共查询到20条相似文献,搜索用时 31 毫秒
1.
This study was carried out to determine the effect of temporary submersion of hypocotyl segments in water on in vitro explant growth and shoot regeneration on MS (Murashige and Skoog, 1962) medium supplemented with 1 mg l−1 BAP (6-benzylaminopurine) and 0.02 mg l−1 NAA (naphthaleneacetic acid) in three flax cultivars. It was observed that water-treated hypocotyl explants gave rise to
the highest values with respect to shoot regeneration percentage, shoot number per hypocotyl, shoot length and total shoot
number per Petri dish, successful rooting and plantlet establishment. This procedure may be applicable for other species cultured
in vitro. 相似文献
2.
Hairy nightshade (Solanum sarrachoides) has the potential to be a model system for the study of plant-pathogen interactions, however, the availability of tissue
culture and transformation methods would strengthen its utility. For the development of tissue culture methods, we investigated,
explant type (cotyledons, hypocotyls, roots), hypocotyl explant origin, cotyledon orientation (abaxial vs. adaxial) in direct
contact with the medium, gelling agents (agar and agargel) and cytokinins (zeatin and 6-benzyladenine) at different concentrations.
Cotyledon explants resulted in the greatest biomass as compared to root and hypocotyl. As for hypocotyl explant origin, explants
proximal to the cotyledons had a significant effect on plant regeneration. However, cotyledon orientation and gelling agent
had no effect on plant regeneration. Medium supplemented with either zeatin or 6-benzyladenine at 1 mg L−1 resulted in significant shoot regeneration. Shoots rooted readily when cultured on a non-hormone based rooting medium. 相似文献
3.
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
4.
Root, hypocotyl and cotyledonary explants of niger (Guizotia abyssinica Cass) CV. Sahyadri were aseptically cultured on Murashige and Skoog's basal medium (MS) containing BAP and kinetin. Multiple shoot regeneration was induced from hypocotyl and cotyledonary explants while root explants produced only callus on MS medium supplemented with BAP. BAP (1 mg l-1) was optimum for shoot regeneration. Regenerated shoots were transferred to MS medium without auxins, with auxins and with increasing concentrations of sucrose for rooting. Complete plantlets were obtained in all cases; however, 0.5 mg l-1 NAA was the best for induction of roots. Ninety-seven per cent of the plantlets survived and completed their life cycle when transferred to natural conditions.Abbreviations BAP
6-benzylamino purine
- NAA
-naphthaleneacetic acid
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid 相似文献
5.
Abstract Callus production, shoot formation via organogenesis and rooting of the regenerated shoots are reported in an Egyptian variety of Pisum sativum L. Calli were initiated from hypocotyl, leaf, root and mature embryo explants when cultured on MS medium containing B5 vitamins and supplemented with 2 mg/l 2,4-D+1 mg/l kin. Among the different types of explants, hypocotyl showed best potential for callus proliferation. Hypocotyl, leaf and immature cotyledon explants were used for shoot organogenesis. The best results of shoot formation were achieved when hypocotyl explants were cultured on MS-medium supplemented with 2 mg/l BA+1 mg/l NAA. However, immature cotyledon explants showed the highest frequency of shoot formation with 1 mg/l BA. Data of in vitro rooting showed that maximum root frequency occurred on culture medium containing half strength of MS salts, 40 g/l sucrose and 2 mg/l NAA. 相似文献
6.
Different vegetative parts of Brassica alboglabra seedlings and mature plants were used as explants in culture.A high frequency (60–100%) of shoot regeneration was obtained from hypocotyl explants, nodal stem segments, internodal segments and shoot apices cultured on Murashige-Skoog basal medium. Addition of 6-benzylaminopurine and kinetin increased the average number of shoots per explant. When detached and transferred to basal medium, the shoots readily developed roots. Regenerated plantlets could be successfully transplanted in soil. 相似文献
7.
The aim of the study was to obtain an efficient system for Carlina acaulis subsp. simplex propagation. The experimental materials were shoot tips, fragments of hipocotyls, cotyledons and roots isolated from 10-day-old
seedlings. The explants were transferred to the proliferation medium supplemented with different types of cytokinin: BA (13.3 μM),
kinetin (13.9 μM) and zeatin (13.7 μM) in combination with NAA (0.54 μM). The best morphogenetic response was observed when
explants were cultured on the BA supplemented medium. The maximum shoot organogenesis frequency was observed for shoot tip
(nearly 94%). On average 8.6 axillary shoots were induced per explant. Multiplication rate increased during the first three
subcultures. The shoots revealed a wide range of morphogenetic responses. Differences were observed in the presence or absence
of hair on the surface of lamina. These changes had epigenetic character and were the effect of changes in DNA methylation,
which is shown by differences in methylation pattern between 18S rRNA and 25S rRNA genes in the analyzed regenerated plants.
Nearly 94% of plantlets were rooted on auxin lacking medium. Addition of auxin (NAA or IAA) increased both the rooting percentage
(100%) and the number of roots per shoot, but their growth was inhibited. Shortening of the auxin exposition time reduced
the number of roots. Moreover, high efficiency (90%) was observed for ex vitro rooting. Plantlets with a large number of roots
survived better than the ones with only a few roots. Plants were able to flower and gave viable seeds. 相似文献
8.
Biochemical changes associated with adventitious shoot regeneration during in vitro culture of hypocotyl explants of Jatropha integerrima were determined. Histological and biochemical studies were undertaken at 7-d intervals, up to four weeks on hypocotyl explants cultured on basal Murashige and Skoog's medium supplemented with 0.5 mg dm-3 N6-benzyladenine and 1.0 mg dm-3 indole-3-butyric acid. Initial cell proliferation occurred within one week of culture; meristemoid differentiation within two to three weeks and shoot development after four weeks. Peak activities of alkaline phosphatase, peroxidase and polyphenol oxidase was observed at day 14 indicating their involvement in the formation of meristematic centers. Protein accumulation and acid phosphatase activity were maximum at day 28. 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.
A high frequency shoot regeneration system for ornamental kale [Brassica oleracea L. var. acephala (D.C.) Alef.] was firstly established from seedling cotyledon and hypocotyl explants. The ability of cotyledon and hypocotyl
to produce adventitious shoots varied depending upon genotype, seedling age and culture medium. The maximum shoot regeneration
frequency was obtained when the explants from cv. Nagoya 4-d-old seedlings were cultured on Murashige and Skoog (MS) medium
supplemented with 3 mg dm−3 6-benzylaminopurine (BA) and 0.1 mg dm−3 naphthaleneacetic acid (NAA). The frequency of shoot regeneration was 65.0 % for cotyledons, 76.1 % for hypocotyls; and the
number of shoots per explant was 4.3 for cotyledons, 8.2 for hypocotyls. Hypocotyl explants were found to be more responsive
for regeneration when compared with cotyledons. Among the 4 cultivars tested, Nagoya showed the best shoot regeneration response.
The addition of 3.0 mg dm−3 AgNO3 was beneficial to shoot regeneration. Roots were formed on the base of the shoots when cultured on half-strength MS medium. 相似文献
11.
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
12.
Summary
In vitro adventitious shoot bud regeneration systems are considered most suitable for Agrobacterium-and biolisticsmediated genetic transformation to obtain transgenic plants. In the present investigation, multiple adventitious
shoot buds could be induced directly from Feronia limonia hypocotyl explants inoculated on Murashige and Skoog (MS) medium containing different growth regulators. During the initial
phase, the hypocotyl segments nearer to the cotyledons responded quickly compared to those closer to the root. The response,
however, was comparable in both the segments in subsequent subculture. Of the various cytokinins, 2.22 μM 6-benzylaminopurine (BA) proved to be more effective compared to kinetin (Kn). The two-way interaction of BA and Kn significantly
influenced shoot regeneration and contributed the most among the interactions studied. The best response, however, was obtained
when 2.22 μM BA and 2.32 μM Kn were combined. Although the effect of auxins like α-naphthaleneactic acid (NAA) combined with cytokinins evoked a significant
responsein terns of number of shoot buds, this response did not supersede the effect of combined cytokinins. Vone of the polyamines
tested induced shoot buds on hypocotyl segments. Adventitious shoots were multiplied on MS medium containing 2.22 μM BA, 6.96 μM Kn, and 0.05 μM NAA. More than 60% of the shoots produced roots when cultured on medium containing one-quarter strength MS salts, 10% suerose,
0.6% agar, and 7.36μM indole-3-butyric acid. The adventious origin of shoot buds showing continuous vascular connections was confirmed through
histological investigations. 相似文献
13.
In this work we report a new method forin vitro chili pepper (Capsicum annuum L.) plant regeneration based on shoot formation from wounded hypocotyls. Chili pepper seeds were surface sterilized and germinated on agar (0.8%) at 25 ± 2°C in the dark. Five factors that may influence shoot regeneration were studied: age of seedlings, hypocotyl wounding site, time elapsed between wounding the hypocotyls and decapitation of seedlings, culture media and cultivars. In order to study the influence of the first three factors on shoot regeneration, the apical, middle or basal hypocotyl regions of seedlings of cv. Mulato Bajio at different stages of development (9, 15, 16, 21 and 28 d old) were wounded with a syringe needle, and the seedlings were cultured on MS semisolid medium without growth regulators at 25 ± 2°C under a 16/8 h light/dark photoperiod (daylight fluorescent lamps; 35 mol m-2 s--1) until decapitation. The seedlings were decapitated (3 mm below the cotyledons) at different times after wounding (0, 2, 4, 10, 12 and 14 d), and each explant was evaluated for bud and shoot formation ( 5 mm in length) at the wounded site after 30 d of incubation. In general, seedlings at the stage of curved hypocotyl (9 d old) wounded in the apical region of hypocotyl were the best explants for shoot regeneration when inoculated on culture medium without growth regulators. Decapitation after wounding also influenced the shoot regeneration efficiency, with 10–14 d being the best period. Up to 90% shoot regeneration in cv. Mulato Bajio was obtained under these conditions. Statistically significant differences were observed for shoot formation among 21 cultivars tested. Regeneration of whole plants was achieved by rooting the shoots with indole-3-butyric acid pulses of 60 mg L–1 for 3 h and then subculturing on MS medium without growth regulators. 相似文献
14.
A micropropagation protocol for shoot culture of sweet viburnum (Viburnum odoratissimum) is described. Nodal explants, initially established on MS medium, were transferred to WPM supplemented with combinations of BA and GA3. Maximum shoot multiplication was observed on explants cultured on medium supplemented with BA concentration higher than 1.1 μM, and 14 μM GA3. Although Stage II medium supplemented with BA concentration higher than 1.1 μM resulted in increased shoot multiplication, it also caused a decrease in shoot length. A negative carry over effect of GA3 on rooting was observed in subsequent Stage III cultures. The presence of GA3 in Stage II medium promoted shoot elongation, but it also caused a decrease in microcutting rooting. For this reason, 0.5 μM BA and 14 μM GA3 were selected for optimum Stage II shoot multiplication. Although 100% microcuttings formed roots when cultured on medium containing 6.0 μM NAA, significant callus formation was observed and ex vitro survival rate was low (49%). Rooting was achieved after 3 weeks with 82% of microcuttings on medium supplemented with 3 μM IBA. The survival rate of plantlets under ex vitro conditions was 100% after 3 weeks. Plants looked healthy with no visually detectable phenotypic variation based on observation of about 30 plants. 相似文献
15.
Summary Tennessee coneflower [Echinacea tennesseensis (Beadle) Small] was regenerated from flower stalks, leaf sections from flowering plants, and hypocotyls and cotyledons from
seedlings. Murashige and Skoog medium (MS) supplemented with naphthaleneacetic acid (NAA) at 0.54 μM and thidiazuron (TDZ) at 22.7 μM yielded the most shoots per leaf explant. NAA and 6-benzylaminopurine concentrations for optimal shoot regeneration from
leaf, flower stalk, cotyledon and hypocotyl explants in MS media were 0.54 and 24.6μM, respectively. All explant types generated shoots; however, those derived from leaves and flower stalks produced the highest
number of shoots per explant and highest percentage of explants with shoots. Explants cultured on media containing high levels
of NAA (5.4–27 μM) formed calluses but no adventitious shoot. Leaf explants responded to a wider range of NAA concentrations than the other
explant types but shoots generated from flower stalks grew the fastest. While all cytokinins tested increased the number of
shoots per explant, the number of shoots in media containing TDZ was increased by nearly threefold. Regenerated shoots from
all explant types cultured on MS medium supplemented with 0.25 μM indole-3-butyric acid initiated roots within 4 wk; NAA was not effective for root induction. All vernalized plantlets developed
into plants that were morphologically identical to the source material. 相似文献
16.
An effective and reproducible protocol for the micropropagation ofKigelia pinnata L through high frequency callus regeneration is described. Seeds were surface sterilized before culturing on Gamborgs basal
medium (B5 medium). After two weeks the cotyledonary node along with a portion of the hypocotyl were carefully excised from
well-developed embryos and subcultured on B5 medium supplemented with different concentrations of 2,4-D and BAR The cultured
cotyledonary node expiants showed callus formation at the base of the lower cut end of the hypocotyl. This callus showed shoot
initiation after two weeks of subculture on the regeneration medium supplemented with various concentrations of BAP alone
or in combination with NAA. The highest number of shoot regeneration occurred on medium containing 5 γM BAP and 0.1 γM NAA.
The optimum rooting of the regenerated shoots was observed on 1/2B5 medium supplemented with 4 γM IBA. Micropropagated plants
were successfully established in soil in field condition with a survival frequency of 100%. 相似文献
17.
Amanda Ferreira da Silva Mendes Luciana Cardoso Cidade Maria Luiza Peixoto de Oliveira Wagner Campos Otoni Walter Dos Santos Soares-Filho Marcio Gilberto Cardoso Costa 《Plant Cell, Tissue and Organ Culture》2009,97(3):331-336
Identification of beta-lactam antibiotics that have negligible effects on plant regeneration is a critical step towards the
establishment of a reliable Agrobacterium-mediated transformation protocol for perennial trees. In the present report, we have evaluated the effects of the novel beta-lactam
antibiotics meropenem and timentin on plant regeneration of a perennial woody fruit plant, Citrus sinensis, in comparison with the commonly used beta-lactam cefotaxime. It was observed that, in contrast to cefotaxime, meropenem
and timentin had a positive or no detrimental effect on the shoot regeneration from epicotyl explants. Residual effects of
the beta-lactams from shoot regeneration medium also affected the subsequent ability of the roots to elongate. The addition
of meropenem and/or timentin in the rooting medium mostly improved or did not affect the rooting ability of the adventitious
shoots. These data indicated that meropenem and timentin can positively replace cefotaxime in Agrobacterium-mediated transformation of C. sinensis. 相似文献
18.
Summary Thirteen soybean genotypes representing maturity groups IV−VI were compared for organogenic responses on three media cultured
under two lighting conditions with hypocotyl sections excised from 7-d-old seedlings. All soybean lines responsed by producing
adventitious shoots on the acropetal end of the hypocotyl explants, confirming genotype-independence of shoot initiation.
Media containing 6-benzyladenine (BA; 5.0–10 μM) induced the greatest numbers of shoots. Histological studies confirmed the adventitious nature of arising shoots by indicative
formation of meristematic zones and shoot primordia from parenchymatous tissues of central pith and plumular trace regions
of the hypocotyl. Incompletely excised cotyledonary buds also contributed to shoot initiation. Degrees of responses were media-dependent
and varied with regard to genotype. Centennial, Epps, and Lyon gave the greatest individual responses. Between cultivars (across
all treatments), the regeneration potential (percentage of explants producing meristem-like structures or shoot primordia)
4 wk after initiation ranged from 47 to 75%. Four wk later, regenerative ability (number of shoots produced per responding
explant) and regeneration efficiency (number of shoots produced per explant plated) yielded 1.4–7.1 and 1.0–5.0 shoots, respectively.
The optimized protocol included initiation on a medium containing 5.0 μM BA for 4 wk, then transfer onto a shoot elongation medium (0.36 μM BA) for 4 wk. For 11 genotypes tested, 66–100% of excised shoots produced roots after 4 wk on media containing 12.5–29.2
μM indole-3-butyric acid. Of 109 regenerants transplanted to soil, 94% survived and no sterility has been observed on those
mature enough to flower. 相似文献
19.
The effects of nitric oxide (NO) on caulogenesis, shoot organogenesis and rhizogenesis from hypocotyl explants of Linum usitatissimum were investigated. Exogenously supplied NO donors, 5 μM sodium nitroprusside (SNP), 2 μM S-nitroso-N-acetylpenicillamine (SNAP) and 2 μM 3-morpholinosydnonimine (SIN-1), significantly promoted shoot differentiation from the
hypocotyl explants of L. usitatissimum excised from its in vitro raised seedlings. Potassium ferrocyanide, a structural analogue of SNP, lacking NO group, did not
promote shoot organogenesis. Likewise, products of NO,
\textNO2 - {\text{NO}}_{2}^{ - } and
\textNO3 - {\text{NO}}_{3}^{ - } supplied as 5 μM NaNO2 and 5 μM NaNO3 did not enhance shoot differentiation. Another source of NO, a mixture of sodium nitrite (SN) provided along with ascorbic
acid (AsA), also caused significant promotion in the average number of shoots per responding explant. SNP also augmented the
rhizogenic response of the microshoots in terms of percentage of responding explants, number of roots per responding explant
and average root length. The NO scavengers, 2-(4-carboxy-phenyl)-4, 4, 5, 5-tetramethylimideazoline-1-oxyl-3-oxide (cPTIO)
or methylene blue (MB), provided along with SNP, SNAP, SIN-1 or SN + AsA, at concentrations equimolar to the optimum concentration
of the donors, reversed the promotory influence, thereby, confirming the role of NO in promotion of in vitro morphogenesis.
However, NO scavengers individually did not affect the observed morphogenic processes. Morphological and histological studies
of hypocotyl segments cultured on BM or BM + SNP for 4, 8 and 12 days demonstrated that SNP enhanced shoot differentiation
by inducing a higher number of shoot primordia, each of which develops into a single shoot. 相似文献
20.
A high frequency in vitro shoot bud differentiation and multiple shoot production protocol from hypocotyl segments of 8 to 10-d-old seedlings of cotton
has been developed. Murashige and Skoog (MS) basal medium with Nitsch and Nitsch vitamins was found to be optimal in shoot
regeneration. A combination of 2 mg dm−3 thidiazuron and 0.05 mg dm−3 naphthaleneacetic acid was the most effective for shoot regeneration (76 %) and an average of 10.6 shoots per responding
explant. Combination of the cytokinins benzylaminopurine and kinetin induced better regeneration response than their individual
treatments. Supplementation of the culture medium with ethylene inhibitor silver nitrate and activated charcoal showed beneficial
effects. Optimal rooting was obtained on half-strength MS medium supplemented with 1 mg dm−3 indolebutyric acid and activated charcoal. Scanning electron micrographs of in vitro cultured explants revealed that shoot primordia were formed de novo. 相似文献