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1.
P. Giridhar Vinod Kumar G. A. Ravishankar 《In vitro cellular & developmental biology. Plant》2004,40(6):567-571
Summary A novel protocol has been developed for inducing somatic embryogenesis from leaf cultures of Decalepis hamiltonii. Callus was obtained from leaf sections in Murashige and Skoog (MS) medium supplemented with α-naphthaleneacetic acid (NAA)+N6-benzyladenine (BA) or 2,4-dichlorophenoxyacetic acid (2,4-D)+BA. Nodular embryogenic callus developed from the cut end of
explants on media containing 2,4-D and BA, whereas compact callus developed on media containing NAA and BA. Upon subsequent
transfer of explants with primary callus onto MS media containing zeatin and/or gibberellic acid (GA3) and BA, treatment with zeatin (13.68μM) and BA (10.65 μM) resulted in the induction of the highest number of somatic embryos directly from nodular tissue. The maturation of embryos
took place along with the induction on the same medium. Embryogenic calluses with somatic embryos were subcultured onto MS
basal medium supplemented with 4.56μM zeatin+10.65 μM BA. After 4wk, more extensive differentiation of somatic embryos was observed. The mature embryos developed into complete
plantlets on growth regulator-free MS medium. A distinct feature of this study is the induction of somatic embryogenesis from
leaf explants of Decalepis hamiltonii, which has not been reported previously. By using this protocol, complete plantlets could be regenerated through indirect
somatic embryogenesis or organogenesis from leaf explants in 12–16 wk. 相似文献
2.
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
3.
G.-Y. Kai L.-M. Dai X.-Y. Mei J.-G. Zheng W. Wang Y. Lu Z.-Y. Qian G.-Y. Zhou 《Biologia Plantarum》2008,52(3):557-560
An efficient in vitro plant regeneration system from leaves of Ophiorrhiza japonica Blume was established for the first time. Callus formation rate was more than 90.4 % from leaf segments on Murashige and
Skoog (MS) supplemented with either α-naphthaleneacetic acid (NAA) alone or in combination with 6-benzyladenine (BA). The
highest shoot regeneration (78.9 %) was achieved on MS medium containing 2.0 mg dm−3 BA and 0.2 mg dm−3 NAA, with an average of 9.4 shoots developed per leaf segment. Shoot regeneration was also improved when the leaf explants
were cultured in MS basal medium supplemented with 0.5 % (m/v) polyvinylpyrrolidone (PVP). The leaf explants from seedlings
with age of about 18–27 d showed the highest shoot regeneration. The regenerated shoots were rooted on half-strength basal
MS medium supplemented with 0.5 mg dm−3 indole-3-butyric acid (IBA), which averagely produced 24.8 roots per shoot. The plantlets were transferred to soil, where
100 % survived after 1 month of acclimatization. 相似文献
4.
Jin Cui Jianjun Chen Richard J. Henny 《In vitro cellular & developmental biology. Plant》2009,45(1):34-43
Plant regeneration through direct somatic embryogenesis in Aeschynanthus radicans ‘Mona Lisa’ was achieved in this study. Globular somatic embryos were formed directly from cut edges of leaf explants and
cut ends or on the surface of stem explants 4 wk after culture on Murashige and Skoog (MS) medium supplemented with N-phenyl-N′-1, 2, 3-thiadiazol-5-ylurea (TDZ) with α-naphthalene acetic acid (NAA), TDZ with 2,4-dichlorophenoxyacetic acid (2,4-D),
or 6-benzylaminopurine (BA) or kintin (KN) with 2,4-D. MS medium containing 9.08 μM TDZ and 2.68 μM 2,4-D resulted in 71%
of stem explants producing somatic embryos. In contrast, 40% of leaf explants produced somatic embryos when induced in medium
containing 6.81 μM TDZ and 2.68 μM 2,4-D. Somatic embryos matured, and some germinated into small plants on the initial induction
medium. Up to 64% of stem explants cultured on medium supplemented with 9.08 μM TDZ + 2.68 μM 2,4-D, 36% of leaf explants
cultured on medium containing 6.81 μM TDZ and 2.68 μM 2,4-D had somatic embryo germination before or after transferring onto
MS medium containing 8.88 μM BA and 1.07 μM NAA. Shoots elongated better and roots developed well on MS medium without growth
regulators. Approximately 30–50 plantlets were regenerated from each stem or leaf explant. The regenerated plants grew vigorously
after transplanting to a soil-less substrate in a shaded greenhouse with more than a 98% survival rate. Three months after
their establishment in the shaded greenhouse, 500 plants regenerated from stem explants were morphologically evaluated, from
which five types of variants that had large, orbicular, elliptic, small, and lanceolate leaves were identified. Flow cytometry
analysis of the variants along with the parent showed that they all had one identical peak, indicating that the variant lines,
like the parent, were diploid. The mean nuclear DNA contents of the variant lines and their parent ranged from 4.90 to 4.99 pg
2C−1, which were not significantly different statistically. The results suggest that the regenerated plants have a stable ploidy
level, and the regeneration method established in this study can be used for rapid propagation of ploidy-stable Aeschynanthus radicans. 相似文献
5.
Summary A protocol for in vitro propagation using direct induction of shoot buds from leaf explants of in vitro-raised shoots of Rosa damascena var. Jwala is reported. The present study is the first report on direct shoot regeneration in scented roses. Elite plants
raised from nodal explants and maintained for over 2yr in vitro on a static liquid shoot multiplication Murashige and Skoog (MS) medium supplemented with 5.0 μM benzyladenine (BA) and 3% sucrose were used. Petioles from fully developed young leaves, obtained after 4 wk of pruning of
old shoots, were found to be ideal for regeneration of shoots. Initially the explants were cultured in an induction medium
[half-strength MS+3% sucrose+6.8μM thidiazuron+0.27 μM α-naphthaleneacetic acid (NAA)+17.7 μM AgNO3] and subsequently transferred to the regeneration medium (MS+2.25 μM BA+0.054 μM NAA) after 7, 14, 21, 28, and 35d. The highest shoot regeneration response (69%) was recorded when shoots were kept in the
induction medium for 21 d and later transferred to regeneration medium. Histological studies revealed direct formation of
shoot buds without the intervening callus phase. In vitro rooting of micro-shoots was accomplished within 2wk on half-strength MS liquid medium supplemented with 10.0 μM IBA and 3% sucrose for 1 wk in the dark and later transferred to hormone-free medium and kept in the light. Plantlets, remaining
in the latter medium for 5–6 wk when transferred to soil, showed 90% survival. 相似文献
6.
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
7.
Summary A method of plant regeneration from hypocotyl segments of Platanus acerifolia Willd, has been developed. Hypocotyl slices were cultured on Murashige and Skoog (MS) basal medium supplemented with a range
of combinations of cytokinins [6-benzyladenine (BA) or kinetin] and auxins [indole-3-butyric acid (IBA), indole-3-acetic acid,
α-naphthaleneacetic acid or 2,4-dichlorophenoxyacetic acid] for adventitious shoot induetion. The highest regeneration frequency
was obtained with MS medium containing 2.0 mg l−1 (8.88 μM) BA and 0.5 mg l−1 (2.46 μM) IBA. Adventitious buds and shoots were differentiated from hypocotyl-derived cellus or directly from the wounded sites within
4–8 wk. The regenerated shoots were elongated and proliferated efficiently on multiplication medium. Complete plantlets were
transplanted to the soil and grew normally in the greenhouse after root formation on rooting medium for 4–6 wk. 相似文献
8.
Antonio J. Díaz-Rondero Lilia Alcaraz-Meléndez 《Plant Cell, Tissue and Organ Culture》1987,10(1):39-45
Experiments were performed to determine optimal conditions for callus induction and plantlet regeneration from damiana (Turnera diffusa, Willd.) leaf explants. MS media [7] and B5 media [4] were used; both of them were supplemented with different combinations of 2,4-dichlorophenoxyacetic, acid (2,4-D) and 6-benzyl-adenine (BA). In all cases the samples were incubated under continous fluorescent light, 100 mol.m-2.sec-1, at 25±2°C for 60 days. Highest callus percentage was obtained in MS media supplemented with 2,4-D 10–5 M plus BA 10-8M. Maximum percentage and multiple shoots and leaves were produced in MS media with BA 10-5M and BA 10-5M plus 2,4-D 10-7M. These explants were transferred for rooting to MS medium without plant growth regulators. Later, rooted plantlets were successfully established in soil. 相似文献
9.
A rapid and reliable micropropagation method was established for Spathoglottis plicata. Nodal and leaf explants dissected from 8-month-old pot-grown seedlings were cultured on charcoal-amended Murashige and Skoog
medium supplemented with 16 combinations of α-naphthaleneacetic acid (NAA) and 6-benzylaminopurine (BA) at concentrations
of 0.54–10.74 μm. Regeneration of protocorm-like bodies (PLBs) and subsequent plantlet development were observed from 98.5% of the nodal explants.
Only 6.5% of leaf explants and occasionally some root segments (dissected from regenerated plantlets) were able to produce
PLBs and then plantlets. The optimum plant growth regulator (PGR) combination for maximal PLB regeneration was 5.37 μm NAA and 0.44 μm BA. The best combination of PGR for plantlet development was 2.69–10.74 μm NAA and 8.88 μm BA. The NAA to BA ratios for maximal PLB induction and plantlet development were 12.2 and 0.3–1.2, respectively. Regenerated
PLBs and plantlets, when cut into pieces of less than 1 mm and subcultured onto the above media, regenerated new PLBs and
plantlets in another 3 months.
Received: 20 February 1997 / Revision received: 27 May 1997 / Accepted: 16 June 1997 相似文献
10.
Pradeep K. Agarwal Rajinder S. Ranu 《In vitro cellular & developmental biology. Plant》2000,36(5):392-397
Summary The in vitro plant regeneration potential of vegetatively propagated geraniums (Pelargonium x hortorum) has been investigated. Using various combinations of growth regulators and a choice of different explants, a regeneration
protocol has been developed to raise in vitro plantlets from young petiole and leaf explants from three different cultivars of geraniums. In all three cultivars, very
young petiole explants exhibited a higher regeneration potential as compared with leaf explants. Regeneration efficiencies
were found to be highly dependent on the cultivar, with cv. Samba showing the highest regeneration potential, followed by
cvs. Yours Truly and then Sincerity. Samba also showed the highest number of shoots from both the petiole [57 shoot buds per
petiole explant in the presence of 3 μM zeatin and 1 μM indole-3-acetic acid (IAA) and leaf explants (43 shoots per leaf explant with 10 μM zeatin and 2 μM IAA). Shoot buds transferred to Murashige and Skoog (MS) medium supplemented with 0.44 μM N6-benzyladenine and 0.11 μM IAA grew vigorously and attained 1–2 cm in length in 3–4 wk. These shoots rooted with 100% efficiency on MS basal medium,
and plants developed that showed normal growth and flowering under greenhouse conditions. 相似文献
11.
Kulkarni Anjali A. Thengane S.R. Krishnamurthy K.V. 《Plant Cell, Tissue and Organ Culture》2000,62(3):203-209
In vitro studies were initiated with Withania somnifera (L.) Dun. for rapid micropropagation of selected chemotypes using nodes, internodes, hypocotyls and embryo explants. Direct
regeneration of shoot buds was observed in MS basal medium supplemented with various concentrations of either benzyladenine
(BA) or thidiazouron (TDZ) depending on the explant. Nodal explants formed multiple shoots both from pre-existing and de novo buds on Murashige and Skoog's medium (MS) containing 0.1–5.0 mg l−1 BA and a ring of de novo shoot buds on MS medium containing 0.2 and 0.3 mg l−1 TDZ. Internodal explants formed shoot buds on MS with 1.0 and 5.0 mg l−1 BA while the hypocotyl explants gave rise to multiple shoots only on MS with 0.5 mg l−1 BA. Isolated embryos gave rise to many shoot buds on MS with 0.2 and 0.3 mg l−1 TDZ. The shoot buds elongated and rooted either on MS medium with 0.01 mg l−1 BA or on half strength MS medium lacking growth regulators, which depended upon the growth regulator used in the shoot bud
induction medium. Except for the embryo-derived plantlets, all other plantlets could be acclimatized with 100% success.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
12.
Summary Procedures for callus induction and subsequent organogenesis in the aquatic plant, water chestnut (Trapa japonica Flerov), were established. Phenolics exuded from explants at the callus-induction stage adversely affect callus growth. For
cotyledonary node-derived callus cultured in Murashige and Skoog (MS) medium (full, half or quarter strength) containing 2,4-dichlorophenoxyacetic
acid (2,4-D) alone or in combination with benzyladenine (BA), the accumulation of phenolics was reduced and callus induction
increased by the addition of 10.8 μM phloroglucinol (PG) to the medium. Ascorbic acid was also effective in reducing phenolic accumulation, but less effective
for callus induction than PG. Half-strength MS medium supplemented with 2.7 μM 2,4-D, 108.0 μM casein hydrolyzate, and 10.8 μM PG supported maximum callus induction. Plant organogenesis was increased by addition of vitamins (0.27 μM biotin and 2.7 μM folic acid) to half-strength MS medium supplemented with 0.27 μM BA. Many shoots developed from the regenerated nodal shoot explants in liquid half-strength MS salts medium supplemented
with 1.08 μM BA and 0.27 μM naphthaleneacetic acid. Individual shoots were excised and cultured in liquid half-strength MS medium supplemented with 5.4
μM IBA and rooted plantlets (108) were transferred and acclimatized in plastic pots. After 3 wk, the plantlets were transplanted
in a water chestnut field and the survival rate was 100%. 相似文献
13.
Plantlet regeneration through shoot formation from young leaf explant-derived callus of Camptotheca acuminata is described. Calli were obtained by placing leaf explants on Woody plant medium (WPM) supplemented with various concentrations
of 6-benzyladenine (BA) and naphthaleneacetic acid (NAA) or 2,4-dichlorophenoxyacetic acid (2,4-D). Callus induction was observed
in all media evaluated. On the shoot induction medium, the callus induced on the WPM medium containing 19.8 μM BA and 5.8
μM NAA was the most effective, providing high shoot regeneration frequency (70.3 %) as well as the highest number of shoots
(11.2 shoots explant−1). The good rooting percentage and root quality (98 %, 5.9 roots shoot−1) were achieved on WPM medium supplemented with 9.6 μM indole-3-butyric acid (IBA). 96 % of the in vitro rooted plantlets with well developed shoots and roots survived transfer to soil. 相似文献
14.
A three-stage procedure for embryogenesis in Trachyspermum ammi was developed from cotyledon and cotyledonary node explants cultured in Murashige and Skoog (MS) liquid medium supplemented
with 0.2 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D). Globular somatic embryos without intervening callus phase developed in 4 wk. The
development of embryos to heart and torpedo stages required second-stage subculture of the explants (along with developing
embryos) in liquid medium with lower concentrations of 2,4-D. Further development of embryos required a third-stage subculture
in hormone-free liquid medium supplemented with 100 mg l−1 casein hydrolysate. Regeneration of complete plantlets occurred after the fully developed somatic embryos were transferred
to solidified half-strength MS medium supplemented with 1 mg l−1 gibberellic acid. 相似文献
15.
So-Young Park Hosakatte N. Murthy Kee-Yoeup Paek 《In vitro cellular & developmental biology. Plant》2002,38(2):168-172
Summary An efficient and rapid in vitro method was developed for regeneration of Phalaenopsis using leaf segments derived in vitro from flower stalk nodes. Leaf segments of four cultivars Tinny Sunshine ‘Annie’, ‘Taisuco Hatarot’, Teipei Gold ‘Golden Star’,
Tinny Galaxy ‘Annie’ cultured on Murashige and Skoog medium supplemented with N
6-benzyladenine (BA; 88,8 μM) and α-naphthaleneacetic acid (NAA; 5,4 μM) produced an average of 10–13 protocorm-like bodies (PLBs) after 12 wk. PLB proliferation was achieved on a modified Hyponex
medium (1 gl−1 6.5N−4.5P−19K+20N−20P−20K+2gl−1 peplone +3% (w/v) potato homogenate +0.05% activated 1 gl−1 charcoal) and an optimal number of 13–18 PLBs developed from single PLB sections of different cultivars. Plantlet development
was also achieved on a modified Hyponex medium. By repeated subculture of PLBs on a proliferation medium, and culturing them
in the plantlet regeneration medium, plantlets could be produced continuously. Approximately 6 mo, were required from the
initiation of culture to the production of plantlets for transplant to community pots. 相似文献
16.
María Laura Vidoz Pablo Klusacek Hebe Yolanda Rey Luis Amado Mroginski 《Plant Cell, Tissue and Organ Culture》2006,86(1):111-115
In vitro protocols for plant regeneration of Arachis correntina through both somatic embryogenesis and organogenesis were developed using immature leaves as explants. Morphologically normal somatic embryos were obtained on culture media composed of 20.70 or 41.41 μM picloram (PIC) with the addition of 0.044 μM 6-benzylaminopurine (BA), resulting in a 33 and 24% of conversion into plants, respectively. The source of explants and the developmental stage of the leaves had a marked effect on somatic embryogenesis. The second folded immature leaves from in vitro growing plants were the most responsive producing up to 30% embryogenesis in MS+41.41 μM PIC. Embryos converted into plants after transfer to MS medium devoid of growth regulators and these plants were successfully acclimatised. Adventitious shoots were obtained on culture media supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) or naphthaleneacetic acid (NAA) with or without 0.044 μM BA, achieving plant regeneration in the induction media. The highest percentage of bud formation was obtained on culture medium composed of␣MS+10.74 μM NAA+0.044 μM BA (12.5%). Roots were formed on all culture media tested. Regenerated plants were transferred to pots and grew well under greenhouse conditions. 相似文献
17.
Summary A method for adventitious shoot induction from petiole explants of Heracleum candicans is reported. Shoot buds were induced on Murashige and Skoog (MS) medium with 4.4μM 6-benzylaminopurine (BA) and 1.1 μM 2,4-dichlorophen-oxyacetic acid (2,4-D). A wound response in the presence of BA and 2,4-D at the time of culture was necessary
for inducing shoot buds. The shoot bud regeneration was significantly influenced by size, type and orientation of explants
on the culture medium. These shoot buds developed into 4–5 cm shoots upon transfer to a medium containing 1.1μM BA and 0.5 μM α-naphthaleneacetic acid (NAA). The regenerated shoots formed rooted plantlets on MS medium supplemented with 4.9 μM indole-3-butyric acid (IBA). About 15 plants were established in the field for further evaluation. 相似文献
18.
Embryogenic callus in Catharanthus roseus was initiated from hypocotyl on Murashige and Skoog’s (MS) medium supplemented with 1.0–2.0 mg dm−3 of 2,4-dichlorophenoxyacetic acid (2,4-D) or chlorophenoxyacetic acid (CPA). Calli from other sources were non-embryogenic.
Numerous somatic embryos were induced from primary callus on MS medium suplemented with naphthalene acetic acid (NAA) within
two weeks of culture. Embryo proliferation was much faster on medium supplemented with 6-benzylaminopurine (BAP). After transfer
to medium with gibberellic acid (GA3, 1.0 mg dm− 3) mature green embryos were developed and germinated well into plantlets on MS liquid medium supplemented with 0.5 mg dm−3 BAP. Later, embryos with cotyledonary leaves were subjected to different auxins treatments for the development of roots.
Before transfer ex vitro, plantlets were cultivated on half strength MS medium containing 3 % sucrose and 0.5 mg dm−3 BAP for additional 2 weeks. Additionally, the effect of liquid medium has been evaluated at different morphogenetic stages. 相似文献
19.
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
20.
The organogenic competence of leaf explants of eleven Carthamus species including C. tinctorius on Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ) + α-naphthaleneacetic
acid (NAA) and 6-benzyladenine (BA) + NAA was investigated. Highly prolific adventitious shoot regeneration was observed in
C. tinctorius and C. arborescens on both growth regulator combinations and the shoot regeneration frequency was higher on medium supplemented with TDZ + NAA.
Nodal culture of nine Carthamus species on media supplemented with BA and kinetin (KIN) individually revealed the superiority of media supplemented with
BA over that of KIN in facilitating a higher shoot proliferation index. Proliferating shoots from axillary buds and leaf explants
were transferred to medium supplemented with 1.0 mg dm−3 KIN or 0.5 mg dm−3 BA for shoot elongation. Elongated shoots were rooted on half-strength MS medium supplemented with 1.0 mg dm−3 each of indole-butyric acid (IBA) and phloroglucinol. The plantlets thus obtained were hardened and transferred to soil. 相似文献