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1.
Summary Cell suspension-derived protoplasts of albino Petunia hybrida were grown for 10 d at the interface between aqueous culture medium (KM8P) and an oxygenated (10 mbar for 15 min) perfluorocarbon liquid, perfluorodecalin. Protoplasts synthesised new cell walls and divided normally at the perfluorodecalin/culture medium interface, with a mean viability after 10 d of > 92.0%. The mean plating efficiency of protoplasts was elevated by 37% (P<0.05) following culture at the perfluorodecalin/medium interface, but was unaltered by perfluorodecalin or oxygen separately. The mean plating efficiency of protoplasts cultured at the interface was further increased to a maximium of 52% above control, in the presence of oxygenated perfluorodecalin and KM8P medium supplemented with the non-ionic, co-polymer surfactant, Pluronic F-68 at 0.01% (w/v). These findings demonstrate the effectiveness of oxygenated perfluorodecalin for promoting protoplast growth, by facilitating oxygen delivery. The finding that Pluronic F-68 further increased the plating efficiency of protoplasts cultured at the perfluorocarbon/aqueous interface suggests that these agents improve growth through separate, but cumulative, mechanisms.Abbreviations ATP
adenosine triphosphate
- PFCs
perfluorochemicals
- STP
standard temperature and pressure 相似文献
2.
An improved protocol for the culture of cassava leaf protoplasts 总被引:2,自引:0,他引:2
P. Anthony M. R. Davey J. B. Power K. C. Lowe 《Plant Cell, Tissue and Organ Culture》1995,42(3):299-302
Viable protoplasts (yield > 1.9 × 107 g–1 fresh weight; mean viability 85±2%, n=5) were isolated from leaves of axenic shoot cultures of Manihot esculenta Crantz. cv. M. Thai 8. Protoplasts were cultured for up to 50 days in liquid, ammonium-free MS medium, overlaying agarose-solidified B5 medium with short glass rods embedded perpendicularly within, and protruding from, the agarose layer. Control protoplasts were cultured identically, but without glass rods. Sustained protoplast division was observed only in the presence of glass rods, where the initial plating efficiency was almost 6-fold greater than control (p < 0.05). The mean final plating efficiency of treated cultures was 1.0±0.2% while, in contrast, significant colony formation was not observed in controls.Abbreviations BA
6-benzyladenine
- CPPU
N-(2-chloro-4-pyridyl)-N'-phenylurea
- MES
2[N-morpholino]ethane sulphonic acid
- MS
Murashige & Skoog (1962)
- NAA
-naphthaleneacetic acid
- IPE
initial plating efficiency
- FPE
final plating efficiency 相似文献
3.
P. Anthony M. R. Davey J. B. Power C. Washington K. C. Lowe 《Plant Cell, Tissue and Organ Culture》1994,38(1):39-43
Image analysis has been used to assess the growth of cell suspension-derived protoplasts of Petunia hybrida cv. Comanche at an interface between aqueous culture medium (KM8P), supplemented with 0.01% (w/v) Pluronic F-68, and oxygenated (10 mbar; 10 min) perfluorodecalin. Protoplasts synthesised a new cell wall and entered normal mitotic division which was sustainable to the cell colony/callus stage. This process was accentuated by the collective and additive effects of oxygen, perfluorodecalin and surfactant media supplements. The mean area (mm3) of protoplast-derived cell colonies after 68 days of growth was increased 35 fold over control (media alone) in the presence of these combined treatments. The new cultural regime, leading to improved cell throughput from protoplasts, is discussed primarily in relation to the role of perfluorodecalin as a gas carrier and possible effects of Pluronic F-68 in stimulating cellular uptake of nutrients and/or growth regulators. Image analysis provides a novel and accurate approach to quantifying cell growth responses.Abbreviations dpi
dots per inch
- FPE
final plating efficiency
- IPE
initial plating efficiency
- KM
Kao & Michayluk (1975)
- PFC
Perfluorocarbon
- UM
Uchimiya & Murashige (1974) 相似文献
4.
T. Dennis Thomas 《In vitro cellular & developmental biology. Plant》2009,45(5):591-598
Protoplast culture and plant regeneration of an important medicinal plant Tylophora indica were achieved through callus regeneration. Protoplasts were isolated from leaf mesophyll cells and cultured at a density
of 5 × 105 protoplasts per gram fresh weight, which is required for the highest frequency of protoplast division (33.7%) and plating
efficiency (9.3%). The first division was observed 2 d after plating and the second division after 4 d. Culture medium consists
of Murashige and Skoog (MS) liquid medium with 4 μM 2,4-D, 0.4 M mannitol and 3% (w/v) sucrose with pH adjusted to 5.8. After 45 d of culture at 25°C in the dark, protoplasts formed colonies consisting of about
100 cells. The protoplast-derived microcalli were visible to the naked eye within 60 d of culture and reached a size of 0.2–0.4 mm
in diameter after 90 d. Calli of 0.2–0.4-mm size were transferred to MS medium supplemented with 2,4-D (4 μM), 3% (w/v) sucrose and 0.8% (w/v) agar, formed friable organogenic calli (7-8 mm size) after 8 wk under incubation in normal light period supplemented with
200 μmol m−2 S−1 of day light fluorescent illumination. The calli were transferred to MS medium supplemented with thidiazuron (TDZ) (1–7 μM)
and naphthalene acetic acid (NAA) (0.2–0.4 μM) for regeneration. The calli developed shoot buds after 3–4 wk, and the frequencies
of calli-forming shoots varied from 5% to 44%. Optimum shoot regeneration occurred on MS medium supplemented with 5 μM TDZ
and 0.4 μM NAA. On this medium, 44% cultures responded with an average number of 12 shoots per callus. Whole plants were recovered
following rooting of shoots in 1/2 MS medium supplemented with 3 μM indole 3-butyric acid. 相似文献
5.
F. H. M. Derks A. Zelcer C. M. Colijn-Hooymans 《Plant Cell, Tissue and Organ Culture》1990,21(3):211-216
Mesophyll protoplasts from in vitro grown plants of a cytoplasmic albino mutant ofLycopersicon esculentum cv. Large Red Cherry were isolated with yields between 0.4 to 4.4 × 106 protoplasts per gram leaf tissue. Success in the culture of these protoplasts was dependent on embedding of the protoplasts in 100 µ1 agarose droplets 0.6% (w/v). A plating efficiency of 4.0% was obtained when the protoplasts were cultured in TM-2 medium with sucrose concentrations of 8.7 to 9.6% (w/v) resulting in an osmotic pressure of 432 to 469 mOsmol kg-1. After 14 days of protoplast culture, microcalli with a diameter of 3 mm were observed. After 3 weeks, macrocalli were obtained which were transferred to regeneration medium. Regeneration of shoot primordia, with a frequency of 19%, was obtained on TM-4 medium supplemented with 1% (w/v) sucrose. The first shoot primordia were visible 10 weeks after protoplast plating. For development of the shoot primordia into shoots it was necessary to increase the sucrose concentration to 6% (w/v). Eight out of eleven regenerants were diploid (2n = 2x = 24); the other three were tetraploid. Efficient regeneration of mesophyll albino protoplasts from tomato opens the way to select at the cellular level for the chloroplast transfers. 相似文献
6.
Plant regeneration from protoplasts isolated from friable embryogenic callus of cassava 总被引:2,自引:0,他引:2
E. Sofiari C. J. J. M. Raemakers J. E. M. Bergervoet E. Jacobsen R. G. F. Visser 《Plant cell reports》1998,18(1-2):159-165
Protoplasts were isolated from friable embryogenic callus (FEC) and from suspensions derived from FEC of cassava genotype
TMS60444. Suspensions yielded the highest number of protoplasts (1.5×106 protoplasts/g fresh weight). Protoplasts plated at a density of 105–106/ml in a medium supplemented with 0.5 mg/l α-naphthaleneacetic acid and 1 mg/l zeatin began dividing after 3 days, and after 30 days this resulted in an absolute plating
efficiency as high as 2.5%. After 2 months of culture, 60% of the developed calli were highly friable and in appearance identical
to the original FEC. The protoplast derived FEC was first purified through two rounds of selection of 3 weeks each before
beeing cultured for regeneration of plants. This was done by culturing the protoplast-derived FEC for 11 weeks on maturation
medium, yielding a maximum of 184 organized embryos per 10.000 initially cultured protoplasts. Most of the organized embryos
were torpedo shaped and matured after they had been isolated from the calli and transferred to fresh medium. Mature embryos
were multiplied by secondary somatic embryogenesis at high efficiency (>90%) on a medium supplemented with 8 mg/l 2,4-dichlorophenoxyacetic
acid. About 30% of the mature secondary somatic embryos developed into shoots after transfer to a medium supplemented with
1 mg/l N6-benzylaminopurine (BAP). Shoots rooted readily on a medium without BAP.
Received: 30 August 1996 / Revision received: 9 June 1997 / Accepted: 1 October 1997 相似文献
7.
F. B. d'Utra Vaz A. V. P. dos Santos G. Manders E. C. Cocking M. R. Davey J. B. Power 《Plant cell reports》1993,12(4):220-225
Enzymatic digestion of newly expanded leaves of glasshouse-grown seedlings of passionfruit released protoplasts which exhibited highest division frequency (38.6%) when plated at a density of 1.5×105 ppts ml–1 in agarose-solidified droplets of KM8P medium containing the antibiotic cefotaxime (250 g ml–1). Cefotaxime was essential for sustained cell division. Protoplast-derived calli were cultured on agarsolidified MS medium with 5.0 mg H NAA, 0.25 mg l–1 BAP and additional vitamins. These calli regenerated shoots on transfer to MS medium with 1.0 mg l–1 BAP. Regenerated shoots were rooted in half-strength MS medium with 3.0 mg l–1 IBA and 0.5 mg l–1 NAA (7 d), followed by sub-culture to MS medium lacking growth regulators. The ability to regenerate plants from protoplasts of passionfruit is discussed in relation to the application of somatic cell techniques for the genetic improvement of this economically important tropical woody plant.Abbreviations B5
medium after Gamborg et al. (1968)
- BAP
6-benzylaminopurine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- d
day
- FDA
fluorescein diacetate
- FPE
final plating efficiency
- f. wt
fresh weight
- h
hour
- 1BA
4-indole-3yl-butyric acid
- IPE
initial plating efficiency
- MES
2-N-morpholinoethane sulphonic acid
- MS
medium after Murashige and Skoog (1962)
- NAA
-naphthaleneacetic acid
- PVP-10
polyvinylpyrrolidone (M. Wt. 10,000)
- rpm
rotations per minute 相似文献
8.
Leaf protoplasts of axenic shoot cultures of Pyrus communis L. cv. Williams' Bon Chretien (syn. Bartlett) underwent cell wall regeneration and division to give multicellular colonies in a modified Murashige and Skoog medium which lacked ammonium ions, but supplemented with 1-naphthaleneacetic acid (NAA), 4-indole-3yl-acetic acid, 6-benzylaminopurine (BAP) and casein hydrolysate. Protoplast-derived colonies gave callus on Murashige and Skoog salts medium with NAA and BAP and exhibited shoot regeneration on half-strength Murashige and Skoog medium supplemented with 0.2 mg 1–1 4-indole-3yl-butyric acid, 2.0 mg 1–1 BAP, 0.2 mg 1–1 gibberellic acid, 50 mg 1–1 casein hydrolysate and 10 mg 1–1 Ca-pantothenate. Following rooting, protoplast-derived plants of pear were transferred to the glasshouse where they completed acclimatization.Abbreviations BAP
6-benzylaminopurine
- FPE
final plating efficiency
- GA3
gibberellic acid
- IAA
4-indole-3yl-acetic acid
- IBA
4-indole-3yl-but yric acid
- IPE
initial plating efficiency
- NAA
1-naphthaleneacetic acid
- f.wt.
fresh weight
- MES
2-N-morpholinoethane sulfonic acid
- MS
Murashige and Skoog (1962)
- %PE
% plating efficiency
- PVP-10
polyvinylpyrrolidone (Av. MW 10,000)
- FDA
fluorescein diacetate 相似文献
9.
An efficient and reproducible protocol is described for the regeneration of Cinnamomum camphora protoplasts isolated from cultured embryogenic suspension cells. Maximum protoplast yield (13.1±2.1×106/g FW) and viability (91.8±3.8%) were achieved using a mixture of 3% (w/v) cellulase Onozuka R10 and 3% (w/v) macerozyme Onozuka R10 in 12.7% (w/v) mannitol solution containing 0.12% (w/v) MES, 0.36% (w/v) CaCl2·2H2O, and 0.011% (w/v) NaH2PO4·2H2O. First divisions occurred 7–10 days following culture initiation. The highest division frequency (24.6±2.9%) and plating efficiency (6.88±0.8%) were obtained in liquid medium (MS) supplemented with 30 g l–1 sucrose, 0.7M glucose, 0.1 mg l–1 NAA, 1.0 mg l–1 BA, and 1.0 mg l–1 GA3. After somatic embryo induction and then shoot induction, the protoplast-derived embryos produced plantlets at an efficiency of 17.5%. Somatic embryos developed into well-rooted plants on MS medium supplemented with 1.0 mg l–1 3-indole butyric acid (IBA). Regenerated plants that transferred to soil have normal morphology. 相似文献
10.
Mesophyll protoplasts were isolated from leaf tissues of a diploid daylily (HemerocallisxRed Magic) by enzymatic digestion with a solution containing 0.5% Pectolyase Y-23, 0.1% Cellulase R-10, 0.1% Driselase, 0.6 M sorbitol and half-strength MS inorganic salts. When cultured on MS medium supplemented with 0.5 mg/l NAA and 0.5 mg/l BA, the protoplasts underwent sustained division to produce multicellular colonies. The optimal plating density for cell division was 0.5 × 105 protoplasts/ml. The highest plating efficiency was obtained in cultures grown in media solidified with 0.2% Gelrite. Under these conditions, formation of colonies occurred from 14% of cultured protoplasts. Calli were recovered from 9 colonies only after the cultures were treated with a conditioned medium. Intact plants were regenerated from protoplast-derived calli through organogenesis.Abbreviations BA
6-benzylaminopurine
- FDA
fluorescein diacetate
- GA3
gibberellic acid
- MS medium
Murashige and Skoog (1962) medium
- NAA
1-naphthaleneacetic acid 相似文献
11.
Plants were regenerated from mesophyll protoplasts of Ipomoea cairica L., a wild relative of sweetpotato (Ipomoea batatas (L.) Lam.), and somatic hybrids between I. cairica L. and sweetpotato cv. Xushu 18 were obtained by PEG-mediated method. I. cairica L. protoplasts were isolated from the leaves of in vitro grown plants and cultured in a modified MS medium containing 0.05 mg l−1 2,4-D and 0.5 mg l−1 kinetin. Nine weeks after plating, the obtained small calluses up to about 2 mm in diameter were transferred to solid MS medium supplemented with 0.05 mg l−1 2,4-D and 0.5 mg l−1 kinetin for callus proliferation. Three weeks after transfer, the calluses were transferred to MS medium supplemented with 0–1.0 mg l−1 IAA and 1.0–3.0 mg l−1 BAP and further to hormone-free MS medium for plant regeneration. The frequencies of calluses forming plants ranged from 6.0% to 41.3% based on the different concentrations of IAA and BAP, and 2.0 mg l−1 BAP gave the highest regeneration frequency of protoplast-derived calluses in I. cairica L.. The regenerated plants, when transferred to soil, showed 100% survival. No morphological variations were observed. Mesophyll protoplasts of I. cairica L. were fused with protoplasts isolated from embryogenic suspension cultures of Xushu 18 by PEG-mediated method. The fused products were cultured with the best protoplast culture system of I. cairica L.. Finally, 114 plants were produced from 63 of the 182 calluses derived from the fused protoplasts, and 46 plants of them were confirmed to be somatic hybrids through peroxidase isozyme, RAPD, morphological and cytological analyses. 相似文献
12.
Treatment of the roots of 24–48 h old seedlings of the forage legumeLotus corniculatus with 1.0% Cellulase YC, and 0.1% Pectolyase Y-23 in 4.2% mannitol solution released protoplasts from the tips of root hairs within 30–40 sec of enzyme incubation. Roots from approximately 1000 seedlings yielded 1.7×105 protoplasts. Ten percent of protoplasts divided to form cell colonies when cultured at 1.0×105 ml–1 in droplets of KM8P medium with 0.6% Sea Plaque agarose. Colonies formed callus on UM agar medium; protoplast-derived tissues produced shoots on B5 medium containing 0.05 mg 1–1 of BAP. Regenerated plants were phenotypically and cytologically normal (2n=2x=24±2), and produced nitrogen fixing root nodules following inoculation withRhizobium. These results confirm the totipotency of protoplasts isolated from specialised epidermal cells of seedling roots ofLotus corniculatus. 相似文献
13.
We developed an efficient plant regeneration system from protoplasts for poplar (Populus alba L.). Protoplasts were isolated from 4-day-old suspension cultures derived from seed-induced calli with a yield of 6.96× 106 cells/g fresh weight cells and then cultured at a concentration of 2.5×105 cells/ml in NH4NO3-free Murashige and Skoog (MS) medium supplemented with 5 μM 2,4-dichlorophenoxyacetic acid (2,4-D), 0.05 μM thidiazuron (TDZ) and 0.5 M glucose as a osmoticum. The plating efficiency of the cultured protoplasts was calculated at 26.5% at day 7 and 31.7% at
day 14. Cell colonies were observed after culturing for 4 weeks. Regenerated colonies were propagated through subculture in
liquid MS medium supplemented with 5 μM 2,4-D. Buds were induced from regenerated calli on MS medium containing 10 μM kinetin or 1 μM TDZ. Regenerated shoots were rooted on half-strength MS medium, and the plantlets were transplanted in soil. Randomly amplified
polymorphic DNA analysis did not detect any DNA polymorphism among the regenerated plants.
Received: 7 March 1997 / Revision received: 16 June 1997 / Accepted: 5 July 1997 相似文献
14.
Summary Successful plant regeneration was achieved for the first time from hairy root-derived protoplasts of Hyoscyamus muticus. High yields (7 × 106 / g fresh weight) of protoplasts were isolated directly from the transformed roots of Hyoscyamus muticus using an enzyme mixture comprising 1 % macerozyme and 2 % cellulase in an osmoticum consisting of 0.2 M CaCl2 and 0.6 M mannitol. Protoplasts were first cultured in liquid NT/PRO I medium and further on semi-solid NT/PRO II agar medium. The procedure permits highly efficient formation of colonies. The plating efficiency varied from 1–9 %. The small individual colonies regenerated easily into shoots and roots at frequencies of 18 % and 70 %, respectively. The time required for the development of small plantlets from protoplasts was 8–11 weeks. The regenerated plants contained rolB from Ri-T-DNA and exhibited an altered phenotype compared to the control plants.Abbreviations BAP
benzylaminopurine
- NAA
naphthaleneacetic acid
- PCR
Polymerase Chain Reaction
- fw
fresh weight 相似文献
15.
Summary Protoplasts were isolated from Agrobacterium rhizogenes A4-transformed cell line of Medicago sativa L. The highest yield of protoplasts (4.2×106 per g fresh weight) was obtained from 12-d-old calluses after being subeultured on fresh medium. The viability of protoplasts
reached over 80%. Protoplasts were induced to undergo sustained divisions when cultured in Durand et al. (DPD) medium supplemented
with 2 mgl−1 (9.05 μM) 2,4-dichlorophenoxyacetic acid, 0,2mgl−1 (0.93 μM) kinetin, 0.3 M mannitol, 2% (w/v) sucrose, and 500 mgl−1 casein hydrolyzate at a plating density of 1.0×105 per ml. An agarose-beads culture method was appropriate for protoplast division of transformed alfalfa. The division frequency
was about 30%. Numerous hairy roots were induced from protocalluses on Murashige and Skoog medium without growth regulators.
Paper electrophoresis revealed that all of the regenerated hairy roots tested synthesized the corresponding opines. This protoplast
culture system would be valuable for further somatic hybridization in forage legumes. 相似文献
16.
Sugar beet protoplasts (Beta vulgaris L.) were isolated from hypocotyl-derived suspension cells and cultured on modified Murashige and Skoog medium supplemented
with 5 μM naphthaleneacetic acid (NAA) and 2 μM 6-benzyl-aminopurine (BAP). Protoplasts were plated at a density 1.0–1.5×105 cm−3 and incubated in either liquid medium or in medium solidified by 1.2% agarose, at 25°C in the dark. Comparison of two methods
of culture unequivocally showed the second to be superior. Immobilizing the protoplast in agarose proved to be essential for
obtaining sustained protoplast division and reproducible colony formation. The plating efficiency after two weeks of culture,
expressed as the percentage of protoplasts which developed to form colonies, reached 40%. Subsequent subcultures of protoplast-derived
callus to regeneration media with different concentrations of BAP (5 μM, 10 μM, 20 μM, 30 μM) resulted in very good callus
proliferation at the three lowest concentrations, although organogenesis was not achieved. 相似文献
17.
Summary Highly viable protoplasts were isolated in large numbers from in vitro-grown leaf and stem tissues of a haploid clone of the apple scion cultivar Golden Delicious (Malus Xdomestica Borkh.). Protoplasts from both sources divided rapidly to give microcallus, when cultured in a modified Kao and Michayluk-based medium. Following two successive subcultures for callusing, shoot buds were regenerated from such calli, on half-strength Murashige and Skoog medium with an increased concentration of group B vitamins and containing 5.0 mg.l-1 6-benzyl-aminopurine and 0.1 mg.l-1 l-naphthaleneacetic acid (for the leaf protoplast-derived calli) or 4-indole-3yl-butyric acid (for stem protoplast-derived calli). The mesophyll protoplast-derived shoots were enfeebled and vitrified, in time with their ultimate death. Conversely, for those shoots deriving from the stem protoplasts, in vitro propagation was successfully achieved. This is the first report on the successful isolation, culture and organogenesis from stem protoplasts of a woody plant genotype.Abbreviations BAP
6-benzylaminopurine
- FPE
final plating efficiency
- IBA
4-indole-3yl-butyric acid
- IPE
initial plating efficiency
- f wt
fresh weight
- KM
Kao and Michayluk (1975)
- MES
2-N-morpholino ethane sulfonic acid
- MPE
intermediate plating efficiency
- MS
Murashige and Skoog (1962)
- NAA
l-naphthaleneacetic acid
- PVP-10
polyvinylpyrrolidone (Av MW 10,000) 相似文献
18.
Protoplasts were isolated from callus tissue of Hibiscus syriacus L. using a solution of 3% Onozuka cellulase, 1% Onozuka macerozyme, and 0.5% hemicellulase. Highest yields of viable protoplasts were obtained from friable, white or yellow callus 8–9 days after subculture on Murashige & Skoog medium with 0.5 mg l-1 2,4-dichlorophenoxyacetic acid and 0.1 mg l-1 kinetin. Protoplasts cultured in thin liquid layers of this medium with mannitol continued dividing for longer than those cultured in droplets or in an agar medium. Cultures were maintained until protoplasts had divided to form groups of more than ten cells. Cell groups developed into callus and continued to grow on an agar medium, but failed to differentiate on a regeneration medium with 2 mg l-1 naphthalene acetic acid and 1 mg l-1 benzylaminopurine. 相似文献
19.
Protoplasts of Cyamopsis tetragonoloba were isolated from leaves of in vitro grown plants. The yield of the protoplasts, their viability and subsequent divisions were greatly influenced by the pH of the media used for isolation and culture of protoplasts. Sustained divisions of the cultured protoplasts were best supported by a modified Kao and Michayluk (1975) nutrient medium containing glucose (0.4 M), NAA (4 mgl–1), 2,4-D (1 mgl–1) and KIN (2 mgl–1 ). The protoplast derived cells developed calli on transfer to Murashige and Skoog (1962) medium supplemented with 1 mgl–1 each of 2,4-D, NAA and KIN. 相似文献
20.
Protoplasts isolated from embryogenic callus of Citropsis schweinfurthii (Engl.) Swing. & M. Kell were cultured in MT (Murashige and Tucker 1969) basal medium containing 5% sucrose supplemented with 0.0, 0.001, 0.01, 0.1 or 1.0 mg l–1 BA, 0, 300, 600 or 900 mg l–1 malt extract and 0.6 M sorbitol. The highest plating efficiency was obtained on MT basal medium containing 5% sucrose supplemented with 0.01 mg l–1 BA and 600 mg l–1 malt extract. MT basal medium containing 5% sucrose and supplemented with 0.01 mg l–1 kinetin was found to be a medium suitable for the development of globular somatic embryos derived from protoplasts into heart-shaped somatic embryos with cotyledon-like structures. The highest percentage of shoot formation was obtained using 0.1 mg l–1 GA3. A complete protoplast to-plant system was developed for C. schweinfurthii, which could facilitate the transfer of nuclear and cytoplasmic genes from this species into cultivated Citrus through protoplast fusion.Abbreviations BA
N6-benzyladenine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- FDA
fluorescein diacetate
- GA3
gibberellin A3
- ME
malt extract 相似文献