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1.
Internode explants collected from in vitro grown shoots of two clones of Fagus sylvatica L. (European beech) and five clones of F. orientalis Lipski (Oriental beech) were used to evaluate their bud regeneration capacity. Adventitious shoot-buds formed on callus,
which developed from internode segments cultured in a Woody Plant Medium supplemented with different concentrations of either
thidiazuron (TDZ) or benzyladenine (BA). After 4 weeks of culture on induction media, the explants were transferred to a proliferation
medium supplemented with 2.2 μM BA, 9.1 μM zeatin and 2.9 μM indole-3-acetic acid (IAA) for another 8 weeks. Medium containing
TDZ was much more efficient than medium containing BA in inducing adventitious buds, the optimal TDZ concentration being 4.5
μM and the optimal BA concentration 17.8 μM. Genotypic variation in shoot regeneration capacity was observed among the two
Fagus species and between clones within each species, with a significant interaction between TDZ concentration and genotype regarding
mean bud number. Thidiazuron induction medium supplemented with a range of individual auxins was investigated, and it was
found that IAA or indole-3-butyric acid at 2.9 μM enhanced the bud forming capacity of explants. Morphogenic response varied
significantly with the position of the internode along the stem. The highest regeneration potential was obtained from apical
internodes, while those distal to the apex were the least productive. Elongated shoots of adventitious origin can be readily
proliferated by axillary branching.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
2.
C. Pugliesi M. Rabaglio F. Cecconi S. Baroncelli 《Plant Cell, Tissue and Organ Culture》1992,28(2):125-128
Different plant explants of Persian buttercup (Ranunculus asiaticus L.) were screened for callus induction and adventitious shoot regeneration on different media to establish totipotent cultures. Murashige & Skoog (MS) medium was used, supplemented with different concentrations of the following growth regulators: kinetin, benzyladenine (BA), naphthaleneacetic acid (NAA) and indoleacetic acid (IAA). Callus was induced and adventitious buds regenerated only from cotyledonary explants after 4–5 weeks. Subculture of the regenerated buds on the same basal medium in presence of gibberellic acid (GA3) and BA produced well-organized shoots. Rooting was obtained by transferring shoots to growth regulator-free MS medium. A high rate of shoot multiplication has been achieved on medium with high concentration of kinetin and long-day photoperiod. Finally the plants were successfully transferred to soil and grown in a greenhouse. 相似文献
3.
Regeneration from Phylloclade Explants and Callus Cultures of Schlumbergera and Rhipsalidopsis 总被引:1,自引:0,他引:1
Phylloclade explants of Schlumbergera and Rhipsalidopsis were cultured in vitro to produce axillary and adventitious shoots. The explants of both species, taken from greenhouse-grown plants, produced only
axillary shoots. There was a pronounced improvement in adventitious shoot formation in phylloclade explants of cultivar CB4
of Rhipsalidopsis by increasing numbers of subcultures of axillary shoots used as donor plants. The axillary shoots generated from the explants
were either subcultured to produce successive generations of axillary shoot cultures or made into phylloclade explants and
tested for adventitious shoot formation at each subculture. The duration of each subculture varied from 6 to 12 weeks. After
the first subculture, sporadic adventitious shoot formation began, and after the third subculture 87% explants of cultivar
CB4 produced adventitious shoots at a frequency of about 12 shoots per explant. In contrast, there was no improvement in regenerative
ability in explants of cultivar Thor-Olga of Schlumbergera up to third subculture. Adventitious shoots could be produced by callus culture too. Cultivar CB4 was highly regenerative,
producing as many as 10 adventitious shoots per square cm of callus. In vitro grown plantlets, when transferred to pots continued to show prolific growth. 相似文献
4.
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
5.
A system has been developed for rapid selection of streptomycin resistant mutants, as adventitious shoots arising from explants of several Solanaceous species. Efficient mutagenesis was achieved by incubating shoot culture-derived leaf strips with 1 or 5 mM nitroso-methylurea, for 90 or 120 min. In Nicotiana tabacum and Lycopersicon peruvianum these treatments resulted in white or variegated adventitious shoots from up to 3.5% of explants placed on medium promoting shoot regeneration. Chlorophyll deficiencies were only observed very rarely in Solanum nigrum. Streptomycin resistant shoots were obtained from leaf explants placed on medium containing 500 mg l-1 streptomycin sulphate, under which conditions explants are bleached and adventitious shoot development suppressed. Green adventitious s shoots appeared at a frequency dependent both on the mutagenic treatment and on the species. The best response was with S. nigrum where >70% of the explants produced streptomycin resistant shoots, most of which retained their resistance on subsequent testing. Maternal inheritance of streptomycin resistance has been confirmed for several N. tabacum and S. nigrum mutants, and there is also evidence for paternal transmission in the latter species. The procedure has been successfully extended to other species, including N. sylvestris and N. plumbaginifolia, and also to obtain spectinomycin resistant mutants.Communicated by R. Hagemann 相似文献
6.
The influences of carbon sources, fructose, glucose, sorbitol and sucrose on shoot proliferation and in vitro rooting of cork oak (Quercus suber L.) were compared at a wide range of concentrations (1–6%, w/v). The highest number of shoots occurred on glucose-containing medium. Nevertheless, we have chosen 3% sucrose which induced a similar rate of proliferation but favoured shoot elongation, permitting an effectively higher number of shoots during transfers. Sorbitol and autoclaved fructose did not stimulate shoot proliferation. Adventitious root formation was strongly dependent on carbohydrate supply. Sorbitol and autoclaved fructose were completely ineffectively on rooting induction. Glucose was the most effective carbon source on rooting promotion followed by sucrose and filter-sterilized fructose. The rooting response induced by fructose was dependent on the sterilizing procedure. The number of adventitious roots produced per shoot increased with increasing glucose and sucrose concentration. The content of reducing sugars in leaves of proliferation cultures and in leaves and roots of rooted plantlets was more dependent on carbon concentration than on glucose or sucrose supplement. The results presented here show that carbohydrate requirements during cork oak micropropagation depend upon the phase of culture. Sucrose (3%) and glucose (4%) were the best carbon sources respectively during proliferation and rooting phases. 相似文献
7.
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
8.
Adult plants are known for recalcitrance when it comes to adventitious organ formation and regeneration. Methods used for
regeneration in explants from seedlings of Campanula carpatica failed to work for explants from adult plants of the same species. The present investigation generated efficient regeneration
methods for mature specimens of four species of Campanula, C. carpatica, C. haylodgensis, C. portenschlagiana and C. poscharskyana. Petiole explants from dark-grown in vitro shoot cultures grown from nodal cuttings of adult plants regenerated successfully
(95%), while explants from light-grown in vitro shoot cultures and greenhouse-grown plants regenerated at 12% and zero percentage,
respectively. Dark-treatment, along with media manipulation with plant growth regulators, further enhanced regenerative capacity
of the explants. A MS-based medium containing 10mg l −1 TDZ and 0.25 mg l−1 NAA was the most efficient regeneration medium. Transgenic shoots from C. carpatica (3%) and C. haylodgensis (1%) and transgenic callus from all species were produced using Agrobacterium tumefaciens, and transformation was confirmed by histochemical and Southern blot analyses. Protocols developed in this study may be useful
for achieving efficient regeneration and transformation of recalcitrant adult plants. 相似文献
9.
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
10.
Mohsen Sanikhani Stefan Frello Margrethe Serek 《Plant Cell, Tissue and Organ Culture》2006,85(1):75-82
In order to optimize shoot regeneration in Kalancho? blossfeldiana, leaf and internode explants of seven cultivars including one inter-specific were studied. The effects of various combinations
of α-naphthalene acetic acid (NAA) (0, 0.57 M) and thidiazuron (TDZ) (0, 0.45, 4.5, 22.5, 67.5 μM) on MS medium were examined.
In all cultivars shoot regeneration frequency and number of shoots per explant were enhanced by increasing TDZ concentration.
Supplementing the media with NAA did not improve shoot regeneration. Maximum regeneration frequency and optimum concentration
of TDZ for shoot regeneration depended significantly on the cultivar. Internode explants, but not leaf explants, of some cultivars,
were able to produce adventitious shoots without treatment with growth regulator. 相似文献
11.
Manuel L. Robert J. L. Herrera F. Contreras Keith N. Scorer 《Plant Cell, Tissue and Organ Culture》1987,8(1):37-48
In vitro plant regeneration of Agave fourcroydes Lem. (Agavaceae) is described. Results suggest that the NO3
-:NH4
+ balance in the culture medium is a key factor controlling callus growth and organogenesis in rhizome cultures. Stem callus showed limited organogenic capacity, but high cytokinin concentrations induced adventitious shoot formation on stem explants. When these shoots were excised and subcultured, new callus formed at their base from which new shoots arose. The shoots from stem explants and rhizome callus formed extensive root systems in vitro and were transferred to pot culture with a 90% survival rate. 相似文献
12.
Ajith Anand Chaluvadi Srinivasa Rao 《In vitro cellular & developmental biology. Plant》2000,36(1):61-64
Summary A protocol is described for rapid multiplication of Piper barberi Gamble (Piperaceae) through shoot tip and nodal explant cultures. Nodal explants with a single axillary meristem showed three
times better response with respect to shoot proliferation when compared to shoot tip explants. The best shoot proliferation
response of nodal explants was observed with a cytokinin combination of N6-benzyladenine (4.43 μM) and kinetin (2.32 μM), with 88% bud break. The number of shoot initials (2.4) produced per nodal explant was twice the number of shoot initials
(1.2) per shoot tip. An average of 6.9±0.58 adventitious shoots were observed from the proximal end of the internodal explants
on Mursashige and Skoog (1962) (Ms) basal medium supplemented with N6-benzyladenine (2.22 μM) and kinetin (0.46 μM). A multiplication rate of 82 shoots per explant could be achieved after 9 wk of subculturing. The in vitro shoots were rooted on one-half and one-quarter MS basal medium. The shoots rooted on one-quarter MS in the dark produced
eight roots with an average root length of 3.36 cm and 98% survival. These plants were transferred to the field with a survival
rate of 75%. 相似文献
13.
Sriskandarajah Sridevy Frello Stefan Serek Margrethe 《Plant Cell, Tissue and Organ Culture》2001,67(3):295-298
Efficient protocols have been developed to induce adventitious shoots in different types of explants of Campanula carpatica Jacq. More than five shoots per explant developed on hypocotyls of 5-week-old seedlings after 2 weeks of culture. Hypocotyls produced twice as many shoots as the cotyledons. TDZ proved to be about 6 times more efficient than BA. NAA had to be added to the regeneration medium to obtain the optimal balance of auxin and cytokinin to induce shoot regeneration. Significant differences were noted between different growth regulator concentrations in their effects on shoot organogenesis. BA induced double the number of callus clumps as TDZ. Incubation of explants in the dark produced about 6 shoots per explant while those in the light produced about 2 shoots per explant. Explants derived from 5-week-old seedlings were five times more regenerative compared to those derived from 15-week-old seedlings. Explants from cv. White Uniform were more organogenic than those from cv. Blue Clip. Root segments were also found to form shoots when treated with CPPU. 相似文献
14.
Primordial shoot explants excised from buds of one Larix decidua tree, about 30 years old, produced more adventitious buds, elongating into shoots, when grown on half strength Litvay medium than when grown on other basal media. Thidiazuron and N6-benzyladenine (BA) were equally effective in adventitious bud induction. In a comparative study of 30-year-old L. decidua, L. leptolepis, L. eurolepis, and L. laricina trees, explants from L. eurolepis and L. decidua produced a high number of cultures with adventitious buds that elongated into shoots; those from L. leptolepis were less productive, and those from L. laricina failed to form adventitious buds. The highest response was obtained with material collected in August and September, and in March and April; the lowest response occurred in explants from the October collection. 相似文献
15.
H. Y. Seo Y. J. Kim T. I. Park H. S. Kim S. J. Yun K. H. Park M. K. Oh M. Y. Choi C. H. Paik Y. S. Lee Y. E. Choi 《In vitro cellular & developmental biology. Plant》2007,43(3):209-214
Sesamum indicum L. was used as an important oil crop in the world. An efficient protocol for in vitro plant regeneration via adventitious shoot formation from deembryonated cotyledon explants isolated from mature seeds of sesame
is developed. Optimal medium for direct adventitious shoot formation was Murashige and Skoog (MS) medium with 22.2 μM 6-benzylaminopurin
(BA) and 5.7 μM indole-3-acetic acid (IAA). Abscisic acid (3.8 μM ABA) and AgNO3 (29.4 μM) were effective in enhancing the frequency of adventitious shoot formation. Preculture of cotyledon explants on
high sucrose concentration (6–9%) for 2 wk and subsequent transfer to 3% sucrose enhanced the frequency of adventitious shoot
induction. Root formation from the adventitious shoots was easily achieved on MS medium containing 2.7 μM of α-naphthalene
acetic acid (NAA). Regenerated plantlets were acclimatized on sand and peat moss (1:1), showing 95% survival with subsequent
flowering and seed set. We established the high-frequency plant regeneration via adventitious shoot formation in S. indicum L. 相似文献
16.
In order to investigate the regeneration of wild beet (Beta maritima) from inflorescence pieces, the effects of growth regulator, genotype, explant source and stage of plant development on adventitious shoot formation and rooting in vitro and subsequent transplanting in the glasshouse were tested. Inflorescence tips produced more adventitious shoots than sub-apical segments and the best micropropagation was achieved on a Murashige and Skoog (MS) medium supplemented with 1.0 mg l–1 BAP. Addition of auxin was not beneficial. The induction rate of adventitious shoots was genotype-dependent and influenced by the stage of plant development. Adventitious shoots were produced from the base of the flower buds, i.e. from the receptacle, not from axils or stalks and only a few buds on inflorescence tip explants produced adventitious shoots. Rooting was increased by using a MS medium with 3% sucrose supplemented with 1.0 mg l–1 NAA. There was no variation in leaf morphology of the transplants. This work shows that inflorescence tips can be used successfully as explants for in vitro multiplication of sugar beet and wild beet.Abbreviations BAP
benzylaminopurine
- IBA
indole-3-butyric acid
- GA3
gibberellic acid
- MS
Murashige and Skoog medium
- NAA
naphthaleneacetic acid
Author for correspondence 相似文献
17.
Jayanand B. Sudarsanam G. Sharma Kiran K. 《In vitro cellular & developmental biology. Plant》2003,39(2):171-179
Summary An efficient and reproducible protocol for the regeneration of shoots at high frequency was developed by using explants derived
from the axillary meristems from the cotyledonary nodes of in vitro-germinated seedlings of chickpea (Cicer arietinum L.). Culture conditions for various stages of adventitious shoot regeneration including the induction, elongation, and rooting
of the elongated shoots were optimized. The medium for synchronous induction of multiple shoot buds consisted of Murashige
and Skoog basal medium (MS) with low concentrations of thidiazuron (TDZ), 2-isopentenyladenine (2-iP), and kinetin. Exclusion
of TDZ and lowering the concentration of 2-iP and kinetin in the elongation medium resulted in faster and enhanced frequency
of elongated shoots. Cultivation of the stunted shoots on MS with giberellic acid (GA3) increased the number of elongated shoots from the responding explants. pH of the medium played a very crucial role in the
regeneration of multiple shoot buds from the explants derived from cotyledonary nodes. A novel rooting system was developed
by placing the elongated shoot on a filter paper bridge immersed in liquid rooting medium that resulted in rooting frequency
of up to 90%. A comprehensive protocol for successful transplantation of the in vitro-produced plants is reported. This method will be very useful for the genetic manipulation of chickpea for its agronomic improvement. 相似文献
18.
Cotyledon explants of Brassica tournefortii L. were excised from germinated seedlings and cultured on Murashige & Skoog's [6] basal medium supplemented with various combinations of cytokinins and auxins, Both cytokinin and auxin were required for induction of shoot organogenesis. Of the three cytokinins tested (in combination with a low concentration of IAA), kinetin was found to be the best for shoot regeneration. On this medium, cotyledonary explants invariably underwent callusing followed by multiple shoot formation. NAA in combination with any of the three cytokinins yielded a reduced number of shoots or none, but favoured good callus growth. Callus so produced also regenerated shoots when subcultured on media containing high concentration of KIN or ZEA and low concentration of IAA. Shoots were rooted during prolonged incubation on the same medium or on MS medium free of growth regulators. Mature plants were grown in the greenhouse. 相似文献
19.
Somatic embryogenesis in Canary Island date palm 总被引:14,自引:0,他引:14
Huong Le Thi Lan Baiocco Michela Huy Bao Phan Mezzetti Bruno Santilocchi Rodolfo Rosati Pasquale 《Plant Cell, Tissue and Organ Culture》1999,56(1):1-7
Shoot regeneration was obtained from leaves of in vitro cultures of wild pear genotypes. The highest regeneration rates, ranging from 40% to 64%, depending on the genotype, were
obtained using leaves wounded by three cuts transversely to the mid-rib, a Quoirin and Lepoivre macro-salt composition, 250
mg l-1 cefotaxime and maintaining the explants in darkness for the first 30 days (induction phase), then transferring them to an
auxin-free medium in light (expression phase). A concentration of 8.8 μM BA induced the highest number of explants to produce
adventitious shoots. TDZ was less effective than BA and induced hyperhydricity in regenerated shoots. The histological studies
revealed that the regenerated shoots originated mainly from callus formed by epidermal and sub-epidermal cells and by cells
of the vascular tissue. The regenerated shoots were micropropagated, rooted and transplanted to the soil.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
20.
S. C. Debnath 《In vitro cellular & developmental biology. Plant》2009,45(2):122-128
An efficient system to regenerate shoots on excised leaves of greenhouse-grown wild lowbush blueberry (Vaccinium angustifolium Ait.) was developed in vitro. The effect of thidiazuron (TDZ) on adventitious bud and shoot formation from apical, medial, and basal segments of the leaves
was tested. Leaf cultures produced multiple buds and shoots with or without an intermediary callus phase on 2.3–4.5 μM TDZ
within 6 wk of culture initiation. The greatest shoot regeneration came from young expanding basal leaf segments positioned
with the adaxial side touching the culture medium and maintained for 2 wk in darkness. Callus development and shoot regeneration
depended not only on the polarity of the explants but also on the genotype of the clone that supplied the explant material.
TDZ-initiated cultures were transferred to medium containing 2.3–4.6 μM zeatin and produced usable shoots after one additional
subculture. Elongated shoots were dipped in 39.4 mM indole-3-butyric acid powder and planted on a peat:perlite soilless medium
at a ratio of 3:2 (v/v), which yielded an 80–90% rooting efficiency. The plantlets were acclimatized and eventually established in the greenhouse
with 75–85% survival. 相似文献