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1.
Summary This is the first communication of direct shoot regeneration from fully developed leaves of potted mature Echinacea purpurea plants. Shoot buds were induced directly on the adaxial surface of mature leaf tissues of E. purpurea 30 d after culture initiation on Woody Plant Medium (WPM) supplemented with various levels of 6-benzyladenine (BA). Maximum
shoot organogenesis, with 12–20 shoots per leaf segment, was obtained with 5% coconut milk and 2.5 mg l−1 (6μM) BA in 30 d. Callus was induced using 0.5 mgl−1 (1μM) α-naphthaleneacetic acid and 2.5 mgl−1 (6μM) BA. The regenerated shoots were rooted on WPM supplemented with 1.5 mgl−1 (3μM) of indole-3-butyric acid, 3% sucrose, and 0.85% agarose. Rooted plants were successfully transferred to soil in pots and
appeared morphologically normal and flowered in a growth chamber. 相似文献
2.
Yuexia Wang Bridget A. Ruemmele Joel M. Chandlee W. Michael Sullivan Jane E. Knapp Albert P. Kausch 《In vitro cellular & developmental biology. Plant》2002,38(5):460-467
Summary Embryogenic callus induction and plant regeneration systems have long been established for creeping bentgrass (Agrostis palustris Huds.), but little research has been reported on optimal medium for embryogenic callus induction and plant regeneration in
velvet bentgrass (Agrostis canina L.), colonial bentgrass (Agrostis capillaries L.), and annual bluegrass (Poa annua L.). The present study compared 14 callus induction media and eight regeneration media for their efficacies on embryogenic
callus induction and plant regeneration in these four species. The embryogenic callus initiation media contained the Murashige
and Skoog inorganic salts and vitamins supplemented with 2,4-dichlorophenoxyacetic acid or 3,6-dichloro-anisic acid and 6-benzyladenine.
l-Proline or casein hydrolyzate was included in some media to stimulate embryogenic callus formation and plant regeneration.
The frequencies of embryogenic callus formation ranged from 0% to 38% and exhibited medium differences within each of the
four species. Callus induction media, plant regeneration media, and genotypes affected plant regeneration rates, which varied
between 0% and 100%. The embryogenic callus induced on Murashige and Skoog medium supplemented with 500 mgl−1 casein hydrolyzate, 6.63 mg l−1 (30 μM) 3,6-dichloro-anisic acid and 0.5–2.0 mg l−1 (2–9 μM) 6-benzyladenine had much higher regeneration rates than those formed on other callus induction media. Embryogenic callus
of annual bluegrass had higher regeneration rates than those of bentgrass species. MSA2D, a media containing 2 mgl−1 (8 μM) 2,4-dichlorophenoxyacetic acid, 100 mgl−1
myo-inositol, and 150 mgl−1 asparagine, was effective in promoting embryogenic callus formation in creeping bentgrass but not in colonial and velvet
bentgrasses and annual bluegrass. 相似文献
3.
Summary Plant regeneration through direct somatic embryogenesis was achieved from root segments derived from in vitro shoots of Rauvolfia micrantha Hook. f. (Apocynaceae) grown for 6 wk in half-strength Murashige and Skoog (MS) medium with 3% sucrose, 100 mgl−1 myo-inositol, and 0.5 mgl−1 α-naphthaleneacetic acid (NAA). The effects of photoperiod and plant growth regulators (PGRs) in half-strength MS medium
were studied for the rapid and maximum induction of somatic embryos. The characteristic globular or heart-shaped stages of
somatic embryogenesis were not found and cotyledonary stage embryos occasionally appeared without the intervention of callus
in total darkness and 16-h photoperiod. Root segments cultured in the medium containing 0.1 mgl−1 NAA and 0.2 mgl−1 6-benzyladenine (BA) under 16-h photoperiod showed the maximum frequency (39%) of embryogenesis. The frequency of embryo
formation was increased to 63% when they were cultured in medium with 0.1 mgl−1 NAA and 0.2 mgl−1 BA in the dark for 4wk, then grown under the 16-h photoperiod. Explants with developing embryos developed into plants after
transfer to half-strength MS medium supplemented with 0.1 mgl−1 BA and 0.05 mgl−1 NAA. The well-developed plants were hardened and most plants (80%) survived and were phenotypically similar to the mother
plants. 相似文献
4.
Summary Callus induction and later plant regeneration were studied in four widely grown garlic (Allium sativum L.) cultivars from Europe. Root segments from in vitro plantlets were used as starting material. In addition to cultivar effects, the effects of auxin and cytokinin levels and
the position of the segments on the root were studied. There were no statistically significant differences among cultivars
for the number of root segments that induced callus in the two series of experiments. The average induction frequency was
34.7% in the first series of experiments. Callus induction on apical root segments was significantly higher compared to callus
induction on non-apical root segments in the second series of experiments. Two months after callus induction, callus lines
were transferred to a regeneration medium consisting of Murashige and Skoog basal medium supplemented with 30gl−1 sucrose and 1 mgl−1 (4.6μM) kinetin. Calluses derived from different experiments were quite uniform with respect to their regeneration potential. Also
it was found that our regeneration system was cultivar-independent. The average shoot regeneration frequency was 17.9% in
the first series of experiments. Highly significant differences were found in the frequency of shoot regeneration among different
callus induction treatments. When the cytokinin 6-(γ,γ-dimethylallylamino)purine (0.1mgl−1∶0.5 μM) was present during callus induction, shoot regeneration ranged from 30.10 to 47.60%. Shoot regeneration from callus induced
on non-apical segments was higher, although not significant, compared to callus induction from apical root segments in the
second series of experiments. All in all, an efficient callus induction and plant regeneration system was developed from both
apical and non-apical segments taken along the entire length of the roots. This system has potential to be used for garlic
transformation. 相似文献
5.
Summary Axillary and terminal buds from suckers of Ananas comosus cv. Phuket were established on Murashige and Tucker-based (MT) medium with 2.0 mgl−1 (9.8 μM) indolebutyric acid, 2.0 mgl−1 (10.74 μM) naphthaleneacetic acid, and 2.0 mgl−1 (9.29 μM) kinetin, followed by multiplication on Murashige and Skoog-based (MS) medium containing 2.0 mgl−1 (8.87 μM) benzyladenine (BA) to provide a continuous supply of axenic shoots. Leaves, excised from such cultured shoots, produced
adventitious shoots from their bases when these explants were cultured on MS medium containing 0.5 mgl−1 (2.26 μM) 2,4-dichlorophenoxyacetic acid (2,4-D) and 2.0 mgl−1 (8.87 μM) BA. Embryogenic callus was produced when leaf explants were cultured on MS medium with 3.0 mgl−1 (12.42 μM) 4-amino-3,5,6-trichloropicolinic acid (picloram). Somatic embryos developed into shoots following transfer of embryogenic
tissues to MS medium with 1.0 mgl−1 (4.44 μM) BA. Cell suspensions, initiated by transfer of embryogenic callus to liquid MS medium with 1.0 mgl−1 (4.14 μM) picloram or 1.0 mgl−1 (4.52 μM) 2,4-D, also regenerated shoots by somatic embryogenesis, on transfer of cells to semisolid MS medium with 1.0 mgl−1 (4.44 μM) BA. All regenerated shoots rooted on growth regulator-free MS medium, prior to ex vitro acclimation and transfer to the glasshouse. These studies provide a baseline for propagation, conservation, and genetic manipulation
of elite pineapple germplasms. 相似文献
6.
An efficient system was developed for direct plant regeneration from in vitro-derived leaf explants of Pistacia vera L. cv. Siirt. The in vitro procedure involved four steps that included (1) induction of shoot initials from the regenerated mature leaf tissue, (2)
regeneration and elongation of shoots from the shoot initials, (3) rooting of the shoots, and (4) acclimatization of the plantlets.
The induction of shoot initials was achieved on an agarified Murashige and Skoog (MS) medium with Gamborg vitamins supplemented
in different concentrations of benzylaminopurine (BA) and indole-3-acetic acid (IAA). The best medium for shoot induction
was a MS medium with 1 mgl−1 IAA and 2 mgl−1 BA. Numerous shoot primordia developed within 2–3 wk on the leaf margin and the midrib region, without any callus phase.
In the second step, the shoot clumps were separated from the leaf explants and transferred to a MS medium supplemented with
1 mgl−1 BA, resulting in a differentiation of the shoot initials into well-developed shoots. The elongated shoots (>3 cm long) were
rooted on a full-strength MS basal medium supplemented with 2 mgl−1 of indole-3-butyric acid in the third stage. Finally, the rooted plants were transferred to soil with an 80% success rate.
This protocol was utilized for the in vitro clonal propagation of this important recalcitrant plant species. 相似文献
7.
Wan-Jun Zhang Jiang-Li Dong Ben-Guo Liang Yong-Sheng Jin Tao Wang 《In vitro cellular & developmental biology. Plant》2006,42(2):114-118
Summary We report a protocol for efficient plant regeneration of four tall fescue (Festuca arundinacea Schreb.) cultivars (‘Surpro’, ‘Coronado’, ‘Summer Lawn’, and ‘Fawn’) via somatic embryogenesis. Calli were initiated from
mature seeds grown on modified Murashige and Skoog (MMS) medium supplemented with 7.0mgl−1 (31.7μM) 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.05 mgl−1 (0.23 μM) kinetin (Kin). Calli were maintained and proliferated by subculture at monthly intervals on MMS medium containing 4.5 mgl−1 (20.4 μM) 2,4-D and 0.2mgl−1 (0.9 μM) Kin. Somatic embryos (SE) were induced from seed-derived calli on SE-induction medium (MMS supplemented with 2.0 mgl−1 2,4-D and 0.2mgl−1 Kin). Plantlets were regenerated from somatic embryogenic calli grown on modified SH medium supplemented with 2 mgl−1 Kin. Using this optimized protocol, 78.6–82.3% of mature seeds of all four cultivars produced SE clusters, of which 93.5–95.3%
regenerated into plants within 10 wk. The regenerants showed no phenotypic abnormalities. 相似文献
8.
Summary An efficient protocol for plant regeneration from stem segments of Murraya koenigii was developed by culturing on Murashige and Skoog (MS) medium supplemented with 2.5 mg l−1 benzyladenine (BA), 25 mgl−1 adenine sulfate, 0.25 mgl−1 indole-3-acetic acid (IAA), and 3% sucrose. The frequency of shoot bud regeneration was higher on similar medium in subsequent
subcultures. The regenerated shoots were rooted on half-strength basal MS medium supplemented with 0.25–0.5 mgl−1 IAA or 1-naphthaleneacetic acid (NAA) within 8–12 d of culture. The maximum percentage of rooting was obtained on MS medium
supplemented with IAA and NAA, each at 0.25 mgl−1. During acclimatization, 95% of rooted plantlets survived were grown normally under greenhouse conditions. 相似文献
9.
Summary The effects of growth regulators on culture response of different pyrethrum (Chrysanthemum cinerariaefolium Vis.) genotypes were investigated. In the genotype Sb/66/107, the presence of 2,4-dichorophenoxyacetic acid (2,4-D) at 2
mgl−1 promoted growth of callus, whereas benzyladenine and 1-naphthaleneacetic acid had no effect. Callus growth was also affected
by the 2,4-D, ranging from 0.5 mgl−1 for genotype Marwanga to 3.0 mgl−1 for Ks/75/336. Among the genotypes, shoots were regenerated from calluses of Sb/66/107, 4331, Marwanga, and MA/70/1013. 相似文献
10.
Dilip K. Das Prasanna Bhomkar N. Shiva Prakash Neera Bhalla-Sarin 《In vitro cellular & developmental biology. Plant》2002,38(5):456-459
Summary A very rapid and efficient regeneration method of Vigna mungo L. has been established using liquid culture. A highly regenerable explant, viz., young multiple shoots obtained by germinating
the seeds in 2 mgl−1 (8.9μM) N6-benzyladenine-supplemented Murashige and Skoog (MS) medium, was used as a source of tissue to initiate the liquid culture.
The liquid medium consisted of half-strength B5 or MS salts supplemented with MS organics, α-naphthaleneacetic acid (0.1 mgl−1, 0.54μM) and N6-benzyladenine (0.5mgl−1, 2.2μM). Transferring the growing tissues to fresh medium every third day resulted in ca. 142% increase in the number of shoot buds produced after 24d. Shoot buds elongated on one-third-strength MS (MS1/3) semisolid medium and plantlets were obtained by transferring the shoots onto MS1/3 semisolid medium supplemented with indolebutyric acid (1 mgl−1, 4.9 μM). 相似文献
11.
Wenhao Dai Zong-Ming Cheng Wayne Sargent 《In vitro cellular & developmental biology. Plant》2003,39(1):6-11
Summary An efficient regeneration and transformation system was developed for two elite aspen hybrid clones (Populus canescens × P. grandidentada and P. tremuloides × P. davidiana). Callus was induced from in vitro leaf explants on modified Murashige and Skoog medium (MSA) and woody plant medium (WPM) containing four different combinations
of cytokinins and auxins. Callus tissues regenerated into shoots on WPM medium supplemented with 2.0 mgl−1 (9.12 μM) zeatin or 0.01 mgl−1 (0.045 μM) thidiazuron. P. canescens × P. grandidentata exhibited the higher callus and shoot production. In vitro leaf explants from the two hybrid clones were cocultivated with Agrobacterium tumefaciens strain EHA105 harboring the binary Ti plasmid pBI121 carrying the uidA gene encoding for β-glucuronidase (GUS) and the npt II gene encoding for neomycin phosphotransferase II. Transformation was confirmed by GUS assays, polymerase chain reaction,
and Southern blot analyses. Agrobacterium concentration, acetosyringone, and pH of the cocultivation medium were evaluated for enhancing transformation efficiency
with the clone P. canescens × P. grandidentata. 相似文献
12.
H. Mercier C. C. J. Vieira R. C. L. Figueiredo-Ribeiro 《Plant Cell, Tissue and Organ Culture》1992,28(3):249-254
Leaf and stem segments of Gomphrena officinalis originated from aseptically grown seedlings were used to initiate cultures. Callus production was obtained on gelled Murashige & Skoog medium supplemented with 6-benzylaminopurine alone (1.0, 5.0 or 10.0 mgl-1) or combined with -naphthalene acetic acid (0.1, 0.5 and 1.0 mgl-1) after 10 to 15 days of culture, and can be transferred to fresh medium every 30 days. The combinations of 5.0 or 10.0 mgl-1 of 6-benzylaminopurine with 0.1 mgl-1 of -naphthalene acetic acid were found to be the best for shoot regeneration. Adventitious shoot formation occurred after 50 to 60 days of culture in leaf and internode stem explants. Nodal segments developed actively growing lateral buds after 30 days of culture. Gelled Murashige & Skoog medium containing 10 mgl-1 of indole-3-butyric acid was considered optimal for the rooting of shoots. Rooted plants transferred to potting soil could be successfully established.Abbreviations BA
6-benzylaminopurine
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- MS
Murashige & Skoog
- NAA
-naphthalene acetic acid 相似文献
13.
Callus cultures were obrained from petiole explants of Carica papaya on MS medium containing 0.5–10.5 μM α-naphthaleneacetic acid (NAA) in combination with 0.5–5 μM benzyladenine (BA). Hard-green calli were transferred to MS medium
containing 100 mgl−1 casein hydrolysate (CH) with specific BA-NAA formulation, where they developed adventitious buds within 2 weeks of culture.
Maximum number of adventitious buds were obtained in 2 μM BA and 0.1 μM NAA. Shoot regeneration occurred from these adventitious
buds by the end of the 4th week. Regenerated shoots were elongated in hormone-free medium and rooted in half-strength MS fortified
with 3 UM NAA and 0.5 μM gibberellic acid (GA3). The regenerants were transferred to soil after acclimatization. 相似文献
14.
Summary An efficient and reproducible protocol for mass propagation of Eclipta alba (L.) Hassk, an important medicinal plant, was standardized by culturing shoot tips and nodal segments taken from in vitro raised plants. Maximum shoot proliferation occurred when the explants were cultured on Murashige and Skoog (MS) medium supplemented
with 1 mg l−1 benzylaminopurine (BAP). The shoot buds formed were further multiplied and maintained on medium containing BAP (0.5 mgl−1) and gibberellic acid (0.5 mgl−1). Rooting was best achieved on MS medium supplement with 1 mg−1 indole-3-butyric acid. Rooted plantlets attained maturity and flowered normally in the field. 相似文献
15.
Summary An efficient in vitro plant regeneration system from cotyledons was established in tetraploid Isatis indigotica Fort. Factors influencing shoot regeneration from cotyledons, including culture medium type, combinations of plant growth
regulators, and sucrose concentrations in the medium, as well as illumination were investigated. Murashige and Skoog's (MS)
medium was found to be best for promoting shoot regeneration, followed by Gamborg's B5 and White's medium. The highest shoot regeneration frequency was achieved from cotyledons cultured on MS medium supplemented
with 2.0 mgl−1 (8.9 μM) 6-benzyladenine and 1.0 mgl−1 (5.4 μM) α-naphthaleneacetic acid (NAA), with 97.9% regeneration, associated with a high number of multiple shoots developed per
explant (8.6 shoots per explant). A sucrose concentration of 3% present in the medium and light conditions were beneficial
for shoot regeneration. The shoots developed were rooted in a half-strength MS medium supplemented with 1.0 mgl−1 (5.4 μM) NAA and successfully transplanted in soil in pots with over 85% survival. The establishment of an efficient plant regeneration
procedure from cotyledons provides a basis for the rapid in vitro multiplication of tetraploid Isatis indigotica Fort., one of the most extensively used medicinal plants in China currently under great shortage. 相似文献
16.
Deepak Prem Subhadra Singh Padam Prakash Gupta Jaivir Singh Gaurav Yadav 《In vitro cellular & developmental biology. Plant》2003,39(4):384-387
Summary Guar (Cyamopsis tetragonoloba L. Taub) is a drought-tolerant multipurpose cash crop. A rapid regeneration system has been developed for four Indian guar
genotypes. Investigations were carried out to assess the effect of different growth regulators and their combinations on a
variety of explants such as the embryo, cotyledons, and cotyledonary nodes for shoot morphogenesis. It was established that
Murashige and Skoog's culture medium containing 6-benzylaminopurine (13.3 μM or 3 mgl−1) in combination with indole-3-acetic acid (11.4 μM or 2mgl−1) with cotyledonary node explants gave the highest frequency of multiple shoot induction. In vitro rooting from cultured shoots was maximal on a half-salt concentration of Murashige and Skoog's culture medium fortified with
indole-3-butyric acid (4.9 μM or 1 mgl−1). In vitro-regenerated plants were grown to pod setting and subsequent maturity in greenhouse conditions. 相似文献
17.
Summary High-frequency embryogenesis systems were established for hybrid yellow-poplar (Liriodendron tulipifera×L. chinense) and hybrid sweetgum (Liquidambar styraciflua×L. formosana) by modifying a medium originally developed for embryogenic yellow-poplar cultures. Embryogenic cultures of both hybrids,
consisting of proembryogenic masses (PEMs), were initiated from immature hybrid seeds on an induction-maintenance medium (IMM)
supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D), benzyladenine (BA), and casein hydrolyzate (CH). For hybrid yellow-poplar,
as many as 2100 germinable somatic embryos per 4000 cells or cell clumps were produced when PEMs were grown in liquid IMM
lacking CH, at a pH that varied with genotype (3.5 or 5.6), followed by size fractionation and plating on semisolid embryo
development medium (DM; IMM lacking 2,4-D and BA) without CH, but supplemented with 4.0 mgl−1 (15 μM) abscisic acid. For hybrid sweetgum, up to 1650 germinable somatic embryos per 4000 cells or cell clumps were produced when
PEMs were grown in liquid IMM without CH, but with 550 mgl−1
l-glutamine, 510 mg l−1 asparagine, and 170 mg l−1 arginine at pH 5.6. Somatic embryos developed from cell clumps on DM without any plant growth regulators or other supplements.
Hundreds of somatic embryos of both hybrids were germinated on DM without CH, transferred to potting mix, and hardened off
in a humidifying chamber for transfer to the greenhouse. 相似文献
18.
Summary Callus induction was observed from hypocotyl, root, and cotyledonary leaf segments, grown on Murashige and Skoog (MS) medium
supplemented with various concentrations and combinations of 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin (KN). Maximum
callusing (100%) was obtained from root and cotyledonary leaf segments grown on MS medium supplemented with a combination
of 2 mg l−1 (9.1 μM) 2,4-D and 0.2 mg l−1 (0.9 μM) KN. The calluses, when subcultured in the same medium, showed profuse callusing. However, these calluses remained recalcitrant
to regenerate regardless of the quality and combinations of plant growth regulators in the nutrient pool. When hypocotyl segments
were used as explants, callus induction was noticed in 91% of cultures which showed shoot regeneration on MS medium supplemented
with 2 mg l−1 2,4-D and 0.2 mg l−1 KN. These shoots were transferred to fresh medium containing various concentrations and combinations of 6-benzyladenine (BA)
and N6-(2-isopentenyl)adenosine (2-iP). Maximum shoot multiplication was observed after 60 d of the second subculture on MS medium
containing 2 mg l−1 (8.9 μM) BA. These shoots were rooted best (87%) on MS medium containing 2 mg l−1 (9.9 μM) indole-3-butyric acid (IBA). The plantlets were transferred to the field after acclimatization and showed 60% survival. 相似文献
19.
Leaf, stem, hypocotyl, cotyledon, root, shoot tip and embryo explants of Capsicum annuum L. cv. mathania were cultured on Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine (BAP) or kinetin (Kin) alone or in
combination with 3-indoleacetic acid (IAA), 3-indolebutyric acid (IBA), α-naphthaleneacetic acid (NAA) or 2,4-dichlorophenoxyacetic
acid (2,4-D). BAP (5.0 mgl−1) in the medium was found to be the best growth regulator for shoot bud differentiation. Shoot buds cultured on 5.0 mgl−1 BAP increased in number but did not elongate. For obtaining complete plantlets, shoot buds were placed on a medium with IBA
or NAA (0.1 mgl−1). Histological evidence revealed direct differentiation of buds from cotyledons. Regenerated plants were normal diploids.
Unorganized callus could not be induced to differentiate shoot buds. 相似文献
20.
Summary Protoplasts were isolated from Agrobacterium rhizogenes A4-transformed cell line of Medicago sativa L. The highest yield of protoplasts (4.2×106 per g fresh weight) was obtained from 12-d-old calluses after being subeultured on fresh medium. The viability of protoplasts
reached over 80%. Protoplasts were induced to undergo sustained divisions when cultured in Durand et al. (DPD) medium supplemented
with 2 mgl−1 (9.05 μM) 2,4-dichlorophenoxyacetic acid, 0,2mgl−1 (0.93 μM) kinetin, 0.3 M mannitol, 2% (w/v) sucrose, and 500 mgl−1 casein hydrolyzate at a plating density of 1.0×105 per ml. An agarose-beads culture method was appropriate for protoplast division of transformed alfalfa. The division frequency
was about 30%. Numerous hairy roots were induced from protocalluses on Murashige and Skoog medium without growth regulators.
Paper electrophoresis revealed that all of the regenerated hairy roots tested synthesized the corresponding opines. This protoplast
culture system would be valuable for further somatic hybridization in forage legumes. 相似文献