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1.
When cotyledonary explants, excised from in vitro germinated seedlings, of pomegranate (Punica granatum L.) were incubated on solid Murashige and Skoog (1962) medium supplemented with 21 μM naptheleneacetic acid (NAA) and 9 μM 6-benzyladenine (BA), 80% of explants developed callus.
A high frequency of shoot organogensis was obtained when explants were incubated on MS medium supplemented with 8 μM BA, 6 μM
NAA, and 6 μM giberrellic acid (GA3). However, adding 24 μM silver nitrate (AgNO3) to this medium markedly enhanced shoot regeneration frequency (63%) and mean number of shoots per explant (11.26) and length
of shoots (2.22 cm). Highest frequency of in vitro rooting, mean number of roots/shoot (4.32), and mean root length (2.71 cm)
were obtained when regenerated shoots were transferred to half-strength MS medium supplemented with 0.02% activated charcoal.
Well-rooted plantlets were acclimatized, and then transferred to soil medium. Moreover, when zygotic embryos of P. granatum, excised from seeds collected at 16 weeks following full bloom, were incubated on MS medium containing 30 g l−1 sucrose, 15% coconut water, 21 μM NAA, and 9 μM BA, they developed the highest frequency of embryogenic callus, clumps with
globular embryos, and mean number of both globular and heart-shaped embryos per callus clump. Subjecting zygotic embryo explants
to six-week dark incubation period was essential for embryogenic callus induction, and these were subsequently transferred
to 16 h photoperiod for further growth and development of somatic embryos. Germination of somatic embryos was observed when
these were transferred to MS medium was supplemented with 60 g l−1 sucrose. 相似文献
2.
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
3.
Muthu Thiruvengadam K. T. Rekha Chang-Hsien Yang Narayanasamypillai Jayabalan Ill-Min Chung 《Plant biotechnology reports》2010,4(4):321-328
An efficient protocol for in vitro organogenesis was achieved from callus-derived immature and mature leaf explants of Momordica charantia, a very important vegetable and medicinal plant. Calluses were induced from immature leaf explants excised from in vitro
(15-day-old seedlings) mature leaf explants of vivo plants (45 days old). The explants were grown on Murashige and Skoog (MS)
medium with Gamborg (B5) vitamins containing 30 g l−1 sucrose, 2.2 g l−1 Gelrite, and 7.7 μM naphthalene acetic acid (NAA) with 2.2 μM thidiazuron (TDZ). Regeneration of adventitious shoots from
callus (30–40 shoots per explant) was achieved on MS medium containing 5.5 μM TDZ, 2.2 μM NAA, and 3.3 μM silver nitrate (AgNO3). The shoots (1.0 cm length) were excised from callus and elongated in MS medium fortified with 3.5 μM gibberellic acid (GA3). The elongated shoots were rooted in MS medium supplemented with 4.0 μM indole 3-butyric acid (IBA). Rooted plants were
acclimatized in the greenhouse and subsequently established in soil with a survival rate of 90%. This protocol yielded an
average of 40 plants per leaf explant with a culture period of 98 days. 相似文献
4.
Olga Molchan Svetlana Romashko Vladimir Yurin 《Plant Cell, Tissue and Organ Culture》2012,108(3):535-539
l-tryptophan decarboxylase (TDC, EC 4.1.1.28) catalyses the formation of tryptamine from tryptophan, and therefore it plays
a role in terpenoid indole alkaloids biosynthesis. In this study, TDC activity and tryptamine accumulation were monitored
in callus cultures of important medicinal plant Vinca minor L. Callus cultures, established from leaf tissues, were incubated on Murashige and Skoog (MS) medium supplemented with 4.4 μM
kinetin and different concentrations (0.44, 1.1, 2.2, 4.4 and 6.6 μM) of naphthaleneacetic acid (NAA), and grown either in
the dark or under 16 h photoperiod. When the basal enzyme activity of TDC was determined in these cultures, it was 0.5–0.7 nmol
tryptamine mg−1 prot. min−1. Moreover, this activity remained linear over time and over protein concentrations, and with optimum pH levels between 6.5
and 7.5, and an optimum temperature of 35°C. The Michaelis–Menten constant (Km) for l-tryptophan was 1.3 mM. TDC cofactor, pyridoxal-5′-phosphate (1 mM), increased the enzyme activity. During later stages of
callus culture growth cycle, an increase in TDC activity was observed, and this activity depended on culture conditions and
age of callus cultures. In addition, TDC activity and tryptamine accumulation in callus cultures were strongly enhanced by
light treatment. 相似文献
5.
Rangan Parimalan Akshatha Venugopalan Parvatam Giridhar G. A. Ravishankar 《Plant Cell, Tissue and Organ Culture》2011,105(3):317-328
Establishment, maintenance, regeneration, and transformation of somatic embryos by both direct and indirect means (callus-mediated)
was achieved for Bixa orellana, a tropical plant whose seeds produce commercially edible ‘annatto pigment,’ which mainly constitutes an apocarotenoid called
bixin. Callus-mediated methodology was found to be efficient in producing a greater number of embryos in a short time. The
maximum of 28 somatic embryos were produced in 16–18 weeks when immature zygotic embryonic stalks were inoculated onto Murashige
and Skoog (MS) medium containing B5 vitamins supplemented with 0.44 μM benzyladenine (BA), 0.054 μM α-naphthaleneacetic acid
(NAA), 2.89 μM gibberellic acid (GA3), 0.02 μM triiodobenzoic acid (TIBA), and 0.011 μM triacontanol (TRIA). Callus initiation from hypocotyl explants was obtained
on MS medium supplemented with 1.07–2.14 μM NAA and 10.2 μM BA. In 3 months, somatic embryos were produced when callus was
inoculated onto MS medium supplemented with 4.44 μM BA, 40 μM AgNO3, and 0.011 μM TRIA. Somatic embryos were efficiently regenerated on MS basal solid and liquid media supplemented with 0.44–4.4 μM
BA, 0.54–2.69 μM NAA, 4.92 μM 2iP, 2.1 μM calcium d-pantothenate, 0.21 μM biotin, 227.7 μM cysteine HCl monohydrate, and 108.6 μM adenine sulfate. Agrobacterium tumefaciens GV 3101 harboring pCAMBIA 1305.2 binary vector-mediated stable transformation of somatic embryos exhibited a transformation
frequency of 2.56%. As somatic embryogenesis in any perennial system is useful in terms of both commercial and scientific
nature, this somatic embryo-based transformation protocol for the commercially important dye-yielding tropical plant B. orellana is useful for its improvement through genetic engineering. 相似文献
6.
Kusampudi Shilpa Chinnasamy Selvakkumar Arun Kumar Senthil Baddireddi Subhadra Lakshmi 《Plant Cell, Tissue and Organ Culture》2010,101(1):105-109
Young leaf explants of Ocimum sanctum L. incubated on solidified Murashige and Skoog (MS) medium supplemented with 2 mg l−1 1-naphthaleneacetic acid (NAA) and 0.2 mg l−1 kinetin (Kn) developed rhizogenic callus. When these were subcultured onto MS medium supplemented with 1.5 mg l−1 2, 4-dichlorophenoxyacetic acid (2, 4-D) and 0.5 mg l−1 NAA, friable rhizogenic callus was observed. Upon transfer of this friable callus onto liquid MS medium containing 4 mg l−1 NAA and 1.3 mg l−1 6-benzyladnine (BA) under continuous agitation at 90 rpm and 16 h photoperiod, roots with an optimum dry weight of 1,460 mg l−1 were obtained. An ethyl acetate extract of these roots exhibited 1, 1–diphenyl-2-picrylhydrazyl (DPPH) radical scavenging
activity. 相似文献
7.
N. Irvani M. Solouki M. Omidi A. R. Zare S. Shahnazi 《Plant Cell, Tissue and Organ Culture》2010,100(3):293-299
Dorema ammoniacum D. Don. (Apiaceae), a native medicinal plant in Iran, is classified as a vulnerable species. Root, hypocotyl, and cotyledon
segments were cultured on Murashige and Skoog (MS) (1962) medium supplemented with either 2,4-dichlorophenyoxyacetic acid
(2,4-D) or naphathalene acetic acid (NAA), at 0–2 mg l−1, alone or in combination with either benzyladenine (BA) or kinetin (KN), at 0–2 mg l−1 for callus induction. The best response (100%) was observed from root segments on MS medium containing 1 mg l−1 NAA and 2 mg l−1 BA. The calli derived from various explants were subcultured on MS medium supplemented with BA (1–4 mg l−1) alone or in combination with NAA or indole-3-butyric acid (IBA), at 0.2 or 0.5 mg l−1 for shoot induction. Calli derived from hypocotyl segments showed significantly higher frequency of plantlet regeneration
and number of plantlets than the calli derived from root and cotyledon segments. Therefore, MS medium supplemented with 2 mg l−1 BA and 0.2 mg l−1 IBA produced the highest frequency of shoot regeneration (87.3%) in hypocotyl-derived callus. The optimal medium for rooting
contained 2.5 mg l−1 IBA on which 87.03% of the regenerated shoots developed roots with an average number of 5.2 roots per shoots within 30 days.
These plantlets were hardened and transferred to the soil. The described method can be successfully employed for the large-scale
multiplication and conservation of germplasm this plant. 相似文献
8.
In vitro mother plants initiated from a mature tree of Sorbus aucuparia, produced numerous propagules on a medium containing 2 μM 6-benzylaminopurine (BAP) and 0.2 μM 1-naphthaleneacetic acid (NAA).
These were rooted on a medium containing 0.25 μM NAA and 0.25 μM indole−3-butyric acid. Adventitious shoots were produced
on excised leaves and internodes on media containing 10 μM thidiazuron and 0.3–1.0 μM NAA. They formed by direct regeneration
in the axils of leaflets of intact leaves. They also developed indirectly, from callus that developed on the rachis of intact
leaves, and the cut ends of petioles and internodes. Somatic embryos were produced on cotyledons of zygotic embryos on medium
containing 1 μM BAP, 1 μM kinetin, 0.5 μM NAA, 500 mg l−1 casein hydrolysate and 250 mg l−1 glutamine. On basal medium, 69% developed cotyledons and 20% germinated after pre-treatment at 4 °C on medium containing
30 g l−1 maltose. 相似文献
9.
Micropropagation has been achieved in a promising larvicidal asteraceous taxon Spilanthes acmella L. using seedling leaf explants. The explants were reared on a variety of growth regulators, namely 2,4-dichlorophenoxyacetic
acid, 1-naphthalene acetic acid, Indole-3-butyric acid, N6-benzyladenine, and kinetin either alone or in combination on Murashige and Skoog’s (MS) medium. The best green and compact
callus was obtained on 1 μM NAA and 10 μM benzyladenine (BA) in 15 d. The callus on subculture to the same but fresh medium
after every 30 d differentiated an average of 12.90 ± 0.32 shoot buds in 50% cultures. Elongation in shoot buds occurred only
if they were transferred to NAA lacking MS+BA medium. An average number of 4.22 ± 0.83 shoots and 15 ± 0.84 shoot buds per
explant were obtained in 70.3% cultures on MS + 10 μM BA in 30 d. One hundred percent excised shoots rooted in MS(1/2) + 0.1 μM
IBA within 2 wk. The plants were gradually hardened and established in soil where they flowered and set viable seeds. The
regenerated plants were morphologically similar to the field grown plants and showed 100% larvicidal activity against malaria
and filarial vectors. 相似文献
10.
Organogenic cultures were induced from zygotic embryo and megagametophyte explants of the Central American cycad species,
Dioon edule. Plant growth medium consisted of B5 major salts, Murashige and Skoog minor salts and organics, 400 mg l−1 glutamine, 100 mg l−1 arginine, 100 mg l−1 asparagine, 60 g l−1 sucrose, 8 g l−1 Difco Bacto agar and was supplemented with kinetin (0 – 13.94 μM) and 2,4-dichlorophenoxyacetic acid (2,4-D) (0 – 9.05 μM)
arranged as a 5×4 factorial in a randomized block design. Callus initiation occurred on a wide range of medium formulations
from megagametophyte explants; however, shoot formation occurred only on medium supplemented with 2.26 μM 2,4-D. In comparison,
callus initiation from explanted zygotic embryos occurred on fewer medium formulations, and adventitious shoot induction occurred
from callus on formulations with 9.29–13.94 μM kinetin + 0.45–9.05 μM 2,4-D. Rooted shoots, derived from megagametophyte and
zygotic embryo cultures, have been regenerated.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
11.
The effects of growth regulators, cold-pretreatment of flower buds, ovule (embryo sac) developmental stage and genotype on
induction of gynogenesis in unpollinated ovule cultures were assessed in niger (Guizotia abyssinica (L. f.) Cass.). Indirect callus-mediated gynogenesis occurred in cvs JNC-6 and Ootacamund when the ovules were cultured on
MS medium supplemented with 30 g l−1 sucrose and 2,4-D either alone (0.5–2.0 μM) or in combination (2.0 μM) with different cytokinins, such as adenine, BA, 2iP
and kinetin (0.5–2.0 μM). An optimum induction of gynogenesis was fostered on medium supplemented with 2.0 μM 2,4-D and 1.0 μM
kinetin. Cold-pretreatment of flower buds had no stimulatory effect, but ovules collected one day before anthesis were most
responsive to gynogenesis. The results showed significant variations in genotypic competence for gynogenesis with cv. Ootacamund
being the most responsive (12.5%) and cv. IGP-76 the least (2.5%). Gynogenic embryos differentiated and matured on media (30 g l−1 sucrose) supplemented with 0.5 μM NAA plus 1.0 μM kinetin, and 0.5 μM ABA, respectively. The haploidy (2n = 1x = 15) of gynogenic plants was confirmed by cytological analysis. 相似文献
12.
The genus Dierama comprises plants with a potential to be developed as ornamentals. D. erectum seeds were decontaminated and germinated on 1/10th strength Murashige and Skoog (Physiol Plant 15:473–497, 1962) (MS) media
without plant growth regulators or sucrose. In an experiment investigating the effects of 6-benzyladenine (BA), meta-Topolin (mT), kinetin (KIN) and zeatin (Z) with or without α-naphthaleneacetic acid (NAA), the highest shoot number per hypocotyl (4.20 ± 0.51)
was obtained from MS medium supplemented with 1.0 μM Z after 8 weeks. This was followed by a combination of 2.0 μM KIN and
2.0 μM NAA with 3.67 ± 0.81 shoots per explant. BA treatments produced 3.20 ± 0.22 shoots per hypocotyl explant when 2.0 μM
was combined with 1.0 μM NAA, while mT gave 3.09 ± 0.99 shoots per explant when 2.0 μM mT was combined with 2.0 μM NAA. Adventitious shoot regeneration was optimised when shoots were grown under a 16-h photoperiod
at 100 μmol m−2 s−1 on MS medium supplemented with 1.0 μM BA. This resulted in an average of 12.73 ± 1.03 shoots per hypocotyl explant. Various
concentrations of ancymidol, activated charcoal and sucrose did not promote in vitro corm formation of this species. Plants
rooted successfully after 8 weeks on MS medium supplemented with 1.0 μM indole-3-butyric acid (IBA) and had an average root
number of 2.73 ± 0.40. After 2 months of acclimatisation, plants had formed corms. The largest corms (of diameter 0.45 ± 0.03 cm)
were produced in plants pre-treated with 0.5 μM IBA. The highest plant survival percentage of 73% was also associated with
this treatment. 相似文献
13.
Nodular callus was induced at a high frequency on young purple red, 5–15 mm long laminae taken from in vitro grown plants of mangosteen. The optimal medium was composed of Murashige and Skoog (MS) nutrients supplemented with 2.22
μM benzyladenine (BA), 2.25 μM thidiazuron (TDZ), 500 mg l-1 polyvinylpyrrolidone (PVP 360 000) and 3% sucrose. A multiplication rate of two–three was obtained by subculture of the nodular
callus at 3–4-week intervals. Plantlet regeneration from the nodules was achieved by transfer to woody plant medium (WPM)
with 500 mg l-1 PVP, 0.4 μM BA and 3% sucrose and overlaying with half strength liquid MS containing 0.32 μM naphthaleneacetic acid (NAA),
0.13 μM BA and 3% sucrose. Elongated shoots were rooted to 100% when wounded at the base of shoot, dipped in 4.4 mM indolebutyric
acid (IBA) solution in the dark for 15 min and cultured on WPM supplemented with 1.11 μM BA, 0.25% activated charcoal, 34.5
μM phloroglucinol (PG) and 3% sucrose.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
14.
Multiple shoot induction and callus regeneration in Sarcostemma brevistigma Wight & Arnott, a rare medicinal plant 总被引:1,自引:0,他引:1
An efficient micropropagation protocol based on multiple shoot induction and callus regeneration has been standardized in
Sarcostemma brevistigma, a rare medicinal plant. The nodal cuttings were cultured on MS medium supplemented with BA (0.5–8 μM) or Kn (0.5–8 μM) alone
or in combination with NAA (0.5–1.5 μM). Maximum multiple shoot induction was observed on MS medium supplemented with 4 μM
BA. On this medium, 100% cultures responded with an average number of 11.3 shoots per explant. However, the average shoot
length was limited to only 0.9 cm on this medium. The addition of 1 μM NAA along with 4 μM BA gave rise to an average number
of 10.9 shoots with an average shoot length of 1.8 cm. Luxuriantly growing callus was obtained on MS medium supplemented with
BA (5 μM) and 2,4-D (2 μM). The callus was subcultured on MS medium supplemented with BA (2–15 μM) or Kn (2–15 μM) alone or
in combination with NAA (0.5–2 μM) for shoot organogenesis. Optimum callus regeneration was obtained on MS medium supplemented
with 10 μM BA and 1 μM NAA. On this medium, 100% cultures responded with an average number of 13.4 shoots per culture. The
shoots obtained via multiple shoot induction and organogenesis were rooted on half-strength MS medium supplemented with NAA
(1–7 μM) or IBA (1–7 μM). IBA was better than NAA in terms of both the percentage of cultures that responded and the average
number of roots per explant. The rooted shoots were successfully transplanted to soil with 86% success. This standardized
protocol will help to conserve this rare medicinal plant. 相似文献
15.
Rocío Elizabeth Escobar-Guzmán Fernando Hernández-Godínez Octavio Martínez de la Vega Neftalí Ochoa-Alejo 《Plant Cell, Tissue and Organ Culture》2009,96(2):181-189
Eight cultivars and two accessions of Physalis ixocarpa Brot. were tested for their capacity to regenerate embryos and plants from anther cultures. Anthers were pretreated at 4°C
for 2 days and then at 35°C for 8 days in the dark while cultured on MS medium supplemented with 0.045 μM 2,4-D + 0.03 mg l−1 vitamin B12 (MS1) or with 2.26 μM 2,4-D + 0.1 mg l−1 vitamin B12 (MS3). Anther incubation proceeded under a 16 h photoperiod at 25 ± 2°C. Embryo formation occurred after 6 weeks of incubation
in these conditions. Androgenetic responses were cultivar- and culture medium-dependent, with the greatest embryo yields recorded
for cv. Chapingo (36.3%) on MS1 medium, and with wild-type 2 (21.8%) on MS3. Further development of regenerated embryos was
promoted on MS medium supplemented with 0.54 μM NAA, 8.88 μM BA and 50 mg l−1 casein hydrolysate. The regenerated plants were cultured on half-strength mineral salts MS medium with 2.85 μM IAA to enhance
root formation. Rooted plantlets were transferred to pots and acclimatized to the greenhouse. Ploidy analysis of regenerated
plants using flow cytometry revealed 72% diploids, 15% haploids and 7% triploids. AFLP analysis of regenerated plants from
anthers of a single parental plant showed different polymorphic patterns indicating their gametophytic origin. 相似文献
16.
A. Ptak A. El. Tahchy G. Wyżgolik M. Henry D. Laurain-Mattar 《Plant Cell, Tissue and Organ Culture》2010,102(1):61-67
The influence of ethylene on in vitro morphogenesis of Leucojum aestivum and galanthamine accumulation was studied. Calli were cultivated on Murashige and Skoog (MS) medium supplemented with 25 μM
4-amino-3,5,6-trichloropicolinic acid (picloram) and 0.5 μM benzyladenine (BA). During incubation under these conditions,
callus cultures produced ethylene (9.5 nL/g fresh weight: F.W.) whereas no ethylene was found in somatic embryos cultivated
on medium supplemented with 0.5 μM α-naphthalene acetic acid (NAA) and 5 μM zeatin. Application of the precursor of ethylene
1-aminocyclopropane-1-carboxylic acid (ACC) increased ethylene production in both cultures, and decreased callus growth by
a factor of 1.2, whereas callus growth was enhanced by a factor of 1.1 in the presence of an inhibitor of ethylene silver
nitrate (AgNO3) or by a factor of 1.2 with an absorbent potassium permanganate (KMnO4). ACC enhanced the induction of somatic embryos and the development of globular embryos. Removal of ethylene by KMnO4 during somatic embryogenesis led to the development of plants with greater length. Silver thiosulphate (STS) induced galanthamine
production in callus cultures (0.1% dry weight), whereas ACC induced galanthamine production in somatic embryo cultures (2%
dry weight). 相似文献
17.
A. V. Loskutov G.-Q. Song K. C. Sink 《In vitro cellular & developmental biology. Plant》2008,44(4):239-245
Callus selection (CS) and the flamingo-bill explant (FB) methods were evaluated for efficacy in transformation for celery.
Agrobacterium tumefaciens strains EHA105 and GV3101, each with the bar gene under the promoters NOS (pGPTV-BAR) or 35S (pDHB321.1), were used. Leaf explants were inoculated and co-cultivated for
2 d in the dark. Calluses emerged on the explants on callus medium (C), Murashige and Skoog (MS) medium + 2,4-Dichlorophenoxyacetic
acid (2,4-D) (2.3 μM) + kinetin (2.8 μM) + timentin (300 mg·l−1). Calluses 4- to 6-wk-old were selected for glufosinate (GS) resistance by a two step method. First, calluses were transferred
to C medium + GS 0.35, 0.5, 1, 2, 5, or 10 mg·l−1; calluses formed only with 0, 0.35 and 0.5 mg·l−1 GS. All growing calluses from 0 and 0.35 mg·l−1 and a few from 0.5 mg·l−1, were divided and placed back on C + GS 0.35–0.5 mg·l−1 for another 5–6 wk. Second, tolerant clones were again divided and placed on C + GS 1–50 mg·l−1. When cultivar XP85 was inoculated with both strains, using pGPTVBAR, 19 glufosinate resistant (GR) callus clones were selected,
but shoots regenerated only for strain EHA105 inoculations. When both of the strains (each with pDHB321.1) were inoculated
on cv. XP166, 3 and 12 GR calluses occurred for EHA105 and GV3101, respectively. Using CS, a total of 34 GR callus clones
were selected, and shoots were regenerated from over 50% of them on Gamborg B5 medium + 6-(γ, γ-dimethylallylamino) purine
2ip (4.9 μM) + naphthaleneacetic acid (NAA; 1.6 μM) and rooted on MS in 5–6 mo total time. Conversely, using FB with inoculation
by GV3101/pDHB321.1 on cv. XP166 yielded putative transgenic celery plants confirmed by polymerase chain reaction (PCR) in
just 6 wk. Transformation of the bar gene into celery was confirmed by PCR for 5 and 6 CS and FB lines, respectively. Southern blot analyses indicated 1–2 copies
in CS lines and 1 copy in FB lines. Herbicide assays on whole plants with 100 and 300 mg·l−1 glufosinate indicated a range of low to high tolerance for lines derived by both methods. The bar gene was found to be Mendelian inherited in one self-fertile CS derived line. 相似文献
18.
Plant regeneration was achieved through direct and indirect somatic embryogenesis in Eucalyptus camaldulensis. Callus was induced from mature zygotic embryos and from cotyledon explants collected from 10, 15, 25, and 30-day-old seedlings
cultured on Murashige and Skoog (MS) basal medium supplemented with different concentrations of naphthaleneacetic acid (NAA).
Maximum callus induction from mature zygotic embryos was obtained on MS basal medium containing 1 mg l−1 NAA. The frequency of callus development varied based on the age of the cotyledon explants 10-day-old explants giving highest
percentage on MS basal medium supplemented with 1 mg l−1 NAA. Callus obtained from mature zygotic embryos gave highest frequency of somatic embryogenesis on MS basal medium containing
0.5 mg l−1 benzyladenine (BA) and 0.1 mg l−1 NAA. Separate age wise culture of the calli, obtained from cotyledons of different ages cultured separately, revealed high
somatic embryogenic potential on callus from 10-day-old cotyledons. Direct somatic embryogenesis too was obtained from hypocotyl
explants without an intervening callus phase on MS basal medium containing 0.5 mg l−1 BA. The effects of abscisic acid (ABA), sucrose, and different strengths of MS medium on somatic embryo maturation and germination
were also investigated. Number of mature somatic embryos increased with lower concentrations (0–1 mg l−1) of ABA while no significant differences were observed at higher concentrations (2–5 mg l−1) of ABA. Compared to basal medium containing lower concentrations of sucrose (1%), the MS medium supplemented with higher
levels of sucrose (4%) showed significantly lower frequency of mature somatic embryos. Basal medium without any dilution gave
the highest number of immature embryos. However, the number of mature embryos was high at higher medium dilutions. 相似文献
19.
In the present study, a simple one medium formulation protocol for callus culture, somatic embryogenesis and in vitro production
of β-carboline alkaloids and diosgenin in Tribulus terrestris L. was developed. Extensive callus induction and proliferation was obtained in leaf explant on Murashige and Skoog (MS) medium
supplemented with 5.0 μM 6 benzyl adenine (BA) and 2.5 μM α-naphthaleneacetic acid (NAA). The embryogenic callus was maintained
on subculture to fresh parental medium at 4-week intervals over a period of 28 months. The frequency of embryo formation was
at a maximum (18.1 ± 0.9 per g of callus) on MS medium containing 5.0 μM BA and 2.5 μM NAA together with 75 mg l−1 casein hydrolysate. Globular embryo developed into torpedo stage embryo under the influence of starvation. The accumulation
of β-carboline alkaloids (harmaline and harmine) and steroidal saponin (diosgenin) in non-embryogenic and embryogenic callus
culture derived from leaf explant was compared with root, leaf, stem, and fruit of the mother plant. The embryogenic callus
accumulated equivalent amounts of harmaline (66.4 ± 0.5 μg/g dry weight), harmine (82.7 ± 0.6 μg/g dry weight), and diosgenin
(170.7 ± 1.0 μg/g dry weight) to that of the fruit of T. terrestris. The embryogenic callus culture of this species might offer a potential source for production of important pharmaceuticals. 相似文献
20.
S. Paul A. Dam A. Bhattacharyya T. K. Bandyopadhyay 《Plant Cell, Tissue and Organ Culture》2011,105(2):271-283
A reproducible protocol for direct and indirect somatic embryogenesis was established in a small aromatic tree, Murraya koenigii. Embryogenic callus was obtained from 90% zygotic embryonic axis (ZE) and 70% cotyledon (COT) explants in Murashige and Skoog
(MS) basal medium supplemented with 8.88 μM 6-benzyladenine (BA) and 2.675 μM α-naphthaleneacetic acid (NAA). Globular somatic
embryos were induced and further matured from such embryogenic callus by subsequent culture on the same basal media containing
thidiazuron (TDZ) (2.27–9.08 μM). The highest frequency of somatic embryos (14.58 ± 0.42) was recovered from ZE-derived callus
after 6 weeks. The age and type of explant and concentration of TDZ played an important role in the development of somatic
embryos. Explants excised from 60-day-old seed differentiated from 96.67% of ZE explants and 86.67% from COT explants when
cultured on MS basal medium supplemented with 4.54 and 9.08 μM TDZ, respectively, after 4 weeks. The best result obtained
for the average frequency of somatic embryos (11.28 ± 0.32) was from ZE explants, which was significantly higher than COT
explants (7.34 ± 0.97). Most of the somatic embryos (above 95%), irrespective of their origin, germinated after 4 weeks in
1/2 MS basal media containing 2.32 μM kinetin (KN) and 1.07 μM NAA. Well-rooted plantlets were successfully acclimatized.
Histological analysis and scanning electron micrographs confirmed the initiation, development, and germination of somatic
embryos from both explants. 相似文献