanensis (Franch.) Hand-Mazz Download PDFInfo
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Abstract
The invention discloses a method for sprouting seeds of polyphylla Smith var.ynynanensis (Franch.) Hand-Mazz, comprising the following steps: mixing wet sand with water content of 60-80% with the seeds of polyphylla Smith var.ynynanensis (Franch.) Hand-Mazz in a ratio of 2-4:1 and placing the mixture under the temperature of 8-15 DEG C for 25-35d; taking out the seeds and soaking the seeds in 50-200mg/L of GA3 solution for 1.5-2.5h; then mixing the wet sand with water content of 60-80% with the seeds in a ratio of 2-4:1, placing the mixture under 18-25 DEG C and keeping the moisture of the wet sand for 55-70d; taking out the seeds and soaking the seeds in 2.5-30mg/L of IAA solution for 1.5-3h; and mixing the wet sand with water content of 60-80% with the seeds in a ratio of 2-4:1, placing the mixture into a plant growing chamber at 20-22 DEG C and keeping the moisture of the wet sand. The method can ensure the seeds of polyphylla Smith var.ynynanensis (Franch.) Hand-Mazz to sprout ahead of time and have high emergence rate.
Description
Technical field
The present invention relates to the method for presprouting of seeds, particularly relate to Yunnan Rhizoma Paridis vernalization kind submethod.
Background technology
Yunnan Rhizoma Paridis is under the jurisdiction of and prolongs the Trilliaceae of section Paris Paris plant in age, mainly is distributed in the southwest of China.Yunnan Rhizoma Paridis is the widest because of Yunnan distributes, and medical value is the highest, by nineteen ninety-five version and version Chinese Pharmacopoeia in 2000 record, formal name used at school is Paris polyphylla Smith var.yunnanensis (Franch.) Hand-Mazz.Motherland's medical science is thought this product bitter, and cold nature is slightly poisonous, returns Liver Channel.Have effects such as clearing heat and detoxicating, swelling and pain relieving, cool liver arresting convulsion.Being usually used in diseases such as malignant boil carbuncle redness, abscess of throat, venomous snake bite, injuries from falls as well, convulsion, is the main composition medicine of patent medicine such as famous Chinese patent drug Yunnan Baiyao, jidesheng sheyao tablets, palace blood is peaceful.Pharmacological research shows that this plant has hemostasis, antitumor, antifertility, immunological regulation, many-sided physiologically active such as resist myocardial ischemia in recent years.Therefore, fully this natural resources of development and utilization has crucial meaning.
Seed dormancy is meant that the seed with vigor is in suitable sprouting condition and can not normally sprouts.It is break-off phenomenon in the development of plants process, perhaps be called " life hidden " phenomenon, concerning plant itself, it is a very important vital movement process, being again a kind of useful biological property, is a kind of biological adaptation to environmental condition and seasonal variety that plant obtains through long-term evolution.
Seed dormancy is the result that the long-term acclimatization of plant changes, can avoid in unsuitable season, sprouting, escape severe environmental conditions such as severe cold, heat, arid with " dormant seed " this form, to keeping the life of plant, profound significance has produced offspring.Seed dormancy also can cause certain difficulty to introducing and planting, has the seed demand of dormancy to take some measures breaking dormancy, and this just brings some extra works for beginning sowing in good time.A lot of traditional Chinese medicinal seeds resting stage reaches 0.5-2, and the wild proterties that has heavy traditional Chinese medicinal seeds resting stage is different in size, and it is extremely irregular to germinate, and this just causes certain difficulty to introducing and planting.
The Yunnan Rhizoma Paridis seed needs one summer of two winters to sprout under field conditions (factors), and a seedling rate of 15 months only is 46.2%, and a large amount of seeds have been lost vitality between very long resting stage.Yunnan Rhizoma Paridis seed deep-sleep has greatly limited the breeding of growing directly from seeds, and has restricted the plant production development of Yunnan Rhizoma Paridis.
Summary of the invention
The objective of the invention is to have hibernation feature, a kind of method that can make the Yunnan Rhizoma Paridis seed have the vernalization of high emergence rate is provided at the Yunnan Rhizoma Paridis seed.
Purpose of the present invention is achieved by following proposal:
A kind of Yunnan Rhizoma Paridis seed accelerating germination method comprises the steps:
Step 1: with the wet sand of water content 60-80% and Yunnan Rhizoma Paridis seed with 2-4: 1 ratio is mixed, 8-15 ℃ of placement 25-35 days;
Step 2: GA3 (gibberellin) solution seed soaking of taking-up seed usefulness 50-200mg/L 1.5-2.5 hour;
Step 3: the wet sand of using water content 60-80% again and seed are with 2-4: 1 ratio is mixed, and places under the 18-25 ℃ of condition, keeps wet husky moisture content 55-70 days;
Step 4: take out IAA (growth hormone) solution seed soaking 1.5-3 hour that seed is used concentration 2.5-30mg/L again;
Step 5: with the wet sand of water content 60-80% and seed with 2-4: 1 ratio is mixed, places 20-22 ℃ of growth chamber again, the maintenance husky moisture content that wets.
A kind of Yunnan Rhizoma Paridis seed accelerating germination method of the present invention is preferably as follows step:
Step 1: mix at 3: 1 with seed with the wet sand of water content 70%, placed in 10 ℃ of refrigerators 1 month;
Step 2: it is clean with flushing with clean water to take out seed, GA3 (gibberellin) solution seed soaking of usefulness 100mg/L 2 hours;
Step 3: use the wet sand of water content 70% to mix at 3: 1 again, place under the 20-22 ℃ of condition, kept wet husky moisture content 2 months with seed;
Step 4: it is clean with flushing with clean water to take out seed, uses IAA (growth hormone) solution seed soaking 2 hours of concentration 25mg/L again;
Step 5: the wet sand with water content 70% mixes with seed at 3: 1, places 20-22 ℃ of growth chamber again, keeps wet husky moisture content.
Described Yunnan Rhizoma Paridis seed is preferably ripe aquatic foods kind, after-ripening 3-5 days.
Wet sand after described wet sand is preferably sterilized.
The Yunnan Rhizoma Paridis seed is sprouted naturally, needs just can emerge in 15 months, and emergence rate is 46.2% only, and seedling-growing time differs, and regularity is low, and etc. weak point, method of the present invention can make the Yunnan Rhizoma Paridis seed sprout in advance, the emergence rate height.
Following experimental example and embodiment are used to further specify but do not limit the present invention.
Experimental example 1 hormone regulating and controlling is to the influence of seed germination
1, experiment purpose: observe the variation of Yunnan Rhizoma Paridis of the present invention germination rate under different hormone kinds and concentration.Screen suitable hormone kind and concentration
2, materials and methods
Seed: the mature seed of the Yunnan Rhizoma Paridis that gather in October, 2004.
Test is the fresh seeds of gathering with planting.Placed 5 days at shady and cool place, and clear water soaked 2 hours, removed kind of a skin, cleans.Choose some groups in 100 mature and plump seeds, soaked 2 hours with the variety classes hormone, then with 1: 3 clean fine sand lamination in the seed culture ware, the husky water content of lamination is 70%.Lamination temperature and time: 0-30 days, 10 ± 1 ℃; 30-150 days, place in 20 ± 1 ℃ of growth chambers.Calculate germination rate in the time of 150 days.
3 results
Table 1
Table 2
Can filter out the seed soaking suitable concn by seed germination rate, the GA3 concentration of 100mg/L and the IAA concentration of 25mg/L, 2.5mg/L can promote seed germination.
The research that experimental example 2 hormone regulating and controllings change embryo rate and endogenous hormones in Yunnan Rhizoma Paridis kind embryo dormancy and the growth course
1 experiment purpose:
Observe the dynamic change of seed embryo rate and endogenous hormones in Yunnan Rhizoma Paridis kind embryo dormancy of the present invention and the growth course.
2 materials and methods
2.1 material
Seed: the mature seed of the Yunnan Rhizoma Paridis that gather in October, 2004.
Kit: adopt enzyme-linked immune analytic method (ELISA), kit is pressed GA in the kit explanation working sample available from China Agricultural University
3, IAA content.
Instrument: the full-automatic multiplex's function of the vigorous Multiskan MK3 of Finland's thunder microplate reader; U.S. Beckman Allegra
TM25R centrifuge refrigerated centrifuge; Switzerland PT1600E refiner; Switzerland plum Teller AX205 balance.
2.2 method
2.2.1 processing method:
Test is the fresh seeds of gathering with planting.Placed 5 days at shady and cool place, and clear water soaked 2 hours, removed kind of a skin, clean, behind the water of the seed epidermis that dries in the shade with 1: 3 clean fine sand lamination in the seed culture ware, the husky water content of lamination is 70%.Lamination temperature and time: 0-30 days, 10 ± 1 ℃; Take out seed, go husky cleaning,, placed in 20 ± 1 ℃ of growth chambers 30-90 days with 2 hours pavilions of GA3 seed soaking of 100mg/L; Take out seed, go husky cleaning,, placed in 20 ± 1 ℃ of growth chambers 90-150 days with the IAA seed soaking of concentration 25mg/L 2 hours.Testing time is after the lamination 0,30,60,90,120,150 day.At each point in time sampling, sample is through preserving (70 ℃) preservation in low temperature refrigerator behind the liquid nitrogen frozen, in order to test.
2.2.2 the mensuration of embryo rate: at different lamination time points, get 30 seeds, central authorities cut in half endosperm along embryo at every turn.Under the binocular anatomical lens, measure the length (mm) of embryo and endosperm with micrometer.
Calculate the embryo rate: embryo rate=embryo length/endosperm is long, asks the average embryo rate of each time point.
2.2.3 infarcted region cardiac muscle dyeing from the apex of the heart entad the end direction heart evenly is cut to 8, be placed in 0.1% the NBT solution, 37 ℃ of gas bath shaking tables shake 10min, blot surface moisture with blotting paper, put in order, take pictures; Cut infarcted region and non-infarcted region and weighing respectively, calculate heart stalk index.The wet quality of heart stalk index=infarcted region/wet whole-heartedly quality.
2.2.3 endogenous hormones GA
3, IAA extraction and mensuration
The extraction of hormone: precision takes by weighing the sample 1.0g of each time point in mortar, adds the 4ml sample extracting solution, grinds to form homogenate in ice bath, and 4 ℃ are extracted 4h, the centrifugal 15min of 1000g down.Supernatant is crossed C
18The glue post, step is: sample on the 80% methyl alcohol balance glue post, collect sample, remove behind the sample and to wash post 100% ether with 100% methyl alcohol and wash post 100% methyl alcohol and wash post and circulate.Sample dries up with nitrogen after will crossing post, removes the methyl alcohol in the extract, is settled to 1ml with sample diluting liquid.
The mensuration of hormone: adopt enzyme-linked immune analytic method (ELISA), kit is pressed GA in the kit explanation working sample available from China Agricultural University
3, IAA content.
2.2.4 all data of statistical disposition are represented with mean.
3 data analysis results show that the regulation and control of exogenous hormone can promote the growth of Yunnan Rhizoma Paridis embryo, and the regulation and control of exogenous hormone simultaneously can impel the raising of Yunnan Rhizoma Paridis Seed Endogenous Hormones, thereby promote the growth of Yunnan Rhizoma Paridis embryo, and the germinating time of Yunnan Rhizoma Paridis is shifted to an earlier date.
Table 3 embryo rate result
Table 4 endogenous hormones result of variations
The screening and the optimization of experimental example 3 Yunnan Rhizoma Paridis seed accelerating germination methods
Determining of 1 alternating temperature stratification ceiling temperature
Alternating temperature stratification is to shorten the seed afterripening process, improves the common method of seed germination rate.The bound temperature of determining alternating temperature stratification is the key in the alternating temperature stratification technology.Definite one side of alternating temperature ceiling temperature will promote the growth of embryo; In addition, to prevent that also too high alternating temperature stratification temperature from suppressing seed germination.
1.1 method
Fresh Yunnan Rhizoma Paridis seed removes exosper, cleans, and 100 every part, wet sand is put in three repetitions, places 5 ℃, 10 ℃, 15 ℃, 20 ℃, 25 ℃, 30 ℃ growth chambers.With average radicle length is index.Experimental period 90 days.
1.2 result
Table 5
As seen from the above table, temperature affects the kind embryonic development of Yunnan Rhizoma Paridis seed significantly.By observing the variation of the average radicle length under the treatment of different temperature behind the 90d, the Yunnan Rhizoma Paridis radicle growth presents the trend of the unimodal curve that afterwards reduces of raising earlier with the rising of handling temperature as can be known.Under 20 ℃ of processing, the Yunnan Rhizoma Paridis radicle growth is the longest, and development degree is the highest; Under 5 ℃ and the 10 ℃ of processing, Yunnan Rhizoma Paridis kind embryonic development is slow, and radicle growth slowly is difficult to break through seed; High temperature treatment (25 ℃ and 30 ℃) is though following Yunnan Rhizoma Paridis seed still can carry out kind of an embryonic development, but compare with 20 ℃, high temperature has suppressed the kind embryonic development, the author thinks that high temperature has improved the allotment that seed sugar and the metaboilic level of fat have promoted the seed nutriment on the one hand, high temperature has also improved the level of metabolic enzyme on the other hand, seed nutrition depletion plants that embryonic development is counter to be suppressed.
Determining of 2 plant hormone kinds and concentration
Plant hormone is to break seed dormancy, promotes the important substance of seed germination, selects suitable hormone types and concentration, can effectively improve seed germination in conjunction with the alternating temperature stratification technology, shortens the latter stage of ripening of seed germination.Different types of plant hormone is selected in this research for use, and each parahormone is to kind of an embryonic development (this sentences radicle long is statistical indicator) influence, to select suitable hormone types and concentration for use under the observation variable concentrations.
2.1 plant hormone select test for use
GA
3Seed soaking concentration is respectively 50,100,200,500mg/L; IAA seed soaking concentration is respectively 0.25,2.5,25,250mg/L; KT seed soaking concentration is respectively 25,50,100,200mg/L.Seed soaking time 24h places 20 ℃ of growth chambers, experimental period 90 days.With average radicle length is index.
2.2 result
Table 6
Table 7
Different types of hormone is to Yunnan Rhizoma Paridis kind embryonic development effect difference.GA
3The seed soaking concentration affects Yunnan Rhizoma Paridis kind embryonic development, and as shown above, the Yunnan Rhizoma Paridis radicle length is along with GA
3The rising of seed soaking concentration manifests the trend that raises and afterwards reduce earlier, shows in the embryo growth course of Yunnan Rhizoma Paridis the GA of suitable concn thus
3(100mg/L) show comparatively significant facilitation.Growth hormone IAA to the influence of the kind embryonic development of Yunnan Rhizoma Paridis and gibberellin roughly the same shows as the Yunnan Rhizoma Paridis radicle length and manifests the trend that raises and afterwards reduce earlier along with the rising of IAA seed soaking concentration; Compared with the control, the IAA of low concentration has promoted the growth (data not shown among the figure) of radicle, the IAA of higher concentration has suppressed the growth of radicle, be that IAA shows as low concentration and the different double effect of high concentration, result of the test shows that the 25mg/LIAA radicle length is longer, and the kind embryonic development is best.This research also finds that basic element of cell division parahormone KT is not so good as contrast to the influence of the kind embryonic development of Yunnan Rhizoma Paridis, and the radicle length of high-concentration and low-concentration KT seed soaking is all not as contrast.Therefore, in promoting Yunnan Rhizoma Paridis seed germination process, should note selecting for use the GA of 100mg/L
3And the IAA of 25mg/L.
3 alternating temperature temperature, GA
3Concentration and IAA concentration three factor orthogonal experiments
According to above-mentioned experimental result, temperature, GA
3Bigger to Yunnan Rhizoma Paridis seed germination facilitation with IAA, suitable alternating temperature temperature, GA are chosen in this research
3Concentration and IAA concentration combination are best vernalization scheme, and by orthogonal experiment this are tested.
Table 8 temperature, GA
3Concentration and IAA concentration three factor orthogonal experiments
Last table is temperature, GA
3Concentration and IAA concentration three factors are to the orthogonal experiments table of Yunnan Rhizoma Paridis seed germination influence.As seen from the above table, temperature, GA
3The growth that the Yunnan Rhizoma Paridis seed base-root in concentration and the equal appreciable impact of IAA concentration.By the extreme difference of calculating and more different factors, in 3 levels of temperature factor, with 20 ℃ of pairing extreme difference maximums, therefore determining 20 ℃ is Yunnan Rhizoma Paridis radicle growth ceiling effect temperature as can be known.By checking different GA
3The extreme difference of concentration and different I AA concentration, 200mg/LGA as can be known
3Promoted the growth of Yunnan Rhizoma Paridis radicle with 25mg/L IAA.Hence one can see that, and best scheme is 20 ℃ of following 200mg/L GA
3With 25mg/L IAA mixed processing.In this research, to handle 7 and be preferred version, the radicle growth length of processing 7 is the longest, shows that thus optimal case is realistic, through crossing check.
Following embodiment all can realize the described effect of above-mentioned experimental example
Embodiment
Embodiment 1:
Step 1: get the aquatic foods kind of Yunnan Rhizoma Paridis maturation, place shady and cool place's after-ripening, or mix with three parts of wet sand and a seed, after-ripening 3-5 days, remove pericarp, clear water is cleaned.Wet sand with water content 70% mixes with seed at 3: 1, places in 10 ℃ of refrigerators 1 month;
Step 2: it is clean with flushing with clean water to take out seed, GA3 (gibberellin) solution seed soaking of usefulness 100mg/L 2 hours;
Step 3: use the wet sand of water content 70% to mix at 3: 1 again, place under the 20-22 ℃ of condition, kept wet husky moisture content 2 months with seed;
Step 4: it is clean with flushing with clean water to take out seed, uses IAA (growth hormone) solution seed soaking 2 hours of concentration 25mg/L again;
Step 5: the wet sand with water content 70% mixes with seed at 3: 1, places 20-22 ℃ of growth chamber again, keeps wet husky moisture content to emerging.
Embodiment 2:
Step 1: get the aquatic foods kind of Yunnan Rhizoma Paridis maturation, place shady and cool place's after-ripening, or mix with three parts of wet sand and a seed, pericarp is removed in after-ripening 5 days, and clear water is cleaned, and uses the wet sand of water content 65% to mix with seed 2.5:1, places in 11 ℃ of refrigerators 28 days;
Step 2: it is clean with flushing with clean water to take out seed, GA3 (gibberellin) solution seed soaking of usefulness 200mg/L 2 hours;
Step 3: use the wet sand of water content 70% to mix at 3: 1 again, place under 20 ℃ of conditions, kept wet husky moisture content 58 days with seed;
Step 4: it is clean with flushing with clean water to take out seed, uses IAA (growth hormone) solution seed soaking 2.5 hours of concentration 25mg/L again;
Step 5: the wet sand with water content 70% mixes with seed at 2.5: 1, places 20 ℃ of growth chambers again, keeps wet husky moisture content to emerging.
Claims (5)
1. a Yunnan Rhizoma Paridis seed accelerating germination method is characterized in that this method comprises the steps:
Step 1: with the wet sand of water content 60-80% and Yunnan Rhizoma Paridis seed with 2-4: 1 ratio is mixed, 8-15 ℃ of placement 25-35 days;
Step 2: take out seed and soaked seed 1.5-2.5 hour with the Gibberellins solution of 50-200mg/L;
Step 3: the wet sand of using water content 60-80% again and seed are with 2-4: 1 ratio is mixed, and places under the 18-25 ℃ of condition, keeps wet husky moisture content 55-70 days;
Step 4: take out growth hormone solution seed soaking 1.5-3 hour that seed is used concentration 2.5-30mg/L again;
Step 5: with the wet sand of water content 60-80% and seed with 2-4: 1 ratio is mixed, places 20-22 ℃ of growth chamber again, the maintenance husky moisture content that wets.
2. the method for claim 1 is characterized in that this method comprises the steps:
Step 1: mix at 3: 1 with seed with the wet sand of water content 70%, placed in 10 ℃ of refrigerators 1 month;
Step 2: it is clean with flushing with clean water to take out seed, soaks seed 2 hours with the Gibberellins solution of 100mg/L;
Step 3: use the wet sand of water content 70% to mix at 3: 1 again, place under the 20-22 ℃ of condition, kept wet husky moisture content 2 months with seed;
Step 4: it is clean with flushing with clean water to take out seed, uses the growth hormone solution seed soaking 2 hours of concentration 25mg/L again;
Step 5: the wet sand with water content 70% mixes with seed at 3: 1, places 20-22 ℃ of growth chamber again, keeps wet husky moisture content.
3. the method for claim 1 is characterized in that in this method:
The concentration of Gibberellins solution is 200mg/L in the step 2; The concentration of growth hormone is 25mg/L in the step 4; Temperature in the step 3 and 5 is 20 ℃.
4. the method for claim 1 is characterized in that Yunnan Rhizoma Paridis seed described in this method is ripe aquatic foods kind, after-ripening 3-5 days.
5. the method for claim 1 is characterized in that the wet husky wet sand after the sterilization that is in this method.
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| CN105409514A (en) * | 2015-11-16 | 2016-03-23 | 曲靖市菱原商贸有限公司 | Fast seedling culture method of paris polyphylla |
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| CN107027624A (en) * | 2017-04-28 | 2017-08-11 | 云南农业大学 | A kind of method for obtaining polygerm Yunnan Paris polyphylla seedling |
| CN107360753A (en) * | 2017-07-20 | 2017-11-21 | 四川省中医药科学院 | A kind of magnificent Paris polyphylla seed fast breeding method |
| CN108307965A (en) * | 2018-03-14 | 2018-07-24 | 上海市药材有限公司 | A kind of vernalization of paris polyphylla seed and method for culturing seedlings |
| CN109197401A (en) * | 2018-08-07 | 2019-01-15 | 永胜县顺源中药材种植有限责任公司 | The method for culturing seedlings of Paris polyphylla |
| CN109220057A (en) * | 2018-10-25 | 2019-01-18 | 天全县营生食用菌种植农民专业合作社 | A kind of method for culturing seedlings of Paris polyphylla |
| CN113141810A (en) * | 2021-04-26 | 2021-07-23 | 三峡大学 | Method for promoting rapid germination of paris polyphylla seeds |
| CN114223471A (en) * | 2021-10-19 | 2022-03-25 | 重庆师范大学 | Rapid seedling method of paris polyphylla |
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