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1.
Pretreatment in thidiazuron improves the in vitro shoot induction from leaves in Curculigo orchioides Gaertn., an endangered medicinal plant 总被引:1,自引:0,他引:1
T. Dennis Thomas 《Acta Physiologiae Plantarum》2007,29(5):455-461
Leaf regeneration via direct induction of adventitious shoots obtained from an endangered medicinal plant, Curculigo orchioides Gaertn. by pretreating with thidiazuron. C. orchioides is an endangered medicinal herb belonging to the family Hypoxidaceae. Direct inoculation of leaf pieces on MS medium supplemented
with various concentrations of BAP (2–8 μM) or TDZ (2–8 μM) alone or in combination with NAA (0.5 and 1.0 μM) produced low
shoot induction both in terms of % response and number of shoots per explant. Hence, leaf explants were pretreated with 15,
25 or 50 μM thidiazuron (TDZ), for 6, 24 or 48 h with the aim of improving shoot regeneration from cultured explants. After
pretreatment, explants were transferred to an agar solidified MS medium that was supplemented with BAP (4 μM), TDZ (6 μM),
BAP (4 μM) + NAA (1.0 μM), TDZ (6 μM) + NAA (0.5 μM). Control explants were incubated directly on the medium without any pretreatment.
The pretreatment of explants with 15 μM TDZ for 24 h significantly promoted the formation of adventitious shoots and the maximum
response was observed on MS medium supplemented with 6 μM TDZ. In this medium, 96 % cultures responded with an average number
of 16.2 adventitious shoots per explant. The percentage of leaf explants producing shoots and the average number of shoots
per explant were significantly improved when TDZ pretreated leaves were cultured onto MS medium supplemented with BAP or TDZ
alone or in combination with NAA. The rooted plantlets were successfully transplanted to soil with 90% success. The present
investigation indicated the stimulatory role of TDZ pretreatment in regulating shoot regeneration from leaf explants of C. orchioides. 相似文献
2.
Epicotyl, petiole, and cotyledon explants derived from 14-d-old seedlings of Albizia odoratissima were cultured on Murashige and Skoog (MS) basal medium supplemented with different concentrations of either 6-benzylaminopurine
(BAP) solely or in combination with 0.5 μM naphthalene-3-acetic acid (NAA). The percentage of shoot regeneration and the number
of shoots regenerated varied significantly depending on the type of explants used, the concentration of plant growth regulators,
and the orientation of explants on the culture medium. The best response in terms of the percentage of shoot regeneration
was obtained from epicotyls cultured horizontally on MS medium supplemented with 5 μM BAP, whereas the highest number of shoots
per responding explant was recorded on medium containing 2.5 μM BAP and 0.5 μM NAA. Successful rooting was achieved by placing
the microshoots onto MS medium containing 25 μM indole-3-butyric acid (IBA) for 24 h first, then transferring to the same
medium without IBA. Of the various substrates tested, vermiculite was the best for plant acclimatization, as 75% of the plants
survived and became established. 相似文献
3.
L. V. Hiregoudar H. N. Murthy J. G. Bhat A. Nayeem B. P. Hema E. J. Hahn K. Y. Paek 《Biologia Plantarum》2006,50(2):291-294
This report describes in vitro shoot induction and plant regeneration from mature nodal explants of Vitex trifolia L. on Murashige and Skoog (MS) medium fortified with benzylaminopurine (BAP), kinetin (KN), thidiazuron (TDZ), adenine (ADE),
and 2-isopentenyladenine (2-iP) (0.25 – 10.0 μM). Multiple shoots differentiated directly without callus mediation within
3 weeks when explants were cultured on medium supplemented with cytokinins. The maximum number of shoots (9 shoots per explant)
was developed on a medium supplemented with 5.0 μM BAP. Shoot cultures was established repeatedly subculturing the original
nodal explant on the same medium. Rooting of shoots was achieved on half strength MS medium supplemented with 0.5 μM naphthaleneacetic
acid (NAA). Rooted plantlets transferred to pots containing autoclaved soil and vermiculite mixture (1:1) showed 90 % survival
when transferred to outdoor. 相似文献
4.
A simple, high frequency, and reproducible method for plant regeneration through direct organogenesis from cotyledonary leaf
explants of Jatropha curcas was developed using Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ) or 6-benzyl
aminopurine (BAP). Medium containing TDZ has greater influence on regeneration as compared to BAP. The induced shoot buds
were transferred to MS medium containing 10 μM kinetin (Kn), 4.5 μM BAP, and 5.5 μM α-naphthaleneacetic acid (NAA) for shoot
proliferation. The proliferated shoots could be elongated on MS medium supplemented with different concentrations and combinations
of BAP, indole-3-acetic acid (IAA), NAA, and indole-3-butyric acid (IBA). MS medium with 2.25 μM BAP and 8.5 μM IAA was found
to be the best combination for shoot elongation. However, significant differences in plant regeneration and shoot elongation
were observed among the genotypes studied. Rooting was achieved when the basal cut end of elongated shoots were dipped in
half strength MS liquid medium containing different concentrations and combinations of IBA, IAA, and NAA for 4 days, followed
by transfer to growth regulators free half strength MS medium supplemented 0.25 mg l−1 activated charcoal. Elongated shoot treated with 15 μM IBA, 5.7 μM IAA, and 11 μM NAA resulted in highest percent rooting.
The rooted plants could be established in soil with more than 90% survival rate. The method developed may be useful in improvement
of J. curcas through genetic modification. 相似文献
5.
An efficient, rapid and reproducible plant regeneration protocol was successfully developed for Cassia angustifolia using nodal explants excised from 14-day-old aseptic seedlings. Of the two cytokinins, 6-benzyladenine (BA) and thidiazuron
(TDZ) evaluated as supplements to Murashige and Skoog (MS) medium, TDZ at an optimal concentration of 5.0 μM was effective
in inducing multiple shoots. The highest rate of shoot multiplication was achieved on MS medium supplemented with 5.0 μM TDZ
and 1.0 μM indole-3-acetic acid (IAA) at pH 5.8. The regenerated shoots when subcultured on hormone free MS medium considerably
increased the rate of shoot multiplication and shoot length by end of fourth subculture passage. Rooting was achieved on the
isolated shoots using MS medium with 60 μM indole -3- butyric acid (IBA) and 1% activated charcoal for 1 week and subsequently
transferring the shootlets to half strength MS liquid media without IBA and activated charcoal. The in vitro raised plantlets
with well-developed shoot and roots were successfully established in earthen pots containing garden soil and grown in greenhouse. 相似文献
6.
Leaf explants of Jatropha curcas cultured on Murashige and Skoog’s (MS) medium supplemented with thidiazuron (TDZ; 0.90 μM) in combination with indole-3-butyric
acid (IBA; 0.98μM) produced adventitious shoot buds directly on the surface of the explants without formation of intervening
callus while shoot bud formation was accompanied with callus formation on medium supplemented with 6-benzylaminopurine (BAP;
13.3 μM) and IBA (2.46 μM). TDZ treatment resulted in more than twice higher rate of shoot bud induction than BAP. Shoot buds
were multiplied and elongated following repeated transfers to medium containing BAP (2.22 μM) and gibberellic acid (GA3; 1.44 μM). The effect of copper sulphate on differentiation of shoot buds from leaf segments was also investigated. Both
shoot induction and multiplication media were supplemented with different levels of CuSO4 (0–5 μM). Significant improvement in shoot bud induction was observed when the concentration of CuSO4 was increased to 10 times the normal MS level. Healthy elongated shoots were rooted on half strength MS medium supplemented
with IBA (2.46 μM). Rooted plantlets were transferred to field and survived. Histological analysis revealed direct formation
of shoot buds from leaf explants. 相似文献
7.
Dormant buds from a mature tree of Populus tremula ‘Erecta’ were incubated on a Murashige and Skoog (MS) medium supplemented with 1.0 μM thidiazuron (TDZ). Induced shoots were
then proliferated on medium of MS or Woody Plant Medium (WPM), or Driver and Kuniyuki Walnut (DKW) supplemented with varying
levels of benzyladenine (BA). Overall, shoots grown on MS medium supplemented with 1.25–2.5 μM BA exhibited the highest frequency
of shoot proliferation (>95%) and more than 60% of responding explants produced more than five shoots per explant. Shoot organogenesis
was induced from both leaf and petiole explants incubated on WPM medium containing BA, or TDZ, or zeatin. Among the different
cytokinins tested, zeatin induced the highest frequency (average 72.1%) of shoot organogenesis. None of explants survived
on media containing no cytokinins within 6–8 weeks following culture. Overall, a higher frequency of shoot regeneration was
obtained from petioles than from leaf explants. The highest frequency of regeneration was achieved when petioles were incubated
on WPM containing 10–20 μM zeatin. Addition of naphthaleneacetic acid (NAA) did not have a significant effect on shoot regeneration
in all treatments. Shoot organogenesis was directly induced from petiole explants without intervening callus. Regenerated
shoots were easily rooted on all tested media supplemented with 0.5 μM NAA. Rooted plants were transferred to potting mix
and grown in the greenhouse. 相似文献
8.
Perumal Venkatachalam Natesan Geetha Narayanasamypillai Jayabalan 《Journal of Plant Biology》1998,41(1):1-8
This study was designed to evaluate the effect of phytohormones on plant regeneration from epicotyl and hypocotyl explants
of two groundnut (Arachis hypogaea) cultivars. Explants cultured on media with auxins and in combination with cytokinin produced high frequency of callus. After
four weeks, callus from these cultures was transferred to medium with cytokinin and reduced auxin, shoot buds regenerated
from the cultures. A high rate of shoot bud regeneration was observed on medium supplemented with 2.0 mg/L BAP and 0.5 mg/L
NAA. Among the different auxins tested, NAA was found to be most effective, producing the highest frequency of shoot buds
per responding cultures. Of the two explants tested, epicotyl was found to be best for high frequency shoot bud regeneration.
Multiple shoots arose on MS medium supplemented with BAP or kinetin (1.0–5.0 mg/L) plus IBA (1.0 mg/L), with maximum production
occurring at 5.0 mg/L. The elongated shoots developed rootsin vitro upon transfer to MS medium supplemented with NAA or IBA (0.5–2.0 mg/L) and kinetin (0.5 mg/L) for 15 days.In vitro produced plantlets, were transferred to soil and placed in a glasshouse developed successfully, matured, and set seeds. 相似文献
9.
Buhara Yucesan Arzu Ucar Turker Ekrem Gurel 《Plant Cell, Tissue and Organ Culture》2007,91(3):243-250
A very efficient and rapid regeneration system via multiple shoot formation was developed for Cichorium intybus L. when leaf explants excised from sterile seedlings were cultured on medium supplemented with different concentrations and
combinations of various plant growth regulators. In a comparison of leaf lamina and petiole explants, lamina explants produced
over three times more shoots than petiole explants, with a mean of 7.5 shoots compared to 2.4. Of the combinations of KIN/IAA,
KIN/NAA, BAP/IAA, or BAP/NAA, 0.5 mg l−1 KIN combined with 0.3 mg l−1 IAA was the most effective, producing a mean of 19.7 shoots per lamina explant while the control treatment involving no plant
growth regulators produced no shoots at all. When either cytokinin was used alone, BAP was found nearly twice more successful
than KIN. However, the most effective treatment of all was the combination of 0.01 mg l−1 TDZ and 1.0 mg l−1 IAA, producing as many as 35.8 shoots per lamina explant. This rate of shoot regeneration is remarkably higher than those
previously reported for C. intybus, most likely due to the highly inductive effect of TDZ, which was tested for the first time in this species. Rooting of the
shoots was readily achieved on medium containing different concentrations of IAA or IBA. IAA was more effective than IBA and
resulted in the highest frequency of shoots that rooted (100%) and mean number of roots per shoot (4.2) when used at 0.5 mg l−1. Hardening off process resulted in a production of more than 80% healthy plantlets. 相似文献
10.
J. Purkayastha T. Sugla A. Paul S. Solleti L. Sahoo 《In vitro cellular & developmental biology. Plant》2008,44(5):442-447
A rapid and efficient method for the large-scale propagation of a highly valuable medicinal plant, Andrographis paniculata Nees, through in vitro culture of nodal explants obtained from 15-d-old aseptic seedling has been developed. High frequency direct shoot proliferation
was induced in nodal explants cultured on Murashige and Skoog’s medium supplemented with 6-benzylaminopurine (BAP). Amongst
the various cytokinins tested (BAP, kinetin, thidiazuron and 2-isopentyl adenine), BAP proved to be the most effective. The
shoot forming capacity of the nodal explants was influenced by the BAP concentration (1–12.5 μM), and the optimal response
was observed at 10 μM BAP, which induced an average of 34 shoots in 94% of the cultures within 4 wk. Significant differences
were recorded in terms of average number of shoots per explant (8.6–34.1) among the different concentrations of BAP investigated.
Concentrations of all cytokinins tested reach a level that can be considered above the optimum level, as marked by a reduced
frequency of shoot proliferation. The multiple shoots obtained on various concentrations of BAP failed to elongate even after
transfer to hormone-free MS medium. Elongation of the induced shoots was achieved on MS basal medium supplemented with 1.0 μM
GA3 within 2 wk. A proliferating shoot culture was established by repeatedly subculturing the original nodal explants on shoot
multiplication medium after each harvest of the newly formed shoots. The explants retained their morphogenic potential even
after three harvests. Therefore, in 90 d, about 60–70 shoots were obtained from a single nodal explant and the nodal explants
from primary shoots further regenerated equivalent number of shoots, depicting their high frequency regeneration potential
in A. paniculata. Rooting was best induced in 94% of shoots cultured on MS medium supplemented with 2.5 μM indole-3-butyric acid (IBA), within
a wk. The plantlets were successfully transferred to soil after hardening with a 92% survival rate. The system is rapid: the
initiation of shoot buds to the transplanting of regenerants to soil is completed in 8–9 wk. 相似文献
11.
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
12.
Landi Sun Suiwen Hou Dali Wu Yingcong Zhang 《In vitro cellular & developmental biology. Plant》2008,44(5):396-400
This study describes a reliable protocol for callus induction and rapid mass propagation of the ecologically important plant,
Zygophyllum xanthoxylon (Bunge) Maxim. The optimum callus induction medium was Murashige and Skoog (MS) supplemented with 4.4 μM 6-benzylaminopurine
(BAP) and 2.7 μM α-naphthalene–acetic acid (NAA), on which the callus induction frequencies from different seedling explants
were all 100%. However, seedling-derived callus did not form regenerated shoots. In order to achieve shoot multiplication,
shoots were developed from cultured plumules, at an average of 3.1 shoots per explant, and the regenerated shoot tips were
further multiplied by subculture. The best shoot multiplication from shoot tips was achieved on MS supplemented with 5.4 μM
NAA and 22.2 μM BAP after 40 d of culture. Seventy-three percent of regenerated shoots formed roots when cultured on MS supplemented
with 8.6 μM IAA after 4 wk of culture. The plants that acclimatized successfully in sand flourished the following year, with
normal morphology and growth characteristics. 相似文献
13.
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
14.
Multiple shoots were induced by culturing nodal explants excised from 1-month-old aseptic seedlings of red pepper (Capsicum annuum L. cv. Pusa Jwala) on Murashige and Skoog (MS) medium supplemented with (0.1–10 μM) thidiazuron (TDZ). The rate of multiple
shoot induction per explant was maximum (14.4 ± 0.06) on MS medium supplemented with 1.0 μM TDZ. Regenerated shoots were elongated
well on growth regulator free MS medium. Adventitious roots were induced two weeks after transfer of elongated shoots to MS
medium supplemented with auxins (IAA, IBA or NAA) in different concentrations. Optimum root formation frequency was obtained
in medium containing 1.0 μM IBA. Ex-vitro rooting was also achieved by pulse treatment with 300 μM IBA for 10 min. Rooted shoots were transplanted in plastic pots
containing garden soil (with 90 % survival rate), where they grew well and attained maturity. Regenerated plants were phenotypically
and cytologically normal. 相似文献
15.
G. G Ning S. P Bai M. Z Bao L. Liu 《In vitro cellular & developmental biology. Plant》2007,43(2):95-100
Using immature embryos and cotyledons as explants, a successful system to culture immature embryos and induce direct regeneration
from cotyledons was established for Prunus mume “Xuemei”. For immature embryo culture, a high frequency of plantlet formation (89.5%) from the embryonic axis was obtained
using half-strength Murashige and Skoog (1/2 MS) medium supplemented with 13.2 μM 6-benzyladenine (BA) and 2.7 μM 1-naphthaleneacetic
(NAA). Shoots formed directly from cotyledons with the embryo axis intact when explants were cultured on 1/2 MS medium containing
2.2 μM BA with different combinations of NAA (2.7, 5.4 μM) and indole-3-butyric acid (IBA) (0, 2.5, 5.0 μM). Better results
were achieved when the embryonic axis was removed from the cotyledons and cultured on 1/2 MS medium supplement with 13.2 μM
BA, 2.7 μM NAA or 2.2 μM BA, 2.2 μM thidiazuron (TDZ), and 2.7 μM NAA, respectively. Regenerated shoots were successfully
rooted on 1/2 MS or Woody Plant medium (WPM) supplemented with 2.5–5.0 μM IBA. The effect of the embryonic axis, BA, and TDZ
on cotyledon regeneration was investigated in detail. Rooted plantlets were transferred to soil successfully. 相似文献
16.
This report deals with micropropagation of the critically endangered and endemic Turkish shrub, Thermopsis turcica using callus, root and cotyledonary explants. Callus cultures were initiated from root and cotyledon explants on MS medium
supplemented with 0.5–20 μM NAA or 2,4-D. The root explants were found to be better in terms of quick responding and callusing
percentages as compared to the cotyledons. Organogenic callus production with adventitious roots and shoots were obtained
on MS medium with only NAA. The calli obtained with NAA, root and cotyledonary explants were cultured with BA and kinetin
(2–8 μM) alone or in combination with a low level (0.5 μM) of 2,4-D or NAA. The best regeneration of shoots from root explants
was observed on hormone-free MS medium. NAA with BA or kinetin in the medium improved shoot induction from the calli obtained
with NAA. Maximum percentage of shoots (93.3%), maximum number of shoots (6.2) and maximun length of shoots (8.22 cm) were
achieved from cotyledonary explants at 4 μM BA and 0.5 μM NAA. The presence of 0.5 μM or higher levels of 2,4-D in shoot induction
medium inhibited the regeneration in T. turcica explants. 83% of in vitro rooting was attained on pulsed-IBA treated shoots. The regenerated plants with well developed shoots and roots were successfully
acclimatized. Application of this study’s results has the potential to conserve T. turcica from extinction. 相似文献
17.
Wenhao Dai Yuanjie Su Cielo Castillo Olivier Beslot 《Plant Cell, Tissue and Organ Culture》2011,104(2):257-262
Shoots were regenerated from in vitro leaf tissues of two genotypes of Viburnum dentatum, a popular shrub species for landscape use. Adventitious shoots were induced when leaf tissues were cultured on woody plant
medium (WPM) supplemented with either benzyladenine (BA) or thidiazuron (TDZ). Effects of cytokinin concentration, indole-3-butyric
acid (IBA), and dark treatment on shoot regeneration were investigated. Dark treatment for the first 4 weeks of leaf explants
cultured in the regeneration medium significantly increased the frequency of regeneration. The highest frequency of shoot
regeneration (70%) for ‘Synnesvedt’ was obtained when leaf tissues were cultured in the medium with 40 μM BA or 8 μM TDZ with
4 weeks dark treatment. The highest frequency of shoot regeneration (90%) for ‘MN34’ was found in the 4 μM TDZ medium with
4 weeks dark treatment. Addition of IBA significantly enhanced shoot regeneration. Ethyl methanesulfonate (EMS) treatment
inhibited callus proliferation, particularly in the early stage of callus recovery; however, no significant difference in
shoot regeneration among different treatments was observed, indicating that the inhibitory effect of EMS was minimal after
calluses re-acquired their capacity to grow and regenerate in the regular medium. Regenerated shoots (>1.5 cm) were rooted
in the half-strength MS medium containing 5-10 μM IBA or naphthalene acetic acid (NAA). Rooted plants were transferred to
the potting medium and grown in the greenhouse. 相似文献
18.
An efficient protocol is described for the rapid in vitro multiplication of an endangered medicinal plant, Tylophora indica (Burm. f.) Merrill, via enhanced axillary bud proliferation from nodal explants collected from young shoots of a two-year-old
plant. The physiological effects of growth regulators [6-benzyladenine (BA), kinetin (Kin) thidiazuron (TDZ), indole-3-acetic
acid (IAA), indole-3-butyric acid (IBA) or α-naphthalene acetic acid (NAA)], ascorbic acid (AA), different strengths of Murashige
and Skoog (MS) medium and various pH levels on in vitro morphogenesis were investigated. The highest number (8.6 ± 0.71) of
shoots and the maximum average shoot length (5.2 ± 0.31 cm) were recorded on MS medium supplemented with 2.5 μM BA, 0.5 μM
NAA and 100 mg/l AA at pH 5.8. Rooting was best achieved on half-strength MS medium augmented with 0.5 μM IBA. The plantlets
regenerated in vitro with well-developed shoot and roots were successfully established in pots containing garden soil and
grown in a greenhouse with a 90% survival rate. The regenerated plants did not show any immediate detectable phenotypic variation.
The described method can be successfully employed for large-scale multiplication and long-term in vitro conservation of T. indica. 相似文献
19.
An efficient and reproducible method for the regeneration of Jatropha curcas plants has been developed. The method employed direct induction of shoot buds from petiole explants, without the formation of an intervening callus using a Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ). The best induction of shoot buds (58.35%) and the number of shoot buds per explant (10.10) were observed when in vitro petiole explants were placed horizontally on MS medium supplemented with 2.27 µM TDZ after 6 weeks. The induced shoot buds were transferred to MS medium containing 10 µM kinetin (Kn), 4.5 µM 6-benzyl aminopurine (BAP) and 5.5 µM α-naphthaleneacetic acid (NAA) for shoot proliferation. The proliferated shoots could be elongated on MS medium supplemented with different concentrations and combinations of BAP, indole-3-acetic acid (IAA), NAA and indole-3-butyric acid (IBA). MS medium supplemented with 2.25 µM BAP and 8.5 µM IAA was found to be the best combination for shoot elongation and 3.01–3.91 cm elongation was achieved after 6 weeks. However, significant differences in plant regeneration and shoot elongation were observed among the genotypes studied. The orientation (horizontal or vertical) and source (in vitro or in vivo) of explants also significantly influenced plant regeneration. The elongated shoots could be rooted on half-strength MS medium supplemented with 2% sucrose, different concentrations and combinations of IBA, IAA and NAA, and 0.25 mg L−1 activated charcoal. Half-strength MS medium supplemented with 2% sucrose, 15 µM IBA, 5.7 µM IAA, 5.5 µM NAA and 0.25 mg L−1 activated charcoal was found to be the best for promoting rooting. The rooted plants could be established in soil with more than 90% survival. 相似文献
20.
A simple, high-frequency and reproducible protocol for induction of adventitious shoot buds and plant regeneration from leaf-disc
cultures of Jatropha curcas L. has been developed. Adventitious shoot buds were induced from very young leaf explants of in vitro germinated seedlings
as well as mature field-grown plants cultured on Murashige and Skoog’s (MS) medium supplemented with thidiazuron (TDZ) (2.27 μM),
6-benzylaminopurine (BA) (2.22 μM) and indole-3-butyric acid (IBA) (0.49 μM). The presence of TDZ in the induction medium
has greater influence on the induction of adventitious shoot buds, whereas BA in the absence of TDZ promoted callus induction
rather than shoot buds. Induced shoot buds were multiplied and elongated into shoots following transfer to the MS medium supplemented
with BA (4.44 μM), kinetin (Kn) (2.33 μM), indole-3-acetic acid (IAA) (1.43 μM), and gibberellic acid (GA3) (0.72 μM). Well-developed shoots were rooted on MS medium supplemented with IBA (0.5 μM) after 30 days. Regenerated plants
after 2 months of acclimatization were successfully transferred to the field without visible morphological variation. This
protocol might find use in mass production of true-to-type plants and in production of transgenic plants through Agrobacterium/biolistic-mediated transformation. 相似文献