CN100400654C - Glasswort tissue culture method and culture medium - Google Patents
Glasswort tissue culture method and culture medium Download PDFInfo
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- CN100400654C CN100400654C CNB2006100031908A CN200610003190A CN100400654C CN 100400654 C CN100400654 C CN 100400654C CN B2006100031908 A CNB2006100031908 A CN B2006100031908A CN 200610003190 A CN200610003190 A CN 200610003190A CN 100400654 C CN100400654 C CN 100400654C
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Abstract
The present invention discloses a tissue culture method for Saliconia europaea L. and a culture medium special for tissue culture of Saliconia europaea L. An inducing and differentiating culture medium for calluses of Saliconia europaea L. is an MS culture medium containing 0.05-0.13 mg/L of TDZ and 0.8-1.2 mg/L of NAA. The tissue culture method for Saliconia europaea L. of the present invention uses seeds, hypocotyledonary axis or roots of the Saliconia europaea L. as explants which are cultured in the inducing and differentiating culture medium for calluses of Saliconia europaea L. to form calluses and to differentiate buds, and then, the differentiated buds are transplanted into a rooting culture medium to root and to form complete plants. The callus formation rate of the method of the present invention is high, and the formation rate of the complete plants is high, which establishes a foundation for verifying the function of salt-resistance genes of the Saliconia europaea L.
Description
Technical field
The present invention relates to a kind of method and special culture media thereof of tissue culture salicornia europaeal.
Background technology
Salicornia europaeal (Saliconia europaea L.) belongs to the Chenopodiaceae salicornia europaeal and belongs to (Saliconia), is one of true halophytes of the salt tolerant the most reported so far.With its generic Salicornia Bigelovii Torr. (Saliconia bigelovvi Torr.) also be the more a kind of halophytes of present research and utilization, but its suitable cloth scope estranged wants much narrow than salicornia europaeal, so its utilization and extention is subjected to the restriction of region.This belongs in the whole world nearly kind more than 50 (L ü ZJ (Lv Zhongjin), Eward PG..The crop irrigated with pure seawater-Salicornia bigelovii Torr.World Agriculture (world agriculture), 2001 (2): 14-16), distribute the most extensive with salicornia europaeal, all produce (Zhao KF (Zhao Kefu) all over the world, Fan H (model sea) .Halophyte resources in theworld that allow seawater irrigation.Chinese Bulletin of Botany (BULLETIN OF BOTANY Vol.), 2000, I7 (3): 282-288).Salicornia europaeal is born in the saltings, the seashore is taken up in the salt lake, in Liaoning of China, all there is distribution (Zhao KF (Zhao Kefu) provinces and regions such as Hebei, Shanxi, Shaanxi, Ningxia, Xinjiang, Shandong, Jiangsu, Feng LT (Feng Litian) .Halophyte resources in China (Chinese halophytes resource). Beijing: Bei jing Scientific Publishers (Beijing Science Press), 2001).Under the seawater irrigation condition, can produce 10-15t/hm
2Biomass, its seed production can reach 1-2t/hm
2(soya bean is 3t/hm
2), oleaginousness is about 30%, protein content can reach 30.2% (O ' leary JW, Glenn EP and Watson MC.Agriculturalproduction of halophytes irrigated with seawater.Plant and Soil, 1985,89:311-321).Unsaturated fatty acids accounting example is very high in its grease, is suitable for eating, and the prospect that therefore develops into oil crops is boundless.Its tender stem is nutritious, can be used as vegetables; Seedcake and stalk can be used as feed; Herb can be used as diuretic(s) (Zhao KF (Zhao Kefu), Feng LT (Feng Litian) .Halophyte resources inChina (Chinese halophytes resource). Beijing: Bei jing Scientific Publishers (Beijing Science Press), 2001).The salicornia europaeal biomass is big, it is the good material of utilizing solid carbon, oxygen supply and conversion energy of saltings, improvement for the saltings ecotope, check winds and fix drifting sand, aspects such as soil conservation, envrionment temperature adjusting have active effect, are the donor of good salt resistant gene and the gedanken experiment material that carries out Mechanism of Salt-tolerant research simultaneously.The research of past to the salicornia europaeal platymiscium mainly concentrates on aspects such as ecology, Physiology and biochemistry and molecular biology.For example the salicornia europaeal nonoculture relies on dynamics research (the Aaron ME.Density-dependent dynamics of SlicorniaEuropaea Monocultures.Ecology of density, 1987,68 (3): 737-741), NaCl is to influence (the Flowers TJ.The effect of sodium chlorideon enzyme activities from four halophyte spieces of Chenopodiaceae.Phytochemistry of Saliconiaramosissima Woods enzymic activity, 1972,11:1881-1886), Salicornia Bigelovii Torr. vacuole Na
+/ H
+Exchange activity (Graham EP, Margaret AD and Karen SS.Increased vacuolar Na
+/ H
+Exchange activity inSaliconia bigelovvi Torr.in response to NaCl.Journal of Experimental Botany, 2002,53 (371): 1055-1065), salinity is to influence (the Moghaieb REA of salicornia europaeal osmoregulation, Hirofumi Saneoka, Kounosuke F.Effect of salinity on osmotic ajustment, glycinebetaine accumulation and the aldehyde dehydrogenase gene expressionin two halophytic plants, Salicornia europaea and Suaeda maritima.PlantScience, 2004,166:1345-1349), the clone of resistant gene of salt and other gene (Cai L (Cai Lun), ZhangFC (Zhang Fuchun), Ma J (Ma Ji) .Cloning and sequence analysis of NHX genes from threespecies of Halophytes from Xinjiang.Plant physiology Communications (Plant Physiology Communications), 2005,41 (3): 383-387; Li JY (Li Jinyao), Zhang FC (Zhang Fu pounds), Ma J (Ma Ji) .Cloning of the orf25 gene of Salicornia europaea L.by a single primer.Chinese Journal of Biotechnology, 2003,19 (1): 120-123) etc.
The tissue culture work of salicornia europaeal is significant.One, salicornia europaeal external regeneration system can be used as the platform of Mechanism of Salt-tolerant research, and for illustrate salt tolerant mechanism on cell levels, the function of checking salicornia europaeal salt-resistant related gene is laid a good foundation on molecular level.Its two, salicornia europaeal have cultivate into sea water vegetable and the health care oil crops potentiality.To promote the cultivation of salicornia europaeal aborning, must cultivate good Cultivar at its multiple practical use.And the somaclonal variation in the tissue culture is one of breeding technique of present widespread use.The foundation of salicornia europaeal external regeneration system (clone) makes the work of this respect become possibility.
Up to now, the tissue culture of salicornia europaeal platymiscium rarely has report.(Chiwon WL such as Chiwon W.Lee, Glenn EP, O ' leary JW.In vitro propagation of Salicornia bigelovii byshoot-tip cultures.Hort.Sci.1992,27 (5): 47) stem apex with Salicornia Bigelovii Torr. (Salicorniabigelovii) is an explant, has realized the numerous soon of Salicornia Bigelovii Torr..This tissue culture for salicornia europaeal provides the data that provides a reference.But the salicornia europaeal of seedling phase does not have branch, and the speed of growth is very slow on substratum, and therefore as if being explant with the stem apex, the source is restricted.
Summary of the invention
The method and the special culture media thereof that the purpose of this invention is to provide a kind of tissue culture salicornia europaeal.
Salicornia europaeal callus induction provided by the present invention and division culture medium are to contain the TDZ of 0.05-0.13mg/L and the MS substratum of 0.8-1.2mg/L NAA.
Described salicornia europaeal callus induction and division culture medium are preferably the thiadiazoles phenylurea (TDZ) that contains 0.1mg/L and the MS substratum of 1.0mg/L naphthylacetic acid (NAA).
The method of tissue culture salicornia europaeal provided by the present invention is to be explant with salicornia europaeal seed, hypocotyl or root, in above-mentioned salicornia europaeal callus induction and division culture medium, cultivate, form callus and break up and sprout, will break up bud and forward the formation whole plant of taking root in the root media to.
Described explant is preferably seed.
Described salicornia europaeal callus induction and division culture medium are preferably the thiadiazoles phenylurea (TDZ) that contains 0.1mg/L and the MS substratum of 1.0mg/L NAA.
Described root media can be the 1/2MS substratum that contains 0.5mg/L IBA and 0.1mg/L KT.
In the aforesaid method, the culture condition of whole tissue culture procedures is 20-28 ℃, secretly cultivates during callus lures tissue to lead, and illumination every day is 8-12 hour between the differentiation phase, intensity of illumination 2500-3500LUX.Described culture condition is preferably 24-26 ℃, and illumination every day is 12 hours in the atomization, intensity of illumination 3000LUX (LX).
The present invention passes through adventitious organogenesis, having set up the external regeneration system of salicornia europaeal, is explant with complete mature seed, on the MS substratum that adds 0.1mg/L TDZ and 1mg/L NAA, form callus at the hypocotyl place after cultivating for three weeks, the average frequency that forms callus is 99%.Callus is cultivated the back differentiation of 3～4 weeks and is sprouted differentiation frequency about 26.7% on the substratum that contains TDZ0.1mg/L and NAA1mg/L.Average every callus produces 0.4 of regeneration bud.The differentiation bud changes the 1/2MS substratum that contains 0.5mg/L IBA and 0.1mg/L KT over to, the formation whole plant of taking root after three weeks after 3 weeks.Method callus rate of formation height of the present invention, whole plant rate of formation height, be further investigation salicornia europaeal Mechanism of Salt-tolerant on cell levels, in the function of checking salicornia europaeal related gene in salt tolerance, coherent signal are transduceed the collaborative approach relevant with other on the molecular level, and cultivate good salicornia europaeal kind (at different purposes such as oil plant, vegetables, medicinal, environment remediation), an ideal research platform is provided.
Description of drawings
Fig. 1 induces callus, differentiation bud that obtains and the complete regenerated plant photo of taking root
Fig. 2 is the relative growth rate point and line chart of callus at CIM-14 and CIM-16
Embodiment
Method among the following embodiment is ordinary method if no special instructions.
The optimization of embodiment 1, salicornia europaeal callus culture condition
1, the selection of explant
With seed, hypocotyl and root segment is the explant induction callus.In order to compare the ability of seed, hypocotyl and root segment evoked callus, it is inoculated in the CIM-16 substratum cultivates, concrete grammar is as follows:
The pericarp of salicornia europaeal seed appearance is removed, at first in 70% ethanol, soaked 30 seconds, changed in 10% the clorox sterilization then over to 15 minutes, then use aseptic water washing four times, part seed is inoculated on the MS substratum, after 25 ℃ of dark 3 weeks of cultivation, its hypocotyl and root segment are cut, be inoculated into CIM-16 substratum (table 1) respectively and go up with evoked callus; Another part seed direct inoculation is gone up with evoked callus to CIM-16 substratum (table 1).Culture condition in the evoked callus process is: temperature (25 ± 1) ℃, dark condition is cultivated down.Each experiment has 6-8 parallel sample, respectively establishes 3 repeating groups.
Table 1 salicornia europaeal callus inducing medium
Annotate: the basic medium in the table 1 is the MS substratum, and pH 5.8.
The result show seed be inoculated in the CIM-16 substratum 3 week the back form faint yellow callus at the hypocotyl place, and very a spot of callus just appears in hypocotyl and root segment after cultivating for 4 weeks, induction frequency is very low (table 2) also.Least significant difference method analysis revealed, there is significant difference in the callus of induce rate of seed, hypocotyl and root segment, shows that seed is the best explant of evoked callus.Different alphabetical a in the table 2, b, c represent LSD multiple comparisons significant difference on 0.05 level.
Table 2 is cultivated the induction frequency of 4 all backs callus
| Explant | Induction frequency (callus number/explant sum) |
| Seed | 0.99±0.006a |
| Hypocotyl | 0.36±0.03b |
| Root segment | 0.10±0.02c |
2, evoked callus Optimum of culture medium
In order to screen best callus inducing medium, the salicornia europaeal seed after the routine sterilization, is inoculated into CIM-1～No. 16 substratum (table 1) with evoked callus.In the whole tissue culture procedures, culture condition is: temperature (25 ± 1) ℃, callus induction carries out under dark condition.
The result shows:
Seed began to sprout in No. 13 substratum of CIM-5～in 2 days, sprouted fully, and had two pieces of oval cotyledons in 5 days.Hypocotyl is expanded gradually after 1 week, and it is plump that cotyledon becomes, a small amount of white callus of formation, not regrowth of root simultaneously in 12 days; The comparatively fine and close callus group of 4 weeks back formation, diameter can reach 3～5mm.Along with the prolongation of incubation time, callusization also takes place in two pieces of cotyledons.
In the CIM-16 substratum, explant forms faint yellow callus at the hypocotyl place after 3 weeks, the faint yellow slightly transparent callus group of 4 weeks back formation.
In the CIM-2 substratum, 2 all backs produce a small amount of white sub-translucent callus at the hypocotyl position, form loose frangible callus group, the slightly general cyan of color after 4 weeks.
And explant does not form callus after cultivating for 4 weeks in CIM-1, CIM-3, CIM-4, CIM-14 and CIM-15 substratum.The CIM-1 substratum is the MS substratum, does not add any hormone, and what obtain is the aseptic seedling of yellow.Explanation will be from salicornia europaeal seed evoked callus, and a certain amount of growth hormone or growth hormone are prerequisites in conjunction with an amount of phytokinin.In the CIM-3 substratum, explant only forms minute quantity white callus after cultivating for 5 weeks; The CIM-4 substratum does not then obtain callus.
In No. 4 substratum of CIM-2～, the sprouting of seed postpones, percentage of germination No. 13 is low than CIM-5～, and along with 2, the increase of 4-D concentration, this trend is obvious further, illustrate that callus is containing 2, after 4～5 weeks of growing on the inducing culture of 4-D, may lose regeneration potential, 2,4-D trends towards suppressing the form generation of plant.As seen, 2 of high density, 4-D is unfavorable for inducing of callus to the toxic effect of explant.CIM-14 and No. 15 substratum only contain hormone TDZ, and the etiolated seedling epicotyl growth that obtains is very active.Illustrate that adding TDZ separately may be not enough to inducing the salicornia europaeal callus.CIM5～13 substratum are easy to the generation (average inductivity more than 98%) of callus induction, but callus is transferred on the division culture medium that contains different hormone combinations, cultivated for 8 weeks after, the bud point does not appear in callus all the time.
The result shows that also the callus of acquisition is broadly divided into three types according to its form and differentiation capability.First type callus is only containing 2, obtains on the substratum of 4-D (CIM-2), and the slightly general green grass or young crops of white, loose frangible, differentiation capability is poor, can grow adventive root (A among Fig. 1) on division culture medium; Second type callus is containing 2, and the substratum of 4-D and a kind of phytokinin (6-BA or KT) obtains on (CIM-5～No. 13), and white is fine and close, the speed of growth is slow, on division culture medium, become green very soon, can differentiate adventive root, but do not obtain regeneration bud (B among Fig. 1); The callus of the third type (C, D among Fig. 1) is easy to induce on the substratum that contains TDZ and NAA (CIM-16), and the callus that obtains is faint yellow slightly transparent, and this callus can obtain regeneration bud (C among Fig. 1) on division culture medium.
3, the optimization of callus differentiation culture based component
In the whole tissue culture procedures, culture condition is: temperature (25 ± 1) ℃, illumination every day 12h, intensity of illumination 30001x.
The salicornia europaeal seed is after the routine sterilization, after in the CIM-16 substratum, cultivating for 5 weeks, the callus that obtains, separated into two parts, after transferring in the CIM-14 substratum (among the MS add TDZ 1mg/L) again or in CIM-16 number (adding the substratum of TDZ 0.1mg/L and NAA 1mg/L among the MS) substratum, cultivating 0,7,14,21 and 28 day, calculate the relative growth rate (RGR) of callus lines according to formula RGR=(FWt one FWo)/FWo at CIM-14, CIM-16, FWt represents to survey periodic callus fresh weight, and FWo represents the original weight of callus.The result as shown in Figure 2, the result shows that the relative growth rate in CIM-16 is greater than CIM-14.
Though the CIM5-13 substratum is easy to the generation (average inductivity more than 98%) of callus induction, and callus is transferred on the division culture medium that contains different hormone combinations, cultivated for 8 weeks after, the bud point does not appear in callus all the time.It has been generally acknowledged that, 2,4-D trends towards suppressing form generation (the Aparna G of plant, Rashid A.TDZ-induced somatic embryogenesis in non-responsive caryopses of rice usinga short treatment with 2,4-D.Plant Cell Tiss Org.Cult, 2004,76:29-33), callus is containing 2, after 4～5 weeks of growing on the inducing culture of 4-D, may lose regeneration potential.Seed is after dedifferentiation substratum (CIM-16) is gone up 4～5 weeks of cultivation, form faint yellow callus cell group, the callus piece of the about 2～3mm of diameter is downcut, and SIM1, the SIM2, SIM3 or the SIM44 kind that change in the table 3 contain in the division culture medium of different concns TDZ.On the MS substratum that adds TDZ 0.1mg/L and NAA 1mg/L, cultivate 3-4 after week, occurred green bud point (E among Fig. 1) on the callus, also have a small amount of bud point to occur on other 3 kinds of substratum successively.The frequency of CIM-16 substratum evoked callus is also very high, reaches as high as 100%, therefore the CIM-16 substratum is defined as inducing the optimal medium of embryo callus subculture.Callus differentiation frequency on the SIM1 substratum is the highest, is about 26.7%, and the regeneration bud that average every callus obtains has 0.4 (table 3).Between 4 kinds of division culture mediums of least significant difference method analysis revealed, there is significant difference in the frequency that SIM1 and SIM2, SIM3, SIM4 regeneration induction bud produce.
The differentiation frequency of table 3 salicornia europaeal callus on different substratum and average regeneration bud number
| The substratum title | Medium component (mg/L) | Callus number/callus the sum of differentiation | The callus number of bud number/differentiation |
| SIM1 | MS+TDZ0.1+NAA1.0 | 0.27±0.003a | 0.4±0.03a |
| SIM2 | MS+TDZ0.5+NAA1.0 | 0.055±0.004b | 0.1±0.06b |
| SIM3 | MS+TDZ1.0+NAA1.0 | 0.043±0.004c | 0.1±0.05b |
| SIM4 | MS+TDZ2.0+NAA1.0 | 0.042±0.003c | 0.1±0.07b |
The differentiation bud transferred on the 1/2MS substratum that adds IBA 0.5mg/L and KT 0.1mg/L again take root, after 21 days, formation whole plant (F among Fig. 1).
For most of dicotyledonss, evoked callus to sprout need to improve phytokinin and growth hormone ratio (Qiu WD (Qiu Wenda) .Tissue culture of horticultural plants (gardening plant tissue culture). Shanghai: Shanghai Scientific; Technical Publishers (Science and Technology of Shanghai press), 1986).Because TDZ has high cytokine activity, the regeneration of bud is had obvious facilitation, this step has promptly obtained the salicornia europaeal regeneration bud under the situation that does not change TDZ and NAA ratio, illustrate that TDZ is very effective to the regeneration of promotion salicornia europaeal bud.
3, the further screening of salicornia europaeal callus induction and division culture medium
With the seed is explant, and seed is inoculated on the 1-3 substratum in the table 4 with evoked callus.Culture condition in the evoked callus process is: temperature (25 ± 1) ℃, dark condition is cultivated down.Each experiment has 6-8 parallel sample, respectively establishes 3 repeating groups.The result is as shown in table 4, show cultivated for 3 weeks after, the induction frequency of No. 1 substratum is that the induction frequency of 98%, No. 2 substratum is that the induction frequency of 99%, No. 3 substratum is 98.7%.
Table 4. salicornia europaeal seed callus inducing culture and induction frequency thereof
| The substratum numbering | Medium component (mg/L) | Induction frequency (callus number/explant sum) |
| 1 | MS+TDZ0.05+NAA0.8 | 98％ |
| 2 | MS+TDZ0.13+NAA1.2 | 99％ |
| 3 | MS+TDZ0.09+NAA0.9 | 98.7％ |
The callus of No. 1 substratum acquisition is inoculated into substratum respectively No. 1, No. 2 substratum, No. 3 substratum and SIM1 substratum, the callus of No. 2 substratum acquisitions is inoculated into substratum respectively No. 1, No. 2 substratum, No. 3 substratum and SIM1 substratum, the callus of No. 3 substratum acquisitions is inoculated into substratum respectively No. 1, No. 2 substratum, No. 3 substratum and SIM1 substratum, temperature (25 ± 1) ℃, illumination every day 12h, intensity of illumination 30001x cultivates 3-4 after week, and the callus of No. 1 substratum acquisition is at No. 1 substratum, No. 2 substratum, it is as shown in table 5 that differentiation frequency in No. 3 substratum and the SIM1 substratum and regeneration bud form frequency:
Differentiation of calli frequency that table 5.1 substratum obtains and regeneration bud form frequency
| The substratum numbering | Medium component (mg/L) | Callus number/callus the sum of differentiation | The callus number of bud number/differentiation |
| 1 | MS+TDZ0.05+NAA0.8 | 23.1％ | 0.27 |
| 2 | MS+TDZ0.13+NAA1.2 | 24.2％ | 0.3 |
| 3 | MS+TDZ0.09+NAA0.9 | 25.1％ | 0.32 |
| SIM1 | MS+TDZ0.1+NAA1.0 | 26.1％ | 0.35 |
The callus that No. 2 substratum obtain in No. 1 substratum, No. 2 substratum, No. 3 substratum and SIM1 substratum differentiation frequency and regeneration bud to form frequency as shown in table 6:
Differentiation of calli frequency that table 6.2 substratum obtains and regeneration bud form frequency
| The substratum numbering | Medium component (mg/L) | Callus number/callus the sum of differentiation | The callus number of bud number/differentiation |
| 1 | MS+TDZ0.05+NAA0.8 | 23.4％ | 0.28 |
| 2 | MS+TDZ0.13+NAA1.2 | 24.7％ | 0.31 |
| 3 | MS+TDZ0.09+NAA0.9 | 25.5％ | 0.33 |
| SIM1 | MS+TDZ0.1+NAA1.0 | 26.3％ | 0.37 |
The callus that No. 3 substratum obtain in No. 1 substratum, No. 2 substratum, No. 3 substratum and SIM1 substratum differentiation frequency and regeneration bud to form frequency as shown in table 7:
Differentiation of calli frequency that table 7.3 substratum obtains and regeneration bud form frequency
| The substratum numbering | Medium component (mg/L) | Callus number/callus the sum of differentiation | The callus number of bud number/differentiation |
| 1 | MS+TDZ0.05+NAA0.8 | 23.7％ | 0.29 |
| 2 | MS+TDZ0.13+NAA1.2 | 25％ | 0.34 |
| 3 | MS+TDZ0.09+NAA0.9 | 25.8％ | 0.38 |
| SIM1 | MS+TDZ0.1+NAA1.0 | 26.5％ | 0.39 |
The induction frequency that shows callus in No. 1 substratum, No. 2 substratum, No. 3 substratum similar at SIM1, and the callus that obtains all has differentiation capability, but the callus that in these 3 kinds of substratum, obtains, all show the higher trend of differentiation frequency in SIM1, illustrate that SIM1 is for optimum division culture medium in the examination substratum.
Claims (3)
1. the method for a tissue culture salicornia europaeal, be to be explant with salicornia europaeal seed, hypocotyl or root, in salicornia europaeal callus induction and division culture medium, cultivate, form callus and break up and sprout, will break up bud and forward the formation whole plant of taking root in the root media to; Described salicornia europaeal callus induction and division culture medium are to contain the TDZ of 0.05-0.13mg/L and the MS substratum of 0.8-1.2mg/L NAA; Described root media is the 1/2MS substratum that contains 0.5mg/LIBA and 0.1mg/L KT.
2. method according to claim 1 is characterized in that: in the described method, the culture condition of whole tissue culture procedures is 20-28 ℃, secretly cultivates during the callus of induce, and illumination every day is 8-12 hour between the differentiation phase, intensity of illumination 2500-3500LUX.
3. method according to claim 2 is characterized in that: described culture condition is 24-26 ℃, secretly cultivates during the callus of induce, and illumination every day is 12 hours between the differentiation phase, intensity of illumination 3000LUX.
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| CN101548643B (en) * | 2009-05-15 | 2011-01-12 | 江苏省农业科学院 | Fast seedling-breeding method of samphire of North American by issue culture |
| CN107197765A (en) * | 2015-09-03 | 2017-09-26 | 邹祥茂 | A kind of soilless culture nursery liquid for extending the broad-leaved epiphyllum florescence and preparation method thereof |
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| CN107027630B (en) * | 2017-05-31 | 2019-03-01 | 甘肃农业大学 | A kind of method that salt sward blade separate living tissue high-efficiency regeneration system is established |
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| CN1524876A (en) * | 2003-02-27 | 2004-09-01 | 中国科学院植物研究所 | Na+/h+ counter rotatable protein of salicornia europaeal , genes encoding same and use thereof |
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| CN1524876A (en) * | 2003-02-27 | 2004-09-01 | 中国科学院植物研究所 | Na+/h+ counter rotatable protein of salicornia europaeal , genes encoding same and use thereof |
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| CN107197765A (en) * | 2015-09-03 | 2017-09-26 | 邹祥茂 | A kind of soilless culture nursery liquid for extending the broad-leaved epiphyllum florescence and preparation method thereof |
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