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1.
Summary The culture conditions for direct, indirect, and repetitive embryogenesis were established forCamellia japonica cv. Elegans and cv. Ville de Nantes. Direct embryo production from leaves averaged 15.3 embryos per responsive leaf on Murashige
and Skoog medium (MS) with 1.0 mg·liter−1 N6-benzyladenine and 0.5 mg·liter−1 2,4-dichlorophenoxyacetic acid. Plantlet production was 7.1 (±1.5) plantlets per leaf. Direct embryo production from
stems averaged 5.7 embryos per shoot, and 2.7 embryos per stem portion, on MS medium supplemented with 1.0 mg·liter−1 N6-benzyladenine and 0.1 mg·liter−1 indolbutyric acid (MS28). Conversion was only obtained after repetitive embryogenesis. Embryogenesis from leaf-derived callus
occurred in all callus after transfer to MS/2–25 medium (half strength MS medium with 25 g·liter−1
D-glucose) (production stage). Plantlet production was 16.3 (±3.6) plantlets per callus. Repetitive embryogenesis increased
embryo population by 2.3- to 3.6-fold every 4 wk. Conversion of secondary embryos was obtained on MS medium supplemented with
2.0 mg·liter−1 N6-benzyladenine, 0.2 mg·liter−1 indolbutyric acid, 5 mg·liter−1 gibberellic acid (MS56). Direct embryo formation from leaves, stems, and cotyledons, and embryogenic callus formation from
leaves were restricted to specific regions of the explant. 相似文献
2.
Sarvesh Adda T. P. Reddy P. B. Kavi Kishor 《In vitro cellular & developmental biology. Plant》1994,30(2):104-107
Summary Induction of somatic embryogenesis, shoot organogenesis, and subsequent plant regeneration in niger seem to be dependent on
genotype, choice of explant, and composition of media growth regulators. Two distinct regeneration protocols have been developed
for somatic embryogenesis and shoot organogenesis. Somatic embryogenesis was induced from epicotyls and cotyledonary explants
(9 to 35%) (but not from hypocotyls and roots) in presence of 2,4-dichlorophenoxyacetic acid, 2,4,5-trichlorophenoxyacetic
acid, and 2,4,5-trichlorophenoxypropionic acid. These embryos matured in MS medium containing Kinetin plus naphthalene acetic
acid (NAA), Kinetin plus Zeatin, and Kinetin plus abscisic acid (ABA). Matured embryos could be germinated on LS and MS basal
media without hormones. Non-embryogenic callus initiated on Linsmaier and Skoog’s (LS) medium from cotyledons of six
different genotypes produced shoots (9 to 32%) on Murashige and Skoog’s (MS) medium fortified with 6-benzylaminopurine
(BAP, 0.5 mg · liter−1), and BAP (1 mg · liter−1) plus NAA (0.1 mg · liter−1). These shoots were rooted with 100% frequency by using indole-3-acetic acid or NAA and transferred successfully to the soil. 相似文献
3.
Kyungchul Han 《In vitro cellular & developmental biology. Plant》1994,30(2):108-112
Summary An efficient and reproducible protocol has been developed for in vitro shoot regeneration from cotyledonary explants derived
by germinating immature ovules ofImpatiens platypetala Lindl. ‘TR6-27-2’. Cotyledonary explants were cultured on a modified Murashige and Skoog (MS) agar-solidified
medium containing 7.5g · liter−1 sucrose, 22.2µ M N6-benzyladenine (BA), and 0.54µM α-naphthaleneacetic acid (NAA). The induction of organogenic tissues occurred after 6 to 8 wk in culture. Exogenous auxin and
cytokinin were essential for the induction of organogenic tissues and survival of explants, and BA was most effective for
the induction of organogenic tissues, compared with other cytokinins tested. The addition of glutamine (500 mg · liter−1) was also important for growth of organogenic tissues after induction and for reducing explant death during culture. The
induction of organogenic tissue was also influenced by the type of cotyledon cultured and the age of the donor seedlings.
On average, eight shoots per explant were induced from organogenic tissues larger than 0.5 cm in diameter 6 to 8 wk after
transfer to a modified MS agar-solidified medium without NAA and BA reduced to 4.44µM. Shoots longer than 0.5 cm in length were successfully rooted 2 to 4 wk after transfer to a basal MS medium containing 30g · liter−1 sucrose. 相似文献
4.
Bud break and multiple shoot formation from tissues of mature trees ofPinus caribaea andPinus kesiya
R. S. Nadgauda N. N. Nagarwala V. A. Parasharami A. F. Mascarenhas 《In vitro cellular & developmental biology. Plant》1993,29(3):131-134
Summary Proliferation of terminal and axillary buds of 20 yr-oldPinus caribaea andP. kesiya trees was obtained on half strength DCR medium supplemented with 0.5 mg·liter−1 6-benzylaminopurine (BA). These sprouts further elongated with the formation of multiple shoots with the ratio of 1:3, on
transfer to medium in which the 0.25 mg·liter−1 BA of the initiation medium was replaced by 0.25 mg·liter−1 kinetin. Rooting was obtained on the same medium. Plantlets thus formed were transferred to perlite:peat:vermiculite mixture
(1:1:1) in polybags (10×5 cm) under 80±5% humidity in a polyhouse. Plantlets ofP. caribaea andP. kesiya were established with 72.5 and 83.3% survival, respectively. 相似文献
5.
Summary This study reports a method for high-frequency shoot organogenesis and plant establishment of Potentilla potaninii Wolf. Hypocotyl and cotyledon explants of P. potaninii were cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of benzyladenine (BA) and α-naphthaleneacetic
acid (NAA) to induce adventitious shoot formation for micropropagation. The highest frequency of adventitious shoot regeneration
was achieved from hypocotyl and cotyledon explants grown on MS medium supplemented with 5.0 mgl−1 BA and 1.0 mgl−1 NAA. The regenerated shoots rooted most efficiently on half-strength MS medium supplemented with 1.0 mgl−1 NAA and 0.5 mgl−1 indole-3-acetic acid or indole-3-butyric acid. The acclimatized plants with normal morphology and growth characters flowered
and set seeds in the following year. 相似文献
6.
Esmaeil Ebrahimie A. A. Habashy M. Mohammadie-Dehcheshmeh M. R. Ghannadha B. Ghareyazie B. Yazdi-Amadi 《In vitro cellular & developmental biology. Plant》2006,42(5):455-460
Summary A rapid and one-step protocol for direct regeneration of shoots from cumin embryo explants has been developed. Embryo explants
with shoot meristems were cultured on shoot regeneration medium for 15–22 d. After embryo culture, shoots were regenerated
from the area adjacent to the region between the cotyledons and embryo axis within 2 wk, without any intermediate callus phase.
Shoot proliferation and elongation were achieved on shoot regeneration medium without subculture. Among the different combinations
of 6-benzylaminopurine, α-naphthaleneacetic acid (NAA), and indole-3-acetic acid (IAA) tested, 0.8 mgl−1 (4.3 μM) NAA in combination with 0.3 mgl−1 (1.71 μM) IAA in the B5 medium resulted in the most efficient direct shoot regeneration. No significant difference was detected for
the number of regenerated explants when different heterogeneous endemic varieties were compared. This plant regeneration procedure
was applicable to different cumin genotypes and regenerated plants were phenotypically normal. 相似文献
7.
Vinayak Singh Namita Singh Chauhan Mohit Singh Asif Idris Raju Madanala Veena Pande Chandra Sekhar Mohanty 《Plant signaling & behavior》2014,9(10)
An in vitro method of multiple shoot induction and plant regeneration in Psophocarpus tetragonolobus (L.) DC was developed. Cotyledons, hypocotyls, epicotyls, internodal and young seedling leaves were used as explants. MS media supplemented with various concentrations of either thidiazuron (TDZ) or N6-benzylaminopurine (BAP) along with NAA or IAA combinations were used to determine their influence on multiple shoot induction. MS media supplemented with TDZ induced direct shoot regeneration when epicotyls and internodal segments were used as explants. TDZ at 3 mg L−1 induced highest rate (89.2 ± 3.28%) of regeneration with (13.4 ± 2.04) shoots per explant. MS media supplemented with BAP in combination with NAA or IAA induced callus mediated regeneration when cotyledons and hypocotyls were used as explants. BAP (2.5 mg L−1) and IAA (0.2 mg L−1) induced highest rate (100 ± 2.66%) of regeneration with (23.2 ± 2.66) shoots per explant. Mature plants produced from regenerated shoots were transferred successfully to the greenhouse. In a comparative study, the phenolics contents of various parts of greenhouse-grown plants with that of in vitro-raised plants showed significant variations. 相似文献
8.
Summary This study describes a protocol for plant regeneration from cultured seedling explants of Arctium lappa. Hypocotyls and cotyledons of A. lappa were induced to form callus by culturing on Murashige and Skoog (MS) medium supplemented with 2.0mg l−1 2,4-dichlorophenoxyacetic acid and 0.5–2.0 mg l−1 benzyladenine (BA). Formation of adventitious buds could be induced from calluses or explants directly by culturing on MS
medium containing 1.0–2.0 mg l−1 α-naphthaleneacetic acid (NAA) and 0.5–2.0 mg l−1 BA. These regenerated shoots were rooted on MS medium with 1.0 mg l−1 indole-3-butyric acid or indole-3-acetic acid in combination with 1.0 mgl−1 NAA. The regenerated plants acclimatized in soil were normal morphologically and in growth characters. They flowered and
set seeds in the following year after acclimatization. 相似文献
9.
Summary Rose (Rose hybrida L.) plants were micropropagated by axillary shoot proliferation method. Maximum number of microshoots per shoot tip explant
were obtained on MS medium supplemented with 5 to 10µM thidiazuron (TDZ). The microshoots formed rooted plants on MS hormone-free medium. No difference in the rooting of microshoots
produced on medium containing TDZ or N6-benzyladenine was observed. The regenerated plants were successfully transplanted to the field and appeared similar to the
parent plant in morphologic features. 相似文献
10.
Plants were regenerated from root explants of Arabidopsis halleri (L.) O’Kane and Al-Shehbaz via a three-step procedure callus induction, induction of somatic embryos and shoot development. Callus was induced from root segments, leaflets and petiole segments after incubation for 2 weeks in Murashige and Skoog medium (MS) supplemented with 0.5 mg/l−1 (2.26 μM) 2,4-D (2,4-dichlorophenoxyacetic acid) and 0.05 mg/l−1 (0.23 μM) kinetin. Only calli developed from root segments continued to grow when transferred to a regeneration medium containing 2.0 mg/l−1 (9.8 μM) 6-γ-γ-(dimethylallylamino)-purine (2ip) and 0.05 mg/l−1 (2.68 μM) α-naphthalenacetic acid (NAA) and eventually 40 of them developed embryogenic structures. On the same medium 38 of these calli regenerated shoots. Rooting was achieved for 50 of the shoots subcultured in MS medium without hormones. The regeneration ability of callus derived from root cuttings, observed in this study, makes this technique useful for genetic transformation experiments and in vitro culture studies. 相似文献
11.
An efficient and rapid plant regeneration system was established for zonal and scented geraniums using leaf discs as explants. Several explants, medium and culture conditions were studied to optimize shoot induction. Leaf discs taken from 4–5 weeks old in vitro grown plants, whatever the genotype, were more effective for shoot regeneration than those taken from greenhouse grown plants. Darkness proved to be a stimulating factor for shoot regeneration and the combination between NAA and two cytokinins gave the best results. Direct shoot regeneration (100%) was obtained from leaf discs of P. capitatum on half-strength MS medium supplemented with 0.5 mg l−1 NAA in combination with 1 mg l−1 of BAP and zeatin in darkness (11.4 shoots per explant). In the same medium and culture conditions, all P. graveolens leaf discs also exhibited direct shoot regeneration (7.3 shoots per explant). For P. x hortorum, 100% of leaf discs underwent shoot regeneration on a MS medium supplemented with 0.2 mg l−1 NAA in combination with 0.5 mg l−1 of BAP and zeatin in darkness (8.8 shoots per explant) or under low light conditions with 0.2 mg l−1 NAA and 1 mg l−1 of BAP and zeatin (7.5 shoots per explant). For this species, the best results for shoot elongation were obtained on half-strength MS medium gelled with Phytagel 0.3% (v/v). Whatever the genotype, all shoots rooted readily when transferred to diluted MS medium (MS/2) containing 1 mg l−1 IAA. Acclimatized plants grew normally and flowered in greenhouse conditions. Flow cytometry analysis made on leaves of acclimatized plants revealed that all the scented geranium plants are similar to mother plants while 71% of P. x hortorum plants which showed strong growth were tetraploid. 相似文献
12.
Callus was initiated from petiole explants of Heracleum candicans on MS medium fortified with BAP and 2,4-D ( 0.5 mg I-1 each). Maximum shoot differentiation from callus occurred on MS medium containing 1 mg I-1 BAP and 0.2 mg I-1 NAA. The regenerated shoots were rooted on MS medium supplemented with 1 mg I-1 IBA. The rooted plants were transferred to the field after successful hardening in pots containing vermiculite. All regenerated plants were diploid with 2n=22 chromosomes in their root tip cells. 相似文献
13.
Plants were regenerated from the in vitro cultured explants of primary leaves of cowpea (Vigna unguiculata L. Walp). Primary leaves, including the intact petiole, were excised from three-day-old seedlings and cultured on Gamborg's B5 basal medium containing 8×10–7 M 2,4,5-trichlorophenoxyacetic acid, 1×10–2 M L-glutamine and 1×10–4 M adenine sulfate. Callus formed at the petiole end. Prolific shoot regeneration occurred when this callus was transferred to B5 basal medium containing 5×10–6 M 6-benzyl-aminopurine (BAP). Regenerated shoots rooted in growth-regulator-free B5 basal medium and were established in soil.Abbreviations BAP
6-benzylaminopurine
- IAA
indole-3-acetic acid
- NAA
1-napthalene acetic acid
- 2,4,5-T
2,4,5-trichloro-phenoxyacetic acid 相似文献
14.
Ajay Kumar Thakur A. Saraswat D. K. Srivastava 《Journal of plant biochemistry and biotechnology.》2012,21(1):23-29
A rapid and efficient protocol is developed for in vitro plantlet regeneration of Populus deltoides clone G48 using petiole explants. The highest frequency of shoot regeneration (74.75%) from petiole was obtained on MS medium supplemented with 0.50?mg/l BAP and 0.20?mg/l IAA. The regenerated shoots started turning brown and necrotic after 10?C15?days in culture. To overcome the browning problem, the explants along with the developing shoot buds were transferred to modified MS medium containing 0.50?mg/l BAP, 0.20?mg/l IAA, 15?mg/l AdS, 0.1% PVP, 100?mg/l casein hydrolysate, 50?mg/l L-glutamine, 250?mg/l (NH4)2SO4 and 0.5% agar. Shoot multiplication and elongation took place on the same medium. Indole-3-acetic acid at 0.10?mg/l was most effective for root regeneration. Using the current protocol, it took 2?months to regenerate plantlets. The in vitro regenerated plantlets were successfully acclimatized and established in greenhouse conditions. This regeneration system using petiole explants provides a foundation for Agrobacterium-mediated genetic transformation of P. deltoides clone G48 for incorporation of various silviculturally important traits. 相似文献
15.
Guang-Zhe Lin Xiao-Mei Zhao Soon-Kwan Hong Yu-Ji Lian 《Plant Cell, Tissue and Organ Culture》2011,106(1):93-103
We have developed a system for the in vitro regeneration of pasqueflowers (Pulsatilla koreana Nakai). The system was based on somatic embryogenesis and shoot organogenesis. Over a growth period of 6 weeks, multiple
shoots were initiated from leaf, petiole, and pedicel explants on Murashige and Skoog (MS) medium containing 0.5 mg l−1 indole-3-acetic acid (IAA) and zeatin (Zn), kinetin (Kin), or 6-benzyladenine (BA). We achieved 100% of adventitious shoot
induced when petiole and pedicel explants were cultured on MS, 0.5–2.0 mg l−1 Zn, and 0.5 mg l−1 IAA. Somatic embryos developed from the explants and generated shoots on MS medium containing 0.25 mg l−1 Zn and 0.5 mg l−1 IAA. Globular and heart-shaped stages of somatic embryos were observed. Histological studies have revealed the stages of
development of somatic embryos. For propagation and growth, the regenerated shoots from organogenic or embryogenic calluses
were transferred to MS medium containing either (1) 1.5 mg l−1 Zn and 0.05 mg l−1 IAA or (2) 1.0 mg l−1 BA and 0.05 mg l−1 IAA. After the length of the shoots reached 3 cm, the shoots initiated by organogenesis as well as those initiated by somatic
embryogenesis were transferred to the root induction medium. After 2 months of culture in half-strength MS with 1.5 mg l−1 α-naphthalene acetic acid (NAA), the rooting ratio was 93%. Finally, the rooted plantlets were acclimatized in a mixture
of mountain soil and perlite. 相似文献
16.
An efficient system for shoot regeneration and Agrobacterium-mediated gene transfer into Brassica napus was developed through the modification of the culture conditions. Different concentrations of benzyladenine (1.5, 3.0 and 4.5 mg dm–3) and thidiazuron (0.0, 0.15 and 0.30 mg dm–3) were evaluated for shoot regeneration of 7, 14 and 21-d-old hypocotyl explants. Maximum shoot regeneration frequency was obtained in 21-d-old explants using 4.5 mg dm–3 benzyladenine and 0.3 mg dm–3 thidiazuron. Under above culture condition, the highest percentage of shoot regeneration frequency was 200 %. Agrobacterium-infected explants grown on the selection medium gave rise to transgenic shoots at a frequency of 11.8 %. Transformed shoots rooted when cultured on a medium supplemented with 2 mg dm–3 of indolebutyric acid and 10 mg dm–3 kanamycin. The rooted plantlets were successfully established in the soil and developed fertile flowers and viable seeds. Evidences for transformation were confirmed by GUS assay and PCR analysis. 相似文献
17.
S. Pandey M. Singh U. Jaiswal V. S. Jaiswal 《In vitro cellular & developmental biology. Plant》2006,42(5):389-393
Summary This study describes a protocol for the regeneration of complete plantlets of Terminalia arjuna from nodal explants of mature trees. Shoot multiplication from nodal explants was achieved by culturing on Murashige and
Skoog (MS) medium containing different concentrations of 6-benzyladenine (BA), thidiazuron or kinetin, or BA in combination
with α-naphthaleneacetic acid (NAA). The best shoot multiplication response was obtained from nodal explants grown
on modified MS (half-strength major salts and Fe-EDTA) medium containing 4.44 μM BA and 0.53 μM NAA. Seasonal variations significantly affected the proliferation potential of nodal explants and best proliferation was
observed from explants collected during April to May. Microshoots were rooted on half-strength MS medium with 4.92 μM IBA. The rooted shoots were acclimatized successfully. 相似文献
18.
Dormant buds from a mature tree of Populus tremula ‘Erecta’ were incubated on a Murashige and Skoog (MS) medium supplemented with 1.0 μM thidiazuron (TDZ). Induced shoots were
then proliferated on medium of MS or Woody Plant Medium (WPM), or Driver and Kuniyuki Walnut (DKW) supplemented with varying
levels of benzyladenine (BA). Overall, shoots grown on MS medium supplemented with 1.25–2.5 μM BA exhibited the highest frequency
of shoot proliferation (>95%) and more than 60% of responding explants produced more than five shoots per explant. Shoot organogenesis
was induced from both leaf and petiole explants incubated on WPM medium containing BA, or TDZ, or zeatin. Among the different
cytokinins tested, zeatin induced the highest frequency (average 72.1%) of shoot organogenesis. None of explants survived
on media containing no cytokinins within 6–8 weeks following culture. Overall, a higher frequency of shoot regeneration was
obtained from petioles than from leaf explants. The highest frequency of regeneration was achieved when petioles were incubated
on WPM containing 10–20 μM zeatin. Addition of naphthaleneacetic acid (NAA) did not have a significant effect on shoot regeneration
in all treatments. Shoot organogenesis was directly induced from petiole explants without intervening callus. Regenerated
shoots were easily rooted on all tested media supplemented with 0.5 μM NAA. Rooted plants were transferred to potting mix
and grown in the greenhouse. 相似文献
19.
An efficient and reproducible method for the regeneration of Jatropha curcas plants has been developed. The method employed direct induction of shoot buds from petiole explants, without the formation of an intervening callus using a Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ). The best induction of shoot buds (58.35%) and the number of shoot buds per explant (10.10) were observed when in vitro petiole explants were placed horizontally on MS medium supplemented with 2.27 µM TDZ after 6 weeks. The induced shoot buds were transferred to MS medium containing 10 µM kinetin (Kn), 4.5 µM 6-benzyl aminopurine (BAP) and 5.5 µM α-naphthaleneacetic acid (NAA) for shoot proliferation. The proliferated shoots could be elongated on MS medium supplemented with different concentrations and combinations of BAP, indole-3-acetic acid (IAA), NAA and indole-3-butyric acid (IBA). MS medium supplemented with 2.25 µM BAP and 8.5 µM IAA was found to be the best combination for shoot elongation and 3.01–3.91 cm elongation was achieved after 6 weeks. However, significant differences in plant regeneration and shoot elongation were observed among the genotypes studied. The orientation (horizontal or vertical) and source (in vitro or in vivo) of explants also significantly influenced plant regeneration. The elongated shoots could be rooted on half-strength MS medium supplemented with 2% sucrose, different concentrations and combinations of IBA, IAA and NAA, and 0.25 mg L−1 activated charcoal. Half-strength MS medium supplemented with 2% sucrose, 15 µM IBA, 5.7 µM IAA, 5.5 µM NAA and 0.25 mg L−1 activated charcoal was found to be the best for promoting rooting. The rooted plants could be established in soil with more than 90% survival. 相似文献
20.
Gaurav Krishna P. Sairam Reddy Pramod W. Ramteke Pogiri Rambabu Sayed S. Sohrab Debashis Rana Parthasarathi Bhattacharya 《Physiology and Molecular Biology of Plants》2011,17(4):375-385
In vitro regeneration of pigeon pea through organogenesis and somatic embryogenesis was demonstrated with pigeon pea cv. JKR105. Embryonic axes explants of pigeon pea showed greater regeneration of shoot buds on 2.5 mg L−1 6-benzylaminopurine (BAP) in the medium, followed by further elongation at lower concentrations. Rooting of shoots was observed on half-strength Murashige and Skoog (MS) medium with 2 % sucrose and 0.5 mg L−1 3-indolebutyric acid (IBA). On the other hand, the regeneration of globular embryos from cotyledon explant was faster and greater with thidiazuron (TDZ) than BAP with sucrose as carbohydrate source. These globular embryos were maturated on MS medium with abscisic acid (ABA) and finally germinated on half-strength MS medium at lower concentrations of BAP. Comparison of regeneration pathways in pigeon pea cv. JKR105 showed that the turnover of successful establishment of plants achieved through organogenesis was more compared to somatic embryogenesis, despite the production of more embryos than shoot buds. 相似文献