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1.
This work describes in vitro morphogenesis and plantlet regeneration from seeds of the naturally polyembryonic tree Syzgium
alternifolium. The basal medium (BM) comprised half strength Murashige and Skoog's (MS) salts, B5 vitamins, 2 mg dm-3 glycine, 250 mg dm-3 ascorbic acid and 20 g dm-3 sucrose. Addition of auxins like indole-3-acetic acid (IAA), 1-naphthalene acetic acid or 2,4-dichlorophenoxyacetic acid
to the basal medium induced formation of roots or callus from the dicotyledonous as well as tricotyledonous seeds. In contrast,
cytokinins like N6-benzyladenine (BA), kinetin, 2-isopentenyl adenine, thidiazuron alone or a combination of BA and auxins induced development
of adventitious shoots. The medium containing 3.0 mg dm-3 BA and 0.5 mg dm-3 IAA induced the highest number of adventitious shoots (32 – 33) from dicotyledonous seed with an average length of 4.1 cm
within 6 weeks of incubation. Rooting of 80 % adventitious shoots was achieved by dipping the shoots in 100 mg dm-3 IAA for 15 min. About 70 % of the rooted shoots were successfully established in pots after hardening.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
2.
. In vitro-grown shoot tips excised from preconditioned stock shoots of 'Troyer' citrange were successfully cryopreserved by encapsulation-dehydration. Optimal survival of cryopreserved shoot tips was achieved when encapsulated shoot tips were dehydrated to 17.1% water content. The sucrose concentration in the preconditioning medium significantly influenced the growth and dry matter percentage of the stock shoots as well as subsequent survival of the cryopreserved shoot tips. Maximal growth of stock shoots was obtained in sucrose concentrations in the range of 0.15 M to 0.29 M, while the dry matter percentage increased as sucrose concentration increased up to 0.44 M. The survival of cryopreserved shoot tips increased from 40% to approximately 80% as the sucrose concentration for stock shoots increased from 0.09 M to 0.22 M or 0.29 M. The benzyladenine concentration in the post-culture medium significantly affected the survival and regrowth of the cryopreserved shoot tips. Survival of the shoot tips was lowest when they were post-cultured on benzyladenine-free medium. However, high benzyladenine concentrations (3-4 µM) induced callus formation. Optimal recovery was obtained in post-culture medium containing 2 µM benzyladenine and 0.05 µM !-naphthalene acetic acid. The extraction of shoot tips from alginate beads greatly improved the regrowth of cryopreserved shoot tips. 相似文献
3.
The addition of 40 mM putrescine (Put) to Murashige and Skoog's (MS) medium resulted in increased shoot multiplication and shoot growth in untransformed plants relative to transformed plants of Cichorium intybus L. Put at a concentration of 40 mM also resulted in flowering in both systems on the 28th day, with elevated titers of endogenous conjugated Put and spermine (Spm) in both untransformed and transformed plants. The addition of 40 µM AgNO3 to untransformed axillary buds of C. intybus L. cultured on MS media resulted in increased shoot multiplication (36.9DŽ.63 shoots per culture) and increased shoot growth (7.82ǂ.76 cm) as compared to transformed ones (11.6ǂ.89 shoots per culture; 3.20ǂ.24 cm). Moreover, cultures treated with 40 µM AgNO3 showed in vitro flowering on the 28th day in both systems, with the endogenous levels of conjugated spermine being higher in untransformed plants than in transformed ones. The morphogenetic response and the endogenous conjugated pool of polyamines were lower following !-DL-difluromethylarginine and !-DL-difluromethylornithine treatments; the addition of put (40 mM) and AgNO3 (40 µM) restored these to normal levels. Under exogenous put feeding, ethylene production was lower in both the untransformed and transformed cultures. We believe that an interplay between polyamine and ethylene biosynthesis is involved in regulating the morphogenetic response in both transformed and untransformed shoots of C. intybus. The response to AgNO3 and Put treatment was not altered by the transformation process. 相似文献
4.
Parthasarathi Bhattacharya Satyahari Dey Nilanjana Das Bimal Chandra Bhattacharyya 《Plant cell reports》1990,9(8):439-442
A procedure for rapid multiplication of Chrysanthemum morifolium RAMAT cv. Birbal Sahni using leaf callus and stem (nodal/internodal) callus as well as node and apical shoots has been developed. Murashige and Skoog's medium (1962) supplemented with 2mg/1 2,4-D yielded good green calli from both leaf and stem segments within 2 weeks. About 1 cm × 1 cm callus regenerated 2–3 shoots after 3 weeks on MS solid medium supplemented with 0.1 mg/l IAA and 0.2 mg/l BAP. Each of the regenerated shoots when transferred to the same shooting medium without agar yielded about 150 new shoots, which in turn regenerated roots after another week in MS half strength or modified White's media (Rangaswamy, 1961). It has been estimated that about 1014 plantlets could be produced in a year from one expiant following the proposed protocol.Abbreviations BAP
6-benzylaminopurine
- IAA
indole-3-acetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- MS
Murashige and Skoog's (1962) medium 相似文献
5.
A liquid culture protocol was developed to regenerate shoots from cotyledons of germinating seeds of jute (Corchorus capsularis L.). Reproducibility of the protocol was tested in three genotypes of jute. Highest bud differentiation rates into normal
shoots (via shoot-forming explants) were obtained on modified Murashige and Skoog's liquid medium containing 2.7 μM α-naphthaleneacetic acid and 4.4 μM benzylaminopurine. Regenerated shoots were excised, and the best root formation could be induced in medium with 2.5 μM indole-3-butyric acid and 1.5% sucrose. Bud primordia were formed directly on the cut surface of the cotyledons. Scanning
electron micrographs and histological studies confirmed the organogenic nature of the regenerated shoots. The physical condition
of the culture medium and the age of the explants played crucial roles in the induction of shoot development using shoots;
2-day-old explants being optimal. Approximately 70% of the shoots were successfully established in soil after hardening.
Received: 20 October 1997 / Revision received: 4 October 1998 / Accepted: 27 October 1998 相似文献
6.
A successful procedure was established for in vitro mass multiplication of Indian rosewood (Dalbergia latifolia Roxb.). In vitro regeneration of plantlets was achieved from callus of shoot tips and shoot segments of over 50-year-old elite trees on Murashige & Skoog's medium containing naphthaleneacetic acid (NAA) and benzylaminopurine (BAP). For rooting, regenerated shoots from the calli were excised and first treated with White's liquid medium or half-strength Murashige & Skoog's medium, supplemented with indole-3-acetic acid, indole-3-butyric acid and naphthaleneacetic acid for 48 h to 72 h. Following this treatment, plantlets were transferred to hormone-free half-strength MS medium. Rooted plantlets were then transferred to pots and grown in the greenhouse.Abbreviations BAP
6-benzylamino pruine
- CH
casein hydrolysate
- CM
coconut milk
- 2, 4-D
dichlorophenoxyacetic acid
- GA
gibberellic acid
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- NAA
naphthaleneacetic acid
- PVP-10
polyvinyl pyrrolidone
- YE
yeast extract 相似文献
7.
Susan H. Woods Gregory C. Phillips John E. Woods Glenn B. Collins 《Plant cell reports》1992,11(5-6):257-261
An efficient protocol has been developed for the in vitro propagation of Mexican Weeping Bamboo through somatic embryogenesis from zygotic embryo explants. Mature seeds and excised embryos were cultured in the light or in the dark on both Murashige and Skoog's and Gamborg's B5 basal media with various supplements. Optimal somatic embryogenesis and plant regeneration were obtained by culture in the dark on Murashige and Skoog's basal medium supplemented with 3 mg/1 2,4-dichlorophenoxyacetic acid, 0.5 mg/1 6-benzylaminopurine and 2.0% sucrose. More than 95% of the germinating somatic embryos developed shoots and roots, and were transferred to soil with 85% success. 相似文献
8.
Induction of single and multiple shoots was obtained from nodal expiants of 60–80 year-old elite trees of rosewood on Murashige and Skoog's basal medium supplemented with 6-benzylaminopurine (1.0 mg 1-1) and -Naphthalene acetic acid (0.05 mg 1-1) or indole acetic acid (0.5 mg 1-1). Multiplication of shoots was obtained on MS (reduced major elements) or Woody Plant Medium supplemented with 6-benzylaminopurine (1.0 mg 1-1) and kinetin (0.5–1.0 mg 1-1). Excised shoots were rooted on half-strength MS with IBA (2.0 mg 1-1) to obtain complete plantlets. The regenerated plantlets have been acclimatized and successfully transferred to the soil.Abbreviations MS
Murashige and Skoog's (1962) medium
- B5
Gamborg (1968) medium
- WPM
Woody plant medium, Lloyd and McCown (1981) medium
- NAA
-naphthalene acetic acid
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- BAP
6-benzyl aminopurine
- KIN
kinetin
- PVP
polyvinyl pyrrolidone
- CH
casein hydrolysate
- ADS
adenine sulphate
- L-Gl
L-glutamine
- L-Arg
L-arginine
- L-Asp
L-asparagine
- PG
phloroglucinol 相似文献
9.
Plant regeneration via somatic embryogenesis was achieved in callus cultures derived from nodal explants of in vitro grown seedlings and excised mature zygotic embryos of three bamboo species on Murashige and Skoog's (MS) basal medium supplemented with 0.5 mg/l kinetin (Kn), 2.0 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D), 10 mg/l adenine sulphate (Ads) and 3% (w/v) sucrose incubated in the light or in the dark. Somatic embryos germinated (95–98%) into normal plants and were transferred to soil with 95% success. In vitro flowering was induced on shoots developed from nodal explants taken from somatic embryo regenerated plants of Bambusa vulgaris, Dendrocalamus giganteus and Dendrocalamus strictus on half-strength MS basal medium supplemented with 0.25 mg/l indole-3-butyric acid (IBA), 0.5 mg/l Ads, 0.5 mg/l gibberellic acid (GA3) and 3% sucrose.Abbreviations BAP
6-benzylaminopurine
- Kn
kinetin
- Ads
adenine sulphate
- IBA
indole-3-butyric acid
- NAA
1-naphthaleneacetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- MS
Murashige and Skoog (1962) basal medium
- GA3
gibberellic acid 相似文献
10.
High frequency bud break and multiple shoots were induced in nodal explants collected between November to February from a 5 year old tree of Morus australis Poir syn. M. acidosa Griff. on Murashige and Skoog's medium supplemented with 6-benzylaminopurine (1.0 mg/1). Incorporation of gibberellic acid (0.3 mg/l) along with BAP (1.0 mg/l) not only induced faster bud break from nodal explants as well as from apical shoot buds, but it also enhanced the frequency of bud break. Nodal explants were more responsive than apical shoot buds. The shoots formed in vitro were multiplied further as nodal segments, and an average multiplication rate of 6-fold per subculture was established within 4–5 months. The shoots were successfully rooted on half-strength MS containing a combination of indole-3-acetic acid, indole-3-butyric acid and indole-3-propionic acid, each at 1.0 mg/1. The plantlets were successfully hardened off and established in natural soil.Abbreviations BAP
6-benzylaminopurine
- GA3
gibberellic acid
- KN
kinetin
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- IPA
indole-3-propionic acid
- MS
Murashige and Skoog (1962) medium
- NAA
1-naphthalene acetic acid 相似文献
11.
S. R. Thengane D. K. Kulkarni V. A. Shrikhande K. V. Krishnamurthy 《In vitro cellular & developmental biology. Plant》2001,37(2):206-210
Summary Regeneration of adventitious shoots from the medicinal plant Nothapodytes foetida (Weight) Sleumer Syn. Mappia foetida (family Ieacinaceceae) has been achieved using different seedling explants. Direct, regeneration of shoot buds was observed
in Murashige and Skoog's (MS) basal medium supplemented with various concentrations of thidiazuron. The optimum levels of
thidiazuron concentrations were 0.91–4.45 μM. Leaf explants formed more shoots followed by hypocotyls or cotyledons. The shoot buds elongated and rooted on MS basal medium
with N6-benzyladenine (0.88–2.22 μM) and indole-3-butyric acid (0.49 μM). 相似文献
12.
Summary Production of microspore-derived embryos from cultured anthers is now a well established technique for the isolation of homozygous lines in many crop plants. We describe here a culture method for embryo induction and plant regeneration from anthers of four sunflower genotypes. For preliminary experiments, anthers of uninucleate microspores were cultured on four types of basal media viz., Murashige and Skoog's MS, Gamborg's B5, Nitsch and Nitsch, and White's W, supplemented with 1.0 mg/l 2,4 dichlorophenoxy acetic acid and 0.5 mg/l 6-benzylaminopurine and 40 g/l sucrose. MS basal medium, being more responsive for embryo induction, was used for further experimentation. To optimise the culture requirement MS basal medium was supplemented with 0.2–2.0 mg/l 2,4 dichlorophenoxy acetic acid and 0.5 and 1.0 mg/l 6-benzylaminopurine. The effect of cold pretreatment, hormone regime and sucrose concentration were tested for embryogenic efficiency. Genotype had a significant effect on the capacity of embryo induction. Addition of silver nitrate (2.5 mg/l), an ethylene inhibitor, stimulated embryo germination. Plantlets were obtained (10–15%) from embryos of only one genotype.Abbreviations 2,4-D
2,4 dichlorophenoxy acetic acid
- NAA
-naphthalene acetic acid
- IAA
indole-3-aceticacid
- BAP
6-benzylaminopurine
- KN
Kinetin
- ABA
abscisic acid
- GA3
gibberellic acid 相似文献
13.
In vitro flowering of bitter melon 总被引:4,自引:0,他引:4
Flowers were formed from shoot tips of bitter melon (Momordica charantia L.) cultured on Murashige and Skoog medium supplemented with 90 mM sucrose, 0.05 mM Fe2+ and 4 µM N6-benzyladenine (BA). The addition of 0.05 mM Fe2+ to the medium prevented chlorosis of the explant and promoted normal flowering. Increasing the ratio of carbon to nitrogen promoted male flower formation but intensively inhibited vegetative growth. The influence of cytokinin on the morphogenesis of the explant was highly notable. Flowers could be formed after a 15- to 20-day exposure to kinetin (Kin) or BA. Kin and BA had opposite effects with regard to the development of the explant. Kin promoted flower formation, especially female, but inhibited branch bud formation. Conversely, BA promoted branch bud formation and also promoted male flower formation when present at a concentration of 1-2 µM, but completely inhibited flower formation at 4-8 µM. Fluorescein diacetate staining and in vitro germination showed that in vitro pollen were of a fairly high viability. 相似文献
14.
We describe an efficient process for the regeneration of Lonicera tatarica plants from cultured stem sections. Induction of multiple shoots was achieved directly from cultured stem cuttings. The highest regeneration rate was achieved on Gamborg's B5 medium supplemented with 4% sucrose and 0.8% Difco bacto-agar in the absence of hormones. Differentiated shoots were elongated for 5-7 days on induction medium supplemented with 0.5 mg/l GA3. Shoot induction and elongation experiments were carried out using original stem explants from either 2-, 6-, or 18-month-old donor plants. The age of the donor plant had no noticeable effect on either process. However, rooting of elongated shoots occurred only with shoots derived from 2-month-old donor plants. Rooting efficiency and proliferation were highest on half-strength WPM medium supplemented with 2 µM indole-3-butyric acid and 0.6% Keylis agar. The plants regenerated from stem explants were morphologically normal, and levels of loganin and secologanin were comparable to those detected in plants grown from seed and maintained through vegetative propagation. 相似文献
15.
Prevalek-Kozlina Branka Kostović-Vranjes Vesna Slade Danko 《Plant Cell, Tissue and Organ Culture》1997,51(2):141-143
A rapid clonal propagation method for Fibigia triquetra (DC.) Boiss. (Brassicaceae), a rare Croatian stenoendemic species,
has been developed. Shoots originated from aseptically germinated seeds were used for culture initiation. The highest multiplication
rate of 9.2 shoots per explant was achieved in a 4-weeks-culture period in the third subculture on half strength Murashige
and Skoog medium supplemented with 0.5 μM 6-benzylaminopurine and 2.9 μM gibberellic acid. Excised shoots were rooted best
with addition of 8.6 μM indole-3-butyric acid on the same basal medium. Rooted plantlets were successfully transferred to
potting soil and acclimatized to outdoor conditions.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
16.
Methyl laurate and 6-benzyladenine promote the germination of somatic embryos of a hybrid rose 总被引:3,自引:0,他引:3
Globular stage somatic embryos were induced in callus cultures of Rosa Heritage 2 Alister Stella Gray on medium containing 13.5 µM 2,4-dichlorophenoxyacetic acid (2,4-D) and developed to the cotyledonary stage on medium containing 9 µM 2,4-D. Cotyledonary-stage embryos were transferred to germination media with or without 1.5 µM 6-benzyladenine (BA) and with or without 44 µM methyl laurate (Mela). BA and Mela both promoted the development of shoots and roots and increased the frequency of bipolar germinations. An average of 56.5% (SEdž.1%) embryos on medium containing both BA and Mela underwent bipolar germinations compared with less than 20% in treatments where either or both were excluded. The effectiveness of BA and Mela was reduced if Mela was included in the development medium or if the concentration of salts and vitamins in the germination media was sub-optimal. There was evidence that growth at one pole of the somatic embryo promoted development at the other. 相似文献
17.
High efficiency shoot regeneration was achieved through leaflet and cotyledon derived calli in Cassia angustifolia - an important medicinal plant. Dark brown compact callus was induced at the cut ends of the explants on Murashige and Skoog's
(MS) medium augmented with 1 μM N6-benzyladenine (BA) + 1 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Such callus pieces on transfer to cytokinins (BA or kinetin)
supplemented medium differentiated shoots within 10 – 15 d. Of the two cytokinins, 5 μM BA was optimum for eliciting morphogenic
response in 83.33 and 70.83 % cultures with an average of 4.16 ± 0.47 and 3.70 ± 0.56 shoots in cotyledon and leaflet derived
calli, respectively. The addition of 0.5 μM α-naphthaleneacetic acid (NAA) to MS + 5 μM BA further elevated the maximum average
number of shoots to 12.08 ± 1.04 and 5.37 ± 0.52 for cotyledon and leaflet calli, respectively. The excised shoots were transferred
to a rooting medium containing either IAA (indole-3-acetic acid), IBA (indole-3-butyric acid) or NAA. Nearly 95 % shoots developed
an average of 5.4 ± 0.41 roots on half strength MS medium supplemented with 10 μM IBA. 相似文献
18.
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
19.
A successful procedure was established for in vitro plant regeneration from callus derived from stem and leaf explants of Centella asiatica on semisolid modified Murashige and Skoog's [7] medium supplemented with 2.0 mg L3 kinetin and 4.0 mg L3 a-naphthaleneacetic acid. The rate of shoot-bud regeneration was the highest (42.8 and 54.3 shoots/culture in stem and leaf derived callus respectively) after 4 weeks of subculture on 4.0 mg L3 6-benzyladenine, 2.0 mg L3 Kn, 0.25 mg L3 a-naphthaleneacetic acid and 20 mg L3 adenine sulfate. Differentiated shoots rooted within 11 days in 1/2 strength MS basal salts supplemented with 0.5 mg L3 indole-3-acetic acid and 2% (w/v) sucrose. About 85% of rooted plantlets were acclimatized and transferred to the greenhouse. 相似文献
20.
Nand Lal Paramvir Singh Ahuja Arun Kumar Kukreja Banibrata Pandey 《Plant cell reports》1988,7(3):202-205
A procedure has been developed for the clonal propagation of Picrorhiza kurroa Royle ex Benth. through shoot tip culture. Murashige and Skoog's medium (1962) supplemented with kinetin (3.0 to 5.0 mg/l) supported rapid proliferation of multiple shoots from the explants. Addition of indole-3-acetic acid (1.0 mg/l) to the kinetin containing medium showed marked improvement in the growth of regenerated shoots. However, presence of IAA in the medium did not alter the frequency of shoot multiplication. Rooting was readily achieved upon transferring shoots onto MS medium containing -naphthaleneacetic acid (1.0 mg/l). Plantlets were successfully transferred to soil.Abbreviations BAP
6-benzylaminopurine
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- Kn
Kinetin
- MS
Murashige and Skoog's (1962) medium
- NAA
-naphthaleneacetic acid 相似文献