CN105265064A - Liquid culture seedling growing method for fern protonemata - Google Patents
Liquid culture seedling growing method for fern protonemata Download PDFInfo
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Abstract
The invention discloses a liquid culture seedling growing method for fern protonemata. The method includes the spore germination step, wherein collected spores are soaked in clean water, after floating objects on the water surface and water are removed, freezing storage is selectively performed for a preset period of time so that spore germination can be promoted, a proper amount of clean water is added, gibberellin is used for processing the spores so that spore germination can be promoted, the processed spores are soaked in water to be cultured, and medicine killing exogenous microbes or inhibiting growth of the exogenous microbes is selectively used in the early culture stage; if bubbles are generated in the spore sedimentation area in the culture process, it shows that the spores are germinated, the protonemata start to grow, and if spore balls float on the water surface in the culture process, predetermined measures should be taken so that the spore balls can subside. The method is simple and easy to implement, it can be effectively avoided that spores which do not germinate mould, the controllability of the liquid culture system of the protonemata is high, the multiplication efficiency is high, and the method can be used for large-scale seedling growing of ferns.
Description
Technical field
The invention belongs to the breeding method of pteridophyte seedling, particularly, relate to a kind of fern protonema liquid culture seedling-cultivating method.
Background technology
Fern is the vascular plant that on the earth, earliest extant occurs.Many ferns have ornamental value because strain shape is graceful; Meanwhile, a large amount of ferns has medicinal or edibility, dark liking by consumers in general.But, because long-term, a large amount of unrestricted predation formulas is excavated, the deterioration of the ecological environment in addition, wild fern resource is seriously damaged, even if in original producton location, a lot of wild fern distribution also sharply reduces, even become extinct, the pteridophyte of natural propagation cannot meet the demand of people.Therefore, the pteridophyte started in production having Important Economic value carries out artificial breeding and cultivation.
At present, pteridophyte artificial breeding mainly adopts the methods such as vegetative reproduction, sporogenesis and tissue cultures.
Vegetative reproduction utilizes the positions such as the root of fern, root-like stock, leaf to produce asexual gemma, or produce new plant by apical meristem.This method is simple, easy, and technical requirement is not high, but proliferation rates is little, need take wild[l strain, and very large to the destruction Of resources, in the long run consider, this method has larger limitation, and the fern seedling breeding method of non-optimal.
Tissue culture technique is one fern mating system fast and effectively, and explant consumption is few, does not damage natural resources, and growth coefficient is high, and in Germ-plasma resources protection, extensive modernization commercial seedling are produced, advantage is remarkable.But, the fern of successfully being bred by tissue cultures is at present less, often kind of fern needs suitable medium and condition of culture, screening operation amount is large, and medium versatility is not strong, and breeding cycle is long, seedling transplanting survival rate is not high, tissue culture system need be set up, equipment, venue cost are higher, and operating technology requires strict, and general production unit is difficult to carry out related work.Therefore, fern tissue cultivating and seedling technology is also subject to a definite limitation.
Sporogenesis forms sporophyte after the spore utilizing fern to produce is sprouted in solid matrix, then obtain seedling by the gametophyte that sporophyte produces.The method has the advantages such as cost is low, technical requirement is not high, easy to operation, breeding amount is large, be particularly suited for generally planting family to use, but this mode of reproduction is subject to the factor impacts such as spore germination rate, sowing media, thickness of sowing, condition of culture, do not sprout spore easily to go mouldy, gametophyte is formed totally unfavorable, and relative complex, loaded down with trivial details is cultivated by sporogenous tissue.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind ofly to be had simple and effectively can avoid not sprouting the method that sporogenesis that spore easily goes mouldy cultivates pteridophyte seedling.
The present invention's adopted technical scheme that solves the problem is:
Fern protonema liquid culture seedling-cultivating method, comprises
Spore germination step: the spore of gathering is added in clean clear water and soaks, after removing the floating thing of the water surface and moisture, optionally the frozen scheduled time is to promote spore germination, add appropriate clean clear water and with gibberellin process to promote spore germination, spore after process is soaked in water to be cultivated, Initial stage of culture optionally use kill or suppress exogenous microbial to grow medicine (as, antibacterial agent is optionally used to have the situation of bacteria breed to occur with process, or optionally use protozoa agent for killing to occur to process the situation having protozoa to grow), have in spore settling zone bubble to produce in incubation and then show that spore is sprouted, protonema starts growth, if having spore agglomerate buoyance lift in incubation in the water surface, predetermined means should be adopted to make the sedimentation of spore agglomerate.
The gibberellin used in the present invention is broad spectrum activity plant growth regulator, widely distributed, diverse in function, can breaking dormancy, and promote seed germination, the gibberellin of debita spissitudo can improve pteridophyta spore germination rate.Usually the spore after gathering is seeded in solid matrix (being generally the soil after process) in prior art and cultivates, sprout, and the mould in upper soll layer is aerobic, heterotroph.Oxygen content in liquid internal environment is much lower compared with soil surface; What the present invention simultaneously adopted is Aquaponic environment, and the content of organic matter extremely low (organic matter not sprouting spores release is little, can reduce the content of organic matter easily and fast in incubation by changing water), cannot meet needed for heterotroph fungus growth.So, can prevent fern seed from sprouting and go mouldy in protonema process of growth.
Wherein, in described spore germination step, frozen temperature is 0 to-20 DEG C, and the frozen time is one thoughtful one month.
Wherein, in described spore germination step, whether the area grown according to fern belongs to winter outdoor temperature and carry out selecting in area below freezing the need of frozen and preserved spore, be specially: be area below freezing when the area that fern grows belongs to winter outdoor temperature, then should carry out frozen operation; If the area that fern grows belongs to winter, outdoor temperature is area above freezing, then can select not carry out frozen operation.
Wherein, in described spore germination step, described floating thing mainly refers to the spore, sporangium and/or other foreign material that float on the water surface.
Wherein, in described spore germination step, the final concentration of described gibberellin is 100 ~ 300ppm, and the described gibberellin processing time is 15 minutes to 24 hours.
Wherein, in described spore germination step; the condition that spore after described process is soaked in when cultivating in water is: 15 ~ 20 DEG C, natural lighting, and culture vessel have exogenous microbial or other kind spore can be avoided to enter culture vessel thus cause pollute covering protection thing.
Wherein, in described spore germination step, described spore is soaked in the process of cultivating in water, needs regularly or aperiodically to add water, to keep the volume of water in culture vessel substantially constant.
Wherein, in described spore germination step, the preferred agricultural streptomycin of described antibacterial agent, the final concentration of described agricultural streptomycin is 0.01 ~ 0.1%, the preferred Tinidazole of described protozoa agent for killing, and the final concentration of described Tinidazole is 0.05 ~ 0.5mg/mL.
Wherein, in described spore germination step, described predetermined means can be any means that can make the sedimentation of spore agglomerate, particularly, realize as stirred by stirring rod or rock the modes such as culture vessel.
Wherein, due to the breeding method that the present invention is a kind of pteridophyte seedling, therefore, the requirement of certain cell chulture water or plant cultivation water should be met to the water used, all to avoid exogenous microbial to pollute as requiring to spore, particularly, the water used in the present invention can be free of contamination clean river, well water or running water, wherein, phreatic water can directly use; River needs to boil more than 30 minutes, to make contained microorganism and spore deactivation; Running water needs to tan by the sun more than 3 days, needs equally to boil more than 30 minutes.And to the clean clear water that the application is particularly limited to, best is aqua sterilisa or the water after boiling process, to avoid living contaminants.
Wherein, described seedling-cultivating method can also comprise:
Protonema incubation step: when spore is cultured to continuation cultivation under natural lighting condition when protonema occurs through described spore germination step, urea and potassium dihydrogen phosphate is added by foliage fertilizer dosage in culture, to promote the growth of protonema, predetermined means should be adopted (as mentioned before in incubation, can the means such as stirring be adopted) to avoid protonema to assemble, be wound agglomerate thus reduce appreciation rate, when protonema content is higher than predetermined value in culture vessel, the mode expanding cultivation is adopted to continue to cultivate.
Fern protonema can carry out photosynthesis, and liquid culture improves intensity of illumination in suitable degree, facilitates protonema growth, can expand cultivation in addition in good time, therefore improve the proliferate efficiency of protonema in incubation.Because protonema carries out cultivating in the glass container of printing opacity, water, fertilising can be changed at any time, once occur going mouldy, microbiological contamination or grow protozoa and cannot process, pollution group can be rejected, cultivation can be expanded according to protonema growing state simultaneously in good time.Therefore, in incubation, controllability is directly seeded in soil much better than compared with spore.
Wherein, the operation that described expansion is cultivated specifically: after stir culture thing, by stoste: new liquid is (1 ~ 5): the volume ratio of 10 accesses new culture vessel, continue to expand cultivation.
Wherein, in described protonema incubation step, protonema content is undertaken by the state of protonema higher than the judgement of predetermined value, as when protonema produces lumps, protonema too high levels can be judged, higher than predetermined value, for improving culture efficiency, the mode expanding cultivation can be adopted to continue to cultivate.
Wherein, described seedling-cultivating method can also comprise:
Gametophyte incubation step, sporophyte incubation step, sporophyte (seedling) transplant step.
Wherein, described gametophyte incubation step is: choose free of contamination humus soil, pH6.0 ~ 7.0, tan by the sun under sunlight, parch, pulverize, remove impurity, get and appropriate fill basin to 2/3 ~ 4/5 of volume as matrix, the humus soil separately got after above-mentioned process is overlying on Culture basin soil table after crossing 50 eye mesh screens, thickness 1 ~ 5cm, and make soil drench in basin by predetermined way, by the protonema sprinkle through described protonema incubation step acquisition in Culture basin soil surface, and ensure protonema more than 80% in flakes, be placed in photoenvironment incubated at room temperature, ground moistening and maintenance should be kept to be unfavorable in the process of cultivating temperature that heterotroph fungi grows (namely, low temperature should be avoided), be cultured to gametophyte and expose soil surface.
Wherein, in described gametophyte incubation step, make the predetermined way of soil drench in basin basin can be placed in pallet, water filling in pallet, thus the object making soil drench in basin can be realized.
Wherein, in described gametophyte incubation step, for increasing seedling yield, by protonema sprinkle after Culture basin, if soil surface still has many places blank---without protonema, again can inoculate without protonema blank space at soil surface in good time, also directly can take the mode repeatedly inoculated, to ensure protonema more than 80% in flakes.
Wherein, described sporophyte incubation step is: after gametophyte exposes soil surface, and keep ground moistening to form fertilized egg to make sperm be combined with ovum, continuation cultivation is until occur sporophyte seedling.
If when inoculating protonema in described gametophyte incubation step, the protonema 100% in Culture basin in flakes, continues cultivation and then can generate about 20-50 strain/cm
2sporophyte seedling.
Wherein, described sporophyte (seedling) transplant step is: after sporophyte grows 3 ~ 4 leaves, carry out first time by 4cm × 4cm seeding row spacing transplant, adopt conventional soil as matrix, indoor cultivation, after first time transplanting 7 ~ 15 days seedlings survive, be transplanted on outdoor seedbed by seeding row spacing 6cm × 10cm, continue to cultivate, grow to after the height (10 ~ 15cm) that meets the requirements until seedling, in autumn or spring in next year being colonizated in production Tanaka by suitable seeding row spacing then.
Wherein, described seedling-cultivating method can also comprise:
Field daily management operation: seedling transplant planting requires routinely to operate after production Tanaka.Note avoiding high light direct projection, apply fertilizer in good time, water, weeding, loosen the soil in time and remove disease seedling, dead seedling, preventing damage by disease and insect.
Wherein, described seedling-cultivating method can also comprise
Spore acquisition step: when spore is ripe (different fern is different in the different regions mature period), when sporophyll becomes yellowish-brown from green (spore distribute before), cut sporophyll, dry, room temperature storage or low temperature (lower than 4 DEG C) storage are to extend spore activity, pulverize sporophyll, cross 100-200 eye mesh screen and sift out spore and sporangium.
Wherein, in described spore acquisition step, after described sporophyll drying, whether the area grown according to fern is belong to winter outdoor temperature to select whether to carry out frozen operation in area below freezing, be specially: the area that fern grows belongs to winter temperature in area below freezing, need frozen about one to three month (cryogenic temperature is about-10 DEG C to-20 DEG C) to improve spore germination rate after sporophyll drying, the area that fern grows belongs to winter temperature in area above freezing, can select not carry out above-mentioned frozen operation.
To sum up, the invention has the beneficial effects as follows:
1, the present invention utilizes liquid environment to cultivate spore promote that it sprouts into protonema, solid matrix can be avoided to go mouldy the adverse effect caused, improved the germination rate of spore by various ways such as frozen and gibberellin.
2, protonema liquid culture system controllability is strong, and proliferate efficiency is high, and the adverse effect simultaneously avoiding solid matrix to go mouldy causing, can obtain a large amount of protonema by expanding cultivation, in scale breeding, meaning is very large.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail, but embodiments of the present invention are not limited thereto.
Breeding method of the present invention can be suitable for each pteridophyte, include but not limited to following cited several concrete pteridophyte, mainly by adopting liquid culture, can effectively prevent from going mouldy in spore germination and protonema process of growth, improve the proliferate efficiency of protonema, strengthen the controllability of protonema cultivating system, following examples give schematically illustrating of different pteridophyte seedling fostering.
Embodiment 1
1. spore acquisition step:
After August, when Matteuccia strthiopteris spore is ripe, when sporophyll becomes yellowish-brown from green (spore distribute before), select the afternoon (taken outdoors need after dew disperses) of continuous sunny, with clean scissors, sporophyll is cut, be placed in totally smooth paper bag, air-dry drying, room temperature storage or low temperature (lower than 4 DEG C) storage are to extend spore activity, above-mentioned obtained sporophyll is fully pulverized, 100-200 eye mesh screen sifts out spore (and sporangium), removes impurity, obtains spore (containing sporangium).Need in refrigerating chamber freezing processing about month (cryogenic temperature is about-20 DEG C) after sporophyll drying, to improve spore germination rate.
2. spore germination step:
By the appropriate pollution-free clean clear water of spore (preferably aqua sterilisa or the water that boiled obtained, to avoid living contaminants) fully to mix and after soaking, remove and float on the spore of the water surface, sporangium, other foreign material and moisture, in-20 DEG C frozen one week to promote spore germination.Add appropriate clean clear water (requiring with aforementioned), with final concentration 100ppm gibberellin process 24 hours, to promote spore germination.
Spore after process is soaked in (water quality requirement: free of contamination clean river, well water, running water in water.Process: phreatic water can directly use; River needs to boil more than 30 minutes, to make contained microorganism, spore deactivation; Running water needs to tan by the sun more than 3 days, same need boil process), spore suspension is placed in glass container, 15 ~ 20 DEG C, cultivate that (covering such as culture vessel available paper covers under natural lighting condition, avoid exogenous microbial or other kind spore to fall into, cause polluting).Note the clean water after often adding process in incubation, keep water volume in culture vessel substantially constant.
If Initial stage of culture has bacterium to grow in a large number, the agricultural streptomycin process of final concentration 0.02% can be added; If have a large amount of protozoa to grow can to add the Tinidazole process of final concentration 0.1mg/mL.If have bubble to produce in spore settling zone in incubation, show that spore is sprouted, protonema starts growth, produce oxygen in a large number bubble time can by spore agglomerate buoyance lift in water meter, suitably to be stirred or other optional predetermined means make spore agglomerate be settled down to the bottom.
3. protonema incubation step:
Cultivate about about one week spore to start to sprout, start after 2 weeks to occur green protonema.Now in culture, urea and potassium dihydrogen phosphate can be added by foliage fertilizer dosage, to promote the growth of protonema.
Often stir in incubation, protonema can be avoided to assemble, be wound agglomerate and reduce appreciation rate.
When (that is, there is agglomerate) during protonema too high levels in culture vessel, cultivation can be expanded, method is: stirred by culture, by stoste: new liquid is (1 ~ 5): the volume ratio of 10 accesses new culture vessel, continue to expand and cultivate, proliferate efficiency can reach more than 5 times.
4. gametophyte is cultivated:
Choose free of contamination humus soil (pH6.0 ~ 7.0), thoroughly tan by the sun under sunlight, parch, pulverize, remove impurity, get and appropriate fill basin to 4/5 of volume as matrix, 50 eye mesh screens crossed by the humus soil separately got after above-mentioned process, tiny for gained humus soil granule is overlying on Culture basin soil table, thickness is about 5cm, then be placed in pallet, water filling in pallet, soil in basin is fully infiltrated.
Mixed by protonema that cultivation obtains, sprinkle, in Culture basin soil table, makes protonema be uniformly distributed, is placed in photoenvironment incubated at room temperature, notes, often to water filling in pallet, keeping ground moistening.Avoid low temperature, to prevent growing of heterotroph fungi.
Cultivate about about 20 days, the visible diameter of soil surface is about the green gametophyte of 1cm.For increasing seedling yield, in gametophyte cultivating process, again can inoculate without protonema blank space at soil surface in good time, also directly can take the mode repeatedly inoculated, to ensure protonema final more than 80% in flakes.
5. sporophyte incubation step:
After gametophyte occurs, keep soil fully moistening, to be combined with ovum to make sperm and to form fertilized egg.Continue to cultivate until start to occur sporophyte seedling.In the present embodiment, protonema 100% in flakes time can generate about 20-50 strain/cm
2seedling.
6. sporophyte (seedling) transplant step:
After sporophyte grows 3 ~ 4 leaves, carry out first time by 4cm × 4cm seeding row spacing transplant, adopt conventional soil as matrix, indoor cultivation.
After first time transplanting 7 ~ 15 days seedlings survive, be transplanted on outdoor seedbed by seeding row spacing 6cm × 10cm, continue to cultivate.(10 ~ 15cm) after the height that meets the requirements is grown to, in autumn or spring in next year being colonizated in production Tanaka by suitable seeding row spacing then until seedling.
7. field daily management operation:
Require routinely to carry out.Note avoiding high light direct projection, apply fertilizer in good time, water, weeding, loosen the soil in time and remove disease seedling, dead seedling, preventing damage by disease and insect.
Embodiment 2
1. spore acquisition step:
After August, when female dragon spore is ripe, when sporophyll becomes yellowish-brown from green (spore distribute before), select the afternoon (taken outdoors need after dew disperses) of continuous sunny, with clean scissors, sporophyll is cut, be placed in totally smooth paper bag, air-dry drying, room temperature storage or low temperature (lower than 4 DEG C) storage are to extend spore activity, above-mentioned obtained sporophyll is fully pulverized, 100-200 eye mesh screen sifts out spore (and sporangium), removes impurity, obtains spore (containing sporangium).Need in refrigerating chamber freezing processing about three months (cryogenic temperature is about-10 DEG C) after sporophyll drying, to improve spore germination rate.
2. spore germination step:
By the appropriate pollution-free clean clear water of spore (preferably aqua sterilisa or the water that boiled obtained, to avoid living contaminants) fully to mix and after soaking, remove and float on the spore of the water surface, sporangium, other foreign material and moisture, in-5 DEG C frozen one month to promote spore germination.Add appropriate clean clear water (requiring with aforementioned), with final concentration 300ppm gibberellin process 5 hours, to promote spore germination.
Spore after process is soaked in (water quality requirement: free of contamination clean river, well water, running water in water.Process: phreatic water can directly use; River needs to boil more than 30 minutes, to make contained microorganism, spore deactivation; Running water needs to tan by the sun more than 3 days, same need boil process), spore suspension is placed in glass container, 15 ~ 20 DEG C, cultivate that (covering such as culture vessel available paper covers under natural lighting condition, avoid exogenous microbial or other kind spore to fall into, cause polluting).Note the clean water after often adding process in incubation, keep water volume in culture vessel substantially constant.
If Initial stage of culture has bacterium to grow in a large number, the agricultural streptomycin process of final concentration 0.01% can be added; If have a large amount of protozoa to grow can to add the Tinidazole process of final concentration 0.05mg/mL.If have bubble to produce in spore settling zone in incubation, show that spore is sprouted, protonema starts growth, produce oxygen in a large number bubble time can by spore agglomerate buoyance lift in water meter, suitably to be stirred or other optional predetermined means make spore agglomerate be settled down to the bottom.
3. protonema incubation step:
Cultivate about about one week spore to start to sprout, start to occur green protonema after 2 weeks.Now in culture, urea and potassium dihydrogen phosphate can be added by foliage fertilizer dosage, to promote the growth of protonema.
Often stir in incubation, protonema can be avoided to assemble, be wound agglomerate and reduce appreciation rate.
When (that is, there is agglomerate) during protonema too high levels in culture vessel, cultivation can be expanded, method is: stirred by culture, by stoste: new liquid is (1 ~ 5): the volume ratio of 10 accesses new culture vessel, continue to expand and cultivate, proliferate efficiency can reach more than 5 times.
4. gametophyte is cultivated:
Choose free of contamination humus soil (pH6.0 ~ 7.0), thoroughly tan by the sun under sunlight, parch, pulverize, remove impurity, get and appropriate fill basin to 2/3 of volume as matrix, 50 eye mesh screens crossed by the humus soil separately got after above-mentioned process, tiny for gained humus soil granule is overlying on Culture basin soil table, thickness is about 1cm, then be placed in pallet, water filling in pallet, soil in basin is fully infiltrated.
Mixed by protonema that cultivation obtains, sprinkle, in Culture basin soil table, makes protonema be uniformly distributed, is placed in photoenvironment incubated at room temperature, notes, often to water filling in pallet, keeping ground moistening.Avoid low temperature, to prevent growing of heterotroph fungi.
Cultivate about about 20 days, the visible diameter of soil surface is about the green gametophyte of 1cm.For increasing seedling yield, in gametophyte cultivating process, again can inoculate without protonema blank space at soil surface in good time, also directly can take the mode repeatedly inoculated, to ensure protonema final more than 80% in flakes.
5. sporophyte incubation step:
After gametophyte occurs, keep soil fully moistening, to be combined with ovum to make sperm and to form fertilized egg.Continue to cultivate until start to occur sporophyte seedling.In the present embodiment, protonema 100% in flakes time can generate about 20-50 strain/cm
2seedling.
6. sporophyte (seedling) transplant step:
After sporophyte grows 3 ~ 4 leaves, carry out first time by 4cm × 4cm seeding row spacing transplant, adopt conventional soil as matrix, indoor cultivation.
After first time transplanting 7 ~ 15 days seedlings survive, be transplanted on outdoor seedbed by seeding row spacing 6cm × 10cm, continue to cultivate.(10 ~ 15cm) after the height that meets the requirements is grown to, in autumn or spring in next year being colonizated in production Tanaka by suitable seeding row spacing then until seedling.
7. field daily management operation:
Require routinely to carry out.Note avoiding high light direct projection, apply fertilizer in good time, water, weeding, loosen the soil in time and remove disease seedling, dead seedling, preventing damage by disease and insect.
Embodiment 3
1. spore acquisition step:
After June, when potted plant venus-hair fern spore is ripe, when sporangium becomes yellowish-brown from green (spore distribute before), select the afternoon (taken outdoors need after dew disperses) of continuous sunny, with clean scissors, sporophyll is cut, be placed in totally smooth paper bag, air-dry drying, room temperature storage or low temperature (lower than 4 DEG C) storage are to extend spore activity, above-mentioned obtained sporophyll is fully pulverized, 100-200 eye mesh screen sifts out spore (and sporangium), removes impurity, obtains spore (containing sporangium).
2. spore germination step:
Fully mix by the appropriate pollution-free clean clear water of spore obtained (preferably aqua sterilisa or the water that boiled, to avoid living contaminants) and after soaking, removal floats on the spore of the water surface, sporangium, other foreign material and moisture.Add appropriate clean clear water (requiring with aforementioned), with final concentration 200ppm gibberellin process 12 hours, to promote spore germination.
Spore after process is soaked in (water quality requirement: free of contamination clean river, well water, running water in water.Process: phreatic water can directly use; River needs to boil more than 30 minutes, to make contained microorganism, spore deactivation; Running water needs to tan by the sun more than 3 days, same need boil process), spore suspension is placed in glass container, 15 ~ 20 DEG C, cultivate that (covering such as culture vessel available paper covers under natural lighting condition, avoid exogenous microbial or other kind spore to fall into, cause polluting).Note the clean water after often adding process in incubation, keep water volume in culture vessel substantially constant.
If Initial stage of culture has bacterium to grow in a large number, the agricultural streptomycin process of final concentration 0.1% can be added; If have a large amount of protozoa to grow can to add the Tinidazole process of final concentration 0.5mg/mL.If have bubble to produce in spore settling zone in incubation, show that spore is sprouted, protonema starts growth, produce oxygen in a large number bubble time can by spore agglomerate buoyance lift in water meter, suitably to be stirred or other optional predetermined means make spore agglomerate be settled down to the bottom.
3. protonema incubation step:
Cultivate about about one week spore to start to sprout, start to occur green protonema after 2 weeks.Now in culture, urea and potassium dihydrogen phosphate can be added by foliage fertilizer dosage, to promote the growth of protonema.
Often stir in incubation, protonema can be avoided to assemble, be wound agglomerate and reduce appreciation rate.
When (that is, there is agglomerate) during protonema too high levels in culture vessel, cultivation can be expanded, method is: stirred by culture, by stoste: new liquid is (1 ~ 5): the volume ratio of 10 accesses new culture vessel, continue to expand and cultivate, increment efficiency can reach more than 10 times.
4. gametophyte is cultivated:
Choose free of contamination humus soil (pH6.0 ~ 7.0), thoroughly tan by the sun under sunlight, parch, pulverize, remove impurity, get and appropriate fill basin to 3/4 of volume as matrix, 50 eye mesh screens crossed by the humus soil separately got after above-mentioned process, tiny for gained humus soil granule is overlying on Culture basin soil table, thickness is about 3cm, then be placed in pallet, water filling in pallet, soil in basin is fully infiltrated.
Mixed by protonema that cultivation obtains, sprinkle, in Culture basin soil table, makes protonema be uniformly distributed, is placed in photoenvironment incubated at room temperature, notes, often to water filling in pallet, keeping ground moistening.Avoid low temperature, to prevent growing of heterotroph fungi.
Cultivate about about 20 days, the visible diameter of soil surface is about the green gametophyte of 1cm.For increasing seedling yield, in gametophyte cultivating process, again can inoculate without protonema blank space at soil surface in good time, also directly can take the mode repeatedly inoculated, to ensure protonema final more than 80% in flakes.
5. sporophyte incubation step:
After gametophyte occurs, keep soil fully moistening, to be combined with ovum to make sperm and to form fertilized egg.Continue to cultivate until start to occur sporophyte seedling.In the present embodiment, protonema 100% in flakes time can generate about 20-50 strain/cm
2seedling.
6. sporophyte (seedling) transplant step:
After sporophyte grows 3 ~ 4 leaves, carry out first time by 4cm × 4cm seeding row spacing transplant, adopt conventional soil as matrix, indoor cultivation.
After first time transplanting 7 ~ 15 days seedlings survive, be transplanted on outdoor seedbed by seeding row spacing 6cm × 10cm, continue to cultivate.(10 ~ 15cm) after the height that meets the requirements is grown to, in autumn or spring in next year being colonizated in production Tanaka by suitable seeding row spacing then until seedling.
7. field daily management operation:
Require routinely to carry out.Note avoiding high light direct projection, apply fertilizer in good time, water, weeding, loosen the soil in time and remove disease seedling, dead seedling, preventing damage by disease and insect.
As mentioned above, the present invention can be realized preferably.
The above; it is only preferred embodiment of the present invention; not any pro forma restriction is done to the present invention; according to technical spirit of the present invention; within the spirit and principles in the present invention; the any simple amendment that above embodiment is done, equivalently replace and improve, within the protection domain all still belonging to technical solution of the present invention.
Claims (10)
1. fern protonema liquid culture seedling-cultivating method, is characterized in that, comprise
Spore germination step: the spore of gathering is added in clean clear water and soaks, after removing the floating thing of the water surface and moisture, optionally the frozen scheduled time is to promote spore germination, add appropriate clean clear water and with gibberellin process to promote spore germination, spore after process is soaked in water to be cultivated, and optionally uses the medicine killing or suppress exogenous microbial to grow at Initial stage of culture; Have in spore settling zone bubble to produce in incubation and then show that spore is sprouted, protonema starts growth, if having spore agglomerate buoyance lift in incubation in the water surface, predetermined means should be adopted to make the sedimentation of spore agglomerate.
2. fern protonema liquid culture seedling-cultivating method according to claim 1, is characterized in that, in described spore germination step, frozen temperature is 0 to-20 DEG C, and the frozen time is one thoughtful one month.
3. fern protonema liquid culture seedling-cultivating method according to claim 1, it is characterized in that, in described spore germination step, whether the area grown according to fern belongs to winter outdoor temperature and carry out selecting in area below freezing the need of frozen and preserved spore, be specially: be area below freezing when the area that fern grows belongs to winter outdoor temperature, then should carry out frozen operation; If the area that fern grows belongs to winter, outdoor temperature is area above freezing, then select not carry out frozen operation.
4. fern protonema liquid culture seedling-cultivating method according to claim 1, is characterized in that, in described spore germination step, the final concentration of described gibberellin is 100 ~ 300ppm, and the described gibberellin processing time is 15 minutes to 24 hours.
5. fern protonema liquid culture seedling-cultivating method according to claim 1; it is characterized in that; in described spore germination step; the condition that spore after described process is soaked in when cultivating in water is: 15 ~ 20 DEG C, natural lighting, and culture vessel has the covering protection thing that exogenous microbial can be avoided to enter culture vessel.
6. fern protonema liquid culture seedling-cultivating method according to claim 1, it is characterized in that, described breeding method also comprises
Protonema incubation step: when spore through described spore germination step be cultured to protonema occur time, urea and potassium dihydrogen phosphate is added by foliage fertilizer dosage in culture, to promote the growth of protonema, predetermined means are adopted to form agglomerate to avoid protonema in incubation, when protonema content is higher than predetermined value in culture vessel, the mode expanding cultivation is adopted to continue to cultivate.
7. fern protonema liquid culture seedling-cultivating method according to claim 6, it is characterized in that, described breeding method also comprises:
Gametophyte incubation step, sporophyte incubation step and seedling replanting step.
8. fern protonema liquid culture seedling-cultivating method according to claim 7, it is characterized in that, described gametophyte incubation step is: the humus soil choosing pH6.0 ~ 7.0, tan by the sun under sunlight, parch, pulverize, remove impurity, get and appropriate fill basin to 2/3 ~ 4/5 of volume as matrix, the humus soil separately got after above-mentioned process is overlying on Culture basin soil table after crossing 50 eye mesh screens, thickness 1 ~ 5cm, and make soil drench in basin by predetermined way, by the protonema sprinkle through described protonema incubation step acquisition in Culture basin soil surface, and ensure protonema more than 80% in flakes, be placed in photoenvironment incubated at room temperature, ground moistening and maintenance should be kept to be unfavorable for the temperature that heterotroph fungi grows in the process of cultivating, be cultured to gametophyte and expose soil surface.
9. fern protonema liquid culture seedling-cultivating method according to claim 8, it is characterized in that, described sporophyte incubation step is: after gametophyte exposes soil surface, and keep ground moistening to form fertilized egg to make sperm be combined with ovum, continuation cultivation is until occur sporophyte seedling;
Described seedling replanting step is: after sporophyte grows 3 ~ 4 leaves, carry out first time by 4cm × 4cm seeding row spacing transplant, adopt conventional soil as matrix, indoor cultivation, after first time transplanting 7 ~ 15 days seedlings survive, be transplanted on outdoor seedbed by seeding row spacing 6cm × 10cm, continue to cultivate, grow to after the height that meets the requirements until seedling, in autumn or spring in next year being colonizated in production Tanaka by suitable seeding row spacing then.
10. fern protonema liquid culture seedling-cultivating method according to claim 1, it is characterized in that, described breeding method also comprises
Spore acquisition step: when spore is ripe, cut filemot sporophyll, dry, room temperature storage or low temperature storage, to extend spore activity, pulverize sporophyll, cross 100-200 eye mesh screen and sift out spore and sporangium.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106613160A (en) * | 2016-11-23 | 2017-05-10 | 云南云投生态环境科技股份有限公司 | Spore breeding method for arsenic hyperaccumulator Pteris cretica |
| CN108575714A (en) * | 2018-05-17 | 2018-09-28 | 湖南省农业环境生态研究所 | A kind of ciliate desert-grass spore germination and intensive seedling production method |
| CN111820123A (en) * | 2019-08-31 | 2020-10-27 | 哈尔滨师范大学 | Artificial insemination method for pteridophyte |
| CN116584385A (en) * | 2022-09-01 | 2023-08-15 | 毕节市中药研究所 | Culture medium and method for inducing germination of pteris animalis spores and effectively culturing protophylls |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0265728A (en) * | 1988-08-31 | 1990-03-06 | Nissha Printing Co Ltd | Adsorbing material of spore, production and use thereof |
| CN102715092A (en) * | 2012-07-06 | 2012-10-10 | 中国热带农业科学院热带作物品种资源研究所 | Method for rapidly reproducing new pteris fern seedlings by using prothallium reproduction approaches |
| CN104904455A (en) * | 2015-05-14 | 2015-09-16 | 河口方舟食品有限公司 | Pteridiumaquilinum spore cultivating method |
-
2015
- 2015-11-17 CN CN201510789978.5A patent/CN105265064B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0265728A (en) * | 1988-08-31 | 1990-03-06 | Nissha Printing Co Ltd | Adsorbing material of spore, production and use thereof |
| CN102715092A (en) * | 2012-07-06 | 2012-10-10 | 中国热带农业科学院热带作物品种资源研究所 | Method for rapidly reproducing new pteris fern seedlings by using prothallium reproduction approaches |
| CN104904455A (en) * | 2015-05-14 | 2015-09-16 | 河口方舟食品有限公司 | Pteridiumaquilinum spore cultivating method |
Non-Patent Citations (1)
| Title |
|---|
| 韩玉林 等: "环境条件对荚果蕨孢子繁殖的影响", 《黑龙江八一农垦大学学报》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106613160A (en) * | 2016-11-23 | 2017-05-10 | 云南云投生态环境科技股份有限公司 | Spore breeding method for arsenic hyperaccumulator Pteris cretica |
| CN108575714A (en) * | 2018-05-17 | 2018-09-28 | 湖南省农业环境生态研究所 | A kind of ciliate desert-grass spore germination and intensive seedling production method |
| CN111820123A (en) * | 2019-08-31 | 2020-10-27 | 哈尔滨师范大学 | Artificial insemination method for pteridophyte |
| CN116584385A (en) * | 2022-09-01 | 2023-08-15 | 毕节市中药研究所 | Culture medium and method for inducing germination of pteris animalis spores and effectively culturing protophylls |
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