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1.
Efficient in vitro regeneration systems for Vaccinium species 总被引:1,自引:0,他引:1
Julia Meiners Melanie Schwab Iris Szankowski 《Plant Cell, Tissue and Organ Culture》2007,89(2-3):169-176
Efficient protocols for shoot regeneration from leaf explants suitable for micropropagation as well as for the development
of transgenic plants were developed for blueberry (Vaccinium corymbosum) and lingonberry (Vaccinium vitis-idaea) cultivars. Nodal segments were used to initiate in vitro shoot cultures of lingonberry cultivar ‘Red Pearl’ and southern
highbush blueberry cultivar ‘Ozarkblue’. In order to develop an optimized regeneration procedure, different types and concentrations
of plant growth regulators were tested to induce adventitious shoot regeneration on excised leaves from micropropagated shoots
of both cultivars. The effect on percentage regeneration and number of shoots per explant was investigated. Results indicated
that zeatin was superior to TDZ and meta-topolin in promoting adventitious shoot formation. A concentration of 20 μM zeatin
was most effective in promoting shoot regeneration in both cultivars, in case of ‘Red Pearl’ along with 1 μM NAA. Shoots were
either allowed to root in vitro on medium containing IBA or NAA or ex vitro in a fog tunnel. IBA was superior to NAA for induction
of root development in vitro in both Vaccinium cultivars. Ex vitro rooting under high humidity was tested with cuttings from mature field-grown plants, from acclimatized
tissue culture derived plants and with unrooted in vitro proliferated shoots planted directly. It was found that in vitro
shoots rooted better under fog than cuttings from the other plant sources and rooting was equivalent to that achieved in vitro. 相似文献
2.
Phillip J. Ainsley Graham G. Collins Margaret Sedgley 《In vitro cellular & developmental biology. Plant》2001,37(6):778-785
Summary Shoot cultures of the paper shell almond (Prunus dulcis Mill.) cultivars ‘Ne Plus Ultra’ and ‘Nonpareil’ were subcultured for 4 wk at 4°C on growth regulator-free basal medium under
low light conditions. Elongated shoots were excised and their response to a range of rooting treatments determined. Various
concentrations of indole-3-butyric acid (IBA) and α-naphthaleneacetic acid were compared over a range of incubation periods
to determine the optimum auxin for root formation. In addition, the effect of shoot base shading, phloroglucinol (PG), and
basal salt composition were examined. The treatment resulting in the best rooting of both cultivars was shoot insertion for
12 h into water-agar (0.6% w/v) with 1.0 mM IBA, followed by 2 wk in basal medium without auxin but with 100.0 μM PG. Explants were maintained under dark conditions for 3 d at the start of the treatment period, then exposed to light. Extending
the darkening period did not improve rooting ability. Whilst half-strength Murashige and Skoog basal medium was suitable for
rooting “Ne Plus Ultra’ shoots, full-strength Almehdi and Parfitt medium resulted in the best rooting of ‘Nonpareil’. Under
these conditions, 60.0% of explants developed adventitious roots. 相似文献
3.
M. J. Prado M. V. Gonzalez S. Romo M. T. Herrera 《Plant Cell, Tissue and Organ Culture》2007,88(1):1-10
The present work reports on a study of plant regeneration carried out with callus from the leaf blades and petioles of field-grown
male adult kiwifruit plants (Actinidia deliciosa (Chev.) Liang and Ferguson). The cultivars used were ‘Tomuri’ and clone A, a selected male plant grown in north western Spain.
The best shoot induction conditions were obtained in ‘Tomuri’ leaf blades cultured in K(h) medium in the presence of 23 μM
Zeatin and 0.1 μM NAA. Under these conditions, more than 80% of organogenic callus induction was observed, with an average
of 14 new shoots in the second subculture. The initial length of the shoots affected shoot elongation, which was accomplished
by culturing isolated shoots in K(h) medium with half-strength salts, supplemented with 0.4 μM Zeatin and 0.1 μM NAA. A possible
detrimental long-term effect of cytokinins on shoot elongation can account for the results, since elongation was not observed
until 1 month of culture in elongation medium. For rooting, shoots (1 cm in length) were basally immersed in a 5 mM IBA solution
for 15 s, and transferred to half-strength K(h) basal medium. Regenerated plants were acclimated in a sterile peat:perlite
substrate for 10 days, and then transferred to soil. AFLP analysis was accomplished with 15 primer combinations from which
13 showed reproducible and well-resolved bands, producing a total of 1321 fragments from which 1281 were polymorphic (97%).
A dendrogram was constructed using both monomorphic and polymorphic bands, showing genetic variation among field-grown plants
and tissue culture-derived regenerants. 相似文献
4.
José Carlos Gonçalves Graça Diogo Maria Teresa Coelho Nieves Vidal Sara Amâncio 《In vitro cellular & developmental biology. Plant》2008,44(5):412-418
Endogenous levels of indole-3-acetic acid (IAA), indole-3-acetylaspartic acid (IAAsp) and indole-3-butyric acid (IBA) were
measured during the first 8 d of in vitro rooting of rootstock from the chestnut ‘M3’ hybrid by high performance liquid chromatography (HPLC). Rooting was induced
either by dipping the basal ends of the shoots into a 4.92-mM IBA solution for 1 min or by sub-culturing the shoots on solid
rooting medium supplemented with 14.8-μM IBA for 5 d. For root development, the induced shoots were transferred to auxin-free
solid medium. Auxins were measured in the apical and basal parts of the shoots by means of HPLC. Endogenous levels of IAA
and IAAsp were found to be greater in IBA-treated shoots than in control shoots. In extracts of the basal parts of the shoots,
the concentration of free IAA showed a significant peak 2 d after either root inductive method and a subsequent gradual decrease
for the remainder of the time course. The concentration of IAAsp peaked at day 6 in extracts of the basal parts of shoots
induced with 14.8-μM IBA for 5 d, whereas shoots induced by dipping showed an initial increase until day 2 and then remained
stable. In extracts from basal shoot portions induced by dipping, IBA concentration showed a transient peak at day 1 and a
plateau between day 2 and 4, in contrast to the profile of shoots induced on auxin-containing medium, which showed a significant
reduction between 4 and 6 d after transferred to auxin-free medium. All quantified auxins remained at a relatively low level,
virtually constant, in extracts from apical shoot portions, as well as in extracts from control non-rooting shoots. In conclusion,
the natural auxin IAA is the signal responsible for root induction, although it is driven by exogenous IBA independently of
the adding conditions. 相似文献
5.
The effects of post harvest application of ethylene, abscisic acid (ABA), indole-3-butyric acid (IBA) treatments or dark storage
on root induction and continued growth of regenerated roots in Pelargonium cuttings were investigated using hydroponics in the greenhouse. Ethylene markedly increased rooting percentage in ‘Greco’
and ‘Surfing’, reduced the number of roots per cutting in ‘Surfing’ and had no effect on the total root lengths in the two
cultivars. Ethylene treatment reduced fresh root mass in ‘Surfing’, increased dry root mass and reduced root water content
in both cultivars. ABA (50 μM) enhanced rooting percentage in ‘Greco’, reduced the number of roots per cutting, reduced total
root lengths and fresh root mass in both cultivars. ABA increased dry root mass and reduced root water content in ‘Surfing’
but this effect was not apparent in ‘Greco’. Storing cuttings in the dark for 4 days had no effect on rooting percentage and
number of roots per cutting in ‘Greco’ and ‘Surfing’. However, dark storage reduced total root lengths in ‘Surfing’ and reduced
fresh root mass in ‘Greco’. Dark storage had no effect on dry root mass and water content in both cultivars. Applying 4 μl l−1 IBA in the rooting solution induced maximum (100%) root induction in ‘Surfing’. However, IBA reduced the number of roots
per cutting in ‘Greco’, reduced total root lengths and fresh root mass in the two cultivars. IBA treatment profoundly increased
and reduced dry root mass and root water content, respectively, in ‘Greco’ and ‘Surfing’. The enhanced root induction observed
after IBA and ABA applications could be ascribed to their influence on ethylene biosynthesis, since ethylene treatment increased
rooting percentage in both cultivars. However, high ABA (100 μM) and IBA (12 μl l−1) levels or dark storage reduced the ability of induced roots to continue growth. We attribute our results to plant stress-response
mechanism and ethylene appears to play an important role in the process of root initiation and root growth in Pelargonium cuttings. 相似文献
6.
Annapurna Dhavala T. S. Rathore 《In vitro cellular & developmental biology. Plant》2010,46(2):180-191
Micropropagation of Embelia ribes was achieved through proliferation of axillary shoots obtained from mature plants. Nodal shoot segments, collected March–May,
exhibited high-frequency (75%) shoot initiation when cultured on Murashige and Skoog (MS) basal medium supplemented with thidiazuron
(TDZ) at 1.13 μM and indole-3-butyric acid (IBA) at 0.49 μM. Subculture of sprouted shoots from the original explants on medium
containing TDZ (1.13 and 0.45 μM) during the first and second subcultures was found essential for further shoot proliferation,
while inhibition of shoot elongation by TDZ could be overcome by transferring shoot cultures onto MS medium containing 6-benzylaminopurine
(BAP; 11.10 μM) for the third subculture. Treating the explants with an antioxidant mixture of 568 μM ascorbic acid, 119 μM
citric acid, and 307 μM glutathione prior to inoculation, coupled with subculture at 2-wk intervals onto fresh medium, both
helped to reduce browning of the explants and facilitated production of five to six shoots/explant. MS medium supplemented
with BAP (4.44 μM) and IBA (0.49 μM) induced shoot multiplication, producing five to six shoots/explant with a shoot length
of 3 to 4 cm over a 4-wk culture period. Shoots of 3 to 4 cm in length exhibited 100% rooting within 4 wk after transfer to
media containing half the nutrient salt concentration of MS medium with 3.69 μM IBA. Ex vitro rooting in the greenhouse from the in vitro shoots treated with 4.93 μM IBA for 30 min exhibited 95% rooting in soilrite™ medium in a 4-wk period. About 85% of micropropagated
plants were established successfully in root trainers. Three-month-old, hardened plants could further be successfully established
in the field. In 1 yr, by using the above protocol, 3,200 plants could be produced from a single shoot and 2,700 could be
established in the field. 相似文献
7.
The influence of the basal medium and different plant growth regulators on micropropagation of nodal explants from mature
trees of lemon cultivars was investigated. Although the basal medium did not affect any of the variables, explants on DKW
medium were greener. Several combinations of 6-benzyladenine (BA) and gibberellic acid (GA) were used to optimise the proliferation
phase. The number of shoots was dependent on the BA and GA concentrations and the best results were obtained with 2 mg l−1 BA and 1 or 2 mg l−1 GA. Explants length was shorter with the higher BA concentrations and, in all genotypes, shoot length was greater with 2 mg l−1 GA. The best results for productivity (number of shoots × the average shoot length) were obtained with 2 mg l−1 BA and 2 mg l−1 GA, although explants with chlorosis and narrow leaves were observed. The presence of BA and GA in the proliferation medium
was essential for the explant multiplication but GA had a greater influence. The transfer of in vitro shoots to rooting media,
containing different concentrations of indole butyric acid (IBA) and indole acetic acid (IAA) produced complete plantlets.
Lemon shoots rooted well in all rooting combinations. The highest rooting percentages were obtained on media containing 3 mg l−1 IBA alone or IBA in combination with 1 mg l−1 IAA and on these media the highest numbers of roots were produced. The average root length was affected significantly by
the IBA and IAA concentrations. Root length was greater when only 3 mg l−1 IBA was used, and in this rooting medium explants had a better appearance, with greener and larger leaves. The success during
the acclimatisation was close to 100% and the plantlets exhibited normal growth in soil under greenhouse conditions. 相似文献
8.
Fotini G. Skiada Katerina Grigoriadou Eleftherios P. Eleftheriou 《Central European Journal of Biology》2010,5(6):839-852
The effects of six basal media on in vitro shoot proliferation of the greek grapevines Vitis vinifera L. cv. ‘Malagouzia’ and ‘Xinomavro’ were investigated. Galzy and Zlenco proved to be the most effective for ‘Malagouzia’
and ‘Xinomavro’, respectively. If only BA was present in the medium, shoot development was poor and the plantlets were chlorotic.
When the medium was supplemented with BA and NAA, growth was enhanced. The best ratio (in μM) of growth regulators was 0.5/0.3
for ‘Malagouzia’, and 0.1/0.03 for ‘Xinomavro’, which resulted in the highest number of microshoots per explant and greatest
proliferation rate. The development of ‘Malagouzia’ and ‘Xinomavro’ explants at 21±2 and 26±2°C was also investigated, revealing
the higher temperature to be more effective. Regarding rooting, 0.5 μM IBA improved root formation at 26°C for ‘Malagouzia’
and 0.5 μM IBA at 21°C for ‘Xinomavro’. Moreover, 0.5 μM IBA resulted in a higher rooting percentage (>95%) and proved to
be more beneficial for the overall morphological appearance of the plantlets of ‘Malagouzia’. After acclimatization, survival
of microshoots cultivated in media with IBA was higher than those in NAA. 相似文献
9.
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
10.
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
11.
J. Y. Choi H. J. Kim C. H. Lee J. M. Bae Y. S. Chung J. S. Shin N. I. Hyung 《In vitro cellular & developmental biology. Plant》2001,37(2):274-279
Summary An efficient and simple plant regeneration system via organogenesis from leaf segments of persimmon (Diospyros kaki Thunb.) cultivars ‘Fuyu’ and ‘Nishimurawase’ has been developed. The regeneration capacity was influenced by the culture
vessels, gelling agents, plant growth regulators, and light conditions. Leaf explants taken from in vitro shoots were cultured on a modified Murashige and Skoog medium (MS1/2N), for 16 wk without transfer to fresh medium. Adventious
shoots appeared after 4 and 8 wk in culture of ‘Nishimurawase’ and ‘Fuyu’ tissues, respectively. The culture of leaf explants
in Erlenmeyer flasks with medium containing 4 g l−1 agar enhanced shoot formation in comparison to media with increased agar concentrations. Optimal shoot regeneration was obtained
with 5 mg l−1 (22.8 μM) zeatin and 0.1 mg l−1 (0.05 μM) indole-3-butyric acid (IBA) for ‘Nishimurawase’, and 10 mg l−1 (45.6 μM) zeatin and 0.1 mg l−1 (0.05 μM) IBA for ‘Fuyn’. Shoot regeneration frequencies in both cultivars were 100%, and shoot numbers per explant reached up to
9.2 for ‘Nishimurawase’ and 2.2 for ‘Fuyu’. Dark incubation during the first 4–5 wk was the most effective condition to successfully
influence shoot regeneration in both cultivars. While dark incubation was essential for adventitious shoot formation by ‘Fuyu’,
it was only slightly beneficial to ‘Nishimurawase’. More than 80% of the regenerated shoots rooted within 4 wk on hormone-free
MS1/2N demium after having been dipped for 30 s in 250 mg l−1 (1.1. mM) IBA solution. 相似文献
12.
Hypocotyl expiants from 22 cultivars ofCatharanthus roseus were cultured on various shoot-inducing media to assess their competence for adventitious shoot formation. The Murashige
and Skoog (MS) media had been supplemented with 14 μM zeatin and 2.5 μM indole-3-butyric acid (IBA), 4.5 μM BA and 0.5 μM
α-naphthaleneacetic acid (NAA), or 14 μM thidiazuron and 2.5 μM IBA. After eight weeks, the expiants from ‘Cooler Raspberry
Red’ showed the greatest frequency of adventitious shoot formation, followed by ‘Cooler Orchid’ and ‘Cooler Treated’. The
highest frequency (86.7%) for ‘Cooler Raspberry Red’ was attained on the medium enhanced with 14 μM zeatin and 2.5 μM NAA.
Excised adventitious shoots were then readily rooted on a half-strength MS basal medium. Afterward, the regenerated plantlets
were transferred to potting soil and grown to maturity in a greenhouse. 相似文献
13.
Adventitious shoot regeneration from leaf explants of southern highbush blueberry cultivars 总被引:1,自引:0,他引:1
Cheng Liu Pete Callow Lisa J. Rowland James F. Hancock Guo-qing Song 《Plant Cell, Tissue and Organ Culture》2010,103(1):137-144
Protocols were developed to optimize adventitious shoot regeneration from four southern highbush blueberry cultivars. Leaf
explants from 6 week-old shoots of the four cultivars were excised and cultured on woody plant medium each containing thidiazuron
(4.54 or 9.08 μM), zeatin (18.2 μM), or zeatin riboside (5.7 or 11.4 μM) either separately or in combination with α-naphthaleneacetic
acid at 2.69 μM. Optimum medium for shoot regeneration was genotype-dependent. Efficient regeneration was obtained at frequencies
of 88.9% for ‘Jewel’, 87.8% for ‘Emerald’, 53.3% for ‘Jubilee’ and 87.8% for ‘Biloxi’. Leaf explants of newly developed shoots
from the cultures having undergone five subcultures had higher regeneration frequencies than those having undergone two subcultures.
Regenerated shoots, 80–100% for each cultivar, rooted in 8 weeks after transplantation to soil. The regeneration systems described
have potential use in genetic transformation of southern highbush blueberry cultivars. 相似文献
14.
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
15.
Filipendula ulmaria (L.) Maxim (meadowsweet) is a medicinal plant that is claimed to have several biological activities, including anti-tumor,
anti-carcinogenic, anti-oxidant, anti-coagulant, anti-ulcerogenic, anti-microbial, anti-arthritic, and immunomodulatory properties.
This report describes, for the first time, an efficient plant regeneration system for F. ulmaria via adventitious shoot development from leaf, petiole, and root explants cultured on Murashige and Skoog’s minimal organics
medium containing different concentrations of thidiazuron (TDZ), benzyladenine, and kinetin either alone or in combination
with different auxins. Relatively extensive/prolific shoot regeneration was observed in all three explant types with TDZ in
combination with indole-3-acetic acid (IAA). Gibberellic acid (GA3), TDZ, and IAA combinations were also tested. The best shoot proliferation was observed among root explants cultured on media
supplemented with 0.45 μM TDZ + 2.85 μM IAA + 1.44 μM GA3. Regenerated shoots were transferred to rooting media containing different concentrations of either IAA, indole-3-butyric
acid (IBA), naphthalene acetic acid, or 2,4-dichlorophenoxyacetic acid. Most shoots developed roots on medium with 2.46 μM
IBA. Rooted explants were transferred to vermiculite in Magenta containers for a 2-wk acclimatization period and then finally
to plastic pots containing potting soil. The plantlets in soil were kept in growth chambers for 2 wk before transferring to
greenhouse conditions. 相似文献
16.
Shoot explants from seedling-derived culture of Eucalyptus ficifoliaF. Muell. cultured on a rooting medium free from indole-3-butyricacid (IBA) develop a root system (Type I) consisting of a fewcomparatively long roots and only small amounts of callus. IBAat 5.0 µM in a rooting medium free from riboflavin inducesthe development, on the shoot explants, of a compact root system(Type II) consisting of callus and many short roots. Riboflavinwhen exposed to light, is able to photo-oxidize IBA; the degreeof photo-oxidation depends on the photon fluence of the lightreceived. The rooting response of the cultures reflects thedegree of photo-oxidation of IBA: concentrations of IBA fromabout 104M to 106M in the rooting medium induceformation of the Type II root system whilst photo-oxidationof the auxin to concentrations of about 108M or lowerinduces the formation of the Type I root system. Thus, exogenousriboflavin and exogenous IBA are linked in a distinct light-induced,riboflavin-mediated change in root morphogenesis. The anatomyof root development in the Type I and Type II root systems wasstudied and factors affecting the development were defined.Characteristics of riboflavin and IBA breakdown in various lightregimes were determined and related to root morphogenesis. Theresults and their implications are discussed. Key words: Auxin photo-oxidation, Riboflavin, Root morphogenesis, Tissue culture 相似文献
17.
María del Socorro Santos Díaz Candy Carranza Álvarez 《In vitro cellular & developmental biology. Plant》2009,45(2):162-170
Two procedures for the in vitro propagation of Encyclia mariae, a threatened Mexican orchid, were developed. In the first procedure, leaves from in vitro germinated seedlings were cultured on Murashige and Skoog medium (MS) supplemented with the range of 2.21–4.4 μM 6-benzylaminopurine
(BA) in combination with 2.69–10.74 μM naphthalene acetic (NAA), 2.07–8.29 μM indole-3-butyric (IBA), or 2.85–11.42 μM indole-3-acetic
acid (IAA) to determine the best medium for the induction of shooting. Maximum direct shoot formation from leaves was observed
on MS containing 22.21 μM BA and 10.74 μM NAA (25 shoots/explant). The second procedure began with the culture of protocorms
on media containing NAA, IBA, or IAA, which induced callus formation with high regenerative potential in the form of protocorm-like-bodies
(PLBs) that eventually differentiated into shoots. The optimal response was attained when these structures were cultured on
medium with 4.14 μM IBA (30 shoots/PLB). To promote the elongation of shoots derived from PLBs, the material was subcultured
onto MS medium containing 22.21 μM BA and 5.37 μM NAA. Through the exploration of the effects of auxins and matrix on the
rooting of shoots, it was determined that the optimal rooting occurred on media supplemented either with 5.71 μM IAA or 4.14 μM
IBA either on agar-gelled medium or in liquid media with coir as the matrix. Rooting was found to be 20% higher in liquid
media than in agar-gelled medium. 相似文献
18.
Summary Carbohydrate type and concentration and their interactive effects on in vitro shoot proliferation of three lingonberry (Vaccinium vitis-idaea ssp. vitis-idaea L.) cultivars (‘Regal’, ‘Splendor’, and ‘Erntedank’) and two V. vitis-idaea ssp. minus (Lodd) clones (‘NL1’ and ‘NL2’) were studied. Nodal explants were grown in vitro on medium with 2 μM zeatin and either glucose, sorbitol, or sucrose at a concentration of 0, 10, 20, or 30 gl−1. The interactive effects of carbohydrate type and concentration and genotype were important for shoot proliferation. The
best response was afforded by sucrose at 20 gl−1 both in terms of explant response and shoot developing potential, although glucose supported shoot growth equally well, and
in ‘NL1’ at 10 gl−1 it resulted in better in vitro growth than sucrose. Carbohydrate concentration had little effect on shoot vigor. The genotypes differed in terms of shoots
per explant, length, and vigor, leaves per shoot, and callus formation at the base of explants; this was manifested with various
types and concentrations of carbohydrate. Changing the positioning of explants on the medium from vertically upright to horizontal
increased the shoot and callus size, but decreased shoot height and leaves per shoot. Proliferated shoots were rooted on a
peat:perlite (1∶1, v/v) medium and the plantlets were acclimatized and eventually established in the greenhouse. 相似文献
19.
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
20.
Eline Kirk Mørk Karin Henriksen Henrik Brinch-Pedersen Kell Kristiansen Karen Koefoed Petersen 《Plant Cell, Tissue and Organ Culture》2012,108(3):501-512
A protocol for adventitious shoot formation in Symphyotrichum novi-belgii was developed after investigating the effects of cultivar and hormone combinations. A Murashige and Skoog medium with 1.0 mg l−1 6-benzyladenine induced adventitious shoot formation in 15 out of 19 cultivars. Addition of 0.1 mg l−1 indole-3-acetic acid or naphthaleneacetic acid increased the total number of shoots per explant, but not the number of shoots
longer than 1 cm. Addition of dichlorophenoxyacetic acid (2,4-D) promoted callus formation, but inhibited shoot elongation.
A transformation system for the two cultivars Victoria Fanny and Victoria Jane was developed by co-cultivation of leaf explants
with Agrobacterium tumefaciens. Three bacterial strains (LBA 4404, A281 and C58) all carrying the binary vector, p35S-GUS-INT, and harbouring the uidA gene coding for β-glucuronidase (GUS) were used. Regeneration of transgenic plants after co-cultivation with A281 was independent
of cultivar, and all explants produced callus followed by indirect shoot formation. In ‘Victoria Fanny’ shoots were formed
faster and without a callus phase after co-cultivation with LBA 4404 or C58. The highest number of potentially transformed
shoots was regenerated after co-cultivation of ‘Victoria Fanny’ leaf explants with LBA 4404. Integration of the transgenes
in the plant genome was confirmed using PCR and Southern blot hybridisation. To verify that the transgenes could be transferred
to offspring, crosses were conducted between three transgenic lines of ‘Victoria Fanny’ and two wild type pollen donors. It
was demonstrated that viable seeds were produced and that the uidA gene was inherited. 相似文献