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1.
Summary A procedure for protoplast isolation and plant regeneration of St. John's wort has been developed to utilize cell-to-cell
variability for optimum production of valuable medicinal compounds. Calluses, induced from hypocotyl segments of St. John's
wort seedlings, were used for protoplast isolation, induction of sustained cell division, and ultimately, plant regeneration.
Callus-isolated protoplasts at a density of 2.0×105 per ml were embedded in 0.6% Na-alginate blocks and cultured in a medium containing modified Murashige and Skoog (MS) salts,
2.5 μM 6-benzylaminopurine (BA), 5.0 μMα-naphthaleneacetic acid (NAA), and 0.5 moll−1 glucose. Protoplast-derived colonies formed compact calluses when transferred onto 0.35% gellan gum-solidified MS medium
supplemented with 2.5 μM BA and 2.5 μM NAA. Shoot organogenesis from the protoplast-derived callus was induced on MS medium supplemented with 5 μM thidiazuron. Complete plantlets were obtained from the regenerated shoots on MS basal medium. A greater than 3-fold variation
of antioxidant activity was observed among the protoplast-derived plantets and chemically distinct germplasm lines were selected
on the basis of phytochemical profiles. The protoplast to plant regeneration protocol developed in this study provides the
foundation for development of novel genotypes with potential expansion of the genetic diversity through somatic hybridization,
and organelle transplantation. 相似文献
2.
Summary Some native species produce seeds with a low frequency of germination accompanied with a period of dormancy. These features
make it difficult to produce new phenotypes through sexual propagation. Maclura tinctoria has been considered an endangered species due to extensive use of its wood and low frequency of seed germination. The objective
of the present study is to establish an in vitro propagation system for this species. Organogenic friable callus formation from nodal segments has been obtained using woody
plant medium (WPM) supplemented with 10.74 μM 1-naphthaleneacetic acid (NAA)+4.43 μM 6-benzylaminopurine (BA). Results indicate that the highest frequency of shoot formation is observed when WPM supplemented
with 4.03 μM NAA+4.43 BA is used. For root formation, the use of WPM medium (pH adjusted to 7.0) supplemented with 23.62 μM indole-3-butyric acid (IBA) and 4.7gl−1 activated charcoal is recommended. For acelimatization, subjecting rooted plantlets to 70%, 50%, and 30% mesh screen, each
successively for a period of 7 d, has resulted in 97% plantlet survival. 相似文献
3.
N. R. Nayak P. K. Chand S. P. Rath S. N. Patnaik 《In vitro cellular & developmental biology. Plant》1998,34(3):185-188
Summary An efficient procedure is outlined forin vitro regeneration of an epiphytic orchid,Cymbidium aloifolium (L.) Sw. using rhizomes developed from seeds. Murashige and Skoog's (1962) medium (MS) containing indole-3-acetic acid (IAA),
indole-3-butyric acid (IBA), or 1-naphthaleneacetic acid (NAA) stimulated growth and proliferation of rhizomes with NAA being
most effective at 5.0 mg.l−1 (27.0 μM). Shoot bud differentiation was induced in the apical portions of the rhizomes on MS medium containing kinetin (Kn) or N6-benzyladenine (BA). The highest frequency of shoot regeneration (91.5%) and the maximum number of shoot buds formed (3.5
shoots/rhizome) were recorded with BA at 1.0 mg.l−1 (4.4 μM). NAA (0.1 mg.l−1, 0.54 μM), whenever added to the medium in conjunction with BA (1.0 mg.l−1, 4.4 μM), slightly enhanced the frequency of shoot bud regeneration (92.6%) and the number of shoot buds formed (5.2 shoots/rhizome).
Moreover, an NAA-BA combination induced rooting in regenerated shoots thereby producing complete plantlets in one step. Shoots
developed on cytokinin-supplemented medium were rooted on MS containing NAA at 1.0 mg.l−1 (5.4 μM). Regenerated plantlets were acclimated and eventually established in a garden. 相似文献
4.
Yongxue Yang Guofeng Liu Manzhu Bao 《In vitro cellular & developmental biology. Plant》2006,42(6):520-524
Summary A protocol of somatic embryogenesis and plant regeneration from petiole segments of Parthenocissus tricuspidata Planch. has been developed. Embryogenic tissue was induced on B5 (Gamborg) basal medium supplemented with 2.25–9.0 μM 2,4-dichlorophenoxyacetic acid, 500 mg l−1 casein hydrolysate (CH), and 0.1 gl−1 activated charcoal. Somatic embryos were induced on B5 medium containing various concentrations of benzyladenine (BA) (4.44,
6.66, and 8.88 μM) and α-naphthaleneacetic acid (NAA) (0, 0.54, and 1.61 μM) plus 500 mg l−1 CH. Ninety percent of normal somatic embryos were converted into plantlets directly on Murashige and Skoog (MS) medium free
of plant growth regulators. Shoots could be induced from abnormal somatic embryos on MS medium containing 4.44 μM BA, 0.05 μM NAA, and 500 mg l−1 CH. Genotypic differences were found in the process of somatic embryogenesis and plant regeneration. Histological analysis
confirmed the process of somatic embryogenesis. Regenerated plantlets with well-developed roots were successfully acclimatized
in greenhouse and all plants showed normal morphological characteristics. 相似文献
5.
I. M. G. Padilla E. Carmona N. Westendorp C. L. Encina 《In vitro cellular & developmental biology. Plant》2006,42(2):193-196
Summary A micropropagation protocol for Pouteria lucuma R. and Pav. var. La Molina, was developed. Shoots from zygotic embryos with a portion of endosperm were established in vitro on Murashige and Skoog (MS) medium with 0.47 μM kinetin (Kin) and 0.54 μM naphthaleneacetic acid (NAA). Multiplication of shoots was accomplished using subapical, shoots. The best axillary-shoot
production was observed on MS basal, medium with 2.2μM, benzyladenine (BA), 0.5 μM NAA, 1.4. μM gibberellic acid (GA3), and 40 mgl−1 adenine sulfate, with the development of up to three axillary shoots per subapical shoot. One hundred percent rooting was
obtained from shoots grown, for 4wk on MS medium with 246 μM indole-3-butyric acid under light conditions. Eighty percent of the microplantlets survived after acclimatization when transplanted
to a substrate previously enriched with beneficial soil bacteria. This study describes, for the first time, arbuscular mycorrhizal
(AM) colonization of this species. Inoculation with AM fungi improved growth and development of lucumo plants and induced
changes to the root morphology. 相似文献
6.
Summary Vegetatively propagated plantlets of six rose cultivars were induced to flower in vitro on media containing full-strength Murashige and Skoog (MS) inorganic salts, Gamborg's B5 organic elements with 400 mg l−1
myo-inositol, and different phytohormone combinations of 6-benzyladenine (BA) with α-naphthaleneacetic acid (NAA); thidiazuron
(TDZ) with NAA; and zeatin (ZT) with NAA. The most efficient flower bud induction (49.1% and 44.1%) was obtained on media
supplemented with 0.5 mg l−1 (2.27 μM) TDZ and 0.1 mg l−1 (0.54 μM) NAA or 0.5 mg l−1 (2.28 μM) ZT and 0.1 mg l−1 (0.54 μM) NAA for cultivar Orange Parade. Scanning electron microscopy (SEM) showed that in vitro flower bud induction occurred mostly between 15 and 30 d in induction medium through the normal flower development processes.
With TDZ and ZT as the best choice for flower induction in all six cultivars tested, different rose cultivars varied in their
responses to phytohormone treatments. Our study also revealed that the total time from original culture and subculture time
before flower induction were two very important factors for in vitro flower induction. Plantlets 156–561 d from original culture and subcultured for 45 d were the best for flower induction.
These authors contributed equally to this work. 相似文献
7.
Summary An efficient protocol was established for in vitro shoot multiplication from nodal explants of Clitoria ternatea on semisolid Murashige and Skoog (MS) basal medium supplemented with 8.9μM 6-benzylaminopurine (BA). Inclusion of 1-naphthaleneacetic acid (NAA) in the culture medium along with BA promoted higher
rates of shoot multiplication than BA alone. The rate of shoot multiplication was maximum (5.21) after 4 wk of culture on
MS basal medium supplemented with 8.9μM BA and 1.34μM NAA. The elongated shoots rooted within 7–8d in half-strength MS basal salts supplemented with 1.34μM NAA and 2% (w/v) sucrose. About 85% of the rooted plantlets were acclimatized and transferred to the greenhouse. 相似文献
8.
Summary This study reports a protocol for successful micropropagation of Decalepis arayalpathra (Joseph and Chandras) Venter. (Janakia arayalpathra Joseph and Chandrasekhran; Periplocaceae), a critically endangered and endemic ethnomedicinal plant in the southern forests of the Western Ghats which is overexploited
for its tuberous medicinal roots by the local Kani tribes. Natural regeneration is rare and conventional propagation is difficult.
Conservation of the species through micropropagation was attempted. The nodal explants of greenhouse-raised plants, were more
desirable than cotyledonary nodal explants of aseptic seedlings. The basal nodes (73%) of 12–16-wk-old greenhouse-grown plants
cultured in Murashige and Skoog (MS) medium containing 12.96 μM 6-benzyladenine (BA), 2.48 μM 2-isopentenyladenine (2-ip) and 2.68 μM α-naphthaleneacetic acid (NAA) formed 16–17 cm long unbranched robust solitary shoots in 8 wk. Cotyledonary nodal explants
cultured in the same medium showed multiple shoot formation and axillary branching. But the shoots were thin, fragile and
not suitable for mass propagation. Single nodes of a solitary shoot subcultured on MS medium containing 2.22 μM BA and 0.24 μM 2-ip together produced 9.8±0.3 nodes from 18.0±0.6 cm long shoots within 5–6 wk. The basal nodes of the shoots so formed
were repeatedly subcultured to increase the stock of propagules while the 2.5–3.0 cm terminal cuttings were used for rooting.
The best root induction (68%) and survival (86%) was achieved on half-strength MS medium supplemented with 1.07 μM NAA. Field-established plants showed uniform growth and phenotypic similarity to parental stock. 相似文献
9.
Summary Goldenseal (Hydrastis canadensis L.), a popular native American medicinal plant, is currently listed as endangered or threatened in over one-third of the
states in which it is listed. The objective of this study was to develop an in vitro culture protocol for Goldenseal. Excise embryos were grown on Gamborg's B-5 medium with 0,1 or 10 μM gibberellic acid (GA3), and supplemented with 30 gl−1 sucrose and 8 gl−1 agar. Germinated embryos provided explants (leaf and root tissue) that were subsequently cultured on various media with combinations
of naphthleneacetic acid (NAA) and benzyladenine (BA). All NAA/BA combinations produced multiple shoots, roots, and callus.
Leaf explants cultured on medium with 1∶10 μM NAA:BA and root explants on medium with 1∶1 μM NAA:BA could be successfully used for mieropropagation. 相似文献
10.
Summary A protocol has been developed for in vitro plant regeneration from cotyledonary nodes of Pterocarpus marsupium Roxb. Multiple shoots were induced from cotyledonary nodes derived from 20-d-old axenic seedlings grown on Murashige and
Skoog (MS) medium containing 2.22–13.32 μM benzyladenine (BA) or 2.32–13.93 μM kinetin alone or in combination with 0.26 μM α-naphthaleneacetic acid (NAA). The highest frequency of responding explants (85%) and maximum number of shoots per explant
(9.5) were obtained on MS medium supplemented with 4.44 μM BA and 0.26 μM NAA after 15 wk of culture. A proliferating shoot culture was established by repeatedly subculturing the orginal cotyledonary
nodal explant on fresh medium after each harvest of the newly formed shoots. Nearly 30% of the shoots formed roots after being
transferred to half-strength MS medium containing 9.84 μM indole-3-butyric acid after 25 d of culture. Fifty percent of shoots were also directly rooted as microcuttings on peat moss,
soil, and compost mixture (1∶1∶1). About 52% plantlets rooted under ex vitro conditions were successfully acclimatized and established in pots. 相似文献
11.
Summary A protocol has been developed for in vitro plant regeneration from cotyledonary nodes of Pterocarpus marsupium Roxb. Multiple shoots were induced from cotyledonary nodes derived from 20-d-old axenic seedlings grown on Murashige and
Skoog (MS) medium containing 2.22–13.32 μM benzyladenine (BA) or 2.32–13.93 μM kinetin alone or in combination with 0.26 μM α-naphthaleneacetic acid (NAA). The highest frequency for shoot regeneration (85%) and maximum number of shoots per explant
(9.5) were obtained on the medium supplemented with 4.44 μM BA and 0.26 μM NAA after 15 wk of culture. A proliferating shoot culture was established by repeatedly subculturing the original cotyledonary
nodal explant on fresh medium after each harvest of the newly formed shoots. Nearly 30% of the shoots formed roots after being
transferred to half-strength MS medium containing 9.84 μM indole-3-butyric acid (IBA) after 25 d of culture. Fifty percent of shoots were also directly rooted as microtuttings on
a peat moss, soil, and compost mixture (1∶1∶1). About 52% of plantlets were successfully acclimatized and established in pots. 相似文献
12.
Summary Segments of male inflorescences of medicinal yam (Dioscorea floribunda) cultured on Murashige and Skoog (MS) medium supplemented with 13.94 μM kinetin (Kn) resulted in the conversion of floral buds into vegetative buds and these later developed into plantlets. Growth
and multiplication of shoots could be obtained by culturing individual shoots in MS modified basal medium, replacing the MS
standard three vitamins with 10.0 mgl−1 thiamine in addition to 13.94 μM Kn. Root induction was also obtained simultaneously from the base of the shoots in the same medium. Such plantlets have been
successfully transferred to potted soil, where they grew normally. Plantlets were also made to develop tubers on MS medium
with 18.91 μM abscisic acid (ABA) and also with 2.68 μM α-naphthaleneacetic acid (NAA) and 40–50 gl−1 sucrose. 相似文献
13.
Irene ávila-Díaz Ken Oyama Carlos Gómez-Alonso Rafael Salgado-Garciglia 《Plant Cell, Tissue and Organ Culture》2009,99(3):335-343
Laelia speciosa is an endangered epiphytic orchid. The effects of various media components on germination of L. speciosa were evaluated. Pods were collected at 4, 7, and 9 months following hand-pollination, and seeds were germinated on Murashige and Skoog (MS)
media with 30 g l−1 sucrose and five concentrations (0.0, 0.04, 0.22, 0.44, and 2.22 μM) of benzyladenine (BA) under light and dark conditions.
Gibberellic acid (GA3; 0.0, 0.29, 1.44, 2.89, 14. 43 and 28.87 μM) with naphthaleneacetic acid (NAA; 0.0, 0.54, 1.34, 2.69, and 5.37 μM) were evaluated
for in vitro subcultivation. MS medium with 30 g l−1 sucrose was effective for germination. The effects of BA and light on germination of L. speciosa seeds differed with pod maturity. All mature seeds germinated using 0.44 μM BA and light. The highest frequency of germinated
seedlings (60%) was obtained using mature seeds grown on MS medium without BA and under light conditions. For subculture,
MS with 30 g l−1 sucrose, 2.69 μM NAA, and 0.29 μM GA3 was effective. Plantlets of 5 cm in length were transplanted to the greenhouse, and a 77.5% of survival rate was obtained.
A successful protocol for micropropagation by seed germination will contribute to the development of a sustainable management
program for L. speciosa. 相似文献
14.
Solange Faria Lua Figueiredo Norma Albarello Vera Regina Campos Viana 《In vitro cellular & developmental biology. Plant》2001,37(4):471-475
Summary A protocol for in vitro propagation of Rollinia mucosa, an important medicinal plant, was developed. The presence of 500 mg l−1 polyvinylpyrrolidone (PVP) during explant excision was important to avoid browning. Axillary buds, adventitious buds, and
shoot cluster proliferation were achieved from epicotyl and hypocotyl explants from nursery-grown seedlings. The highest direct
organogenesis percentage from hypocotyl explants was obtained upon culture of explants on Murashige and Skoog medium supplemented
with 2.2 μM benzyladenine (BA) plus 2.32 μM kinetin. Epicotyl explants display highest regeneration frequency on a medium containing 8.8 μM BA and 0.54 μM naphthaleneacetic acid. Gibberellic acid was necessary for shoot elongation. Root induction was observed when shoots were
pretreated with activated charcoal for 7 d in the dark before culture on Woody Plant Medium supplemented with 49.21 μM indolebutyric acid for 10 d. Root development was observed when 20 g l−1 sucrose was used. Rooted plantlets were acclimatized and grown in the greenhouse. 相似文献
15.
Yan Ma David H. Byrne Jing Chen 《In vitro cellular & developmental biology. Plant》1996,32(2):103-108
Summary Several rose species (Rosa rugosa, R. wichuraiana, R. setigera, R. laevigata, R. banksiae, R. roxburghii, R. odorata) and interspecific hybrids were cultured to determine the appropriate concentrations of nutrients and growth regulators for
shoot proliferation and root initiation. Cultured shoot tips and lateral buds from different genotypes proliferated multiple
shoots on a basal medium [Murashige and Skoog (MS) salts, vitamins, glycine, sucrose, and agar] supplemented with 0 μM to 17.8 μM (4 mg·l−1) 6-benzyladenine (BA) and 0 μM to 0.54 μM (0.1 mg·l−1) naphthalene, acetic acid (NAA). The ability of the explants to proliferate shoots and initiate roots was affected by the
genotype, the nodal position of the explant, the strength of the MS basal salts, and the growth regulators used. The buds
nearest the apex exhibited the slowest rate of development. Most species had the highest shoot proliferation when cultured
on basal MS medium supplemented with 8.9 μM (2 mg·l−1) BA, but the degree varied by species. Root development was enhanced by lowering the concentration of MS salts. With difficult-to-root
species, rooting was improved by supplementing the media with 11.4 μM (2 mg·l−1) indole-3-acetic acid (IAA) or by giving them a 7-d dark treatment at 10°C. 相似文献
16.
C. P. Kaviraj G. Kiran R. B. Venugopal P. B. Kavi Kishor Srinath Rao 《In vitro cellular & developmental biology. Plant》2006,42(2):134-138
Summary This study reports a protocol for plant regeneration from cultured explants of green gram [Vigna radiata (L.) Wilezek] via somatic embryogenesis. Somatic embryos were induced from nature cotyledons of var., TAP-7 and Pusa Baisaki
when cultured on Murashige and Skoog (MS) medium fortified with different concentrations of 2,4-dichlorophenoxyacetic acid,
α-naphthaleneacetic acid (NAA) and 2,4,5-trichlorophenoxyacetic acid singly or in combination with 2.22–8.88 μM N
6-benzylaminopurine (BAP) or 2.32–9.38 μM kinetin. The type and concentration of auxin and plant genotype influenced the frequency of somatic embryogenesis. NAA was
the most effective auxin for somatic embryo induction. The well-developed, cotyledonary shaped embryos of var. TAP-7 germinated
into plantlets at a frequency of 56.6% on MS medium supplemented with 1.88 μM abscisic acid and 6.66 μM BAP. Regenerated plants were transferred to soil and grown to maturity with 90% survival. The protocol described here offers
a good potential for genetic improvement using gene transfer techniques and the production of synthetic seeds of V. radiata. 相似文献
17.
Summary Totipotent callus of Cypripedium formosanum, an endangered slipper orchid species, was induced from seed-derived protocorm segments on a quarter-strength Murashige and
Skoog medium containing 4.52 μM 2,4-dichlorophenoxyacetic acid and 4.54 μM 1-phenyl-3-(1,2,3-thiadiazol-5-yl)-urea (thidiazuron). This callus proliferated well and was maintained by subculturing on
the same medium. On average, 13 protocorm-like bodies could be obtained from a piece of 4 mm callus after being transferred
to the medium with 4.44 μM N6-benzyladenine after 8 wk of culture. The regenerated protocorm-like bodies formed shoots and roots on medium containing 1
g l−1 activated charcoal and 20 g l−1 potato homogenate. After 24 wk of culture on this medium, well-developed plantlets ready for potting were established. 相似文献
18.
P. S. Sha Valli Khan E. Prakash K. R. Rao 《In vitro cellular & developmental biology. Plant》2002,38(2):186-190
Summary A protocol has been developed for plantlet regeneration from seed callus of Bixa orellana L. Seeds demonstrated a high percentage of callus induction (63±7.3%) and a high yield (356±14.7 mg per seed) of white friable
callus on Murashige and Skoog (MS) medium containing 5.0 μM l-naphthaleneacetic acid (NAA) and 2.5μM N
6-benzyladenine (BA) within 6 wk of culture in the dark. Callus induction frequency was greater under 24h dark as compared
to 16h light/8h dark photoperiod or 24h light photoperiod. Increased myo-inositol (MI: 200mgl−1) and addition of ascorbic acid (AA: 200 mgl−1) to the culture medium positively improved callus induction frequency and growth. Shoot differentiation from white friable
seed callus was best using 10.0 μM BA and 5.0 μM NAA, where the highest percentage of calluses forming shools (74.9±4.8%), the highest number of shoots per callus (six or
seven) and the highest shoot-forming index (5.0) were obtained within 6 wk. Shoots elongated to 4 cm within 4 wk of transfer
onto MS medium devoid of growth regulators. Shoots were rooted using half-strength MS medium containing 5.0 μM indole-3-butyric acid (IBA). About 85% of these plants were established in pots containing pure garden soil and organic manure
after 3 wk of hardening. Regenerated plants were morphologically uniform with normal leaf, shape and growth patterns. These
plants are currently being screened for the presence of agronomically useful genetic variants. 相似文献
19.
Chin-Wen Ho Wei-Ting Jian Hui-Chun Lai 《In vitro cellular & developmental biology. Plant》2006,42(3):240-246
Summary
In vitro seedlings of Lilium × formolongi Hort. evs. Norikula, RaiZen No. 1, RaiZen No. 3, RaiZen Early, and Bailansa were used to induce callus by variously modified
Murashige and Skoog (MS) media, using protocols for flask culture and bioreactor culture. Green embryogenic callus proliferated
from roots near the base of bulblets of five varieties on media containing 0.53–5.3 μM α-naphthaleneacetic acid (NAA), and 28 cell lines were obtained by subcultures on the same medium. For flask culture, the
fresh weight (FW) of embryogenic cell clumps doubled every 4 wk on MS basal salts supplemented with 0.53°M NAA and 30 g l−1 sucrose. The maximum frequency of somatic embryos that developed into plantlets was 76.67±17% when plated onto solid MS basal
medium without plant growth regulators (PGRs). Among the treatments using four types of bioreactors, the best cell growth
and regeneration rate (74±0.14%) of somatic embryos was in a modified 2–1 bioreactor. Cells incubated in the other three bioreactors
furned brown and died. Histological study revealed that regeneration was by somatic embryogenesis. The regenerants showed
normal growth and flowering after 8–9 mo, in the field. A cell line of cv. Norikula has been subcultured in MS basal salts
containing 0.53 μM NAA every 2 mo. for 6 yr. The cell aggregates became more synchronous and many typical embryogenic cells with dense cytoplasm
were observed under a light microscope. The long-term embryogenic cells plated on MS basal medium still gave rise to numerous
somatic embryos and converted into plantlets. 相似文献
20.
D. J. Williams K. H. Al-Juboory R. M. Skirvin 《In vitro cellular & developmental biology. Plant》1998,34(4):289-292
Summary The objective of this study was to evaluate the ability ofHosta Golden Scepter (GS) ovary explants to generate adventitious shootsin vitro. Ovaries were transversely cut into halves and transferred to petri dishes containingHosta initiation medium supplemented with naphthaleneacetic acid (NAA) at 2.5 μM and N6-benzyladenine (BA) at 10 μM. GS produced adventitious shoots from the ovary base via organogenesis. The number of adventitious shoots regenerated from
callus increased linearly with repeated subculturing on Murashige and Skoog (MS) medium supplemented with 2.5 μM NAA and 10 μM BA. The number of multiple shoots developing from callus (15.8), shoot tip (8.4), leaf (6.7), and root (4.3) occurred on
MS medium supplemented with 2.5 μM NAA and 20–30 μM BA. There were significant differences in the number of shoots regenerated from shoot tips and callus on MS medium with 50
and 100 mgmyo-inositol per l. Similarly, there were significant differences in the number of axillary shoots and adventitious shoots produced
with 20 g/l sucrose treatment. 相似文献