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1.
Factors influencing reliable shoot regeneration from leaf explants of rapeseed (Brassica napus L.) were examined. Addition of AgNO3 to callus induction medium was significantly effective for shoot regeneration in all three genotypes initially tested. When
48 genotypes subsequently were surveyed, a large variation of shoot regenerability was observed, ranging from 100 to 0% in
frequency of bud formation and from 7.5 to 0 in the number of buds per explant. A significant correlation (r=0.84) was observed between the frequency of bud formation and the number of buds per explant. The shoot regenerability from
leaf explants was not related to that from cotyledonary explants (r=0.28). Histological observations showed that an organized structure developed from calluses produced at vascular bundle tissues
after 7 days of culture on callus induction medium, and they developed shoot apical meristems one week after transfer onto
shoot induction medium. Regenerated plantlets were obtained 2 months after the initiation of culture and they normally flowered
and set seeds. No alterations of morphology or DNA contents were observed in regenerated plants and their S1 progenies. 相似文献
2.
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
3.
Direct plant regeneration from leaf explants of Plumbago species and its genetic fidelity through RAPD markers 总被引:1,自引:0,他引:1
G R ROUT 《The Annals of applied biology》2002,140(3):305-313
In vitro plant regeneration was achieved from leaf explants of Plumbago rosea and Plumbago zeylanica on Murashige & Skoog (1962) medium supplemented with 1.5 mg litre?1 6‐benzylaminopurine, 0.25 mg litre?1 indole‐3‐acetic acid, 50 mg litre?1 adenine sulfate and 3% (w/v) sucrose. The shoot initials developed within 2–3 wk on the leaf margin as well as from the wounds of the leaf. High frequency shoot‐bud regeneration was achieved on similar medium in subsequent subcultures. The semi‐mature leaves produced more shoot‐buds as compared to the younger leaves. Mature leaves did not show any response for shoot bud initiation. More than 85% of the semi‐mature explants produced shoot‐buds per leaf explant within 4 wk of culture. Shoots rooted easily on medium having half‐strength basal Murashige & Skoog (1962) medium supplemented with 0.25 mg litre?1 indole‐3‐butyric acid and 2% (w/v) sucrose; 84–92% of the in vitro rooted plantlets survived in the greenhouse. The regenerated plantlets appeared morphologically similar to the mother plants. No variation was detected among the regenerated plants by the use of Randomly Amplified Polymorphic DNA (RAPD) markers. This method might be useful for assessing plant improvement programmes. 相似文献
4.
5.
Houcheng Zhou Ming Li Xia Zhao Xiucai Fan Aiguang Guo 《Plant Cell, Tissue and Organ Culture》2010,101(1):79-87
A complete protocol for adventitious shoot regeneration was developed from the leaves of peach rootstock ‘Nemaguard’(Prunus persica × P. davidiana) grown in vitro. Shoot explants were cultured in vitro in Murashige and Skoog medium supplemented with 3.55 μM 6-benzyladenine
and 7.38 μM indole-3-butyric acid (IBA). Non-expanded leaves along with their petioles from 3-week-old in vitro-grown shoots
were used as explants. Regeneration percentage was influenced by plant growth regulators, basal medium, explant type, dark
period, and gelling agents. Optimal regeneration was observed with leaf explants wounded by transverse cuts twice along the
midrib and first incubated with abaxial surfaces facing upward in the dark for 3 weeks, and then transferred to the light
and cultured with the adaxial side in contact with regeneration medium, as seen on 1/2 MS, woody plant medium or Schenk and
Hildebrandt medium supplemented with 9.08 μM thidiazuron, 0.54 μM IBA and 0.25% agar. This produced the highest regeneration
percentage at 71.7% and a mean of 5.74 ± 3.24 shoots on 1/2 MS medium. Adventitious shoots were rooted (98.3–100%) and rooted
plantlets survived after acclimatization to the greenhouse. 相似文献
6.
In vitro anther-derived monoploids (2n=x=12) of Solanum phureja were compared for shoot regeneration from leaf and stem explants under various environmental conditions. Monoploids from the same or different diploid clones varied for frequency and earliness of shoot regeneration and number of shoots formed per explant. Leaf explants regenerated at higher frequencies than stem explants. Explants from stock plantlets subcultured at a 2- or 4-week interval regenerated earlier and at a higher frequency than those from plantlets subcultured at longer intervals. Regeneration frequency and number of shoots per explant were greater when explants were incubated at 20°C compared to 25°C. Explants from stock plantlets maintained under a 16 h as opposed to an 11 h photoperiod exhibited increased shoot regeneration; however, neither photoperiod nor the maintenance temperature of the stock plantlets influenced regeneration frequency. Genotypic differences were observed for the frequency of chromosome doubling among regenerated shoots whereas temperature treatments had no influence on chromosome doubling.Abbreviations BA
benzyladenine
- GA3
gibberellic acid
- IAA
indole-3-acetic acid
- NAA
-naphthale-neacetic acid 相似文献
7.
Anne -Claire van Altvorst Svetla Yancheva Hans Dons 《Plant Cell, Tissue and Organ Culture》1995,40(2):151-157
Adventitious shoot formation was studied with leaf, stem and axillary bud explants of carnation (Dianthus caryophyllus L.). The shoot regeneration procedures were applicable for a wide range of cultivars and shoot regeneration percentages were high for all explant types. Using axillary bud explants, shoot regeneration efficiency was independent of the size of the bud and of its original position in the plant. In contrast, shoot regeneration from stem and leaf explants was strongly dependent on their original position on the plant. The most distal explants (just below the apex) showed the highest level of shoot regeneration. The adventitious shoot primordia developed at the periphery of the stem segment and at the base of leaf explants. In axillary bud, stem and leaf explants, shoot regeneration originated from node cells, located at the transition area between leaf and stem tissue. Moreover, a gradient in shoot regeneration response was observed, increasing towards the apical meristem.Abbreviations BA
benzyladenine
- NAA
naphthaleneacetic acid 相似文献
8.
L. Burdyn C. Luna J. Tarracó P. Sansberro N. Dudit A. Conzález L. Mrocinski 《In vitro cellular & developmental biology. Plant》2006,42(3):235-239
Summary Adventitious bud regeneration from leaf and internode explants of Aloysia polystachya was achieved. Shoots from nodal segments grown in vitro were cut into pieces and used as sources of explants. Organogenesis was induced from both explants cultured on quarter-strength
Murashige and Skoog (MS) semisolid medium (plus sucrose 5 g l−1) containing different combinations of 6-benzyladenine (BA) and α-naphthaleneacetic acid (NAA) under 116 μmol m−2 s−1 photosynthetic photon flux density (PPFD), 14-h photoperiod, and at a temperature of 27±2°C. The type of explant markedly
influenced organogenesis and growth of the regenerated shoots. The regeneration frequencies were higher with leaf explants,
while the number of shoots formed per responsive explant was greater with internode explants. However, the growth of regenerated
shoots from internodes was seriously affected by vitrification. The number of shoots produced per responsive leaf explant
increased from one to seven as the percentage of leaf explants producing shoots increased from 20 to more than 80%. NAA at
0.05 μM in combination with BA at 0.5μM induced the highest regeneration rate (87±8.8%) after 20 d of culture, yielding 5.9±0.8 shoots per responsive leaf explant.
Histological examination confirmed the occurrence of direct organogenesis. The regenerated shoots from the best induction
treatment were transferred to a fresh medium without plant growth regulators for 30 d. Finally, the elongated shoots were
rooted by pre-treatment in an aqueous solution of NAA at 500 μM for 2 h and transferred to 1/4 MS. All plantlets raised in vitro were phenotypically normal and successfully hardened to ex vitro conditions. An experimental field plot with 2-yr-old in vitro-regenerated plants was established. 相似文献
9.
Two Phalaenopsis orchids, Phalaenopsis amabilis and Phalaenopsis ‘Nebula’, were used to test the effects of induction period (30, 45 and 60 days), subculture period (30, 45 and 60 days),
and explant length (1, 1.5 and 2 cm) on direct somatic embryogenesis from different regions (leaf tip, adaxial side, abaxial
side and cut end) of leaf explants from in vitro grown seedlings. The results showed that the cut end had a highest competence
to form embryos than the other regions of the leaf explants from both orchids. In addition, the suitable culture conditions
were 60 days for induction period in darkness, 45 days for subculture period in light and 1 cm for explant length. Besides,
the combinations of N6-benzyladenine (BA) and naphthaleneacetic acid were tested on their effects on plantlet conversion and further development
of leaf-derived embryo. It was found that 0.5 mg/l of BA showed the highest response on plantlet conversion rate and the lowest
browning rate of explants. In this communication, the embryo structures and development were proved by scanning electron microscopy. 相似文献
10.
Adventitious shoot buds were induced from leaf and stem explants of Bacopa monnieri on Murashige and Skoog medium supplemented with benzyladenine or kinetin. The source of the explants as well as different
gelling agents in the medium were found to influence shoot induction and eventual shoot growth. The best response was obtained
in leaf explants taken from shoot cultures grown in medium supplemented with 2 μM benzyladenine and gelled with 0.2% gelrite. A transverse section of the leaf explant incubated in this medium showed several
shoot primordia emerging from the leaf surface. This system exhibited a potential for repeated harvesting of the shoots from
the original leaf explant as the latter continued to expand and regenerate new shoots, upon repeated periodical subculturing
onto fresh medium. However, the callusing response of the plant was very low. Qualitative TLC studies of the regenerated shoots
revealed a phytochemical profile similar to that of the field grown-plants.
Received: 20 March 1998 / Revision received: 1 December 1998 / Accepted: 12 December 1998 相似文献
11.
Effects of age and orientation of the explant on callus induction and de novo shoot regeneration from cotyledonary leaf segments of Jatropha curcas were studied. The callus induction and shoot regeneration capacity of cotyledonary leaf segments were found significantly related to the age of the explants and their orientation in culture medium. The youngest explant, derived from the cotyledonary leaf of germinated seed induced the highest regeneration response as compared to one- and two-week-old explants. A gradient response with age of the explant was observed in percentage of callus induction, shoot regeneration from callus and the number of shoots per regenerating callus. The explants cultured with their abaxial side in medium showed significantly higher regeneration response. The youngest explant was found to be most amenable to Agrobacterium-mediated transformation as compared to older explants. The fact that callus induced from the edges of the explant followed by de novo shoot induction, and strong transient gus expression observed in the edges of the explant are significant for routine Agrobacterium-mediated transformation and generation of stable transgenic plants in J. curcas. 相似文献
12.
The influence of light regime, explant position and orientation on direct embryo formation from leaf explants of two Phalaenopsis, P. amabilis and P. Nebula, were investigated to optimize the protocol for regenerating of this orchid. When explants were cultured in light,
direct embryogenesis was retarded in both species. Embryos showed whitish to pale green in color and larger size than those
cultured in darkness. Furthermore, light regime induced explant browning, embryo necrosis and eventually low plantlet conversion
rate. Sixty days of culture in darkness is the most suitable duration for direct embryo induction. Explant orientation also
significantly affected direct embryo formation, and explants placed adaxial-side-up on culture medium had higher embryogenic
response than abaxial-side-up orientation. In both species, the cut end had highest embryogenic competence than other parts
of the explant. Moreover, when the leaf explant was cut transversely into two segments, the leaf basal segment had higher
embryogenic competence than the leaf tip segment. 相似文献
13.
Influence of petiole and lamina on adventitious shoot initiation from leaf explants of Paulownia fortunei 总被引:2,自引:0,他引:2
Adventitious shoot bud differentiation occurred preferentially from the petiolar cut ends of leaf explants of Paulownia fortunei cultured on Murashige and Skoog medium supplemented with 4 μmα-naphthaleneacetic acid and 20 μm benzyladenine. The details of plantlet regeneration and successful transplantation to soil have been reported earlier. We
now show that besides medium supplementation with auxin and cytokinin, the presence of lamina and petiole in the explant influence
shoot bud induction. Explants with the basal half of the lamina and the entire petiole were much more responsive than those
with whole lamina and petiole. A dual-culture-medium technique which permitted incubation of the two ends of excised petioles
under two different phytohormone regimes was devised. Our data suggest that some of the diffusible factors from the lamina
may be phytohormones, and that the establishment of an endogenous phytohormone gradient in the explants may affect shoot bud
differentiation in this culture system.
Received: 7 April 1998 / Revision received: 8 April 1998 / Accepted: 17 April 1998 相似文献
14.
15.
Plant regeneration from seedling explants of green bean (Phaseolus vulgaris L) via organogenesis 总被引:1,自引:0,他引:1
Chandra I. Franklin Tony N. Trieu Robert A. Gonzales Richard A. Dixon 《Plant Cell, Tissue and Organ Culture》1991,24(3):199-206
Green bean (Phaseolus vulgaris L.) plants were regenerated from 3-day old seedling explants via organogenesis. The explants contained a cotyledon and a small portion (2–3 mm) of embryonic axis split in half. Explants were cultured on a defined medium containing glutamine as the sole nitrogen source. A ring of meristematic tissue was produced at the base of the axillary bud located at the cotyledonary node. The meristematic tissue was produced only if the axillary bud was present together with the cotyledon in the explant. Buds and shoots developed from the meristematic ring. Selected shoots produced roots when excised from the cluster of buds and transferred to root induction medium. Rooted shoots (plantlets) grew well and produced viable seeds when grown in the greenhouse. Histological studies revealed the origin of buds from the peripheral layers of the meristematic ring.Production of buds and shoots was a continuous process, so that new shoots could be removed from the explant for plantlet production every 10–14 days. With the cultivar Dark Red Kidney, an average of 49 buds and 8 shoots were regenerated per explant by 30 days after culture initiation. Sixty-seven percent of the shoots produced roots, and 90–95% of the plantlets survived greenhouse acclimatization to produce healthy plants. 相似文献
16.
Summary Factors affecting in vitro shoot production and regeneration of Cercis yunnanensis Hu et Cheng were investigated by comparing various growth regulators and explant types. For optimum shoot production from
axillary buds, Murashige and Skoog (MS) media containing 6-benzyladenine, either alone or in combination with a low concentration
of thidiazuron, resulted in the greatest number of shoots formed per explant (>3). Explants (2 mm long) containing one axillary
bud placed in directcontact with the medium yielded the most shoots per bud (1.6) when grown on growth regulator-free medium.
Root formation on 70–80% of shoot explants was accomplished using either indole-3-butyric acid or α-naphthaleneacetic acid
in the medium, with significantly more roots formed on explants possessing and apical bud than those without the bud. Direct
shoot organogenesis from leaf explants occurred on MS medium containing 10–30 μM thidiazuron, with up to 42% of leaf explants producing shoots. 相似文献
17.
Plantlet regeneration via organogenesis was achieved in callus cultures derived form mature leaves, stems and leaves, petioles and roots of young seedling of Psoralea corylifolia on Murashige and Skoog medium supplemented with 2.5–3.0 mg L-1 BA, 1.0 mg L-1 NAA and 3% (w/v) sucrose. The rate of shoot bud regeneration was positively correlated with the concentration of hormones in the nutrient media. Shoot buds regenerated more readily from juvenile explants (seedling source) as compared to the mature explants. Addition of adenine sulphate (5 mg L-1) to the culture medium increased the growth of shoot buds. Optimum responses were obtained in hypocotyl and leaf explants using NAA in combination with BA, the highest rate of shoot bud regeneration being in hypocotyl explants. Rooting was readily achieved on the differentiated shoots on MS basal media without growth regulators. Regenerated plantlets were successfully established in the greenhouse. 相似文献
18.
Summary Explants derived from adventitious buds, rhizomes, stems, and leaves of a medicinal plant, Polygonatum cyrtonema, were studied for plantlet regeneration, and only adventitious bud explants were able to be regenerated into plantlets. Regeneration
was also accompanied by the formation of rhizome-like tissue, the medicinal portion of the plant. The optimum hormone combination
for plantlet regenertion was 4.44 μM benzyladenine plus 2.26 μM 2,4-dichlorophenoxyacetic acid, at which new adventitious buds were obtained from 96.6% of the adventitious bud explants,
with an average of 5.2 buds per explant. The best medium for root induction was half-strength Murashige and Skoog medium with
4.57 μM α-naphthaleneacetic acid, as 92% of regenerated buds rooted. Regenerated plantlets were successfully transferred to a greenhouse
with 86% survival. Histological observation indicated that new adventitious buds originated from the superficial meristematic
cell cluster of the granular callus induced from adventitious bud explants via organogenesis. 相似文献
19.
Juliana Lischka Sampaio Mayer Giulio Cesare Stancato Beatriz Appezzato-Da-Glória 《Plant Cell, Tissue and Organ Culture》2010,103(3):411-416
The present study describes the direct regeneration of protocorm-like bodies (PLBs) in leaf explants of the tropical species
Oncidium flexuosum. The explants were inoculated in a solid, modified Murashige and Skoog (MS) medium with different concentrations of the growth
regulator thidiazuron (TDZ) and with or without 2,4-dichlorophenoxyacetic acid (2,4-D) and naphthalene acetic acid (NAA),
and kept away from light or in a 16-h photoperiod. The presence of auxins, 2,4-D, and NAA inhibited the formation of PLBs.
The highest frequency of explants that regenerated PLBs (80%) was obtained when they were maintained in a culture medium containing
1.5 μM TDZ under dark conditions. In the same culture medium but under a 16-h photoperiod, 95% of the leaf explants presented
necrosis. Therefore, darkness was crucial for the regeneration of PLBs in O. flexuosum leaf explants, which is in disagreement with the literature. PLBs developed from the division of epidermal and subepidermal
cells mainly on the adaxial side of the apex region of the explant. Plants with well-developed leaves and roots grew after
the PLBs were transferred to growth regulator-free medium under a 16-h photoperiod. 相似文献
20.
Summary
In vitro propagation of Quassia amara L. (Simaroubaceae) was attempted using mature and juvenile explants. Attempts to establish in vitro culture using leaf and internode explants from a plant more than 15yr old were unsuccessful due to severe phenolic exudation.
Plant regeneration through direct and indirect somatic embryogenesis was established from cotyledon explants. Murashige and
Skoog (MS) medium with 8.9 μM N6-benzyladenine (BA) and 11.7 μM silver nitrate induced the highest number (mean of 32.4 embryos per cotyledon) of somatic embryos. Direct somatic embryogenesis
as well as callus formation was observed on medium with BA (8.9–13.3 μM). Semi-mature pale green cotyledons were superior for the induction of somatic embryos. Embryos developed from the adaxial
side as well as from the point of excision of the embryonic axis. More embryos were developed on the proximal end compared
to mid and distal regions of the cotyledons. Subculture of callus (developed along with the somatic embryos on medium with
BA alone) onto medium containing 8.9 μM BA and 11.7 μM silver nitrate produced a mean of 17.1 somatic embryos. Primary somatic embryos cultured on MS medium with 8.9 μM BA and 11.7μM silver nitrate produced a mean of 9.4 secondary somatic embryos. Most of the embryos developed up to early cotyledonary stage.
Reduced concentration of BA (2.2 or 4.4 μM) improved maturation and conversion of embryos to plantlets. Ninety percent of the embryos converted to plantlets. The optimized
protocol facilitated recovery of 30 plantlets per cotyledon explant within 80d. Plantlets transferred to small cups were subsequently
transferred to field conditions with a survival rate of 90%. 相似文献