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1.
Summary
In vitro regeneration of plants via somatic embryogenesis through cell suspension culture was achieved in horsegram. Embryogenic calluses
were induced on leaf segments on solid Murashige and Skoog (MS) medium with 9.0 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Differentiation of somatic embryos occurred when the embryogenic calluses were transferred
to liquid MS medium containing 2,4-D. Maximum frequency (33.2%) of somatic embryos was observed on MS medium supplemented
with 7.9 μM 2,4-D. Cotyledonary-torpedo-shaped embryos were transferred to liquid MS medium without growth regulators for maturation
and germination. About 5% of the embryos germinated into plants, which grew further on solid MS medium. The plants were hardened
and established in soil. Effects of various auxins, cytokinins, carbohydrates, amino acids, and other additives on induction
and germination of somatic embryos were also studied. A medium supplemented with 7.9 μM 2,4-D, 3.0% sucrose, 40 mg l−1
L-glutamine, and 1.0 μM abscisic acid was effective to achieve a high frequency of somatic embryo induction, maturation, and further development. 相似文献
2.
A protocol was developed for the induction, maturation and germination of somatic embryos from leaf tissue of jojoba [Simmondsia chinensis (Link) Schneider]. Explants were placed on their adaxial sides in Petri dishes and maintained in darkness on half-strength
Murashige and Skoog basal medium (MS/2). Combinations of 2,4-dichlorophenoxyacetic acid (1.35–4.52 μM) with 6-benzylaminopurine
(1.33–4.43μM) and 2 synthetic cytokinins, N-(2-chloro-4pyridyl)-N′-phenylurea (1.21–4.03μM) or (E)-6-[3-(trifluoromethyl)-but-2-enylamino]
purine (1.11–3.71μM) resulted in formation of embryogenic cultures and somatic embryos. After two 30-day subcultures, embryogenic
cultures were transferred onto MS/2 medium supplemented with different auxins and cytokinins. Somatic embryo maturation, germination
and plantlet formation were achieved using 1-naphthaleneacetic acid (3.75μM) or indole-3-butyric acid (3.44μM) in combination
with BA (0.44 or 1.33μM) or F3iP (0.37 or 1.11μM). Histology confirmed each stage of development.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
3.
A. Vasudevan N. Selvaraj A. Ganapathi S. Kasthurirengan V. Ramesh Anbazhagan M. Manickavasagam C. W. Choi 《In vitro cellular & developmental biology. Plant》2008,44(4):300-306
Enhanced numbers of multiple shoots were induced from shoot tip explants of cucumber. The effects of amino acids (leucine,
isoleucine, methionine, threonine, and tryptophan) and polyamines (spermidine, spermine, and putrescine) along with benzyladenine
(BA) on multiple shoot induction were investigated. A Murashige and Skoog (MS) medium containing a combination of BA (4.44 μM),
leucine (88 μM), and spermidine (68 μM) induced the maximum number of shoots (36.6 shoots per explant) compared to BA (4.44 μM)
alone or BA (4.44 μM) with leucine (88 μM). The regenerated shoots were elongated on the same medium. Elongated shoots were
transferred to the MS medium fortified with BA (4.44 μM), leucine (88 μM), and putrescine (62 μM) for root induction. Rooted
plants were hardened and successfully established in soil with a 90% survival rate. 相似文献
4.
Konstantin V. Kiselev Anna V. Turlenko Yuri N. Zhuravlev 《Plant Cell, Tissue and Organ Culture》2009,99(2):141-149
A somatic embryogenesis protocol for plant regeneration of northern red oak (Quercus rubra) was established from immature cotyledon explants. Embryogenic callus cultures were induced on Murashige and Skoog medium
(MS) containing 3% sucrose, 0.24% Phytagel™, and various concentrations of 2,4-dichlorophenoxyacetic acid (2,4-d) after 4 weeks of culture in darkness. A higher response (66%) of embryogenic callus was induced on 0.45 μM 2,4-d. Higher numbers of globular- (31), heart- (17), torpedo- (12), and cotyledon-stage (8) embryos per explant were obtained
by culturing embryogenic callus on MS with 3% sucrose, 0.24% Phytagel™, and devoid of growth regulators after 8 weeks culture
in darkness. Continuous sub-culturing of embryogenic callus on medium containing 2,4-d yielded only compact callus. Desiccation of embryos for 3 days in darkness at 25 ± 2°C followed by cold storage at 4°C in
darkness for 8 weeks favored embryo germination and development of plantlets. Cotyledon-stage embryos subjected to desiccation
and chilling treatment cultured on MS with 3% sucrose, 0.24 Phytagel™, 0.44 μM 6-benzylaminopurine (BA), and 0.29 μM gibberellic
acid germinated at a higher frequency (61%) than with 0.44 μM BA alone and control cultures. Germinated plantlets developed
a shoot and root, were acclimatized successfully, and maintained in a growth room for plantlet development. 相似文献
5.
A simple and efficient system was developed for rapid somatic embryogenesis from leaf explants of Merwilla plumbea, a traditional but threatened medicinal plant in South Africa. Friable embryogenic callus (FEC) was obtained from leaf explants
on embryogenic callus induction medium containing agar-solidified Murashige and Skoog (MS) salts and vitamins, 8.3 μM picloram,
2.3 μM thidiazuron (TDZ) and 20 μM glutamine. FEC was subsequently incubated in embryogenic callus proliferation medium containing
4.5 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 4.1 μM picloram for 7 days before it was transferred to liquid somatic embryo
medium (SEML) containing MS medium supplemented with 0.4 μM picloram and 0.9 μM TDZ. In SEML supplemented with 150 mg L−1 haemoglobin, 5.4–35.6 somatic embryos per settled cell volume of 500 mg FEC were obtained. These embryos were at globular
to cotyledonary developmental stages. Embryo maturation, germination and plant formation rate was 94.4% following transfer
of SEs to half-strength MS medium supplemented with 1.4 μM gibberellic acid. Plantlets transferred into soil acclimatized
in the misthouse and established successfully in the greenhouse (100%). This is the first report on induction of Merwilla plumbea somatic embryogenesis. The protocol developed offers controlled vegetative propagation by alleviating extinction threats,
ensures germplasm conservation and provides a system for physiological, biochemical, molecular and cellular studies of embryo
development. 相似文献
6.
P. I. P. Perera D. M. D. Yakandawala V. Hocher J-L. Verdeil L. K. Weerakoon 《Plant Cell, Tissue and Organ Culture》2009,96(2):171-180
The effect of growth regulators on induction of androgenesis in coconut was investigated using seven different growth regulators
at various concentrations and combinations. Three auxins (1-naphthalene acetic acid—NAA, indoleacetic acid—IAA, picloram)
and three cytokinins (2-isopentyl adenine-2-iP, kinetin, zeatin) were tested either alone or in combination with 2,4-dichlorophenoxyacetic
acid (2,4-D), using modified Eeuwens Y3 liquid medium as the basal medium. Among the tested auxins, 100 μM NAA in combination
with 100 μM 2,4-D enhanced the production of calli/embryos (123) whereas IAA and picloram showed negative and detrimental
effects, respectively, for androgenesis induction over 100 μM 2,4-D alone. Kinetin and 2-iP enhanced the production of calli/embryos
when 100 μM 2,4-D was present in the culture medium. Both cytokinins at 10 μM yielded the highest frequencies of embryos (113
and 93, respectively) whereas zeatin (1 or 2.5 μM) had no impact on microspore embryogenesis. When calli/embryos (produced
from different treatments in different experiments) were sub-cultured in somatic embryo induction medium (Y3 medium containing 66 μM 2,4-D), followed by maturation medium (Y3 medium without growth regulators) and germination medium (Y3 medium containing 5 μM-6-benzyladenine—BA and 0.35 μM gibberellic acid—GA3), plantlets were regenerated at low frequencies (in most treatments ranging from 0% to 7%). 相似文献
7.
Seabuckthorn (Hippophae rhamnoides) is a multipurpose small tree with unique berries of high nutritional and pharmaceutical values. A clonally propagated plant
originating from a 20-year-old tree of H. r. rhamnoides × mongolica hybrid cultivar Julia and seedling offspring of this cultivar were investigated regarding induction of shoot organogenesis
in leaf explants and in roots of intact seedlings, and induction of direct somatic embryogenesis in explants from shoot tissue.
The highest percentage of leaf explants showing shoot organogenesis was achieved (juvenile explants, 65%; adult explants,
75%) when incubated in Murashige and Skoog (MS) medium supplemented with either 4.5 μM of the phenylurea cytokinin thidiazuron
(TDZ) or 2.25 μM TDZ plus 2.2 μM 6-benzyladenine (BA), for juvenile and adult explants, respectively, both supplemented with
0.53 μM α-naphthaleneacetic acid (NAA). Juvenile explants developed on average 18 shoots per explant in the MS medium supplemented
with 4.5 μM TDZ, a four fold increase over those incubated on the medium supplemented with 2.25 μM TDZ and 2.2 μM BA. Adult
leaf explants grown on medium containing 2.25 μM TDZ and 2.2 μM BA medium produced 12 shoots per explant, while those grown
on medium containing 4.5 μM TDZ produced 5 shoots per explant. Shoot organogenesis was observed in roots of intact seedlings
pre-cultured on plain medium lacking nutrients (PM) or woody plant medium (WPM) salts and then grown on WPM salts supplemented
with 4.4 μM BA, 0.29 μM gibberrelic acid (GA3), and 57.0 μM indoleacetic acid (IAA). The number of shoots formed on each seedling
root system was ten fold higher when the pre-culture was in WPM medium indicating a promoting effect of mineral nutrients
in the pre-culture medium. Somatic embryogenesis was induced in both juvenile and adult leaf explants in 65 and 78% of the
explants, respectively, in MS-based medium supplemented with 2.0 μM N-(2-Chloro-4-pyridyl)-N
1-phenylurea (CPPU), 0.53 μM NAA and varying concentrations of BA. There was an interaction effect between MS salt strength
and BA concentration. The most effective medium for inducing somatic embryogenesis in juvenile explants contained half strength
MS salts and 2.2 μM BA and full strength MS salts and 13.2 μM BA for adult explants. 相似文献
8.
Ai Hua Chen Jing Li Yang Yu Da Niu Chuan Ping Yang Gui Feng Liu Chang Yeon Yu Cheng Hao Li 《Plant Cell, Tissue and Organ Culture》2010,102(3):357-364
We developed a new protocol for highly efficient somatic embryogenesis and plantlet conversion of Schisandra chinensis. Friable embryogenic callus was induced from cotyledonary leaves and hypocotyls of germinated zygotic embryos on Murashige
and Skoog (MS) agar medium containing 2,4-dichlorophenoxyacetic acid (2,4-D). Preculture of zygotic embryos on 2,4-D-containing
medium increased embryogenic callus induction efficiency. The highest embryogenic callus induction frequency of 56.7% was
obtained from shoot apical meristem-containing hypocotyl explants from 1-week-old germinated embryos on MS medium containing
4.0 mg l−1 2,4-D. Embryogenic callus proliferation, somatic embryo (SE) formation, and subsequent plantlet conversion occurred under
optimal culture conditions. The effects of MS medium strength, sucrose, gibberellic acid (GA3), and 6-benzyladenine (BA) on SE formation and plantlet conversion were evaluated. Low MS medium strength (1/4 to 1/2) was
necessary for SE formation, and the optimal sucrose concentration was 2.0%. Supplementing medium with GA3 negatively impacted SE formation and subsequent development. BA significantly increased the number of SEs and the plantlet
conversion capacity. One-third-strength MS medium with 1.0% sucrose and 0.5 mg l−1 BA produced the highest number of SEs (309 embryos from 9 mg embryogenic callus) and the highest frequency of plantlet conversion
from germinated SEs (52.6%). When transplanted to soil, 90% of the regenerated plants developed into normal plants. 相似文献
9.
Mi Jin Jeong Hyun Jin Song Dong Jin Park Ji Yun Min Jin Seong Jo Bo Min Kim Hak Gon Kim Yong Duck Kim Ru Mi Kim Chandrakant S. Karigar Myung Suk Choi 《Plant Cell, Tissue and Organ Culture》2009,98(1):59-65
An efficient plant regeneration protocol for shoot organogenesis from Hovenia dulcis callus cultures was established. Induction of organogenic callus was achieved on Murashige and Skoog (MS) medium supplemented
with 4.65 μM kinetin and 4.5 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Further differentiation of organogenic callus into
primordia, shoot-like structures, and plantlets was achieved on MS medium supplemented with 0.23 μM gibberellic acid (GA3) and 0.46 μM kinetin. Numerous abnormal shoots developed upon transfer of callus to MS medium containing cytokinins, and
these failed to grow further into whole plantlets. However, transfer of ‘abnormal’ shoots to a fresh MS medium lacking cytokinins
resulted in growth of normal shoots. Elongated shoots subsequently were rooted in basal MS medium, and whole plantlets were
established in a soil mix. Analysis of regenerated plants using random amplified polymorphic DNA (RAPD) confirmed the genetic
stability of these regenerant plantlets. 相似文献
10.
Guohua Ma Jaime A. Teixeira da Silva Jinfeng Lü Xinhua Zhang Jietang Zhao 《Plant Cell, Tissue and Organ Culture》2011,105(3):355-361
An efficient propagation and regeneration system via direct shoot organogenesis for an endangered species, Metabriggsia ovalifolia, was established. High activity cytokinins [6-benzyladeneine (BA) and thidiazuron (TDZ)] and low activity auxins [α-naphthaleneacetic
acid (NAA), indole-3-butyric acid (IBA) and indole-3-acetic acid (IAA)] could directly induce adventitious shoots from leaf
or petiole explants within 5 weeks. Cytokinins (TDZ or BA) combined with auxin (NAA) in the induction media induced more adventitious
shoots than when auxins or cytokinins were used alone. Adventitious shoots could be induced and also mass-propagated on media
containing 2.5–5.0 μM TDZ (or BA) and 0.25–0.5 μM NAA. Adventitious roots differentiated at the proximal end of shoots on
rooting media containing half-strength MS salts and 0.5 μM IBA, 0.5 μM NAA, 0.1% activated charcoal or no plant growth regulators.
Over 90% of plantlets survived following acclimatization and transfer to a potting mixture (1:1, sand:vermiculite) in basins. 相似文献
11.
K. Samuel D. Debashish B. Madhumita G. Padmaja Siva Ram Prasad V. Bhaskara Ramana Murthy P. S. Rao 《In vitro cellular & developmental biology. Plant》2009,45(4):466-473
The propagation of Givotia rottleriformis Griff. is difficult as a result of long seed dormancy associated with poor seed germination. The present study was undertaken
to develop a protocol to overcome seed dormancy by culture of zygotic embryo axes and then develop an efficient method for
micropropagation of Givotia. Best germination frequency (78.3%) was achieved from mature zygotic embryo axes isolated from acid-scarified fresh seeds
when cultured on Murashige and Skoog (MS) medium (half-strength major salts) with 28.9 μM gibberellic acid (GA3). Efficient plant conversion was achieved by transfer of 10-d-old germinated embryos to MS medium (half-strength major salts)
supplemented with 1.2 μM kinetin (KN) and 0.5 μM indole-3-butyric acid (IBA). However, acid scarification of 1-yr-old seeds
decreased the germination frequency of zygotic embryo axes in comparison to those obtained from non-acid-scarified seeds which
germinated (96.2%) and converted into plants (80.3%) on MS basal (half-strength major salts) medium. Multiple shoot bud induction
was achieved by culture of shoot tips derived from in vitro germinated seedlings on MS medium with 0.5 μM thidiazuron for 4 wk, and the shoots elongated after transfer to a secondary
medium with 1.2 μM KN. A maximum number of 7.8 shoots per explant with an average shoot length of 3.2 cm was achieved after
two subcultures on this medium. The in vitro regenerated shoots rooted (41.5%) on half-strength MS medium with 0.5 μM IBA. The in vitro generated seedlings and micropropagated plants were established in soil with a survival frequency of 70% and 60%, respectively. 相似文献
12.
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
13.
Summary Suspension cultures initiated from calluses derived from seedling leaf explants of Acacia sinuata (Lour.) Merr. produced somatic
embryos. Embryogenic callus was induced on semisolid MS (Murashige and Skoog, 1962) medium supplemented with 4.52 μM 2,4-dichlorophenoxyacetic
acid (2,4-D) and 2.22 μM 6-benzylaminopurine. A high frequency of somatic embryos was induced in MS liquid medium supplemented
with 4.52 μM 2,4-D and 10% coconut water. Further studies on ontogeny of somatic embryos showed that the cells destined to
become somatic embryos divided into spherical proembryos. Subsequent development led to the formation of globular, heart,
torpedo-shaped and cotyledonary-stage embryos. The conversion of somatic embryos occurred on auxin-free MS medium. Effects
of various auxins, cytokinins, carbohydrates and amino acids in enhancing productin, of somatic embryos were studied. MS medium
supplemented with 87.64 mM sucrose and 342.46 μM glutamine promoted higher somatic embryo production whereas cytokinin had
no effect and led to recallusing of embryos. About 8–10% of embryos converted into plants. 相似文献
14.
Y. W. Kim R. Newton J. Frampton K.-H. Han 《In vitro cellular & developmental biology. Plant》2009,45(4):400-406
Embryogenic suspensor mass (ESM) was established from immature seeds of Fraser fir. The initiation frequency of ESM was dependent
on genotype, collection time, medium, and plant growth regulators (PGR) used. The ESM initiation potential was higher with
seeds collected in late June (clone 16-273, 4.7%) or early July (clone 16-45, 2.2%) and decreased as the zygotic embryos matured.
Excised proembryo stage of zygotic embryos was most appropriate to initiation of ESM. Most of the ESM arose from the seeds
that were at the proembryo stage. From the four different culture media we compared, seven ESM lines were obtained: two lines
from Murashige and Skoog (MS) medium with 4.4 μM benzyladenine (BA), one from Schenk and Hildebrandt (SH) medium with 4.5 μM
thidiazuron (TDZ), and four from SH with 4.4 μM 6-benzyladenine. However, only one ESM line from clone 16-273 (June 24, SH+TDZ)
could be proliferated in subsequent culture. Different concentrations of l-glutamine and casein hydrolysate (CH) in the medium were also compared for their effect on ESM proliferation. The highest
proliferation rate (1.16-fold) was obtained from SH medium supplemented with 250 mg/L CH and 3.42 mM l-glutamine. In contrast, the lowest rate was noted when 1,000 mg/L CH plus 3.42 mM l-glutamine (0.17-fold) was added to the medium. As for somatic embryo maturation, the highest number of mature precotyledonary
(100.1/g−1 FW ESM) or cotyledonary (64.3/g−1 FW ESM) somatic embryos was obtained on a medium containing 20 or 80 μM abscisic acid, 10% polyethyleneglycol, 4% maltose,
and 0.3% gellan gum. For germination of the somatic embryos, the cotyledonary somatic embryos derived from maturation medium
were transferred on half-strength Litvay medium containing 0.3% gellan gum. The somatic plantlets were recovered from the
germination medium and transferred to soils. 相似文献
15.
Md. Mahabubur Rahman Muhammad Nurul Amin Futoshi Ishiguri Shinso Yokota Rubaiyat Sharmin Sultana Yuya Takashima Kazuya Iizuka Nobuo Yoshizawa 《Plant biotechnology reports》2009,3(3):259-266
A plantlet regeneration protocol was developed on pot-grown mature plants of Elaeocarpus robustus Roxb. cv. Dwarf from nodal and leaf explants. The best yield of adventitious shoots was achieved from the leaf-derived calli
in a modified MS (MMS1, half strength of major salts, full strength of minor salts, and vitamins) medium containing 4.0 μM BA + 4.0 μM Kn + 0.5 μM
NAA + 15% coconut water (CW). The shoot multiplication rate was amplified about twofold per culture after the addition of
15% CW to the medium. The rate of shoot multiplication reached maximum at the 5th subculture, and it maintained this rate
throughout the 3 subsequent subcultures. The best rooting in vitro was investigated by subculturing the microcuttings in an
MMS2 (half strength of both major salts and minor salts and full strength of vitamins) medium containing 1.0 μM IBA in the dark
for one initial week at 30°C, followed by subculturing them in a plant-growth regulator (PGR)-free medium in the light. The
plantlets raised in vitro were successfully established under ex vitro conditions. 相似文献
16.
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
17.
In vitro somatic embryogenesis from cell suspension cultures of cowpea [Vigna unguiculata (L.) Walp]
We report, an efficient protocol for plantlet regeneration from the cell suspension cultures of cowpea through somatic embryogenesis. Primary leaf-derived, embryogenic calli initiated in MMS [MS salts (Murashige and Skoog 1962) with B5 (Gamborg et al. 1968) vitamins] medium containing 2,4-Dichlorophenoxyacetic acid (2,4-D), casein hydrolysate (CH), and l-Glutamic acid-5-amide (Gln). Fast-growing embryogenic cell suspensions were established in 0.5 mg l–1 2,4-D, which resulted in the highest recovery of early stages of somatic embryos in liquid MMS medium. Embryo development was asynchronous and strongly influenced by the 2,4-D concentration. Mature monocotyledonary-stage somatic embryos were induced in liquid B5 medium containing 0.1 mg l–1 2,4-D, 20 mg l–1 l-Proline (Pro), 5 M Abscisic acid (ABA), and 2% mannitol. B5 medium was found superior for the maturation of somatic embryos compared to MS and MMS media. The importance of duration (5 d) for effective maturation of somatic embryos is demonstrated. A reduction in the 2,4-D level in suspensions increased the somatic embryo induction and maturation with decreased abnormalities. Sucrose was found to be the best carbon source for callus induction while mannitol for embryo maturation and maltose for embryo germination. Extension of hypocotyls and complete development of plantlet was achieved in half-strength B5 medium supplemented with 3% maltose, 2500 mg l–1 potassium nitrate, and 0.05 mg l–1 thidiazuron (TDZ) with 32% regeneration frequency. Field-established plants were morphologically normal and fertile. This regeneration protocol assures a high frequency of embryo induction, maturation, and plantlet conversion. 相似文献
18.
An efficient micropropagation system via direct shoot organogenesis from hypocotyl segments of Embelia ribes Burm F. was developed. A high frequency (84%) of adventitious shoot induction was obtained on Murashige and Skoog (MS) medium
supplemented with additives (283.85 μM ascorbic acid [AA], 118.96 μM citric acid [CA], 142.33 μM cysteine, and 684.22 μM glutamine)
and 1.13 μM of thidiazuron (TDZ) after 4 weeks following culture. Further development of shoot primordia into well-grown shoots
of 4–5 cm in length was achieved by sub-culturing explants along with shoot primordia on MS medium supplemented with 0.44 μM
benzyl adenine (BA) and 0.49 μM indole butyric acid (IBA) for three sub-culture periods with an interval of 15 days between
them. The highest shoot multiplication was obtained when explants were incubated on MS medium supplemented with 2.2 μM BA
and 0.49 μM IBA in 4 weeks. All in vitro developed shoots, 3–4 cm in length, rooted when grown on half-strength MS basal medium
along with 2.47 μM IBA within 4 weeks. Moreover, 100% of shoots developed roots when these were treated with 4.93 μM IBA for
20 min and then transferred to pots containing soilrite mix and grown in the greenhouse. In vitro and ex vitro rooted plants
showed a survival of 85 and 95% respectively, during hardening in the greenhouse for a 6-week period. 相似文献
19.
T. Dennis Thomas 《In vitro cellular & developmental biology. Plant》2007,43(5):442-448
An efficient and reproducible method for inducing a large number of bulblets from rhizome explants of Curculigo orchioides Gaertn., an endangered medicinal herb, has been developed. The rhizome pieces, measuring about 1 × 1 cm (length × width),
were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of the cytokinins 6-benzylaminopurine,
kinetin, and thidiazuron (TDZ) alone or in combination with 1-naphthalene acetic acid or indole-3-butyric acid (IBA). Of the
three cytokinins used, TDZ at 7 μM gave the maximum response, with 82% of the cultures responding with an average number of
15.4 bulblets per explant. The addition of auxins with cytokinin considerably increased the response. The optimum induction
occurred on MS medium supplemented with 7 μM TDZ and 0.5 μM IBA. On this medium, 88% of the cultures responded with an average
number of 21.4 bulblets per explant. Experiments were also carried out to investigate the role of the sugars sucrose, mannose,
and glucose along with 7 μM TDZ and 0.5 μM IBA. The results indicate that sucrose and mannose at particular concentrations
have critical roles in promoting in vitro bulblet induction. The maximum result was observed on MS medium supplemented with 7 μM TDZ, 0.5 μM IBA, and 200 mM mannose.
On this medium, 97% of the cultures responded with an average number of 26.8 bulblets per culture. Several secondary bulblets
developing from the leaf blades of primary bulblets were produced when the latter were transferred to MS basal medium for
further development. Out of the 45 bulblets transferred to soil, 40 survived. This protocol can be used for the rapid micropropagation
of this endangered medicinal herb. 相似文献
20.
In vitro regeneration of Acacia mangium via organogenesis 总被引:1,自引:0,他引:1
Plant regeneration of Acacia mangium was achieved through organogenesis in callus cultures. Calli were induced from five types of explants (embryo axes and cotyledons
of mature zygotic embryos as well as leaflets, petioles and stems of seedlings) of A. mangium on MS (Murashige and Skoog, 1962) basal medium containing 9.05 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 13.95 μM kinetin (KT). Green or green purple compact nodules containing clusters of meristematic centers were induced
in these calli after transfer to MS basal medium containing 1.14–22.75 μM thidiazuron (TDZ) and 1.43–2.86 μM indole-3-acetic acid (IAA). A combination of 4.55 μM TDZ and 1.43 μM IAA promoted the
highest percentage of calli to form nodules, in 8–11% of calli derived from cotyledons, embryo axes, leaflets or petiole and
in 4% of calli derived from stems. Twenty-two percent of the nodules formed adventitious shoots on MS basal medium containing
0.045 μM TDZ. Shoots were elongated on MS medium containing 0.045 μM TDZ supplemented with 7.22 μM gibberellic acid. The medium
containing 10.75 μM NAA and 2.33 μM KT promoted rooting of 10% of the elongated shoots. Plantlets grew up well in the green house.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献