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1.
The initiation and development of somatic embryos and organogenic shoots and corm-like structures (CLSs) from petiole-derived
calli of Amorphophallus rivieri Durieu were observed histologically. The petioles were cultured on Murashige and Skoog (MS) medium supplemented with 5.37 μM
α-naphthaleneacetic acid (NAA) and 4.44 μM N6-benzylaminopurine (6-BA) for callus induction. The shoot and corm organogenesis occurred from the compact calli when they
were transferred to a medium containing 0.54 μM NAA and 4.44 μM 6-BA. A combination of 13.57 μM 2,4-dichlorophenoxyacetic
acid (2,4-D) and 8.88 μM 6-BA or 24.18 μM NAA and 6.66 μM 6-BA was optimum for induction of somatic embryos, which failed
to produce plantlets because of their structural abnormalities. Shoot regeneration predominantly happened through organogenesis
although somatic embryogenesis infrequently occurred. The subepidermal cells of the compact callus converted to competent
cells and started divisions, which resulted in formation of the meristemoids. The meristemoid cells continued division to
develop into bud primordia. Subepidermal cells could also form the globular structures. Subsequently, these globoids developed
into CLSs from which plantlets regenerated during subculture. Meanwhile, the CLSs were capable to form cormels, which could
be a promising way for the propagation of A. rivieri. 相似文献
2.
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
3.
This report deals with micropropagation of the critically endangered and endemic Turkish shrub, Thermopsis turcica using callus, root and cotyledonary explants. Callus cultures were initiated from root and cotyledon explants on MS medium
supplemented with 0.5–20 μM NAA or 2,4-D. The root explants were found to be better in terms of quick responding and callusing
percentages as compared to the cotyledons. Organogenic callus production with adventitious roots and shoots were obtained
on MS medium with only NAA. The calli obtained with NAA, root and cotyledonary explants were cultured with BA and kinetin
(2–8 μM) alone or in combination with a low level (0.5 μM) of 2,4-D or NAA. The best regeneration of shoots from root explants
was observed on hormone-free MS medium. NAA with BA or kinetin in the medium improved shoot induction from the calli obtained
with NAA. Maximum percentage of shoots (93.3%), maximum number of shoots (6.2) and maximun length of shoots (8.22 cm) were
achieved from cotyledonary explants at 4 μM BA and 0.5 μM NAA. The presence of 0.5 μM or higher levels of 2,4-D in shoot induction
medium inhibited the regeneration in T. turcica explants. 83% of in vitro rooting was attained on pulsed-IBA treated shoots. The regenerated plants with well developed shoots and roots were successfully
acclimatized. Application of this study’s results has the potential to conserve T. turcica from extinction. 相似文献
4.
《Plant science》1987,53(3):257-262
Conditions were developed for the isolation, culture and regeneration of mesophyll protoplasts of the tree legume, Pithecellobium dulce Benth. The presence of 2,4-dichlorophenoxyacetic acid (2,4-D) was essential to induce initial cell divisions and addition of naphthaleneacetic acid (NAA) improved the response. Sustained division and cell colony formation were achieved from the protoplasts cultured in a modified KM8P medium containing 2,4-D (2.3 μM), NAA (3 μM) and benzyladenine (BA) (2.3 μM). Dilution of the osmotica included in the protoplast culture medium was necessary to induce sustained proliferation of the protoplast-derived cells. Differentiation of shoots from the protoplast-derived calli occurred on Murashige and Skoog (MS) medium supplemented with BA (5 μM) and indole-3-acetic acid (1 μM). Omission of 2,4-D from the culture medium, after the initial 2 weeks of protoplast culture, was obligatory to induce shoot morphogenesis. 相似文献
5.
El Bahri Trabelsi Selima Naija Nedra Elloumi Zina Belfeleh Monji Msellem Rachida Ghezel Sadok Bouzid 《Acta Physiologiae Plantarum》2011,33(2):319-324
Somatic embryogenesis of olive Olea europaea (L.) ‘Chetoui’ was studied using cell suspension cultures initiated from mature leaf-derived calli. Calli were developed
on half-strength MS medium supplemented with 10 μM NAA and 2.25 μM 2i-P in the dark. Different combinations of three plant
growth regulators (2,4-D, NAA and zeatin) were tested to determine cell proliferation and somatic embryogenesis induction
and differentiation. Embryogenic suspension cultures were established in olive-modified medium for embryogenesis (OMe) containing
2.5 μM 2,4-D and 2.5 μM zeatin. Pre-globular and globular embryos were induced from mature olive tissue in liquid medium.
In addition, the nitrogen form as inorganic (reduced; (NH4)2SO4 or oxidized; KNO3) and organic (CH) was used separately or in combination to improve the cell growth and proliferation. The most effective
growth rate and cell proliferation were obtained with the medium containing inorganic and organic nitrogen forms. 相似文献
6.
Daniela Lopes Paim Pinto Ana Maria Rocha de Almeida Mailson Monteiro Rêgo Maurecilne Lemes da Silva Evelyn Jardim de Oliveira Wagner Campos Otoni 《Plant Cell, Tissue and Organ Culture》2011,107(3):521-530
Mature zygotic embryos of three genotypes of Passiflora edulis Sims, including ‘FB-100’, ‘FB-200’, and ‘FB-300’ were incubated on a Murashige and Skoog (MS) (1962) medium supplemented with different concentrations (18.1–114.8 μM) of 2,4-diclorophenoxyacetic acid (2,4-D) and 4.4 μM of
6-benzyladenine (BA). MS basal medium and MS with BA induced germination of P. edulis embryos. The highest frequencies of embryogenic calli were observed when explants were incubated on MS medium supplemented
with 72.4 μM 2,4-D and 4.4 μM BA for ‘FB-200’, which showed the highest potential for embryogenic callus formation. Cytological
and histological analyses of pro-embryogenic callus revealed two distinct cell types: thin-walled, small, isodiametric cells
with large nuclei and dense cytoplasm, typical of intense metabolic activity; and elongated and vacuolated cells, with small
nuclei and less dense cytoplasm. Differentiation of somatic embryos was promoted on MS medium supplemented with activated
charcoal and indole-3-acetyl-l-aspartic acid (IAA-Asp) either with or without 2,4-D. However, no conversion of somatic embryos into plantlets was observed. 相似文献
7.
Jameel M. Al-Khayri Feng H. Huang Teddy E. Morelock Tahani A. Busharar 《In vitro cellular & developmental biology. Plant》1992,28(2):64-66
Summary A system for the regeneration of spinach (Spinacia oleracea L.) from mature dry seed explants has been established. The response of two commercial spinach cultivars, ‘Grandstand’ and
‘Baker’, was examined. Callus proliferation was most prominent on MS medium supplemented with 9.3 μM of 6-furfurylaminopurine (kinetin) and 3.39 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Adventitious shoot formation was observed within 8 wk after callus was transferred
onto regeneration medium. Shoot regeneration was best from callus induced on 9.3 μM kinetin and 4.56 μM 2,4-D. The regeneration medium contained 9.3 μM kinetin, 0.045 μM 2,4-D, and 2.89 μM gibberellic acid (GA3). Shoots were rooted on hormone-free medium, and plants grown in a greenhouse showed normal phenotype. This system is beneficial
in rapid propagation of spinach plants, particularly when only a limited number of seeds are available. 相似文献
8.
Qin-Mei Wang Feng-Zhan Gao Xiang Gao Fan-Yu Zou Xin Sui Meng Wang Yue-Jun Hui Li Wang 《Plant Cell, Tissue and Organ Culture》2012,109(2):191-200
An efficient in vitro micropropagation system for Clivia miniata Regel was developed using basal tissues of young petals and young ovaries as explants. For callus induction, explants were
incubated on Murashige and Skoog (MS) medium containing either 2.22 μM 6-benzyladenine (BA) and 4.52 μM 2,4-dichlorophenoxyacetic
acid (2,4-D) or 4.44 μM BA, 5.37 μM α-naphthaleneacetic acid (NAA), and 9.05 μM 2,4-D. Moreover, callus was induced from young
ovaries when these were incubated on MS medium containing 8.88 μM BA, 10.74 μM NAA, and 9.05 or 18.10 μM 2,4-D. Subsequently,
callus was transferred to MS medium supplemented with kinetin (KT) and NAA for shoot organogenesis. Frequency of shoot regeneration
from petal-derived callus was highest when callus was transferred to medium containing 2.69 μM NAA with either 9.29 or 13.94 μM
KT. Shoot regeneration frequency from ovary-derived callus was highest when this callus was transferred to medium containing
9.29 μM KT and 10.74 μM NAA. Overall, different explant types exhibited different organogenic capacities wherein, young petals
had higher shoot regeneration frequencies than young ovaries. The highest rooting frequency (98.25 ± 3.04%) was obtained when
shoots were transferred to half-strength MS medium without plant growth regulators. Regenerated plantlets were transplanted
to soil mix and acclimatized, yielding a 96.80% survival frequency. Only 0.6% of regenerated plantlets exhibited morphological
changes. The diploid status (2n = 22) of regenerated plantlets was determined using chromosome counts of root-tips. Moreover, inter-simple sequence repeats
were used to assess the genetic fidelity of regenerated plantlets. Overall, regenerated plants shared 90.5–100.0% genetic
similarities with mother plants and 89.0–100.0% similarities with each other. 相似文献
9.
Plantlet regeneration through shoot formation from young leaf explant-derived callus of Camptotheca acuminata is described. Calli were obtained by placing leaf explants on Woody plant medium (WPM) supplemented with various concentrations
of 6-benzyladenine (BA) and naphthaleneacetic acid (NAA) or 2,4-dichlorophenoxyacetic acid (2,4-D). Callus induction was observed
in all media evaluated. On the shoot induction medium, the callus induced on the WPM medium containing 19.8 μM BA and 5.8
μM NAA was the most effective, providing high shoot regeneration frequency (70.3 %) as well as the highest number of shoots
(11.2 shoots explant−1). The good rooting percentage and root quality (98 %, 5.9 roots shoot−1) were achieved on WPM medium supplemented with 9.6 μM indole-3-butyric acid (IBA). 96 % of the in vitro rooted plantlets with well developed shoots and roots survived transfer to soil. 相似文献
10.
P. Jha C. B. Yadav V. Anjaiah V. Bhat 《In vitro cellular & developmental biology. Plant》2009,45(2):145-154
An efficient in vitro plant regeneration protocol through somatic embryogenesis and direct shoot organogenesis has been developed for pearl millet
(Pennisetum glaucum). Efficient plant regeneration is a prerequisite for a complete genetic transformation protocol. Shoot tips, immature inflorescences,
and seeds of two genotypes (843B and 7042-DMR) of pearl millet formed callus when cultured on Murashige and Skoog (MS) medium
supplemented with varying levels of 2,4-dichlorophenoxyacetic acid (2,4-D; 4.5, 9, 13.5, and 18 μM). The level of 2,4-D, the
type of explant, and the genotype significantly effected callus induction. Calli from each of the three explant types developed
somatic embryos on MS medium containing 2.22 μM 6-benzyladenine (BA) and either 1.13, 2.25, or 4.5 μM of 2,4-D. Somatic embryos
developed from all three explants and generated shoots on MS medium containing high levels of BA (4.4, 8.8, or 13.2 μM) combined
with 0.56 μM 2,4-D. The calli from the immature inflorescences exhibited the highest percentage of somatic embryogenesis and
shoot regeneration. Moreover, these calli yielded the maximum number of differentiated shoots per callus. An efficient and
direct shoot organogenesis protocol, without a visible, intervening callus stage, was successfully developed from shoot tip
explants of both genotypes of pearl millet. Multiple shoots were induced on MS medium containing either BA or kinetin (4.4,
8.8, 17.6, or 26.4 μM). The number of shoots formed per shoot tip was significantly influenced by the level of cytokinin (BA/kinetin)
and genotype. Maximum rooting was induced in 1/2 strength MS with 0.8% activated charcoal. The regenerated plants were transferred
to soil in pots, where they exhibited normal growth. 相似文献
11.
A short-term regeneration system from leaf-base-derived callus of wheat (Triticum aestivum L.) was developed. Embryogenic callus formation and shoot regeneration were achieved from the first basal segments of 3–4-day-old seedlings. Callus formation frequency as well as plantlet regeneration frequency was dependent on the composition of basal medium and the concentration of 2,4-dichlorophenoxyacetic acid (2,4-D). MS medium with 2,4-D 4.5–9.0 mol l–1 was optimal for the culture of wheat leaf base. Effects of different combinations of plant growth regulators, which were added in either callus induction medium or shoot regeneration medium, were tested. Adding of BAP in callus induction medium shortened the time of shoot emergence but could not improve the producing of embryogenic calli and green plantlets. Optimal ratio of 2,4-D, BAP and NAA gave similar regeneration frequency to control. Existence of cytokinins in regeneration medium had no effect on increasing the regeneration frequency. The regenerants could grow to normal, fertile plants after they were transferred into soil. 相似文献
12.
We investigated the optimal levels of growth regulators, culture media, and pH on callus growth and organogenesis of in-vitro
cultured ‘Kyoho’ grapes. Calli were induced by culturing leaf blades on an MS basal medium supplemented with 1 mg/IL BA and
0.01 mg/L 2,4-D. In addition, calli originating from the exocarp and mesocarp of grape fruits devel-oped on MS media supplemented
with 0.1 mg/L IAA, NAA, or 2,4-D, or with 0.2 mg/L BA. In testing the potential for plant regeneration from shoot tips on
various media, we found that the Nitsch medium, with I mg/L BA, was optimal for caulogenesis. The type of shoot development
depended on the pH of the medium, with vigorous multiple-shoot devel-opment occurring at pH 6.0, and single shoots forming
at pH 5.0. Finally, we were able to obtain rooted seedlings from the regenerated shoots that had been cultured on 1/4-strength
Nitsch medium supplemented with 0.03 mg/L NAA. 相似文献
13.
Efficient in vitro regeneration systems for Vaccinium species 总被引:1,自引:0,他引:1
Julia Meiners Melanie Schwab Iris Szankowski 《Plant Cell, Tissue and Organ Culture》2007,89(2-3):169-176
Efficient protocols for shoot regeneration from leaf explants suitable for micropropagation as well as for the development
of transgenic plants were developed for blueberry (Vaccinium corymbosum) and lingonberry (Vaccinium vitis-idaea) cultivars. Nodal segments were used to initiate in vitro shoot cultures of lingonberry cultivar ‘Red Pearl’ and southern
highbush blueberry cultivar ‘Ozarkblue’. In order to develop an optimized regeneration procedure, different types and concentrations
of plant growth regulators were tested to induce adventitious shoot regeneration on excised leaves from micropropagated shoots
of both cultivars. The effect on percentage regeneration and number of shoots per explant was investigated. Results indicated
that zeatin was superior to TDZ and meta-topolin in promoting adventitious shoot formation. A concentration of 20 μM zeatin
was most effective in promoting shoot regeneration in both cultivars, in case of ‘Red Pearl’ along with 1 μM NAA. Shoots were
either allowed to root in vitro on medium containing IBA or NAA or ex vitro in a fog tunnel. IBA was superior to NAA for induction
of root development in vitro in both Vaccinium cultivars. Ex vitro rooting under high humidity was tested with cuttings from mature field-grown plants, from acclimatized
tissue culture derived plants and with unrooted in vitro proliferated shoots planted directly. It was found that in vitro
shoots rooted better under fog than cuttings from the other plant sources and rooting was equivalent to that achieved in vitro. 相似文献
14.
Callus induction and in vitro plantlet regeneration systems for safflower (Carthamus tinctorius L.) cv. Bhima using root,
hypocotyl, cotyledon and leaf explants were optimized by studying the influence on organogenesis of seedling age, media factors,
growth regulators and excision orientation. Supplementation of the medium with an auxin: cytokinin ratio < 1 enhanced the
growth rate of callus cultures; however, for 2,4-D the ratio was > 1.34–11.41 μM concentrations of growth regulators (IAA,
NAA, BA and Kinetin) in the medium were found effective for callus induction and regeneration in all explants. The calli could
be maintained over 32 months. BA (4.43 μM) combined with casein hydrolysate (10 mg l-1) yielded the highest rate of shoot
production on hypocotyl (3–6) and cotyledon (5–7) explants and cotyledonary derived callus (4–8). More shoots were produced
on explants cut from the most basal region of cotyledons from 5 to 7-day-old seedlings than from older seedlings or more distal
cut sites. Apolar placement of explants, inhibited shoot regeneration. The shoot regeneration potential remained upto 7 months
in calli developed on NAA + BA. Of three media tested, MS was superior to SH-M and B5. Rooting of shoots was not efficient;
42% of the shoots were rooted on MS medium containing sucrose (7–8%) + IAA (2.8–5.7 μM). Capitula induction was observed in
both callus mediated shoots on cotyledons and shoots on rooting medium with sucrose, IAA, NAA and IBA. Well developed plantlets
were transferred to the field with a 34% success rate.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
15.
Hypocotyl expiants from 22 cultivars ofCatharanthus roseus were cultured on various shoot-inducing media to assess their competence for adventitious shoot formation. The Murashige
and Skoog (MS) media had been supplemented with 14 μM zeatin and 2.5 μM indole-3-butyric acid (IBA), 4.5 μM BA and 0.5 μM
α-naphthaleneacetic acid (NAA), or 14 μM thidiazuron and 2.5 μM IBA. After eight weeks, the expiants from ‘Cooler Raspberry
Red’ showed the greatest frequency of adventitious shoot formation, followed by ‘Cooler Orchid’ and ‘Cooler Treated’. The
highest frequency (86.7%) for ‘Cooler Raspberry Red’ was attained on the medium enhanced with 14 μM zeatin and 2.5 μM NAA.
Excised adventitious shoots were then readily rooted on a half-strength MS basal medium. Afterward, the regenerated plantlets
were transferred to potting soil and grown to maturity in a greenhouse. 相似文献
16.
Wenhao Dai Yuanjie Su Cielo Castillo Olivier Beslot 《Plant Cell, Tissue and Organ Culture》2011,104(2):257-262
Shoots were regenerated from in vitro leaf tissues of two genotypes of Viburnum dentatum, a popular shrub species for landscape use. Adventitious shoots were induced when leaf tissues were cultured on woody plant
medium (WPM) supplemented with either benzyladenine (BA) or thidiazuron (TDZ). Effects of cytokinin concentration, indole-3-butyric
acid (IBA), and dark treatment on shoot regeneration were investigated. Dark treatment for the first 4 weeks of leaf explants
cultured in the regeneration medium significantly increased the frequency of regeneration. The highest frequency of shoot
regeneration (70%) for ‘Synnesvedt’ was obtained when leaf tissues were cultured in the medium with 40 μM BA or 8 μM TDZ with
4 weeks dark treatment. The highest frequency of shoot regeneration (90%) for ‘MN34’ was found in the 4 μM TDZ medium with
4 weeks dark treatment. Addition of IBA significantly enhanced shoot regeneration. Ethyl methanesulfonate (EMS) treatment
inhibited callus proliferation, particularly in the early stage of callus recovery; however, no significant difference in
shoot regeneration among different treatments was observed, indicating that the inhibitory effect of EMS was minimal after
calluses re-acquired their capacity to grow and regenerate in the regular medium. Regenerated shoots (>1.5 cm) were rooted
in the half-strength MS medium containing 5-10 μM IBA or naphthalene acetic acid (NAA). Rooted plants were transferred to
the potting medium and grown in the greenhouse. 相似文献
17.
Renata Garcia Georgia Pacheco Erica Falcão Gabriela Borges Elisabeth Mansur 《Plant Cell, Tissue and Organ Culture》2011,106(1):47-54
Passiflora suberosa is used in popular medicine, improvement programs, and as an ornamental plant. The goal of this study was to establish efficient
protocols for plant regeneration and callus induction from nodal, internodal and leaf segments excised from in vitro-grown
plants. The different morphogenetic responses were modulated by the type and concentration of plant growth regulators, according
to the basal medium and light conditions. Shoot formation occurred through three pathways: (1) development of preexisting
meristems, (2) direct organogenesis, and (3) indirect organogenesis. Development of preexisting meristems was observed from
nodal segments (1 shoot/explant) in response to α-naphthaleneacetic acid (NAA), picloram (PIC), and 2,4-dichlorophenoxyacetic
acid (2,4-D), using two basal media (MS and MSM). Direct organogenesis in this species was obtained for the first time in
this work, through shoot development from internodal segments in the presence of 6-benzyladenine (BA). The highest regeneration
rates were achieved on MSM medium, regardless of the BA concentration. Indirect organogenesis was achieved from all explant
types on media supplemented with BA, used alone or in combination with NAA. The highest regeneration efficiency was obtained
from internodal segments cultured on MSM medium plus 44.4 μM BA. Compact, friable, or mucilaginous non-morphogenic calluses
were induced by thidiazuron, PIC, 2,4-D, and NAA. High-yielding friable calluses obtained on MSM medium supplemented with
28.9 μM PIC are being used for the establishment of suspension cultures and further analysis of the production of bioactive
compounds. 相似文献
18.
A novel protocol for callus-mediated shoot regeneration was established for an important medicinal and ornamental plant native
to South China, Curcuma kwangsiensis, using shoot base sections excised from seedlings in vitro as explant sources. The frequency of callus formation reached
91% for explants cultured on MS medium containing 1.4 μM TDZ, 4.4 μM BA and 2.3 μM 2,4-D. 8.2 shoots per callus was achieved
on MS medium supplemented with 1.4 μM TDZ, 17.8 μM BA and 2.7 μM NAA. Single shoots transferred into MS medium free of plant
growth regulator rooted well. Regenerated plants acclimatized ex vitro at 100%, and grew vigorously under shaded greenhouse
conditions. 相似文献
19.
A haploid callus line from anther cultures of the Asiatic hybrid lily ‘Connecticut King’ was maintained for a long term. The
survival and growth of the haploid calluses were affected by auxins of picloram, α-naphthaleneacetic acid (NAA) and 2,4-dichlorophenoxyacetic
acid (2,4-D) and temperatures of 25, 15 and 7 °C during culture. Picloram was more suitable for maintenance of the haploid
calluses, whereas NAA and 2,4-D led to root and shoot formation from the haploid calluses. The best temperature for maintenance
was 25 °C. About 90% of cells in calluses were maintained in haploid level during 60 weeks of subculture, and about 80% of
cells were haploid in the calluses maintained over 2 years with the MS medium containing 4 μM picloram in the dark at 25 °C.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
20.
Wan-Jun Zhang Jiang-Li Dong Ben-Guo Liang Yong-Sheng Jin Tao Wang 《In vitro cellular & developmental biology. Plant》2006,42(2):114-118
Summary We report a protocol for efficient plant regeneration of four tall fescue (Festuca arundinacea Schreb.) cultivars (‘Surpro’, ‘Coronado’, ‘Summer Lawn’, and ‘Fawn’) via somatic embryogenesis. Calli were initiated from
mature seeds grown on modified Murashige and Skoog (MMS) medium supplemented with 7.0mgl−1 (31.7μM) 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.05 mgl−1 (0.23 μM) kinetin (Kin). Calli were maintained and proliferated by subculture at monthly intervals on MMS medium containing 4.5 mgl−1 (20.4 μM) 2,4-D and 0.2mgl−1 (0.9 μM) Kin. Somatic embryos (SE) were induced from seed-derived calli on SE-induction medium (MMS supplemented with 2.0 mgl−1 2,4-D and 0.2mgl−1 Kin). Plantlets were regenerated from somatic embryogenic calli grown on modified SH medium supplemented with 2 mgl−1 Kin. Using this optimized protocol, 78.6–82.3% of mature seeds of all four cultivars produced SE clusters, of which 93.5–95.3%
regenerated into plants within 10 wk. The regenerants showed no phenotypic abnormalities. 相似文献