CN101194594A - Breeding method for alsophila spinulosa - Google Patents
Breeding method for alsophila spinulosa Download PDFInfo
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- CN101194594A CN101194594A CN200710179691.6A CN200710179691A CN101194594A CN 101194594 A CN101194594 A CN 101194594A CN 200710179691 A CN200710179691 A CN 200710179691A CN 101194594 A CN101194594 A CN 101194594A
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- 241001196073 Alsophila spinulosa Species 0.000 title claims abstract description 21
- 238000009395 breeding Methods 0.000 title abstract description 14
- 238000000034 method Methods 0.000 claims abstract description 31
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 17
- 239000001963 growth medium Substances 0.000 claims abstract description 9
- 235000011202 Angiopteris lygodiifolia Nutrition 0.000 claims description 48
- 241000723185 Cyathea Species 0.000 claims description 48
- 239000006870 ms-medium Substances 0.000 claims description 18
- 238000005286 illumination Methods 0.000 claims description 17
- 239000002609 medium Substances 0.000 claims description 17
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 7
- 229920000053 polysorbate 80 Polymers 0.000 claims description 7
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 claims description 6
- 230000000422 nocturnal effect Effects 0.000 claims description 5
- 239000007787 solid Substances 0.000 claims description 5
- 239000011573 trace mineral Substances 0.000 claims description 5
- 235000013619 trace mineral Nutrition 0.000 claims description 5
- 239000000470 constituent Substances 0.000 claims description 4
- 230000000249 desinfective effect Effects 0.000 claims description 2
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 claims description 2
- 241000196324 Embryophyta Species 0.000 abstract description 10
- 230000001488 breeding effect Effects 0.000 abstract description 10
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- RTKIYNMVFMVABJ-UHFFFAOYSA-L thimerosal Chemical compound [Na+].CC[Hg]SC1=CC=CC=C1C([O-])=O RTKIYNMVFMVABJ-UHFFFAOYSA-L 0.000 description 3
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- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
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- Y02P60/216—
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- Cultivation Of Plants (AREA)
Abstract
The invention discloses a breeding method for Alsophila spinulosa, in particular to a breeding method for plants (Alsophila spinulosa(Wall.ex Hook.)Tryon). The breeding method for Alsophila spinulosa which is provided by the invention is that spores of Alsophila spinulosa are vaccinated in an MS culture medium and cultivated after disinfection, and are replanted and managed to finally gain spore seedlings. Using small amount of spore material, the invention can quickly breed large numbers of spore bodies which can become seedlings for about 70 days, and the planting survival rate can reach more than 80%. The invention overcomes the shortcomings of the current Alsophila spinulosa breeding technique of long breeding period, high cost and low efficiency, and starts a new path of breeding Alsophila spinulosa by people quickly and enables industrial manufacture of Alsophila spinulosa on a large scale, thereby satisfying demands of Alsophila spinulosa from the markets and simultaneously reducing damage to wild resources and environment. In addition, the invention provides an excellent experiment system for starting researches in the aspects of the genetic breeding of species and for protecting the biology and the like, and the invention has extensive application prospect.
Description
Technical field
The present invention relates to the propagation method of plant, particularly a kind of propagation method of spinulose tree fern (Alsophila spinulosa (Wall.exHook.) Tryon).
Background technology
Whole world Cyatheaceae (Cyatheaceae) plant have approximately 6 belong to 500 surplus kind, originate in the torrid zone and semi-tropical mountain region.China is located in the northern fringe on Cyatheaceae plant distribution ground, and kind is few, and at present known only has 2 to belong to 14 kinds and 2 mutation, mainly is distributed in southwest and South China.The Cyatheaceae plant of most kinds all has tree-shaped upright stalk, is the arbor shape, and large-scale pinnate leaf submanifold is born in the top of stem, forms umbrella, so be called tree fern again.According to the fossil record, come across Mesozoic Jurassic Period morning or Triassic period in evening the earliest Cyatheaceae plant on the earth history, once extensively distributed in mid-term in Mesozoic Era, very flouring.Afterwards because geology transition and climatic variation, the particularly influence of quaternary glacier phase, a large amount of kind extinctions, also shrink significantly the area, the last torrid zone and suitable " sanctuary " of these environment of subtropics of only remaining in is called as botanic " living fossil " for these reasons.Effects such as this section plant is worth except having higher ornamental plantation, and is also pharmaceutically acceptable, and that its root-like stock has is clearing heat and detoxicating, wind dispelling wets; The trunk of China orchidist Cyatheaceae plant commonly used is as the carrier of the orchid that grows nonparasitically upon another plant; In addition, the trunk of its some kind also can prepare pen container, as pen container tree (Sphaeropteris lepifera) etc.
At present the quantity of spinulose tree fern is very rare, and only the low altitude area is preserved preferably the border of evergreen broad-leaved forest and just seen growth on the south subtropics, the Central Asia, mainly is distributed in ground such as Guizhou, Sichuan, Guangdong and Taiwan in China, is listed in the first-grade state protection plant.Its main cause in imminent danger be the spore life-span short, growth cycle is long, spore germination, gametophytic formation are strict to environmental condition.And because it has higher viewing and admiring and medical value, and market demand is bigger, but the artificial propagation culture technique of Cyatheaceae plant lags behind, and causes wild spinulose tree fern resource to be seriously damaged.
At present, mainly studied Cyathea dregei (Finnie ﹠amp both at home and abroad; Van Staden.Multiplication of the tree fern Cyathea dregei.HortScience, 1987,22 (4): 665), Cyathea-Australis (Goller K, Rybczynski JJ.In-vitro culture used forwoody fern Cyathea-Australis.Acta Societatis BotanicorumPoloniae, 1995,64:1-17), spinulose tree fern (Alsophila spinulosa (Hook.) Tryon) (KharePB, Behera SK.Studies on reproductive biology of a threatened treefern, Cyathea spinulosa Wall.ex Hook.Current Science, 2005,89:173-177; Flourish all one's life, Su Chengduan, Xu Zhenglan, He Liming, the research of spinulose tree fern tissue culture, Plant Physiology Communications, 1985, (1): 38; Cheng Zhiying, Zhang Fenglei, Lan Qinying, Xu Zaifu, Tao Guoda, quick breeding of spinulose tree fern and quality saving Study on Technology, Yunnan plant research, 1991,13 (2): 181-188) with Chinese spinulose tree fern (Alsophila costularisBak.) (Cheng Zhiying, Liu Daohua, the tissue culture of China spinulose tree fern, Plant Physiology Communications, 1992,28 (3): method for tissue culture 210-211), but the propagation method that spinulose tree fern fast-germination, short-term are emerged yet there are no play-by-play.
Summary of the invention
A kind of propagation method that the purpose of this invention is to provide spinulose tree fern (Alsophila spinulosa (Wall.ex Hook.) Tryon).
The propagation method of this spinulose tree fern that is provided (Alsophila spinulosa (Wall.ex Hook.) Tryon) is a spore of cultivating spinulose tree fern in following solid culture medium, obtains the sporophyte seedling; The constituent of described solid culture medium is identical with the MS medium, and wherein macroelement and trace element concentration are the 1/8-1 of MS medium, and the concentration of molysite and organic principle is identical with the MS medium, and pH is 5.5-6.0.
In the described solid culture medium, macroelement and trace element concentration specifically can be 1/2 of MS medium, and pH is 5.8.
Described spinulose tree fern spore with containing 3-6g/100ml NaClO and the disinfection of 0.1-0.3g/100ml Tween-80 solution, if material pollutes easily, can soak 10-15s with 70% alcohol before all are handled before cultivating.
The described time of disinfecting is 4.5-6min.
The described solution disinfection processing 5min that is specially with containing 5g/100mlNaClO and 0.2g/100ml Tween-80 that disinfects.
The condition of culture of described spinulose tree fern spore is: under 22-28 ℃, the dark earlier 12-72h that cultivates goes to it then and continues under the illumination to cultivate, and intensity of illumination is 20-80 μ molm
-2S
-1, light application time is that the condition of culture of the described spinulose tree fern spore of 8-16h/d is specially under 25 ℃, and the dark earlier 24h that cultivates goes to it then and continues under the illumination to cultivate, and intensity of illumination is 50 μ molm
-2S
-1, light application time is 12h/d.
Above the propagation method of described spinulose tree fern spore also comprise the step of transplanting described sporophyte seedling.
The management condition of described transplanting process shaded for the autumn in summer, kept day temperature 21-26 ℃ winter in spring, and nocturnal temperature 10-15 ℃, air humidity 70-90% executed one time the 1/2MS medium in every month behind the slow seedling 20-35d.
Propagation method of the present invention is suitable for spinulose tree fern (Alsophila spinulosa (Wall.ex Hook.) Tryon).
The present invention is a material with the spore, utilizes tissue culture technique, in conjunction with the proprietary cultivation management measure to pteridophyte, has set up the RAISING ALSOPHILA SPINULOSA FROM SPORES method again.Utilize method of the present invention, with a spot of spore material, can go out a large amount of sporophytes by fast breeding, average 7-9 days spores can be sprouted, 25-35 days formation gametophytes, 68-97 days formation sporophyte seedlings.Compare with other existing cultural method, the inventive method has that fast-germination, short-term are emerged, cultivation period is lacked, characteristics with low cost.The inventive method has remedied that existing spinulose tree fern propagation technique cultivation period is long, cost is high and inefficient deficiency; opened up the new way of an artificial spinulose tree fern of breeding fast; make the large-scale industrial production of spinulose tree fern become possibility; thereby satisfy the demand of market, can reduce its wild resource and environment damage simultaneously spinulose tree fern.In addition, the present invention also provides good test system for the research of carrying out aspects such as this species genetic breeding and conservation biology, is with a wide range of applications.
Description of drawings
Fig. 1 is the gametophyte of spinulose tree fern.
Fig. 2 is the test-tube plantlet (sporophyte seedling) of spinulose tree fern.
Fig. 3 is the spinulose tree fern seedling of field planting.
Embodiment
Employed experimental technique is conventional method if no special instructions among the following embodiment.
Consisting of of the medium of MS described in the experiment:
The composition of MS medium
| Title | Content (mg/L) |
| Macroelement NH 4NO 4 KNO 3 CaCl 2·2H 2O MgSO 4·7H 2O KH 2PO 4 | 1650 1900 440 370 170 |
| Elements K I H 3BO 3 MnSO 4·4H 2O ZnSO 4·7H 2O Na 2MoO 4·2H 2O GuSO 4·5H 2O CoCl 2·6H 2O | 0.83 6.2 22.3 8.6 0.25 0.025 0.025 |
| Molysite Na 2-EDTA FeSO 4·7H 2O | 37.3 27.8 |
| Organic principle nicotinic acid puridoxine hydrochloride (VB 6) nicotinic acid thiamine (VB 1) the glycine inositol | 0.5 0.5 0.1 2.0 100.0 |
The breeding of embodiment 1, spinulose tree fern (Alsophila spinulosa (Wall.ex Hook.) Tryon)
One, 1/2MS culture medium preparation
Constituent in the 1/2MS medium is identical with the MS medium, and wherein, macroelement and microelement concentration are 1/2 of MS medium, and the concentration of molysite and organic principle is identical with the MS medium, and agar 7g/L, pH are 5.8.Boil dissolving back branch and be filled in the 100ml triangular flask, 121 ℃ then, high pressure steam sterilization 20min.
Two, the sterilization of spinulose tree fern spore
Get ripe spinulose tree fern (Alsophila spinulosa (Wall.ex Hook.) Tryon) spore 5mg and place in the 1.5ml centrifuge tube, splash into sterile water, fully vibration makes into suspension, leave standstill 1h after, the centrifugal 2min of 4000r/min abandons supernatant; Splash into the thimerosal that about 1.0ml contains 0.2g/100ml Tween-80 and 5g/100mlNaClO then in centrifuge tube, to the spore sterilization that carries out disinfection, behind the 5min, the centrifugal 1min of 4000r/min abandons supernatant; Use aseptic water washing 4-5 time at last, obtain aseptic spore suspension.
Three, the inoculation of aseptic spore and cultivation
Under aseptic condition, earlier with the surface wettability of sterile water with the 1/2MS medium of above-mentioned preparation, make its surface form the very thin moisture film of one deck (water that collects in the bottom during with the inclination triangular flask can not allow dropper suct continuous water droplet and be advisable), with dropper the aseptic spore suspension that step 2 obtains evenly is seeded on the medium then.After the inoculation, under 25 ℃, the dark earlier 24h that cultivates goes to it then and continues under the illumination to cultivate, and intensity of illumination is 50 μ molm
-2S
-1(light source is a fluorescent lamp), light application time is 12h/d.
Four, the transplanting of spinulose tree fern sporophyte seedling and management
Cultivate after 7 days, spore begins to sprout, and germination rate is that 82%, 25 day (starting at the same day from inoculation) forms gametophyte (Fig. 1), and 68 (starting at the same day from inoculation) sky begins to form the sporophyte seedling.To about 88-90 days (starting at the same day from inoculation), the high 3-4cm of seedling placed the natural lighting lower refining seedling 6 days with triangular flask, opened the bottle cap hardening again 6 days.Press from both sides out test-tube plantlet (Fig. 2) gently with tweezers then, clean medium after, be transplanted in the plain sand of 100 order dusting covers screening, and to place cultivation indoor cultivation, cultivation temperature be 25 ℃ that intensity of illumination is 50 μ molm
-2S
-1(light source is a fluorescent lamp), light application time is 12h/d.(composition of culture matrix and mixed proportion are: turfy soil: fine sand: fertilizer=4: 2: 1) in after 1 month its field planting being coiled to the cave, put hot-house culture, Xia Qiu shades, and keeps day temperature 21-26 ℃ winter in spring, nocturnal temperature 10-15 ℃, air humidity 70-90%.Slow seedling was executed the 1/2MS medium one time in every month after 1 month, and through about 120 days, seedling is long to the 5-10cm height, and well developed root system, carries out basin in the plant division, field planting seedling (Fig. 3), average planting survival rates 86%.
The breeding of embodiment 2, spinulose tree fern (Alsophila spinulosa (Wall.ex Hook.) Tryon)
One, 1/8MS culture medium preparation
The constituent of 1/8MS medium is identical with the MS medium, and wherein, macroelement and microelement concentration are 1/8 of MS medium, and the concentration of molysite and organic principle is identical with the MS medium, and agar 6.5g/L, pH are 5.5.Boil dissolving back branch and be filled in the 100ml triangular flask, 121 ℃ then, high pressure steam sterilization 20min.
Two, the sterilization of spinulose tree fern spore
Get ripe spinulose tree fern spore 2mg, place in the 1.5ml centrifuge tube, splash into sterile water, fully vibration makes into suspension, leave standstill 1h after, the centrifugal 2min of 4000r/min abandons supernatant; Splash into the thimerosal that about 1.0ml contains 0.1g/100ml Tween-80 and 3g/100mlNaClO then in centrifuge tube, to the spore sterilization that carries out disinfection, behind the 4.5min, the centrifugal 1min of 4000r/min abandons supernatant; Use aseptic water washing 4-5 time at last, obtain aseptic spore suspension.
Three, the inoculation of aseptic spore and cultivation
Under aseptic condition, earlier with the surface wettability of sterile water with the 1/8MS medium of above-mentioned preparation, make its surface form the very thin moisture film of one deck (water that collects in the bottom during with the inclination triangular flask can not allow dropper suct continuous water droplet and be advisable), with dropper the aseptic spore suspension that step 2 obtains evenly is seeded on the medium then.After the inoculation, under 22 ℃, the dark earlier 12h that cultivates goes to it then and continues under the illumination to cultivate, and intensity of illumination is 20 μ molm
-2S
-1(light source is a fluorescent lamp), light application time is 8h/d.
Four, the transplanting of spinulose tree fern sporophyte seedling and management
After cultivating 7d (starting at the same day from inoculation), spore begins to sprout, and germination rate is 91%, and 35d (starting at the same day from inoculation) forms gametophyte (Fig. 1), and 97d begins to form the sporophyte seedling.To 148-156 days (starting at the same day from inoculation), the high 3-4cm of seedling placed natural lighting lower refining seedling 10d with triangular flask, opens bottle cap hardening 10d again.Press from both sides out test-tube plantlet (Fig. 2) gently with tweezers then, clean medium after, be transplanted in the plain sand of 100 order dusting covers screening, and to place cultivation indoor cultivation, cultivation temperature be that 22 ℃ of intensities of illumination are 20 μ molm
-2S
-1(light source is a fluorescent lamp), light application time is 8h/d.(composition of culture matrix and mixed proportion are: turfy soil: fine sand: fertilizer=4: 2: 1) in behind the 40d its field planting being coiled to the cave, put hot-house culture, Xia Qiu shades, and keeps day temperature 21-26 ℃ winter in spring, nocturnal temperature 10-15 ℃, air humidity 79-90%.Executed the 1/2MS medium one time in every month behind the slow seedling 25d, when treating seedling length to 5-10cm height and well developed root system, basin in the plant division, field planting sporophyte seedling (Fig. 3), average planting survival rates 70%.
The breeding of embodiment 3, spinulose tree fern (Alsophila spinulosa (Wall.ex Hook.) Tryon)
One, full dose MS culture medium preparation
Macroelement is all identical with the MS medium with iron salt concentration with trace element, and the concentration of molysite and organic principle is identical with the MS medium, and agar 8g/L, pH are 6.0.Boil dissolving back branch and be filled in the 100ml triangular flask, 121 ℃ then, high pressure steam sterilization 20min.
Two, the sterilization of spinulose tree fern spore
Get ripe spinulose tree fern spore 7mg, place in the 1.5ml centrifuge tube, splash into sterile water, fully vibration makes into suspension, leave standstill 1h after, the centrifugal 2min of 4000r/min abandons supernatant; Splash into the thimerosal that about 1.0ml contains 0.3g/100ml Tween-80 and 6g/100mlNaClO then in centrifuge tube, to the spore sterilization that carries out disinfection, behind the 6min, the centrifugal 1min of 4000r/min abandons supernatant; Use aseptic water washing 4-5 time at last, obtain aseptic spore suspension.
Three, the inoculation of aseptic spore and cultivation
Under aseptic condition, earlier with the surface wettability of sterile water with the MS medium of above-mentioned preparation, make its surface form the very thin moisture film of one deck (water that collects in the bottom during with the inclination triangular flask can not allow dropper suct continuous water droplet and be advisable), with dropper the aseptic spore suspension that step 2 obtains evenly is seeded on the medium then.After the inoculation, under 28 ℃, the dark earlier 72h that cultivates goes to it then and continues under the illumination to cultivate, and intensity of illumination is 80 μ molm
-2S
-1(light source is a fluorescent lamp), light application time is 16h/d.
Four, the transplanting of spinulose tree fern sporophyte seedling and management
After cultivating 9d, spore begins to sprout, and germination rate is 80%, and 32d forms gametophyte (Fig. 1), and 90d begins to form the sporophyte seedling.To 122-130 days (starting at the same day) from inoculation, during the high 3-4cm of seedling, triangular flask is placed natural lighting lower refining seedling 8d, open bottle cap hardening 8d again.Press from both sides out test-tube plantlet (Fig. 2) gently with tweezers then, clean medium after, be transplanted in the plain sand of 100 order dusting covers screening, and to place cultivation indoor cultivation, cultivation temperature be 28 ℃ that intensity of illumination is 80 μ molm
-2S
-1(light source is a fluorescent lamp), light application time is 16h/d.(composition of culture matrix and mixed proportion are: turfy soil: fine sand: fertilizer=4: 2: 1) in behind the 30d its field planting being coiled to the cave, put hot-house culture, Xia Qiu shades, and keeps day temperature 21-26 ℃ winter in spring, nocturnal temperature 10-15 ℃, air humidity 79-90%.Executed the 1/2MS medium one time in every month behind the slow seedling 25d, when treating seedling length to 5-10cm height and well developed root system, basin in the plant division, field planting sporophyte seedling (Fig. 3), average planting survival rates 74%.
Experimental result shows, utilizes the inventive method breeding spinulose tree fern, and average 7-9d days spores can be sprouted, and forms gametophyte in 25-35d days, forms the sporophyte seedling in 68-97 days, and the sporophyte seedling transplanting survival rate can reach 86%.
Claims (10)
1. the propagation method of a spinulose tree fern is a spore of cultivating spinulose tree fern in following solid culture medium, obtains the sporophyte seedling; Constituent in the described solid culture medium is identical with the MS medium, and wherein macroelement and trace element concentration are the 1/8-1 of MS medium, and the concentration of molysite and organic principle is identical with the MS medium, and pH is 5.5-6.0.
2. method according to claim 1 is characterized in that: in the described medium, macroelement and trace element concentration are 1/2 of MS medium, and pH is 5.8.
3. method according to claim 1 and 2 is characterized in that: described spinulose tree fern spore is before cultivating, with containing 3-6g/100ml NaClO and the disinfection of 0.1-0.3g/100ml Tween-80 solution.
4. method according to claim 3 is characterized in that: the described time of disinfecting is 4.5-6min.
5. method according to claim 4 is characterized in that: the described solution disinfection 5min that contains 5g/100mlNaClO and 0.2g/100ml Tween-80 for using that disinfects.
6. method according to claim 1 is characterized in that: the condition of culture of described spinulose tree fern spore is: under 22-28 ℃, the dark earlier 12-72h that cultivates goes to it then and continues under the illumination to cultivate, and intensity of illumination is 20-80 μ molm
-2S
-1, light application time is 8-16h/d.
7. method according to claim 6 is characterized in that: the condition of culture of described spinulose tree fern spore is under 25 ℃, and the dark earlier 24h that cultivates goes to it then and continues under the illumination to cultivate, and intensity of illumination is 50 μ molm
-2S
-1, light application time is 12h/d.
8. method according to claim 1 is characterized in that: the propagation method of described spinulose tree fern spore also comprises the step of transplanting described sporophyte seedling.
9. method according to claim 8, it is characterized in that: the management condition of described transplanting process shaded for the autumn in summer, kept day temperature 21-26 ℃ winter in spring, nocturnal temperature 10-15 ℃, air humidity 70-90% executed one time the 1/2MS medium in every month behind the slow seedling 20-35d.
10. according to claim 1 or 8 described methods, it is characterized in that: described spinulose tree fern is spinulose tree fern (Alsophilaspinulosa (Wall.ex Hook.) Tryon).
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| Application Number | Priority Date | Filing Date | Title |
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| CN200710179691.6A CN101194594B (en) | 2007-12-17 | 2007-12-17 | Breeding method for alsophila spinulosa |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200710179691.6A CN101194594B (en) | 2007-12-17 | 2007-12-17 | Breeding method for alsophila spinulosa |
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| CN102934550A (en) * | 2012-11-13 | 2013-02-20 | 雷学军 | Spinulose tree fern propagation and transplant method |
| CN104322225A (en) * | 2013-11-30 | 2015-02-04 | 钟诚 | Artificial propagation method of cyathea spinulosa |
| CN105145365A (en) * | 2015-09-22 | 2015-12-16 | 云南省农业科学院花卉研究所 | Green globular body tissue culture propagation method for Alsophila costularis |
| CN110432154A (en) * | 2019-09-19 | 2019-11-12 | 玉林师范学院 | A kind of cultural method of spinulose tree fern tissue |
| CN110786243A (en) * | 2019-12-03 | 2020-02-14 | 厦门市园林植物园 | Propagation method for tissue culture of penholder tree |
| CN111837775A (en) * | 2019-08-31 | 2020-10-30 | 哈尔滨师范大学 | Culture medium and method for breeding Alsophila sinensis by using culture medium |
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| CN102934550A (en) * | 2012-11-13 | 2013-02-20 | 雷学军 | Spinulose tree fern propagation and transplant method |
| CN102934550B (en) * | 2012-11-13 | 2014-10-22 | 雷学军 | Spinulose tree fern propagation and transplant method |
| CN104322225A (en) * | 2013-11-30 | 2015-02-04 | 钟诚 | Artificial propagation method of cyathea spinulosa |
| CN105145365A (en) * | 2015-09-22 | 2015-12-16 | 云南省农业科学院花卉研究所 | Green globular body tissue culture propagation method for Alsophila costularis |
| CN105145365B (en) * | 2015-09-22 | 2016-12-07 | 云南省农业科学院花卉研究所 | A kind of tissue culture propagation method of China Caulis cyatheae spinulosae green spheroids approach |
| CN111837775A (en) * | 2019-08-31 | 2020-10-30 | 哈尔滨师范大学 | Culture medium and method for breeding Alsophila sinensis by using culture medium |
| CN110432154A (en) * | 2019-09-19 | 2019-11-12 | 玉林师范学院 | A kind of cultural method of spinulose tree fern tissue |
| CN110786243A (en) * | 2019-12-03 | 2020-02-14 | 厦门市园林植物园 | Propagation method for tissue culture of penholder tree |
| CN112021178A (en) * | 2020-09-01 | 2020-12-04 | 中国科学院植物研究所 | Method for promoting germination of Alsophila spinulosa spores by using multiwalled carbon nanotubes |
| CN112021178B (en) * | 2020-09-01 | 2021-10-22 | 中国科学院植物研究所 | Method for promoting germination of Alsophila spinulosa spores by using multiwalled carbon nanotubes |
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