共查询到20条相似文献,搜索用时 62 毫秒
1.
Xiaohuan Wang Zhenhua Gao Yunzhen Wang Ray A. Bressan Stephen C. Weller Xia Li 《In vitro cellular & developmental biology. Plant》2009,45(4):435-440
An in vitro regeneration system with a 100% efficiency rate was developed in peppermint [Mentha x piperita] using 5- to 7-mm-long second internode stem segments of 3-wk-old stock plants. Shoots developed at sites of excision on
stem fragments either directly from the cells or via primary calluses. The optimal medium for maximum shoot initiation and
regeneration contained Murashige and Skoog (MS) salts, B5 vitamins, thidiazuron (TDZ, 11.35 μM), ZT (4.54 μM), 10% coconut
water (CW), 20 g l−1 sucrose, 0.75% agar, adjusted to pH 5.8. A frequency of 100% shoot initiation was achieved, with an average of 39 shoots
per explant. This regeneration system is highly reproducible. The regenerated plants developed normally and were phenotypically
similar to Black Mitcham parents. 相似文献
2.
A reproducible protocol for clonal propagation of Spilanthes acmella has been established. Routinely, the cultures were established in spring (January–April) season because of the highest aseptic
culture establishment and high frequency shoot proliferation. Incorporation of 5 μM N6-benzyladenine (BA) to Murashige and Skoog (MS) basal medium showed 100% bud-break and promoted multiple shoot proliferation
in cultures. Interestingly, a higher concentration of BA (7–15 μM) promoted stunted shoots with pale leaves while a lower
concentration (1–3 μM) resulted in shoots with long internodes and excessive adventitious root proliferation from all over
their surface. For recurrent shoot multiplication, single node segments from in vitro-developed shoots were excised and cultured
on MS + BA (5 μM) medium where 20.3-fold shoot multiplication was achieved every 5 weeks. Finally, these shoots were successfully
rooted on half-strength MS medium (major salts reduced to half-strength) with 50 g l−1 sucrose, with a frequency of 100%. Transplantation survival of micropropagated plants was 88.9%. Additionally, accumulation
of scopoletin, a phytoalexin, was revealed for the first time in the uninfected leaves of Spilanthes. Further, the quantitative estimation by HPLC with a fluorescence detector showed that the amounts of scopoletin content
(0.10 μg g−1 DW) in the leaves of micropropagated plants are comparable to those of field-grown mother plants. The study thus signifies
the effectiveness of in vitro methodology for true-to-type plant regeneration of Spilanthes and their later utility for biosynthesis and constant production of scopoletin throughout the year. 相似文献
3.
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
4.
The effect of plant growth regulators (PGRs), gelling agents, sucrose and heat shock on shoot multiplication, shoot growth,
rooting and subsequent survival of Chlorophytum borivilianum Sant. et Fernand was evaluated. Benzyladenine (BA) was found to be better cytokinin over kinetin (KIN) for shoot multiplication. Sucrose
concentrations from 116–290 mM in the basal medium (BM) promoted shoot multiplication. Heat shock (50 °C, 1 h) also promoted
shoot multiplication at these sucrose concentrations on both BM medium and BM supplemented with 5.0 μM BA. Beneficial effect
of sucrose was also observed on rooting of shoots on BM as well as BM supplemented with 5.0 μM indole-3-butyric acid (IBA).
Phytagel as a gelling agent was found to be more effective for shoot proliferation and growth compared to agar. Amongst various soil
mixtures tested, higher survival of plants was observed in soil containing Vermicompost. It was interesting to note that a maximum plant survival (> 95 %) was observed when plants were directly transferred to
net-house (irradiance reduced to 50 % with green net, without humidity and temperature control) than poly-house (with humidity
and temperature control). Random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analysis of regenerated
plants showed genetic similarity to mother plant. 相似文献
5.
B. Kozomara B. Vinterhalter Lj. Radojević D. Vinterhalter 《In vitro cellular & developmental biology. Plant》2008,44(2):142-147
A procedure for the micropropagation of Chimonanthus praecox (L) Link, wintersweet, has been developed using buds from adult trees excised in spring. Shoot cultures established on Murashige
and Skoog (1962) medium supplemented with 0.5 mg l−1 6-benzyladenine (BAP) and 0.1 mg l−1 indole-3-butyric acid (IBA) were difficult to maintain in vitro through extended periods of time due to browning of the medium, shoot and leaf necrosis, and hyperhydricity. A treatment
combining the use of 0.1% w/v activated charcoal and addition of a double phase agar-solidified/liquid medium improved propagation, enabling a successful
in vitro propagation scheme to be developed. Optimal shoot multiplication occurred on medium containing 0.5 mg l−1 BAP, and rooting on medium with 2.0 mg l−1 IBA for 7 d, followed by transfer to hormone-free medium. Rooted plantlets were easily acclimated in a glasshouse and replanted
and cultured outdoors. 相似文献
6.
Federico Antonio Gutiérrez Miceli Adan Domínguez Estudillo Miguel Abud Archila Teresa del Rosario Ayora Talavera Luc Dendooven 《In vitro cellular & developmental biology. Plant》2008,44(1):33-39
Renealmia mexicana (Klotzsch ex. Petersen) is a tropical plant found in southern México with an ornamental value and a potential source of curcuminoids.
Its distribution in Chiapas has decreased because of deforestation and low propagation and germination rate, so a protocol
for in vitro propagation was developed. An orthogonal experimental design of L9 (34) in triplicate was used to investigate the effect of 6-benzyl adenine (BA), indole butyric acid (IBA), silver nitrate (AgNO3), and sucrose on shoot, root, and leaf development of plantlets grown in vitro. Plantlets with well-developed shoots and roots were transferred to pots containing a mixture of peat moss and agrolite for
hardening before transfer to soil. The Murashige and Skoog (Physiol. Plant. 15:473–497, 1962) mineral medium (MS) supplemented
with 4.4 μM BA, 2.5 μM IBA, 11.7 μM AgNO3y and 5.5% (w/v) sucrose gave most shoots, 8.9 μM BA, 2.5 μM IBA, 17.7 μM AgNO3 and 5.5% (w/v) sucrose most roots, and 8.9 μM BA, 4.9 μM IBA, 11.7 μM AgNO3 and 3.0% (w/v) sucrose most leaves, although other combinations were statistically equivalent in each case. Sucrose was the factor that
most explained the variation in the promotion of shoots, roots, and leaves. The protocol developed resulted in up to 100%
survival when plantlets were transferred to soil using AgNO3, confirming that hardening of plantlets in vitro using hormonal stimulation was a suitable strategy to improve acclimatization. 相似文献
7.
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
8.
Summary We describe an in vitro propagation protocol for Zingiber petiolatum (Holttum), I. Theilade, a rare species from the southern part of Thailand. Fruits were surface-sterilized and seeds germinated
on Murashige and Skoog medium (MS) medium supplemented with 3% sucrose. Three-month-old seedlings were used as initial plant
material for in vitro propagation. Terminal buds of the plants were inoculated on MS medium containing 6-benzylaminopurine (BA; 2.2–35.5 μM) alone or in combination with 1-naphthaleneacetic acid (0.5 μM). Eight weeks after inoculation, the cultures were transferred to MS medium without plant growth regulators for 4wk. The
cultures transferred from MS medium with 17.8 μM BA revealed the highest shoot induction rate of 6.1±0.7 shoots per explant. Rooting was spontaneously achieved in MS medium
without plant growth regulators. Rooted plants were successfully transplanted to soil. 相似文献
9.
Diwakar Aggarwal Anil Kumar M. Sudhakara Reddy 《Plant Cell, Tissue and Organ Culture》2010,102(1):45-52
An efficient shoot organogenesis system has been developed from mature plants of selected elite clones of Eucalyptus tereticornis Sm. Cultures were established using nodal explants taken from freshly coppice shoots cultured on Murashige and Skoog medium
containing 58 mM sucrose, 0.7% (w/v) agar (MS medium) and supplemented with 2.5 μM benzyladenine (BA) and 0.5 μM α-naphthaleneacetic
acid (NAA). Shoot organogenesis was achieved from leaf segments taken from elongated microshoots on MS medium supplemented
with 5.0 μM BA and 1.0 μM 2,4-dichlorophenoxyacetic acid (2,4-D). The addition of cefotaxime to the medium promoted shoot
differentiation, whereas carbenicillin and cephalexin inhibited shoot differentiation. Maximum shoot bud organogenesis (44.6%)
occurred in explants cultured on MS medium supplemented with 5.0 μM BA, 1.0 μM 2,4-D and 500 mg/l cefotaxime. Leaf maturity
influenced shoot regeneration, with maximum shoot organogeneisis (40.5%) occurring when the source of explants was the fifth
leaf (14–16 days old) from the top of microshoot. Shoot organogenic potential also varied amongst the different clones of
E. tereticornis. Random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) analyses indicated clonal uniformity of
the newly formed shoots/plants, and these were also found to be true-to-type. 相似文献
10.
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
11.
Rishi K. Tyagi Rajni Goswami Rajkumari Sanayaima Rakesh Singh Rajesh Tandon Anuradha Agrawal 《In vitro cellular & developmental biology. Plant》2009,45(6):721-729
Cardamom (Elettaria cardamomum Maton) has great commercial value as a spice crop in India. A one-step protocol for direct regeneration of plants and in vitro conservation by slow growth method has been developed. A maximum of 6.5 shoots/culture were obtained in 2 mo or 15.1 shoots/culture
in 4 mo on Murashige and Skoog (Physiol Plant 15:473–497, 1962) medium (MS) + 5 μM benzylaminopurine gelled with 0.7% agar (micropropagation medium). Rooting also occurred simultaneously
on the same medium. Using one shoot tip or nodal explant, about 30,375 plants can be regenerated in a year on the micropropagation
medium. In vitro conservation by slow growth method was achieved on 1/2 MS (major salts) + 5 μM BAP + 0.7% agar (conservation medium); about
70% of the cultures survived up to 18 mo at 25 ± 2°C. Successful regrowth of plants on micropropagation medium was obtained
by culturing nodal explants excised from 18-mo-old conserved plants. Some 96% of the plants survived the hardening treatment
and grew normally in a greenhouse. If 24 cultures are conserved on the conservation medium, it is possible to regenerate at
least 750 plants by using explants derived from 70% of the surviving shoots and culturing the same in micropropagation medium
for 4 mo. These plants may be used for planting or as a source of explants for the next conservation cycle. On the basis of
20 random amplified polymorphic DNA and 13 inter-simple sequence repeat primers analyses, no significant reproducible variation
was detected among the in vitro-conserved plants compared with the mother plants. 相似文献
12.
Branka Vinterhalter Dijana Krstić Milošević Teodora Janković Jelena Milojević Dragan Vinterhalter 《Central European Journal of Biology》2012,7(4):690-697
Gentiana dinarica Beck, rare and endangered species of Balkan Dinaric alps, was in vitro propagated (micropropagated) from axillary buds of plants collected at Mt. Tara, Serbia. G. dinarica preferred MS to WPM medium, with optimal shoot multiplication on MS medium with 3% sucrose, 1.0 mg l−1 BA and 0.1 mg l−1 NAA. Rooting was not clearly separated from shoot multiplication since BA did not completely inhibit root initiation. Spontaneous
rooting on plant growth regulator-free medium occurred in some 30% of shoot explants. Rooting was stimulated mostly by decreased
mineral salt nutrition and a medium with 0.5 MS salts, 2% sucrose and 0.5–1.0 mg l−1 IBA was considered to be optimal for rooting. Rooted plantlets were successfully acclimated and further cultured in peat-based
substrate. 相似文献
13.
Diane E. Darlington Chiu-Yueh Hung Jiahua Xie 《Plant Cell, Tissue and Organ Culture》2009,99(2):157-165
Agrobacterium
tumefaciens strain LBA4404 containing the plasmid pBI121, carrying the reporter gene uidA and the kanamycin resistance gene nptII, was used for gene transfer experiments in selenium (Se)-hyperaccumulator Astragalus racemosus. The effects of kanamycin on cell growth and division and acetosyringone on transformation efficiency were evaluated. The
optimal concentration of kanamycin that could effectively inhibit cell growth and division in non-transgenic tissues was 50 mg l−1 and thus all putative transgenic plants were obtained on induction medium containing 50 mg l−1 kanamycin. The verification of transformants was achieved by both histochemical GUS assay and PCR amplification of nptII gene. Southern blot analysis was performed to further confirm that transgene nptII was stably integrated into the A. racemosus genome. A transformation frequency of approximately 10% was achieved using this protocol, but no beneficial effect from the
addition of acetosyringone (50 μM) was observed. This transformation system will be a useful tool for future studies of genes
responsible for Se-accumulation in A. racemosus. 相似文献
14.
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
15.
M. Frabetti P. Gutiérrez-Pesce E. Mendoza-de Gyves E. Rugini 《In vitro cellular & developmental biology. Plant》2009,45(2):129-134
An efficient protocol for in vitro propagation of the valuable ornamental and medicinal plant Bush germander (Teucrium fruticans L.) was developed through axillary shoot proliferation. A Murashige and Skoog agar medium supplemented with benzylaminopurine
(6.6 μM), α-naphthaleneacetic acid (0.053 μM), and sucrose (3%) significantly improved the production of multiple shoots directly
from nodal segment explants, resulting in an average of 2.8 shoots per segment with an average of 6.8 nodes per shoot that
would be potential newly formed explants. The new shoots were developed without a marked decrease in the average height of
the shoots. Shoots treated with 2.5 μM indole-3-butyric acid showed the highest average root number (7.9) and the highest
percentage of rooting (94%). Plantlets were hardened off and transferred to jiffy pots for acclimatization under greenhouse
conditions, resulting in a 100% survival rate. 相似文献
16.
Summary An efficient in vitro plant regeneration system from cotyledons was established in tetraploid Isatis indigotica Fort. Factors influencing shoot regeneration from cotyledons, including culture medium type, combinations of plant growth
regulators, and sucrose concentrations in the medium, as well as illumination were investigated. Murashige and Skoog's (MS)
medium was found to be best for promoting shoot regeneration, followed by Gamborg's B5 and White's medium. The highest shoot regeneration frequency was achieved from cotyledons cultured on MS medium supplemented
with 2.0 mgl−1 (8.9 μM) 6-benzyladenine and 1.0 mgl−1 (5.4 μM) α-naphthaleneacetic acid (NAA), with 97.9% regeneration, associated with a high number of multiple shoots developed per
explant (8.6 shoots per explant). A sucrose concentration of 3% present in the medium and light conditions were beneficial
for shoot regeneration. The shoots developed were rooted in a half-strength MS medium supplemented with 1.0 mgl−1 (5.4 μM) NAA and successfully transplanted in soil in pots with over 85% survival. The establishment of an efficient plant regeneration
procedure from cotyledons provides a basis for the rapid in vitro multiplication of tetraploid Isatis indigotica Fort., one of the most extensively used medicinal plants in China currently under great shortage. 相似文献
17.
An efficient, rapid and large scale propagation of a multipurpose herb, Ocimum basilicum through in vitro culture of nodal segments with axillary buds from mature plants has been accomplished. Among the cytokinins,
6-benzyladenine (BA), thidiazuron (TDZ), kinetin (Kin) and 2-isopentenyl adenine (2-iP) tested as supplements to Murashige
and Skoog (MS) medium, 5.0 μM BA was optimum in inducing bud break. The highest rate of shoot multiplication was achieved
on half-strength MS medium supplemented with 2.5 μM BA and 0.5 μM indole-3-acetic acid (IAA) combination. The shoots regenerated
from TDZ supplemented medium when subcultured to hormone-free MS medium considerably increased the rate of shoot multiplication
and shoot length by the end of third subculture. For rooting, MS medium supplemented with 1.0 μM indole-3-butyric acid (IBA)
proved to be better than that supplemented with IAA or α-naphthalene acetic acid (NAA). The in vitro raised plantlets with
well developed shoots and roots were successfully established in earthen pots containing garden soil and were grown in greenhouse
with 90% survival rate. Chlorophyll a and b, carotenoids and net photosynthetic rate were measured in leaves during ex vitro
acclimatization at 0, 7, 14, 21 and 28 days. Firstly these parameters showed a decreasing trend but subsequently increased
after 7 days of acclimatization. These findings indicate that the adaptation of micropropagated plants to ex vitro conditions
is more extended in time than generally accepted. 相似文献
18.
Hypocotyls explants of 1-week-old Ruta gravoelens L. seedlings showed high regeneration ability on Murashige and Skoog’s medium supplemented with 5.2 mM H2PO4
−, 250 mM adenine sulphate, 3.0% sucrose and 8.87 μM BA. Increasing phosphate and adenine sulphate concentrations had pronounced
effects on organogenesis and shoot growth compared to the stander MS medium. Multiple shoots were successfully maintained
for 60 days in static liquid medium without any hyperhydricity phenomena. Uncountable shoot buds and shoots were developed
on the leaf surface which was in contact with the medium. The growth index increased linearly during the culture course (R
2 = 0.992–0.999) and the highest GI (30.43) was achieved on medium enriched with 90 mM nitrogen and 4.5% sucrose. Medium supplemented
with 6.0% sucrose and 60 mM N yielded the highest biomass (4.37 g per flask). However, medium with the stander N content and
6.0% sucrose developed the maximum percentage of biomass (6.67%). The highest percentage of furanocoumarins (11.46 mg g−1 dry weight) calculated as xanthotoxin was produced in medium enriched with 90 mM N and 4.5 or 6.0% sucrose, whereas the maximum
rutin (73.5 mg g−1 dry weight) was on medium supplemented with 90 mM N and 4.5% sucrose. Developing system seems to be used for secondary metabolites
production because the shoot rooting did not appear. So, all energy is devoted for shoots multiplication, their growth and
secondary metabolites production. 相似文献
19.
Yaser Hassan Dewir Nisha Singh Shakira Shaik Ashley Nicholas 《In vitro cellular & developmental biology. Plant》2010,46(1):41-46
The present study reports a simple protocol for indirect shoot organogenesis and plant regeneration of Sutherlandia using rachis and stem segments. Different concentrations (0.0–68.08 μmol l−1) of thidiazuron (TDZ) were used for callus induction and shoot organogenesis. The highest percentage of callus formation
(97.5%) and the highest percentage of explants forming shoots (88.8%) were obtained from rachis explants cultured onto Murashige
and Skoog (MS) medium (Murashige and Skoog, Physiol. Plant. 15:473–495, 1962) supplemented with 45.41 μmol l−1 TDZ. Scanning electron microscopy demonstrated the early development of adventitious shoots derived from callus cultures.
Shoot clusters were further developed and grown in MS hormone-free medium. The presence of l-canavanine was determined by thin-layer chromatography and confirmed after column fractionation using silica gel and nuclear
magnetic resonance spectroscopy. Individual shoots were rooted on different concentrations and combinations of MS salt strength
and IBA. Half-strength MS salt medium supplemented with 24.6 μmol l−1 IBA was optimal for root induction in which 78% of shoots were rooted. The in vitro plants were successfully acclimatized in a growth chamber with a 90% survival rate. 相似文献
20.
Sushma Tamta Lok Man S. Palni Vijay K. Purohit Shyamal K. Nandi 《In vitro cellular & developmental biology. Plant》2008,44(2):136-141
An efficient and reproducible method for the regeneration of multiple shoots of brown oak (Quercus semecarpifolia Sm.) has been developed in which a part of the petiolar tube containing a primary shoot is used as the explant. Explants
derived from in vitro grown seedlings were cultured either on Murashige and Skoog or Woody Plant medium (WPM) containing different concentrations
of benzyladenine (BAP) throughout the range of 1–20 μM. WPM supplemented with 20 μM BAP was found to be best for adventitious
shoot induction and for the multiplication of individual shoots. In-vitro-produced shoots were rooted using a two-step method. Firstly, shoots were cultured on WPM containing indolebutyric acid (IBA)
at either 50 or 100 μM for 24 or 48 h. Secondly, the shoots were transferred to plant-growth-regulator-free half-strength
WPM. The second step not only considerably improved the rooting percentage but also minimized the formation of basal callus.
The most effective first-step treatment was found to be 100 μM IBA for 24 h, which initiated rooting at a frequency of 100%.
Well-rooted plants were transferred to plastic cups containing nonsterile, sieved soil and farmyard manure, hardened under
greenhouse conditions, and then successfully established in pots. This procedure is suitable for use in large-scale production
of plants and may have potential application in additional oak species. 相似文献