共查询到20条相似文献,搜索用时 15 毫秒
1.
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
2.
Jianming Gao Jinzhi Li Caibo Luo Liang Yin Shengchun Li Guangxiao Yang Guangyuan He 《Molecular biology reports》2011,38(2):1413-1417
To establish a successful in vitro plant regeneration system in Alternanthera philoxeroides (Mart.) Griseb, an orthogonal design was used to investigate the effects of three factors (plant growth regulators, explant
types and dark treatment in initial-stage), each having three levels. The effects of these factors and levels on callus induction
and shoot regeneration were quantitatively evaluated by analysis of variance. The experimental results showed that the callus
induction was significantly affected by 2, 4-dichlorophenoxyacetic acid (2, 4-D), and shoot differentiation from subcultured
pieces of callus was enhanced mostly by dark treatment in initial-stage. The optimal conditions for callus induction are obtained
from the stem explants cultured on semi-solid Murashige and Skoog (MS) medium plus 2.2 μM BA and 2.2 μM 2, 4-D, with 20 days
dark treatment in initial-stage. The highest frequency of shoot regeneration is obtained from the calli cultured on semi-solid
MS medium plus 8.8 μM BA, without dark treatment in initial-stage. 相似文献
3.
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
4.
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
5.
A novel protocol for callus-mediated shoot regeneration was established for an important medicinal and ornamental plant native
to South China, Curcuma kwangsiensis, using shoot base sections excised from seedlings in vitro as explant sources. The frequency of callus formation reached
91% for explants cultured on MS medium containing 1.4 μM TDZ, 4.4 μM BA and 2.3 μM 2,4-D. 8.2 shoots per callus was achieved
on MS medium supplemented with 1.4 μM TDZ, 17.8 μM BA and 2.7 μM NAA. Single shoots transferred into MS medium free of plant
growth regulator rooted well. Regenerated plants acclimatized ex vitro at 100%, and grew vigorously under shaded greenhouse
conditions. 相似文献
6.
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
7.
Attempts were made to study the effect of thidiazuron (TDZ) on adventitious shoot induction and plant development in Paulownia tomentosa explants derived from mature trees. Media with different concentrations of TDZ in combination with an auxin were used to
induce adventitious shoot-buds in two explant types: basal leaf halves with the petiole attached (leaf explant) and intact
petioles. Optimal shoot regeneration was obtained in leaf explants cultured on induction medium containing TDZ (22.7 or 27.3 μM)
in combination with 2.9 μM indole-3-acetic acid (IAA) for 2 weeks, and subsequent culture in TDZ-free shoot development medium
including 0.44 μM BA for a further 4-week period. The addition of IAA to the TDZ induction medium enhanced the shoot-forming
capacity of explants. The caulogenic response varied significantly with the position of the explant along the shoot axis.
The highest regeneration potential (85–87%) and shoot number (up to 17.6 shoots/explant) were obtained in leaf explants harvested
from the most apical node exhibiting unfolded leaves (node 1). An analogous trend was also observed in intact petiole explants,
although shoot regeneration ability was considerably lower, with values ranging from 15% for petioles isolated from node 1
to 5% for those of nodes 2 and 3. Shoot formation capacity was influenced by the genotype, with regeneration frequencies ranging
from 50% to 70%. It was possible to root elongated shoots (20 mm) in basal medium without growth regulators; however, rooting
frequency was significantly increased up to 90% by a 7-day treatment with 0.5 μM indole-3-butyric acid, regardless of the
previous culture period in shoot development medium (4 or 8 weeks). Shoot quality of rooted plantlets was improved not only
by IBA treatment but also by using material derived from the 4-week culture period. Regenerated plantlets were successfully
acclimatized in the greenhouse 8 weeks after transplanting. 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.
Azza A. Tawfik Mohamed F. Mohamed 《In vitro cellular & developmental biology. Plant》2007,43(1):21-27
Nodular meristematic callus was induced on the basal cut surface of apical shoot explants of salvia cultured on Murashige
and Skoog (MS) medium supplemented with 4.5, 13.5, or 22.5 μM thidiazuron (TDZ). Cultures were incubated in the dark for 1 wk
and then transferred to light conditions for 4 wk. A higher percentage of explants developing callus was observed on medium
containing either 4.5 or 13.5 μM TDZ, although explants on 4.5 μM developed larger calluses. The callus was maintained on
medium containing 4.5 μM TDZ and 0.45 mM ascorbic acid. Shoot differentiation, after each of three successive maintenance
passages, was induced from callus grown on medium containing either 4.4 or 8.8 μM benzyladenine (BA). A greater number of
shoots were harvested from callus differentiated on BA (4.4 or 8.8 μM) medium with 0.45 mM ascorbic acid added. Shoots developed
roots on MS medium supplemented with 4.9 μM of indole-3-butyric acid. The addition of ascorbic acid to the shoot differentiation
medium enhanced rooting, number of roots per shoot, and survival rate. Approximately 75% in vitro plantlets were acclimatized to ex vitro conditions. Histological investigations confirmed both adventitious meristem initiation during the callus induction phase,
and subsequent organogenic shoot development on the differentiation medium. The novel protocol for the meristematic callus
induction and plant regeneration in this study may be useful for biotechnological applications for salvia improvement via
genetic transformation or mutagenesis and in vitro propagation approaches. 相似文献
10.
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
11.
Summary Protocols for both axillary bud proliferation and shoot organogenesis of Euphorbia pulchurrima Winter RoseTM were developed using terminal buds and leaf tissues. Greenhouse-grown terminal buds were placed on Murashige-Skoog (MS) basal
medium supplemented with various concentrations of either benzlyaminopurine (BA) or thidiazuron (TDZ). Explants produced the
greatest number of axillary buds on media containing between 2.2 and 8.8 μM BA. The number of explants that produced axillary buds increased with increasing BA concentration. TDZ at concentrations
between 2.3 and 23.0 μM caused hyperhydricity of shoots and were not effective in promoting shoot proliferation. The most calluses and shoots were
produced from leaf midvein sections from in vitro grown plants placed on the medium containing 8.8–13.3 μM BA and 17.1 μM indole-3-acetic acid (IAA) for 1 mo. before transferring to the medium containing only BA. Adventitious buds were produced
only from red-pigmented callus, and explants that produced callus continued to produce adventitious shoots in the presence
of IAA. Five-mo.-old shoots derived from shoot culture or organogenesis rooted readily in artificial soil with or without
treatment with indolebutyric acid, and were acclimatized in the greenhouse. 相似文献
12.
Nodular callus was induced at a high frequency on young purple red, 5–15 mm long laminae taken from in vitro grown plants of mangosteen. The optimal medium was composed of Murashige and Skoog (MS) nutrients supplemented with 2.22
μM benzyladenine (BA), 2.25 μM thidiazuron (TDZ), 500 mg l-1 polyvinylpyrrolidone (PVP 360 000) and 3% sucrose. A multiplication rate of two–three was obtained by subculture of the nodular
callus at 3–4-week intervals. Plantlet regeneration from the nodules was achieved by transfer to woody plant medium (WPM)
with 500 mg l-1 PVP, 0.4 μM BA and 3% sucrose and overlaying with half strength liquid MS containing 0.32 μM naphthaleneacetic acid (NAA),
0.13 μM BA and 3% sucrose. Elongated shoots were rooted to 100% when wounded at the base of shoot, dipped in 4.4 mM indolebutyric
acid (IBA) solution in the dark for 15 min and cultured on WPM supplemented with 1.11 μM BA, 0.25% activated charcoal, 34.5
μM phloroglucinol (PG) and 3% sucrose.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
13.
Blanca P. Martínez-Bonfil Guadalupe Salcedo-Morales Alma R. López-Laredo Elsa Ventura-Zapata Silvia Evangelista-Lozano Gabriela Trejo-Tapia 《Plant Cell, Tissue and Organ Culture》2011,107(2):195-203
Castilleja tenuiflora is a medicinal plant that grows in pine–oak woods primarily in southern and central Mexico. It is highly valued for its medicinal
properties, which have been attributed to aucubin-like iridoids. In the present study, we developed an efficient protocol
for in vitro shoot proliferation and ex vitro rooting of C. tenuiflora. Using a colorimetric method, we determined total iridoid contents of various different tissues of propagated plants. The
shoots were induced from nodal explants cultured on Murashige and Skoog (MS) (1962) medium supplemented with indole-3-butyric
acid (IBA) (0 and 0.5 μM) and different concentrations of thidiazuron (TDZ), 6-benzyladenine (BA), or kinetin (KIN) (0–20 μM).
Of the cytokinins tested, KIN was more effective for shoot induction than TDZ or BA, and the highest shoot proliferation rate
was achieved with 5 μM KIN (4 shoots per explant). Plantlets were rooted on MS medium, nutrient solution, or potting mix,
alone or in combination with auxins. The best responses (100% rooting efficiency) were obtained by dipping shoots in half-strength
MS medium containing 7.5 μM IBA before transfer to potting mix. On average, each shoot formed 9 roots of 39.3 ± 3.8 mm in
length after 21 days. These roots appeared to be more functional than those that developed in nutrient solution, and were
associated with a high survival rate (95%) during acclimatization and cultivation in a greenhouse, where flowering occurred
after 4 months. Propagated plants accumulated iridoids, thus representing a potential source of pharmacologically useful compounds. 相似文献
14.
Manickam V.S. Elango Mathavan R. Antonisamy R. 《Plant Cell, Tissue and Organ Culture》2000,62(3):181-185
The development of stem callus mediated plant regeneration system for Withania somnifera is described. Maximum callus proliferation was obtained on Murashige and Skoog medium supplemented with 2.26 μM 2,4-D. Three-week-old,
white, friable callus was used for shoot regeneration. The maximum shoot regeneration (6.2 ± 0.34 shoots/explant) was achieved
in four weeks when callus was cultured on MS medium fortified with 4.44 μM BA and 0.57 μM IAA. Regenerated shoots were excised
and multiplied (8.4 ± 0.43 shoots/explant) on MS medium supplemented with 4.44 μM of BA. Multiple shoots were divided into
single shoots and were rooted (5.1 ± 0.49 rootlets/shoot) on half strength MS medium supplemented with 9.84 μM of IBA. After
a hardening phase of 3 weeks the plantlets were transferred to the field.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
15.
L. Venkatachalam R. Thimmaraju R. V. Sreedhar N. Bhagyalakshmi 《In vitro cellular & developmental biology. Plant》2006,42(3):262-269
Summary Direct shoot and cormlet regeneration from leaf explants were obtained in triploid dessert banana cultivar Nanjanagud Rasabale
(NR) that is classified under the group ‘Silk’ and has the genotype AAB. The response for both cormlet and direct shool formation
was observed only in leaf explants obtained from shoots cultured in liquid medium but not in similar explants obtained from
shoots grown on gelled medium. Shoot initiation occurred after a sequential culture of leaf (sheath) explants on modified
Murashige and Skoog (MS) medium supplemented with different growth regulators. In the sequence, the leaf explants were cultured
first on medium with a high level (22.4 μM) of benzyladenine (BA), second on indolc-3-butyric acid (IBA) supplemented medium, and third on reduced BA medium under incubation
in the dark. The highest adventitious shoot regeneration in 24% of the explants, with the number of shoots ranging from 2
to 3 per explant, occurred in the explants incubated at the first step in medium with 22.4 and 0.198 μM IBA. Further growth and complete shoot formation occurred under incubation in a 16-h photoperiod. While keeping the culture
conditions constant and replacing BA with picloram (0.83–20.71 μM) in the initial step, adventious origin of cormlets occurred in 12% of the explants. However, when rhizome explants (also
obtained from shoots grown in liquid medium) were cultured with various growth regulators in the first step, medium containing
2,4,5-trichlorophenoxyacctic acid (7.82 μM) produced friable callus that re-differentiated into roots only. Physical forms of the medium, ie.e. agar-gelled or liquid,
imparted specific effects on the extent of multiplication of leaf-regenerated shoots with no differences in morphology and
growth patterns when compared to those of meristem-derived plants. 相似文献
16.
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
17.
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
18.
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
19.
Kottackal Poulose Martin A. K. Pradeep Joseph Madassery 《Acta Physiologiae Plantarum》2011,33(4):1141-1148
Trichopus zeylanicus subsp. travancoricus (known as Arogyapacha), an endangered ethnomedicinal plant of the Western Ghats of South India, serves as the major source
of the commercial drug Jeevani. The present study established a long-term high frequency in vitro propagation protocol for Arogyapacha. Callus obtained from
the branch–petiole explants cultured on Murashige and Skoog (MS) medium with 4.5 μM 2,4-dichlorophenoxyacetic acid upon subculture
to medium with different concentrations of 6-benzyladenine (BA) either alone or in combination with an auxin favoured shoot
morphogenesis. Medium with 13.3 μM BA alone facilitated high frequency shoot bud (mean of 93.2) formation. Medium with lower
concentrations of BA (4.4, 6.6 and 8.8 μM) alone or in combination with lower concentration of α-naphthaleneacetic acid (NAA)
or indole-3-butyric acid (IBA) favoured better shoot growth than 13.3 μM BA containing medium, but with reduced number of
shoot buds. Subsequent cultures on medium with lower concentrations of BA and also on MS basal media facilitated shoot formation
as well as growth of shoots. The shoot regeneration potential showed no decline up to 5 years. Culture of the in vitro-derived
whole branch–leaf explants on MS basal medium developed shoots directly from the node. On medium with 19.6 μM IBA, the whole
branch–leaf explants induced nodular callus from the node, which developed shoots later. Subsequent cultures on medium with
BA exhibited high frequency shoot formation. The transfer of shoots after 10–15 days culture on half-strength MS medium containing
2.7 μM NAA to half-strength basal medium induced a mean of 11.3 roots. Field survival of plantlets relied on the soil mix:
a 1:4 ratio of sand and red-soil exhibited the highest plantlets survival (86.6%). RAPD profile of the source plant and plants
regenerated from calli after 4 years showed no polymorphism. The established plantlets with morpho-floral features similar
to that of the source plants flowered normally and set fruits. 相似文献
20.
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