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1.
J. B. Kim C. J. J. M. Raemakers E. Jacobsen R. G. F. Visser 《Plant Cell, Tissue and Organ Culture》2006,86(2):233-238
In Alstroemeria high frequencies of compact embryogenic callus (CEC) induction (40%) and friable embryogenic callus (FEC) induction (15%) were obtained from nodes with axil tissue cultured first on a Murashige and Skoog (MS) medium supplemented with 10 μM thidiazuron and 0.5 μM indole-3-butyric acid and after that on a Schenk and Hildebrandt (SH) medium supplemented with 9.1 μM 2,4-dichlorophenoxy acetic acid and 2.2 μM benzylaminopurine (BA). Both types of callus were maintained on modified MS medium supplemented with 20.8 μM picloram. CEC and FEC formed somatic embryos and subsequently plants when transferred to MS medium supplemented with 2.2 μM BA. Plants were produced after 12 weeks (CEC) or after 16 weeks (FEC) of culture. Regenerated plants were established in the greenhouse and flowered normally. 相似文献
2.
Plant regeneration in Arachis pintoi was obtained via two developmental pathways: organogenesis and somatic embryogenesis. Organogenic callus cultures were initiated
from pieces of leaf on MS medium supplemented with NAA or 2,4-D in combination with BA, KIN or 2iP. The most suitable combination
for plant regeneration through organogenesis was an initial medium composed of 10 mg/l NAA+1 mg/l BA followed by transfer
of the callus to a shoot induction medium (MS+1 mg/l BA). Rooting of regenerated shoots was readily achieved by culture on
MS+0.01 mg/l NAA. Embryogenic callus cultures were initiated from pieces of leaf on MS medium supplemented with PICL in combination
with KIN, ZEA, BA or 2iP, and the most suitable combinations were 20 mg/l PICL+1 mg/l BA or 2iP. When pieces of embryogenic
callus were subcultured on MS+1 mg/l BA, somatic embryos were differentiated and developed further into well-developed plants
in MS+1 g/l AC followed by MS medium devoid of plant growth regulators.
Received: 29 April 1999 / Revision received: 24 November 1999 / Accepted: 18 December 1999 相似文献
3.
A procedure was developed for plant regeneration of Hybanthus enneaspermus, a rare ethnobotanical herb from the Deccan peninsula in India, through seed-derived callus. Seeds demonstrated a high induction
frequency (69.4±2.8%) and a high yield (364.4±2.5 mg) of light-yellow friable callus on Murashige and Skoog's (MS) medium
containing 2.6 μm NAA and 2.2 μm BA within 4 weeks of incubation. After 1 year of subculture, yellow friable and light-green compact calli types were established
from initial light-yellow friable callus. Shoot differentiation was achieved from light-green compact callus, but not from
yellow friable callus. Shoot differentiation resulted when light-green compact callus was transferred to MS medium supplemented
with 8.8 μm BA and 2.6 μm NAA; the highest percentage of calli forming shoots (66.6±4.8%) and the highest number of shoots (8.9±0.3) were achieved
in this medium. Differentiated shoot buds elongated to 4–5 cm within 4 weeks. The addition of casein hydrolysate (500 mg/l)
and more potassium phosphate (1.86 mm) to the culture medium enhanced shoot differentiation. Rooting was achieved on the shoots using half-strength MS medium containing
4.8 μm IBA. About 70% of the plants were established in pots containing pure garden soil after 2 weeks of hardening. The regenerated
plants were morphologically uniform and exhibited normal seed set.
Received: 23 July 1998 / Revision received: 18 November 1998 / Accepted: 26 November 1998 相似文献
4.
Embryogenic calli were initiated from embryonic explants of Pinus roxburghii using female gametophytes containing immature pre-cotyledonary embryos. Zygotic embryos were collected at different developmental
stages and cultured on various media. Initiation of embryogenic calli was achieved in pre-cotyledonary zygotic embryos of
a 0.1-mm to 1.2-mm embryonal head on Douglus fir cotyledon revised medium (DCR) medium supplemented with 2,4-D or NAA and
BA. Embryogenic callus development was initiated from the suspensor region of immature embryos. The method of immature embryo
culture was significant as rapid embryogenic callus development occurred in megagametophytes where the suspensor was stretched
onto the medium from the cut micropylar end. Sixty embryogenic lines were established from 2500 explants cultured during one
season. A pro-embryo with six to eight meristematic cells and suspensor of six to ten long, vacuolated cells dominated the
early phase of the callus development. Cleavage polyembryony occurred in proliferating callus, constituting a method of multiplication
of these somatic embryos. Somatic embryos developed to stage-I and stage-II embryos on DCR medium supplemented with 5 μM 2,4-D or 10 μM NAA.
Received: 30 June 1999 / Revision received: 15 November 1999 / Accepted: 3 December 1999 相似文献
5.
Friable embryogenic callus and somatic embryo formation from cotyledon explants of African marigold (Tagetes erecta L.) 总被引:3,自引:0,他引:3
Embryogenic callus and somatic embryos were induced from cotyledonary explants of African marigold (Tagetes erecta L.). Cotyledons were first cultured on MS medium supplemented with 2.0 mg l–1 2,4-D and 0.2 mg l–1 kinetin. After 5 weeks, calli were transferred to MS medium supplemented with 0.02 mg l–1 thidiazuron where compact embryogenic callus developed. Friable embryogenic callus developed when the compact embryogenic
callus was transferred to medium containing 2,4-D and subcultured every 2 weeks. Friable embryogenic callus has been maintained
for more than 2 years without losing the capacity to generate embryos. Embryo development was obtained when friable embryogenic
callus was transferred to MS medium supplemented with 3 mg l–1 ABA and 60 g l–1 sucrose. The addition of 10–30 mM
l-glutamine improved embryo development.
Received: 13 May 1997 / Revision received: 24 February 1998 / Accepted: 28 March 1998 相似文献
6.
Somatic embryogenesis and plant regeneration from embryogenic suspension cultures of perennial ryegrass 总被引:1,自引:0,他引:1
Ousama M. Faizzaghmout William A. Torello 《In vitro cellular & developmental biology. Plant》1990,26(4):419-424
Summary Embryogenic callus induced from mature caryopses of perennial ryegrass (Lolium perenne L.) were placed in liquid half-strength Murashige and Skoog (MS) basal medium and supplemented with 6.0 mg/l 2,4-dichlorophenoxy
acetic acid (2,4-D), 3 g/l (w/v) casein hydrolysate (CH), and B5 vitamins, to initiate fast-growing highly embryogenic cell
suspension cultures. Newly initiated suspension cultures contained a high level of large non-embryogenic cells (NE) with relatively
few embryogenic (E) cells. Cell types were separated by discontinuous Percolls gradients or by filtering the newly initiated
cultures through 31-μm nylon mesh. The growth conditions of the E cell were optimized by testing various media components
including 2,4-D and sucrose, and subculture diluton ratio. Optimal shoot formation occurred after pretreatment of the embryogenic
cells on solidified callus maintenance medium supplemented with 60 mg/l cefotaxime for 4 weeks prior to transfer to regeneration
medium Regeneration media consisted of half-strength MS basal medium supplemented with B5 vitamins, 0.5 mg/l fluridone, and
0.5 mg/l BA. Most plants regenerated were albino with only a few green plants.
Journal Paper number MAES 2959 of the Massachusetts Agricultural Experiment Station. 相似文献
7.
Hairy roots of Panax ginseng were obtained after root disks were infected with wild-type strain Agrobacterium rhizogenes 15834. Three lines of hairy roots with different pigmentation were selected. Embryogenic callus was induced on Murashige
and Skoog medium containing 1.0 mg/l 2,4-D. The frequency of embryogenic callus formation was 37.4% in a line with red pigmentation.
Somatic embryo development from embryogenic callus was efficiently achieved by lowering the concentration of 2,4-D (0.5 mg/l).
After the germination of somatic embryos on medium with 10 mg/l GA3, the embryos were transferred to 1/2-MS medium without GA3. The transformed ginseng plantlets had an actively growing root system with abundant lateral roots. The phenotypical alteration
of transformed ginseng plants might be valuable character for increasing root yield.
Received: 27 March 1999 / Revision received: 18 May 1999 / Accepted 8 July 1999 相似文献
8.
Regeneration of garlic callus as affected by clonal variation, plant growth regulators and culture conditions over time 总被引:3,自引:0,他引:3
A long-term regeneration system for garlic (Allium sativum L.) clones of diverse origin was developed. Callus was initiated on a modified Gamborg's B-5 medium supplemented with 4.5 μM 2,4-D and maintained on the same basal medium with 4.7 μM picloram+0.49 μM 2iP. Regeneration potential of callus after 5, 12 and 16 months on maintenance medium was measured using several plant growth
regulator treatments. The 1.4 μM picloram+13.3 μM BA treatment stimulated the highest rate of shoot production. Regeneration rate decreased as callus age increased, but healthy
plantlets from callus cultures up to 16-months-old were produced for all clones. Regeneration of long-term garlic callus cultures
could be useful for clonal propagation and transformation.
Received: 24 September 1998 / Revision received: 27 January 1999 / Accepted: 26 February 1999 相似文献
9.
Embryogenic calli were induced from leaf explants of coffee (Coffea canephora) on McCown's woody plant medium (WPM) supplemented with 5 μM N6–(2-isopentenyl)-adenosine (2-iP). These calli were co-cultured with Agrobacterium tumefaciens EHA101 harboring pIG121-Hm, containing β-glucuronidase (GUS), hygromycin phosphotransferase (HPT), and neomycin phosphotransferase
II genes. Selection of putative transgenic callus was performed by gradual increase in hygromycin concentration (5, 50, 100 mg/l).
The embryogenic calli surviving on medium containing 100 mg/l hygromycin showed a strong GUS-positive reaction with X-Gluc
solution. Somatic embryos were formed from these putative transgenic calli and germinated on WPM medium with 5 μM 2-iP. Regenerated
small plantlets with shoots and roots were transferred to medium containing both 100 mg/l hygromycin and 100 mg/l kanamycin
for final selection of transgenic plants. The selected plantlets exhibited strong GUS activity in leaves and roots as indicated
by a deep blue color. GUS and HPT genes were confirmed to be stably integrated into the genome of the coffee plants by the
polymerase chain reaction.
Received: 14 December 1998 / Revision received: 12 March 1999 / Accepted: 24 March 1999 相似文献
10.
F. A. Redway V. Vasil D. Lu I. K. Vasil 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1990,79(5):609-617
Summary Immature embryos, inflorescences, and anthers of eight commercial cultivars of Triticum aestivum (wheat) formed embryogenic callus on a variety of media. Immature embryos (1.0–1.5 mm long) were found to be most suitable for embryogenic callus formation while anthers responded poorly; inflorescences gave intermediate values. Immature embryos of various cultivars showed significant differences in callus formation in response to 11 of the 12 media tested. No significant differences were observed when the embryos were cultred under similar conditions on MS medium with twice the concentration of inorganic salts, supplemented with 2,4-D, casein hydrolysate and glutamine. Furthermore, with inflorescences also no significant differences were observed. Explants on callus formation media formed two types of embryogenic calli: an off-white, compact, and nodular callus and a white compact callus. Upon successive subcultures (approximately 5 months), the nodular embryogenic callus became more prominent and was identified as aged callus. The aged callus upon further subculture, formed an off-white, soft, and friable embryogenic callus. Both the aged and friable calli maintained their embryogenic capacity over many subculture passages (to date up to 19 months). All embryogenic calli (1 month old) from the different callus-forming media, irrespective of expiant source, formed only green shoots on regeneration media that developed to maturity in the greenhouse. There were no significant differences in the response of calli derived from embryos and inflorescences cultured on the different initiation media. Also, the shoot-forming capacity of the cultivars was not significantly different. Anther-derived calli formed the least shoots. Aged and friable calli on regeneration media also formed green shoots but at lower frequencies. Plants from long-term culture have also been grown to maturity in soil.Florida Agricultural Experiment Station Journal Series No. R-00494 相似文献
11.
High-frequency plant regeneration through callus initiation from mature embryos of maize (<Emphasis Type="Italic">Zea Mays</Emphasis> L.) 总被引:10,自引:0,他引:10
An efficient maize regeneration system was developed using mature embryos. Embryos were removed from surface-sterilized mature seeds and sliced into halves. They were used as explants to initiate callus on induction medium supplemented with 4.0 mg l–1 2,4-dichlorophenoxyacetic acid (2,4-D). The induction frequency of primary calli was over 90% for all inbred lines tested. The primary calli were then transferred onto subculture medium supplemented with 2.0 mg l–1 2,4-D. Following two biweekly subcultures, embryogenic calli were formed. Inclusion of a low concentration (0.2 mg l–1) of 6-benzylaminopurine (BA) in the subculture medium significantly promoted the formation of embryogenic callus. The addition of silver nitrate (10 mg l–1) also supported an increased frequency of embryogenesis. The embryogenic callus readily formed plantlets on regeneration medium supplemented with 0.5 mg l–1 BA. The regenerated plantlets were transferred to half-strength Murashige and Skoog medium supplemented with 0.6 mg l–1 indole-3-butyric acid to develop healthy roots. The regenerated plantlets were successful on transfer to soil and set seed. Using this system, plantlets were regenerated from seven elite maize inbred lines. The frequency of forming green shoots ranged from 19.8% to 32.4%. This efficient regeneration system provides a solid basis for genetic transformation of maize.Abbreviations BA 6-Benzylaminopurine - 2,4-D 2,4-Dichlorophenoxyacetic acid - IBA Indole-3-butyric acid - KT KinetinCommunicated by M.C. Jordan 相似文献
12.
Oldenlandia umbellata L., commonly known as “Indian madder”, is an ancient Indian herb valued as a source of red color dye and various medicinal
products. In this study, successful protocols have been developed for induction of somatic embryogenesis and organogenesis
in O. umbellata. Emerging young leaves, shoot apices, and stems were used as explants, grown on Murashige and Skoog (MS) media supplemented
with various auxins, including indole acetic acid, indole butyric acid, napthaleneacetic acid (NAA), and 2,4-Dichlorophenoxyacetic
acid, each at levels ranging between 0.1 and 0.5 mg/l, cytokinins, including benzyladenine (BA) and kinetin, each at concentration
ranging between 0.5 and 5 mg/l, with and without coconut milk (CM) at levels of 0.5–5%. For callus induction, NAA at 2.5 mg/l
was optimal; while, for rapid embryogenic callus induction, 0.2 mg/l NAA, 0.5 mg/l BA, and 0.1% CM induced the highest frequency
(95.86%). Shoots developed upon transfer of embryogenic calli to MS medium containing 1.5 mg/l BA, 0.3 mg/l NAA and 1% CM.
For root induction, 0.3 mg/l NAA and 1.0% CM promoted highest and earliest rooting.
C. Rajasekaran contributed equally to this work. 相似文献
13.
Plant regeneration from callus protoplasts of the forage legume Astragalus adsurgens Pall. 总被引:1,自引:0,他引:1
A reproducible release of viable protoplasts was obtained from friable calli of Astragalus adsurgens. Protoplasts underwent sustained divisions and formed cell colonies when cultured in either liquid or agarose-solidified
Kao and Michayluk (1975) protoplast medium (KM8P) supplemented with 1.5 mg/l 2,4-D, 0.5 mg/l BA and 0.5 M glucose. Compared to liquid culture, agarose bead culture improved division frequency effectively, the two culture systems
showing a plating efficiency of 0.8±0.5% and 6.5±0.7%, respectively. Upon transfer to Murashige and Skoog (1962) medium (MS)
with 1–2 mg/l BA, alone or in combination with NAA or 2,4-D at 0.1 mg/l, the protoplast-derived calli produced complete plantlets
through somatic embryogenesis. The maximum percentage of calli producing somatic embryos (52.5± 2.2%) occurred on MS medium
containing 0.1 mg/l NAA and 1 mg/l BA, whereas the maximum number of calli regenerating plantlets (64.7±6.2) was obtained
on MS medium with 0.1 mg/l NAA and 2 mg/l BA.
Received: 25 April 1997 / Revision received: August 1997 / Accepted: 2 September 1997 相似文献
14.
Somatic embryogenesis and plant regeneration in cultured immature inflorescences of Setaria italica 总被引:1,自引:0,他引:1
Young inflorescence explants of Setaria italica in culture showed high capacity for regenerating plantlets through somatic embryogenesis. Embryogenic callus formation was initiated from the explants cultured on Murashige and Skoog's medium with 2 mg/l 2,4-D and 0.2–0.5 mg/l KT or BAP, but it was better for the maintenance of embryogenic growth to subculture the calli on the medium with 2,4-D and KT/BAP and on the medium with 2 mg/l 2iPA and 0.2 mg/l NAA alternately. A number of plantlets were regenerated when embryogenic calli were transferred onto the same basic medium but with 2 mg/l BAP and 0.5 mg/l NAA. Plant regeneration capacity has been maintained in some embryogenic calli during fourteen months of subculture.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- NAA
naphthaleneacetic acid
- IAA
3-indoleacetic acid
- 2iPA
N6-(2-isopentenyl) adenosine
- BAP
6-benzylaminopurine
- KT
kinetin
- CH
casein hydrolysate 相似文献
15.
Moso bamboo [Phyllostachys heterocycla var. pubescens (Mazel ex J. Houz.) Ohwi] is one of the most important forest crops in China and the rest of Asia. Although many sympodial bamboo tissue culture protocols have been established, there is no protocol available for plantlet regeneration as indicated by callus induction for monopodial bamboos, such as Moso bamboo. In the present report, embryogenic callus induction, embryoid development, and germination were established for Moso bamboo from zygotic seed embryos. Callus was initiated from zygotic embryos after 10–20 d culture on MS media supplemented with 4.0 mg/L 2, 4-D and 0.1 mg/L zeatin (ZT). About 50% of the explants produced calli, and nearly 15% of the calli were found to be embryogenic in nature. These embryogenic calli can be subcultured for proliferation in the Murashige and Skoog media (MS) supplemented with 0.5–2.0 mg/L 2, 4-D. These calli were found to have maintained their capacity for regeneration even after one year of subculture. The viable somatic embryoids regenerated in medium containing 5.0–7.0 mg/L ZT. Nearly 5% of the calli were found capable of regenerating into plantlets directly in MS medium containing 5.0–7.0 mg/L ZT. Root growth was more pronounced when the plantlets were transferred to medium containing 2.0 mg/L NAA. After 30 days of subculture, the plantlets were transferred to a greenhouse. 相似文献
16.
Efficient plant regeneration was achieved from callus derived from immature-cotyledon explants of oleaster (Elaeagnus angustifolia L.). Calli were obtained on MS media containing 3% sucrose and different concentrations of TDZ. The highest rate of green,
compact and nodular callus was formed on MS medium supplemented with 1 mg/l of TDZ. Shoot organogenesis was achieved when
the callus was transferred onto MS media containing 3% sucrose and BA alone (05–4 mg/l) or BA (0.5 and 1 mg/l) combined with
NAA or IAA (0.5 and 1 mg/l). Maximum organogenesis was obtained with 1 mg/l BA in combination with 0.5 mg/l NAA. Rooting of
the shoots was achieved on MS medium supplemented with 0.2 mg/l IBA. Regenerated plantlets were acclimatized and successfully
transplanted to soil. 相似文献
17.
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
18.
A system for rapid plant regeneration through somatic embryogenesis from shoot tip explants of sorghum [Sorghum bicolor (L.) Moench] is described. Somatic embryogenesis was observed after incubation of explants in dark for 6–7 weeks through
a friable embryogenic callus phase. Linsmaier and Skoog medium supplemented with 2,4-dichlorophenoxyacetic acid (2 mg l−1) and kinetin (0.1 mg l −1) was used for induction of friable embryogenic calli and somatic embryos. Germination of somatic embryos was achieved about
5 weeks after transfer onto Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine (2 mg l−1) and indole-3-acetic acid (0.5 mg l −1) under light. Seeds from in vitro-regenerated plants produced a normal crop in a field trial, and were comparable to the crop grown with the seeds of the mother
plant used to initiate tissue culture. The simplicity of the protocol and possible advantages of the system for transformation
over other protocols using different explants are discussed.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
19.
The turf-type bermudagrasses are genetically variable and do not respond uniformly to tissue culture and plant regeneration
protocols. We evaluated the callus induction response of two explant types, young inflorescences and nodes, from multiple
genotypes including triploid TifSport, TifEagle, and Tift97-4 and tetraploid Tift93-132, Tift93-135, Tift93-156 and Tift93-157
on MS medium supplemented with 1–1.5 mg l−1 2,4-D + 0.01 mg l−1 BA + 1.16 g l−1 proline. Four types of callus were observed. Type I was fluffy, soft, and white non-embryogenic callus, common to all cultures.
Type II was globular, transparent, and hard, but sticky callus, which was pre-embryogenic and could be selected for subculture.
Type III callus was transparent, compact, and embryogenic. Type IV callus was opaque white and compact. Both Type III and
Type IV calluses were embryogenic and regenerative. A combination of gelling agents in the medium (2 g l−1 Gelrite and 5 g l−1 agar) improved callus quality and increased the rate of compact callus formation during subculture. Embryogenesis from compact
callus was observed in TifEagle, TifSport and Tift93-132, and shoots were regenerated on MS medium with 0.1 mg l−1 2,4-D + 0.5–4.0 mg l−1 BA. Low intensity light treatment (30 μmol m−2 s−1 of cool white fluorescence) to callus before regeneration greatly enhanced regeneration frequency from 6.7% to 40% in recalcitrant
TifSport. 相似文献
20.
The study was carried out to establish in vitro culture conditions for plant regeneration of tef, Eragrostis tef (Zucc.) Trotter. Mature seeds of two Ethiopian varieties, DZ-01-354 and DZ-01-196, were used to initiate callus cultures
on Murashige and Skoog (MS) medium with different auxins. Four- and 8-week-old calli induced on a medium with 2.0 mg/l 2,4-dichlorophenoxyacetic
acid (2,4-D) were subcultured onto various media to induce somatic embryogenesis. Compact, nodulated, embryogenic callus was
observed after transfer onto MS-callus proliferating (CP) medium. Embryogenic tissue appeared on soft and amorphous callus
and developed into somatic embryos during a subsequent subculture to MS embryo-promoting (EP) media. Various growth regulator
combinations were tested in CP and EP media to obtain a high efficiency of somatic embryo formation. The highest frequency
of calli forming somatic embryos (56.1–68.3%) was observed when CP media with 2.0 or 4.0 mg/l 2,3,5-triiodobenzoic acid were
employed and then cultures were transferred to EP media with 0.5 mg/l 2,4-D and 0.5 mg/l kinetin followed by 0.5 mg/l indole-3-acetic
acid and 0.5 mg/l N6-benzyladenine. Plant development from somatic embryos was obtained on MS medium supplemented with 1.0 mg/l gibberellic acid.
On average, 71.2% of calli displaying somatic embryos converted into plants. Regenerated plants were successfully transferred
to soil. Neither chlorophyll-deficient plants nor morphological variants were found among regenerants. All regenerated plants
were fertile.
Received: 9 May 1997 / Revision received: 25 September 1997 / Accepted: 3 January 1998 相似文献