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1.
In vitro propagation of northern red oak (Quercus rubra) shoots was successful from cotyledonary node explants excised from 8-wk-old in vitro grown seedlings. Initially, four shoots per explant were obtained on Murashige and Skoog (MS) medium supplemented with 4.4 μM
6-benzylaminopurine (BA), 0.45 μM thidiazuron (TDZ), and 500 mg l−1 casein hydrolysate (CH) with a regeneration frequency of 64.7% after 3 wk. Subculturing explants (after harvesting shoots)
to fresh treatment medium significantly increased shoot bud regeneration (16.6 buds per explant), but the buds failed to develop
into shoots. A higher percentage (73.3%) of the explants regenerated four shoots per explant on woody plant medium (WPM) supplemented
with 4.4 μM BA, 0.29 μM gibberellic acid (GA3), and 500 mg l−1 CH after 3 wk. Explants subcultured to fresh treatment medium after harvesting shoots significantly increased shoot regeneration
(16 shoots per explant). Shoot elongation was achieved (4 cm) when shoots were excised and cultured on WPM supplemented with
0.44 μM BA and 0.29 μM GA3. In vitro regenerated shoots were rooted on WPM supplemented with 4.9 μM indole-3-butyric acid. A higher percentage regeneration response
and shoot numbers per explant were recorded on WPM supplemented with BA and GA3, than on MS medium containing BA and TDZ. Lower concentrations of BA and GA3 were required for shoot elongation and prevention of shoot tip necrosis. Each cotyledonary node yielded approximately 20
shoots within 12 wk. Rooted plantlets were successfully acclimatized. 相似文献
2.
Protocols for in vitro propagation of non-toxic variety of J. curcas through axillary bud proliferation and direct adventitious shoot bud regeneration from leaf segments have been established.
Shoot bud proliferation from axillaries was assessed on an initial basal Murashige and Skoog (MS) salt medium supplemented
with different concentrations of benzyladenine (BA), kinetin and thidiazuron (TDZ) followed by subculture to medium with 4.4-8.9
μM BA. Regardless of the concentration of BA in the subculture medium, shoot multiplication rate was optimum (10–12.3) with
primary culture on medium supplemented with 2.3–4.5 μM TDZ. Efficient adventitious shoot regeneration from leaf tissues was
achieved with culture on medium with 8.9–44.4 μM BA + 4.9 μM indole-3-butyric acid (IBA) followed by transfer to medium supplemented
with 8.9 μM BA + 2.5 μM IBA. Similarity index between toxic Indian variety and the non-toxic variety based on 435 RAPD markers
was 96.3%. Crossing studies followed by phorbol ester quantitation revealed that outcrosses with toxic J. curcas do not affect the phorbol ester content of seeds borne on the non-toxic variety. 相似文献
3.
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
4.
Murugesan Dhandapani Doo Hwan Kim Seung-Beom Hong 《In vitro cellular & developmental biology. Plant》2008,44(1):18-25
High-frequency plant regeneration of C. roseus cv. ‘little bright eye’ via somatic embryogenesis and organogenesis from five out of six explants was standardized. Two factors
were found to be important for regeneration: (1) the type of explants, and (2) the combination and concentrations of plant
growth regulators. The highest regeneration percentage through somatic embryogenesis was obtained from mature zygotic embryo
in MS medium supplemented with 7.5 μM of thidiazuron (TDZ). The mature embryo also regenerated efficiently via organogenesis
in MS medium supplemented with either 2.5 μM TDZ or 5.3 μM α-naphthalene acetic acid (NAA) and 2.2 μM 6-benzylaminopurine
(BA). Hypocotyl and cotyledon did not induce somatic embryogenesis and organogenesis in TDZ-containing medium but gave a maximum
percentage of shoots in MS medium supplemented with 5.3 μM NAA and 2.2 μM BA. Stem nodes and meristem tips showed better regeneration
via organogenesis in the medium supplemented with NAA and BA and in lower concentrations of TDZ. 相似文献
5.
A new reliable protocol for the induction of adventitious shoot formation and plant regeneration from apple callus has been
developed. High regeneration frequency was obtained with this method in four different genotypes (Jork9, M26, Gala and McIntosh)
and callus maintained regeneration ability for several months. The procedure consists of inducing vegetative shoot apices,
excised from in vitro shoots, for 20 days in darkness on an MS medium without glycine, supplied with 17.8 μM BA, 2.7 μM NAA
and 250 mg l−1 cefotaxime. The explants are then transferred to a fresh auxin-free medium and given light. Histological studies revealed
that all the regenerated shoots originated from callus. Regenerated shoots were multiplied, rooted and successfully established
in soil.
Received: 2 April 1999 / Revision received: 10 November 1999 / Accepted: 15 November 1999 相似文献
6.
Benzyladenine Induced Somatic Embryogenesis and Plant Regeneration of Leptadenia reticulata 总被引:1,自引:1,他引:0
K.P. Martin 《Biologia Plantarum》2004,48(2):285-288
Plant regeneration through indirect somatic embryogenesis was attempted from leaf, internode, node and shoot-tip derived callus
of Leptadenia reticulata. Somatic embryos at the highest frequency was induced on Murashige and Skoog (MS) medium supplemented with 8.87 μM benzyladenine
(BA) and 2.46 μM indole-3-butyric acid (IBA). From different explants, only shoot-tip and node explant derived calli induced
somatic embryos. Transfer of the embryogenic callus to suspension cultures of the same concentration of growth regulators
facilitated the development of embryos. Suspension cultures with reduced concentration of BA (2.22 μM) either alone or in
combination with 0.49 μM IBA fostered maturation of embryos. Half-strength MS solid medium with 1.44 μM GA3 and BA (0.22 or 0.44 μM) facilitated conversion of embryos into plantlets at higher rate compared to that on with BA alone.
About 77 plantlets were recovered from 10 mg callus. Plantlets transferred to small cups and subsequently to field survived
in 80 %. All the plantlets established in the field exhibited morphological characters similar to that of the mother plant.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
7.
L. Xu U. Najeeb R. Raziuddin W. Q. Shen J. Y. Shou G. X. Tang W. J. Zhou 《In vitro cellular & developmental biology. Plant》2009,45(5):610-618
Wetland species mat rush (Juncus effusus L.) is an important economic plant, but no information is available regarding plant regeneration, callus induction, and its
proliferation from in vitro seed grown plantlets. The present study investigates the effects of growth regulator combinations and medium innovation on
tissue culture system of five mat rush varieties. Addition of N6-benzyladenine (BA) and 2,4-dichlorophenoxyacetic acid (2,4-D)
in Murashige and Skoog (MS) medium showed significantly positive effect on callus proliferation, plant regeneration, and its
multiplication compared to the medium devoid of BA. The highest callus induction frequency (80.95%, 90.48%, 75.40%, 70.83%,
and 83.33%) was observed in MS medium containing 0.5 mg L−1 (2.2 μM) BA in Yinlin-1, Nonglin-4, Gangshan, Taicao, and Taiwan green, respectively. Various growth regulator combinations
with successive subculture (medium replacement) were found essential to develop organogenic calluses and to regenerate shoots.
The combination of 0.1 mg L−1 BA (0.4 μM) and 2 mg L−1 2,4-D (9.0 μM) in MS medium was found best for callus proliferation for all the varieties under trial. The plant regeneration
required two steps involving successive medium replacements as well as optimal hormonal balances. Successful plant regeneration
(over 70%) was observed only by transferring the organogenic callus from regeneration medium I [MS medium containing 0.5 mg
L−1 BA (2. μM) and 1.0 mg L−1 kinetin (KT; 4.6 μM)] to the regeneration medium II [MS medium containing 0.5 mg L−1 BA (2.2 μM), 1.0 mg L−1 KT (4.6 μM) and 3.0 mg L−1 indoleacetic acid (IAA; 17.1 μM)]. Our results confirmed the importance of the ratio of auxin (IAA) to cytokinin (BA and
KT) in the manipulation of shoot regeneration in J. effusus L. The maximum plant survival frequency and multiplication rates (90.97% and 5.40 and 94.23% and 8.25) were recorded in the
presence of 0.5 mg L−1 BA (2.2 μM) in the 1/2 MS multiplication medium for the varieties of Nonglin-4 and Taicao, respectively. About 100% survival
rate was also observed for all the varieties in soil conditions. The efficient plant regeneration system developed here will
be helpful for rapid micropropagation and further genetic improvement in J. effusus L. 相似文献
8.
Pumpkin ash (Fraxinus profunda (Bush) Bush) is at risk for extirpation by an exotic insect, the emerald ash borer (EAB). Pumpkin ash is limited to wetland
areas of the Eastern United States, and has been listed as an endangered species because of EAB activity. Pumpkin ash provides
many benefits to the ecosystem, and its wood is used in the manufacturing industry. In vitro regeneration provides an integral
tool for the mass propagation and genetic transformation of pumpkin ash to combat EAB. Therefore, a plant regeneration protocol
was developed for pumpkin ash. Aseptically extracted hypocotyls formed adventitious shoots following 4 weeks on Murashige
and Skoog (MS) medium supplemented with 0–22.2 μM 6-benzyladenine (BA) and 0–6.8 μM thidiazuron (TDZ) then transferred for
an additional 4 weeks on MS medium with Gamborg B5 vitamins plus 0.2 g L−1 glycine (B5G) containing 6.7 μM BA, 1 μM indole-3-butryic acid (IBA), and 0.29 μM gibberellic acid (GA3). As adventitious shoots developed, these were transferred to a MSB5G medium with 13.3 μM BA, 1 μM IBA, and 0.29 μM GA3 for shoot elongation. Elongated shoots were successfully micropropagated using MSB5 medium with 10 μM BA and 10 μM TDZ. Adventitious
root formation was as high as 94% using woody plant medium supplemented with 4.9 μM IBA with shoots cultured for 10 days in
the dark followed by culture under a 16-h photoperiod. Acclimatization to the greenhouse was successful and normal plant growth
was observed. This protocol will provide a means for genetic transformation for EAB resistance and mass propagation for conservation. 相似文献
9.
This paper describes an efficient in vitro micropropagation of Artemisia vulgaris using shoot tip and nodal explants. Among the various growth regulators tested, MS medium and B5 vitamins supplemented with BA (4.44 μM) and KN (2.32 μM) combination was found to yield a better response than BA (4.44–13.32 μM)
or KN (0.46–13.92 μM) alone in the medium. BA and KN combinations produced a maximum of 23.3 shoots per explant with 99.8%
shooting frequency. Multiple shoots raised were elongated on MS medium containing 0.44 μM BA and 1.44 μM GA3. Rooting was highest (98.2%) on MS medium containing 8.56 μM IAA. Rooted plantlets were successfully transferred to plastic
cups containing autoclaved garden soil, farmyard soil and sand (2:1:1) for hardening. After 65 days, the plantlets were transferred
to Botanical Evaluation Garden and maintained. The survival rate of plantlets varied under acclimatization. Plants looked
healthy with no visually detectable phenotypic variations. This is the first report on plant regeneration via organogenesis
of A. vulgaris. 相似文献
10.
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
11.
Phillip A. Wadl Adam J. Dattilo Lisa M. Vito Robert N. Trigiano 《Plant Cell, Tissue and Organ Culture》2011,106(3):513-516
Pityopsis ruthii is an endangered herbaceous perennial species from the United States. In vitro multiplication of this species can be valuable
for germplasm conservation. Flower receptacles of P. ruthii were cultured on Murashige and Skoog medium (MS) supplemented with 11.4 μM indole-3-acetic acid (IAA) in combination with
2.2, 4.4 or 8.8 μM 6-benzyladenine (BA). Shoots were visible within 14–28 days and three plants were successfully rooted on
MS medium supplemented with 5.7 μM IAA. A two tailed t-test for paired-variates revealed that shoot regeneration on MS medium
amended with 11.4 μM IAA and 2.2 μM BA was significantly higher (P < 0.05) than on other treatments. Leaf explants were also cultured on MS not supplemented with growth regulators or supplemented
with 11.4 μM IAA in combination with 0, 2.2, 4.4 or 8.8 μM BA. Shoots were visible within 21–35 days and one plant was successfully
rooted on MS medium supplemented with 5.4 μM NAA. Shoot regeneration on MS medium augmented with 11.4 μM IAA and 2.2 μM BA
was significantly higher (P < 0.05) than the other treatments according to analysis of variance (ANOVA) with a rank transformation. Hyperhydricity and
rooting of shoots was problematic for explants derived from flower receptacles and leaf tissue, but viable plants were regenerated
using both explants sources indicating the potential role for micropropagation in the ex situ conservation of the species. 相似文献
12.
M. A. K. Azad S. Yokota F. Begum N. Yoshizawa 《In vitro cellular & developmental biology. Plant》2009,45(4):441-449
Somatic embryogenesis and subsequent plant regeneration were established from hypocotyl and internode explants collected from
in vitro-grown seedlings and in vitro-proliferated shoots, respectively. Somatic embryogenesis was significantly influenced by the types of auxin and cytokinin.
Friable calluses with somatic embryos developed well in Murashige and Skoog basal (MS) medium supplemented with 0.8–8.8 μM
6-benzylaminopurine (BA) and 2.0–8.0 μM 2,4-dichlorophexoxyacetic acid (2,4-D) or α-naphthaleneacetic acid (NAA). The maximal
frequency of embryogenic callus and somatic embryo formation were obtained when the MS medium was amended with 8.8 μM BA and
4.0 μM 2,4-D. The best embryo germination occurred in a hormone-free 1/2-MS medium. The highest percentage of shoot proliferation
was observed in embryogenic calluses in MS medium containing 2.0 μM BA and 1.0 μM NAA. In vitro-grown shoots were rooted in MS medium with 0.5–2.0 μM indole-3-butyric acid. Regenerants were transferred to vermiculite and
successfully established under an ex vitro environment in garden soil. 相似文献
13.
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
14.
Xiaojiao Han Hongqiang Yang Kaixuan Duan Xinrong Zhang Haizhou Zhao Shuzhen You Qianqian Jiang 《Plant Cell, Tissue and Organ Culture》2009,96(1):29-34
The effects of sodium nitroprusside (SNP) on the multiplication, regeneration and rooting of Malus
hupehensis Rehd. var. pinyiensis Jiang in tissue culture have been investigated. The results showed that the multiplication of plantlets was promoted significantly
by applying 20 μM SNP to the Murashige and Skoog (MS) medium containing 2.0 μM 6-benzylaminopurine (BA) and 1.0 μM zeatin
(ZT). Multiplication of plantlets from the 1st subculture was more sensitive to SNP than that from the 4th or 7th subculture.
The differentiation and regeneration of adventitious shoots from leaves or cotyledons increased significantly when 20–30 μM
SNP was supplied to the medium MS containing 25 μM BA, 2.5 μM α-naphthaleneacetic acid (NAA) and 2.5 μM ZT. Adventitious shoots regeneration frequency from cotyledons was higher than that
from leaves at the presence of SNP. The rooting of plantlets was promoted by SNP significantly and the best result for rooting
was achieved in the half-strength MS medium containing 75 μM SNP. In addition, adventitious roots without callus distributed
at the base of shoots when SNP was supplied. 相似文献
15.
P. I. P. Perera V. R. M. Vidhanaarachchi T. R. Gunathilake D. M. D. Yakandawala V. Hocher J. L. Verdeil L. K. Weerakoon 《Plant Cell, Tissue and Organ Culture》2009,99(1):73-81
Coconut is a cross pollinating palm, propagated only by seeds. Tissue culture is the only vegetative propagation method available
for coconut. Consistent callogenesis was obtained by culturing unfertilised ovaries at -4 stage in CRI 72 medium containing
100 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.1% activated charcoal. Callusing was improved by application of 9 μM thidiazuron
(TDZ). Embryogenic calli were subcultured onto somatic embryogenesis induction medium containing 66 μM 2,4-D. Stunted growth
was observed in the somatic embryos after subculture onto CRI 72 medium containing abscisic acid (ABA). Maturation of somatic
embryos could be achieved in Y3 medium without growth regulators. Conversion of somatic embryos was induced by adding gibberellic acid (GA3) to conversion medium containing 5 μM 6-benzyladenine (BA) while 2-isopentyl adenine (2iP) increased the frequency of plant
regeneration. A total of 83 plantlets was produced from 32 cultured ovaries. 相似文献
16.
Vinod Kumar Ashwani Sharma Bellur Chayapathy Narasimha Prasad Harishchandra Bhaskar Gururaj Parvatam Giridhar Gokare Aswathanarayana Ravishankar 《Acta Physiologiae Plantarum》2007,29(1):11-18
Direct shoot bud induction and plant regeneration was achieved in Capsicum frutescens var. KTOC. Aseptically grown seedling explants devoid of roots, apical meristem and cotyledons were inoculated in an inverted
position in medium comprising of Murashige and Skoog (Physiol Plant 15:472–497, 1962) basal medium supplemented with 2-(N-morpholine) ethanesulphonic acid buffer along with 2.28 μM indole-3-acetic acid, 10 μM silver nitrate and either of 13.31–89.77 μM
benzyl adenine (BA), 9.29–23.23 μM kinetin, 0.91–9.12 μM zeatin, 2.46–9.84 μM 2-isopentenyl adenine. Profuse shoot bud induction
was observed only in explants grown on a media supplemented with BA (26.63 μM) as a cytokinin source and 19.4 ± 4.2 shoot
buds per explant was obtained in inverted mode under continuous light. Incorporation of polyamine inhibitors in the culture
medium completely inhibited shoothoot bud induction. Incorporation of exogenous polyamines improved the induction of shoot
buds under 24 h photoperiod. These buds were elongated in MS medium containing 2.8 μM gibberellic acid. Transfer of these
shoots to hormone-free MS medium resulted in rooting and rooted plants were transferred to fields. This protocol can be efficiently
used for mass propagation and presumably also for regeneration of genetically transformed C. frutescens. 相似文献
17.
Angela Carra Maurizio Sajeva Loredana Abbate Mirko Siragusa Francesco Sottile Francesco Carimi 《Plant Cell, Tissue and Organ Culture》2012,109(2):373-381
A new technique to regenerate caper plants (Capparis spinosa L. subsp. rupestris) starting from flower explant is reported. In vitro plant regeneration was attempted using stigma, anthers and unfertilized
ovules of unopened flowers collected in the field. Plant regeneration was achieved from unfertilized ovules on MS medium supplemented
with 88 mM sucrose and 13 μM 6-benzyladenine (BA). New individuals obtained from unfertilized ovules were used as source material
for micropropagation and multiple shoots were obtained on MS medium supplemented with the adeninic cytokinin BA and the auxin
indole-3-butyric acid (IBA). Explants obtained in micropropagation step were used for rooting step under several treatments.
The best results (100% of rooted explants) were obtained when explants were dipped for 10 min in 50 μM IBA solution and successively
maintained in growth regulator free medium. New plants were vigorous, of good quality and presented phenotypic characters
similar to mother plants. Furthermore genetic stability of regenerants was verified through flow cytometric analysis and two
different DNA-based techniques. 相似文献
18.
Kaitlin J. Palla Paula M. Pijut 《In vitro cellular & developmental biology. Plant》2011,47(2):250-256
A plant regeneration protocol was developed for white ash (Fraxinus americana L.). Hypocotyls and cotyledons excised from embryos were cultured on Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine
(BA) plus thidiazuron (TDZ), and compared for organogenic potential. Sixty-six percent of hypocotyl segments and 10.4% of
cotyledon segments produced adventitious shoots, with a mean number of adventitious shoots per explant of 3.5 ± 0.9 and 2.5 ± 1.5,
respectively. The best regeneration medium (52% shoot formation; 47% shoot elongation) for hypocotyls was MS basal medium
containing 22.2 μM BA plus 0.5 μM TDZ, producing a mean of 3.9 ± 0.4 adventitious shoots. Adventitious shoots were established
as proliferating shoot cultures following transfer to MS medium with Gamborg B5 vitamins supplemented with 10 μM BA plus 10 μM
TDZ. For in vitro rooting, woody plant medium with indole-3-acetic acid (IAA) at 0, 2.9, 5.7, or 8.6 μM in combination with 4.9 μM indole-3-butyric
acid (IBA) was tested for a 5- or 10-d dark culture period, followed by culture under a 16-h photoperiod. The best rooting
(78% to 81%) of in vitro shoots was obtained with a 5 d dark culture treatment on medium containing 2.9 or 5.7 μM IAA plus 4.9 μM IBA, with an average
of 2.6 ± 0.4 roots per shoot. Rooted plants were successfully acclimatized to the greenhouse. This adventitious shoot regeneration
and rooting protocol will be used as the basis for experimental studies to produce transgenic white ash with resistance to
the emerald ash borer. 相似文献
19.
An efficient protocol was developed for micropropagation of an economically important timber-yielding multipurpose tree, Pterocarpus marsupium Roxb. Multiple shoots were induced from cotyledonary nodes (CNs) derived from 18-d-old axenic seedlings on Murashige and
Skoog (MS) medium supplemented with thidiazuron (TDZ) (0.1–10 μM). The highest shoot regeneration frequency (90%) and maximum
number (15.2 ± 0.20) of shoots per explant was recorded on MS medium amended with 0.4 μM TDZ. Continuous presence of TDZ inhibited
shoot elongation. In the primary medium, TDZ-initiated cultures were transferred to the secondary medium supplemented with
another cytokinin, 6-benzyladenine (BA), for shoot growth and elongation. Maximum (90%) shoot elongation with an average shoot
length of 5.4 ± 0.06 cm was observed at 5 μM BA. To further enhance the number of shoots per explant, mother tissue was repeatedly
subcultured on fresh shoot induction medium after each harvest of newly formed shoots. Thus, by adopting this strategy, an
average of 44 shoots per explant could be obtained. About 65% of in vitro regenerated shoots produced a maximum number (4.4 ± 0.2) of roots per shoot by a two-step culture procedure employing pulse
treatment and subsequent transfer of treated shoots to a low concentration of 0.2 μM indole-3-butyric acid along with phloroglucinol
(3.96 μM). The in vitro-raised plantlets were successfully acclimatized first under culture room conditions, then to greenhouse with 70% survival
rate. 相似文献
20.
Summary Tissue culture and plant regeneration protocols for the salt marsh plants Juncus roemerianus Scheele and Juncus gerardi Loisel, were developed. J. roemerianus callus was induced from mature seeds cultured on Murashige and Skoog (MS) medium supplemented with 2.22 μM 6-benzylaminopurine (BA), 5.37 μM α-naphthaleneacetic acid (NAA), 2.26 μM 2,4-dichlorophenoxyacetic acid (2,4-D), and 50 ml l−1 coconut water (callus induction medium). The callus was subcultured on MS medium containing 2.22 μM BA, 5.37 μM NAA, and 9.05 μM 2,4-D for callus maintenance. Shoot regeneration occurred 2 wk after transferring the callus onto shoot regeneration medium,
which consisted of MS medium containing BA or thidiazuron. A high frequency of shoot regeneration was obtained when the medium
contained 13.3 μM BA. Regenerated shoots were transferred to MS medium supplemented with 10.7 μM NAA for root production. Rooting did not occur in the shoots regenerated on the thidiazuron-containing media. The callus
induction medium for J. roemerianus was also effective in inducing callus of J. gerardi from young inflorescences. The same medium was also used for callus maintenance. Shoot regeneration occurred 10 d after transferring
the callus onto MS medium supplemented with 0.44 μM BA and 0.57 μM indole-3-acetic acid. Root regeneration occurred after transferring the shoots onto MS medium plus 0.44 μM BA and 14.8 μM indole-3-butyric acid. The regenerated plants of both J. roemerianus and J. gerardi grew vigorously in potting soil in the greenhouse. J. roemerianus regenerants also grew well in a saltwater-irrigated field plot. Tissue culture-produced plants of J. roemerianus and J. gerardi can be used for planting in created or restored wetlands. 相似文献