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1.
Summary
Salvia valentina Vahl and Salvia blancoana Webb & Heldr subsp. mariolensis Figuerola, two endemic species of Salvia from the Mediterranean coastal region of Spain, were successfully gegenerated in vitro from adult plants using two explant types (apical and nodal segments). Maximum shoot proliferation for both species was obtained
with nodal explants: for S. blancoana on Murashige and Skoog medium supplemented with 6-γ-γ-dimethylallylaminopurine at 1 mg 1−1 (4.9 μM). and for S. valentina on the same medium with kinetin at 1–2 mg 1−1 (4.6–9.3 μM). The influence of apical dominance, and the explant viability in culture were found to be the main differences between the
two species during the shoot multiplication phase. Rooting of shoots was low, specially for S. valentina. For both species, rooting was achieved in Murashige and Skoog medium without growth regulators. In general the addition
of the auxins indole 3 acetic acid or indole-3-butyric acid did not improve the rooting or, in the case of naphthaleneacetic
acid, had an inhibitory effect. In the best rooting media, rooting shoots increased in length. The rooted plantlets were acclimated
to ex vitro conditions and a survival percentage > 70% was obtained under greenhouse conditions. This work was carried out as an ex situ conservation method for these Spanish endemic species. 相似文献
2.
Melissa E. Heatley Roberta H. Smith 《In vitro cellular & developmental biology. Plant》1996,32(2):115-118
Summary
Arachis hypogaea L. peanut, has been a difficult species to manipulate in tissue culture. Lack of a reliable and quick regeneration method
for peanuts has proven to be one of the hindrances in the application of transformation protocols to the crop. A protocol
to initiate shoot apex elongation and rooting of these shoots is described. This protocol was successful with two peanut cultivars.
Shoot apices were isolated from germinated seedlings and placed on Murashige and Skoog salts containing N6-benzyladenine for shoot initiation. Once shoot elongation occurred, the explant was transferred to a rooting medium containing
Murashige and Skoog salts and only one plant growth regulator, α-naphthalene acetic acid. In as few as 3 weeks, the explants
began to root and could be transferred to soil. Forty-five percent of explants isolated from germinating peanut seeds would
root on this medium. Elongation and rooting of the shoot apices were not hindered by the addition of an antibiotic to the
medium, indicating that the regeneration method could be useful inAgrobacterium tume-faciens-mediated transformation protocols. 相似文献
3.
K. Chengalrayan M. Gallo-Meagher 《In vitro cellular & developmental biology. Plant》2001,37(4):434-439
Summary Sugarcane (Saccharum spp. hybrid cv. CP 84-1198) embryogenic calluses were induced from young leaves cultured on modified Murashige and Skoog
basal medium supplemented with 13.6 μM 2,4-dichlorophenoxyacetic acid. Five concentrations, 0.5, 1.0, 2.5, 5.0, and 10.0 μM, of five different growth regulators, 6-benzylaminopurine, kinetin, 6-γ,γ-(dimethylallylamino)purine, zeatin, and thidiazuron,
were tested with or without 22.5 μM α-naphthaleneacetic acid to compare their ability to induce regeneration from embryogenic callus. After 4 wk on medium, the
percentage of shoot meristem induction was evaluated, and after 10 wk the total number of shoots produced, as well as the
percentage of shoots greater than 1 cm in length, was obtained. Although it had the lowest percentage of elongated shoots,
medium containing thidiazuron alone performed better than all other growth regulators tested, with the highest percentage
of shoot induction and the largest number of shoots, particularly at a concentration of 2.5 μM. 相似文献
4.
Margarita Velcheva Zehava Faltin Aliza Vardi Uri Hanania Yuval Eshdat Oded Dgani Nachman Sahar Avihai Perl 《In vitro cellular & developmental biology. Plant》2010,46(6):477-484
A system for genetic transformation and subsequent plant regeneration via indirect organogenesis from callus was developed
for Aloe vera. Young seedlings served as primary explants. Callus cultures were established on Murashige and Skoog (1962) medium supplemented
with 3 mg l−1 benzylaminopurine and 2 mg l−1 indole acetic acid. A protocol was developed to switch from the differentiated stage, using in vitro shoots or young regenerated plants, back to the de-differentiated stage of the callus and vice versa. Long-term maintenance
of this callus paved the way for genetic manipulation of Aloe vera. Calluses were bombarded with a plasmid containing uidA and hpt genes, both under the control of the 35S promoter. Dithiothreitol and gibberellic acid were found to play a major role in
reducing tissue necrosis following bombardment. Transformed shoots were regenerated under stepwise selection in hygromycin-containing
liquid medium supplemented with different antioxidants. Amberlite XAD-4 resin was embedded into alginate beads and added to
the selection medium. Amberlite was best for adsorbing different phenolic compounds and blocking explant necrosis. Shoot initiation
occurred after transfer of the transformed cells to Murashige and Skoog medium supplemented with 2.0 mg l−1 thidiazuron and 0.1 mg l−1 indole butyric acid. Murashige and Skoog medium supplemented with 1 mg l−1 zeatin riboside promoted shoot elongation. Rooting and plant development were obtained on Murashige and Skoog basal medium
supplemented with 15 mg l−1 hygromycin lacking growth regulators. The transgenic nature of the regenerated plants was verified by histochemical GUS assay
and Southern blot hybridization. 相似文献
5.
T. R. Marks 《Plant Growth Regulation》1996,20(1):57-60
This paper reports that rhizogenesis in woody plant species in vitro was mediated through the basipetal transport of auxin from the shoot apex. This can directly induce roots in easy-to-root species such as Betula pendula, but was dependent upon an interaction with exogenous auxin in more difficult-to-root species such as Daphne cneorum, and to a lesser extent in Quercus robur. Shoot apex removal reduced rhizogenesis in Quercus, and inhibited it in Daphne, even in the presence of exogenous auxin, whereas rooting in Betula was unaffected. That basipetally transported auxin modulates rhizogenesis was demonstrated by the inhibition of root induction in Betula shoots by the auxin transport inhibitor 2,3,5-triiodobenzoic acid (TIBA), and by the substitution of indole-3-acetic acid (IAA) for a bud in Betula internodal sections.Abbreviations IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- TIBA
2,3,5-triiodobenzoic acid
- MS
Murashige and Skoog medium
- WPM
woody plant medium 相似文献
6.
Kottackal Poulose Martin Chun-Lai Zhang Adrian Slater Joseph Madassery 《Plant Cell, Tissue and Organ Culture》2007,88(1):51-59
Shoot necrosis and plant death occurred during large-scale propagation of Musa cultivars, viz. Grande Naine (AAA), Dwarf Cavendish (AAA), Nendran (AAB) and Quintal Nendran (AAB). The timing and frequency of necrosis
was influenced by the cultivar. Grande Naine and Dwarf Cavendish showed necrosis after the 7th subculture at frequencies of 27 and 29%, whilst at the rooting stage (after seven transfers of shoot multiplication) the
frequencies were 18 and 19% respectively. In Nendran and Quintal Nendran, necrosis occurred after the 5th subculture at frequencies of 38 and 40%, and at the rooting stage (after five transfers of shoot multiplication) the frequencies
of necrosis were 26 and 27% respectively. Several methods were tested for alleviating shoot necrosis, including shortening
the culture period, altering the media salt strength, use of various plant growth regulators, different levels of sucrose,
fructose, silver nitrate, and increasing the concentration of calcium chloride. Only the addition of calcium chloride proved
effective in reducing shoot necrosis. Transfer of shoots showing early signs of necrosis to Murashige and Skoog medium with
an addition of 50–100 mg l-1 calcium chloride facilitated recovery of more than 90% of the shoots. The same addition to shoot multiplication medium for
2–4 transfers after the 6th and 4th subcultures respectively, for bananas and plantains alleviated shoot necrosis up to the 12th subculture in all cultivars. This also prevented necrosis during subsequent rooting. The prevention of loss of cultures by
shoot necrosis by the addition of calcium chloride at the shoot multiplication stage, and thereby the reduction of production
cost, will be beneficial to the banana micro-propagation industry. 相似文献
7.
Anin vitro propagation protocol for a leguminous liana,Bauhinia vahlii, has been established. In the first experiment, cotyledonary nodes fromin-vitro-germinated seedlings were cultured on various basic media (Murashige and Skoog medium, Woody Plant medium, B5, and 1/2 Murashige and Skoog medium) containing 1.0M thidiazuron. Shoot proliferation (96.20%) and multiplication (5.55 shoots/explant) was best when cultured on Murashige and Skoog medium. The second experiment compared responses to benzylaminopurine, kinetin, zeatin and thidiazuron. Murashige and Skoog medium supplemented with 1.0M thidiazuron proved most effective for both shoot proliferation and shoot multiplication. The effect of cytokinin type and concentration and their interaction was found to be significant (P<0.001) for explant proliferation, shoot number and length. Subsequent rooting (55.14%) of the regenerated shoots was achieved on half-strength Murashige and Skoog medium supplemented with IM 1- naphthaleneacetic acid. Successful transfer of regenerants to soil has been accomplished, and efforts are being made to gradually transfer them to field conditions. 相似文献
8.
A. V. Raghu S. P. Geetha Gerald Martin Indira Balachandran P. N. Ravindran 《In vitro cellular & developmental biology. Plant》2006,42(6):584-588
Summary A protocol was developed for rapid clonal propagation of the important medicinal climber, Tinospora cordifolia, through in vitro culture of mature nodal explants. Shoots were initiated on both Murashige and Skoog (MS) medium and woody plant medium (WPM)
supplemented with 2.32 μM kinetin (KIN). Of the two basal media tested, WPM was found to be superior to MS medium for the induction of multiple shoots.
Among the cytokinins tested, N6-benzyladenine (BA) was more effective than KIN for axillary shoot proliferation. KIN was superior to BA in terms of shoot
elongation. An average multiplication rate of 6.3 shoots per explant was obtained with WPM supplemented with 8.87 μM BA. Shoot clumps harvested from this medium were transferred to WPM supplemented with 2.22 μM BA and 4.65 μM KIN for shoot elongation. Elongated shoots were rooted in half-strength MS medium supplemented with 2.85 μM indole-3-acetic acid (IAA). Rooted plantlets were successfully transferred to sand and established with 80% survival. 相似文献
9.
Paula S. Campos M. Salomé S. Pais 《In vitro cellular & developmental biology. Plant》1996,32(3):184-189
Summary Shoot propagation ofPersea indica (L.) K. Spreng was achieved using seedling axillary buds cultured on MS (Murashige and Skoog, 1962) medium with 1 mg/l (2.8
μM) N6-benzyladenine (BA). Forty percent of the obtained shoots did not elongate, but showed bud proliferation, which was maximal
(three axillary buds per shoot) at the end of the seventh subculture. Sixty percent of the shoots elongated, did not show
bud proliferation, and formed calluses at their base. Successful rooting (84.6%) was achieved dipping the base of each elongated
shoot in 3 g/l (16.11 mM) indolebutyric acid (IBA) for 1–2 s, and transferring to half strength MS medium without growth regulators. These shoots
presented an acclimatization success of 100%. Results suggest that micropropagated elongated shoots ofP. indica can be adequately used in reforestation programs. 相似文献
10.
A highly efficientin vitro regeneration methodology for mature chutese tallow tree (Sapium sebiferum Roxb.) has been developed. Shoot segments cultured on MS medium supplemented with 7.5 µM NAA produced light green callus. Optimum shoot differentiation resulted when callus was transferred to MS medium with 1 µM BA and 0.25 µM NAA. Shoot forming ability of callus was higher on MS medium compared to B5, half-MS or WPM. A continuous shoot harvest system at four-week intervals was established. Shoot yield continued for six months without loss of vigour. Regenerated shoots were rooted by culturing on half strength agar-gelled MS medium containing 1 µM IBA. Rooted plantlets were transferred to 1:1 soil vermiculite mixture and acclimatized with 67 % survival rate. Fully acclimatized plants were planted in the field, and performance is being evaluated.Abbreviations 2, 4- D
2, 4-dichlorophenoxyacetic acid
- NAA
1 - napthaleneacetic acid
- BA
benzyladenine
- Kn
kinetin
- 2-ip
6 - (, -dimethylallylamino)-purine
- IBA
indole-3-butyric acid
- WPM
woody plant medium (1980)
- MS
Murashige and Skoog (1962) medium
- B5
Gamborg et al's medium (1968) 相似文献
11.
Summary A rapid shoot multiplication protocol was established for an important medicinal plant, Vitex negundo L., belonging to the family Verbenaceae, using Murashige and Skoog medium, achieved by shoot multiplication as well as callus regeneration. Shoot multiplication
was induced by different concentrations of 1-phenyl-3-(1,2,3-thiadiazol-5-yl)-urea (TDZ), Benzyladenine and 6-furfuryl amino
purine separately along with 10% (v/v) coconut water. Green organogenetic callus was obtained by the combined effect of 0.5–2.15
μM TDZ and 1.7 μM indole-3-acetic acid (IAA) along with 1% polyvinylpyrrolidone (PVP), and produced the maximum number of shoots when subcultured
onto medium containing 2.7 μM TDZ alone. Elongation of in vitro shoots was observed in MS medium containing 2.4 μM gibberellic acid and rooting was induced by the combined effect of 1.71 μM IAA and 1.62 μM α-naphthalene acetic acid. 相似文献
12.
María Teresa Herrera Margarita Cacho M. Purificación Corchete Jorge Fernandez-Tarrago 《Plant Cell, Tissue and Organ Culture》1990,22(3):179-182
Shoot tips from seedlings of Digitalis thapsi L. were cultured on Murashige and Skoog's medium and the effect of various auxins (2,4-D, NAA and IAA) were analyzed alone or in combination with cytokinis (BA and kinetin). Shoot multiplication and direct rooting of the new shoots were obtained after four weeks of culture in MS medium without hormones, but callus formation and the appearance of abnormal phenotypes were frequent. The addition of auxins to the cultures prevented the formation of callus but not the appearance of variant phenotypes. Both drawbacks could be avoided by combination of NAA or IAA with BA or kinetin. The best results for shoot multiplication and direct rooting were obtained with 0.5 mg l-1 NAA and 0.1 or 0.5 mg l-1 kinetin.Abbreviations BA
6-benciladenine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- IAA
indole-3-acetic acid
- Kin
kinetin
- NAA
naphtalene acetic acid
- MS
Murashige and Skoog 相似文献
13.
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
14.
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
15.
Adaucto B. Pereira-Netto 《In vitro cellular & developmental biology. Plant》1996,32(4):253-256
Summary
Hancornia speciosa fruit is highly desired for the juice and ice cream industry in tropical regions. A rapid reduction in germination ability
ofH. speciosa seeds has been a problem for its large-scale cultivation. This paper describes anin vitro technique that may lead to an alternative propagation method forH. speciosa. Shoot apices and nodal segments from aseptically germinated young embryos were cultivatedin vitro on. Murashige and Skoog (1962) medium supplemented with growth regulators. Shoot multiplication was maintained by sequential
subculture of shoot tips and nodal segments. N6-benzyladenine was the most effective cytokinin for the induction of shoot growth. N6-furfurylamino-purine, at various concentrations, yielded multiplication rates sevenfold lower than the highest multiplication
rate found with N6-benzyladenine. Increased root initiation rate and root elongation was observed with the presence of γ-(indole-3) butyric
acid in the half-strength Murashige and Skoog culture medium, especially at 10μM. N6-benzyladenine strongly inhibited rooting, even in the presence of γ-(indole-3) butyric acid. Thein-vitro-raised rooted plantlets were acclimatized to greenhouse environment through progressive reduction in relative humidity and
later transplanted to the field. 相似文献
16.
Summary A procedure for plant regeneration, flower and plant formation from petiolar and inflorescence nodal explants of culantro
is discribed. Leaf petioles were excised from young leaves of non-flowering plants while nodal explants were excised from
the inflorescence. Explants were cultured in Murashige and Skoog (MS) medium alone or supplemented with 0.5μM naphthaleneacetic acid (NAA) and 0.9, 1.8, 4.5, or 9 μM thidiazuron (TDZ). All explants produced multiple shoots. In addition, nodal explants formed flowers. Shoot number, flower
number and shoot length were influenced by TDZ and NAA. Rooted shoots from both types of explants were transferred to soil
where plants were successfully established. 相似文献
17.
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
18.
Shoot apical meristems were used to establish regenerative axillary bud cultures of 9 muscadine grape cultivars. Meristems taken from 10 cm long shoots had less contamination (3%) and a higher survival rate (94%) than those from shorter or longer shoots. Of media tested, MS, 1/2 MS, and C2D resulted in equivalent shoot proliferation rates, whereas, WPM produced stunted shoots. When pooling results for 3 cultivars, 5, 10 and 20 M BA and 5 M TDZ produced the highest average number of shoots per cultured apex (3.4–3.8). However, shoots produced with TDZ were stunted and did not root well. For rooting of shoots directly in potting mix, a rooting powder pretreatment significantly increased the number of roots per shoot but did not affect percent rooting or root length. For rooting in vitro, 1 M NAA significantly increased all parameters measured. Although more shoots rooted in vitro than in vivo (77% vs. 46%), the latter was judged preferable since acclimatized plants were produced in less time and a major culture step was eliminated. Significant differences among cultivars were noted for measured responses in all experiments.Abbreviations BA
benzyladenine
- Kin
kinetin
- MS
Murashige & Skoog (medium)
- NAA
naphthaleneacetic acid
- TDZ
thidiazuron
- WPM
woody plant medium 相似文献
19.
An efficient plant regeneration system for Mucuna pruriens L. (DC.) using cotyledonary node explants
Summary The purpose of this study was to develop an efficient micropropagation system for Mucuna pruriens, an important medicinal plant in India. A range of cytokinins was investigated for multiple shoot regeneration with cotyledonary
node explants from 7-d-old aseptic seedlings. Of all the cytokinins, 6-benzyladenine (BA), kinetin (KIN) and 2-isopentenyl
adenine (2-iP) tested in Murashige and Skoog medium (MS), BA was the most effective and 5.0 μM was found to be optimum for inducing maximum shoots. Medium types, medium strength and pH were also investigated for induction
and proliferation of shoots. The highest efficiency of shoot proliferation was observed in 5.0 μM BA and 0.5 μM α-naphthalene acetic acid (NAA) in half-strength MS medium at pH 5.8. The best condition for rooting was half-strength MS
medium solidified with agar and with 2.0 μM indole-3-butyric acid (IBA). After rooting, the plantlets were transferred to plastic pots filled with sterile soilrite where
90% grew and all exhibited normal development. 相似文献
20.
Shoot proliferation has been achieved in Garcinia mangostana L. using seed explants. Maximum mean number of shoots per explant (16.8) was obtained from cultures on Murashige and Skoog medium supplemented with 40 mM 6- benzyladenine, and 2.5 mM -naphthaleneacetic acid and kept at 30 °C under an 8 hour photoperiod. Cultures on the same medium but supplemented with 2 g l-1 activated charcoal produced fewer shoots. However, growth of these shoots was more organized and 75% rooting was obtained. Woody Plant Medium was not a suitable medium for shoot proliferation. Ex vitro establishment was best obtained on planting medium consisting of sand, soil and organic material (3:2:1).Abbreviations BA
6-benzyladenine
- IBA
indole-3-butyric acid
- NAA
-naphthaleneacetic acid
- MS
Murashige & Skoog (1962) basal medium
- WPM
Woody Plant basal medium (Lloyd & Mc Cown 1980) 相似文献