共查询到20条相似文献,搜索用时 62 毫秒
1.
Protocols for in vitro plant multiplication from somatic tissues and production of artificial seeds through encapsulation of nodes were developed for Aristolochia tagala Cham., a rare and valuable medicinal plant, as a measure of conservation and as a prerequisite for genetic transformation procedure. A maximum number of adventitious shoots were regenerated from leaf-derived callus on Murashige and Skoog (MS) medium containing 6-benzylaminopurine (BAP; 2 μM), α-naphthaleneacetic acid (NAA; 0.5 μM), and phloroglucinol (PG; 10μM). Nodes collected from in vitro established shoot cultures were encapsulated in 3 % (m/v) sodium alginate and 1 % (m/v) calcium chloride. Multiple shoots were successfully regenerated from the encapsulated nodes cultured on MS medium supplemented with 3 μM BAP and 0.5 μM kinetin (KIN). Regenerated shoots from callus and artificial seeds were successfully rooted and acclimated to greenhouse conditions. Since roots of A. tagala are primarily used in traditional medicine, a protocol for regenerating roots directly from the leaf derived callus was also developed. Maximum root length was obtained when the callus was cultured in MS medium supplemented with KIN (1 μM), indole acetic acid (IAA; 0.5 μM), NAA (0.1 μM), and PG (10 μM). Biochemical parameters were studied in calli grown with and without PG in the medium to establish a correlation between these parameters and shoot morphogenesis. An increment of antioxidant enzymes (peroxidase and catalase) and metabolites (sugars and proteins), and a decrease in the amount of polyphenol oxidase was observed in the calli which were grown in the presence of PG. 相似文献
2.
C. Kiranmai V. Aruna S. Karuppusamy T. Pullaiah 《Journal of plant biochemistry and biotechnology.》2008,17(1):99-101
Callus cultures were established from seedling explants of Pergularia daemia (Forsk) Chiov on Murashige and Skoog (MS) medium supplemented with different concentrations of auxins. Optimal callus developed from leaf explants on MS medium supplemented with 2,4-D (2 mg l?1) + 2iP (0.1 mg l?1), was used for morphogenesis. Adventitious shoots were regenerated (70%) from the calli on MS medium supplemented with NAA (0.1 mg l?1)+ BAP (2 mg l?1). Individual shoots were rooted on half strength MS medium supplemented with 0.1 mg l?1 IBA. Plantlets with well developed roots were successfully transferred to soil and 50% of the transferred plants survived. 相似文献
3.
Compact, friable and embryogenic calli were initiated from immature inflorescences and young leaf bases of one week old seedlings of Paspalum scrobiculatum cultured on MS medium supplemented with 2,4-D. A stable, embryogenic suspension culture was initiated from these calli and maintained in a liquid version of the same MS medium. Embryogenic calli and somatic embryos were obtained by plating suspension culture cells onto semi-solid medium containing 2,4-D. Complete, normal plantlets developed on 2,4-D free medium at a high frequency from somatic embryos. NAA and BAP in the medium promoted plant development.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- NAA
1-naphthaleneacetic acid
- BAP
6-benzylaminopurine
- ABA
Abscisic acid
- MS
Murashige and Skoog (1962)
- CM
Coconut milk 相似文献
4.
Present report is the first direct evidence of azadirachtin production in androgenic haploid cultures of Azadirachta indica, a woody medicinal tree. Anther cultures at early-late-uninucleate stage of microspores were established on MS medium with BAP (5 µM), 2,4-D (1 µM) and NAA (1 µM) containing 12% sucrose. The calli, induced, were further multiplied on 2,4-D and Kinetin media. Shoots, differentiated on BAP (2.2 µM) + NAA (0.05 µM) medium, were elongated on MS + BAP (0.5 µM) and multiplied on MS + BAP (1 µM) + CH (250 mg/l). Thereafter, the shoots were rooted on ¼ MS + IBA (0.5 µM). Cytological analysis of the calli and regenerants have confirmed their haploid status with the chromosome number as 2n = x = 12. The haploid cell lines and leaves from in vitro grown plantlets were analyzed for azadirachtin by RP-HPLC and mass spectroscopy. Maximum azadirachtin (728.41 µg/g DW) was detected in calli supporting best shoot proliferation while least (49 µg/g DW) was observed in an undifferentiated line from maintenance medium. This study has brought us a step closer to develop genetically pure lines that could serve as new and attractive alternative ways of homogeneous controlled production of high value compounds, round the year, independent of geographical and climatic barrier.Key words: anther culture, Azadirachta indica, azadirachtin, haploid plants, RP-HPLC 相似文献
5.
N. Bhagya K. R. Chandrashekar Anitha Karun U. Bhavyashree 《Journal of plant biochemistry and biotechnology.》2013,22(4):474-482
A protocol for the regeneration of a large number of plantlets via indirect shoot organogenesis and somatic embryogenesis has been developed from the stem and leaf explants of Justicia gendarussa Burm. f. The callus was efficiently induced from the explants using Murashige and Skoog (MS) medium supplemented with α-Naphthalene acetic acid (NAA) + Benzyl amino purine (BAP) (1.0?+?0.1 mg/l). The highest number of plantlets through indirect shoot organogenesis was obtained when the callus was subcultured to MS medium with BAP + NAA (0.1?+?1.0 mg/l). The maximum number of plantlets via somatic embryos was obtained in the medium with BAP + NAA (1.0?+?0.1 mg/l) for stem derived calli and Kinetin (Kn) + NAA (2.0?+?0.1 mg/l) for leaf derived calli. The in vitro developed shoots were rooted well in half strength MS medium supplemented with 0.5 mg/l of Indole-3-acetic acid (IAA). The in vitro regenerated plantlets were hardened using a mixture of sterile sand:soil:manure (1:1:1). The present study is the first report on the regeneration of plants through somatic embryogenesis from stem and leaf derived calli of J. gendarussa. 相似文献
6.
Androgenic haploids of the neem tree (Azadirachta indica A. Juss.) were produced by anther culture at the early- to late-uninucleate stage of pollen. Haploid formation occurred via callusing. The best medium for inducing callusing in the anther cultures was Murashige and Skoog's basal medium (MS) (9% sucrose) supplemented with 1 microM 2,4-D, 1 microM NAA and 5 microM BAP, while anther callus multiplied best on MS medium supplemented with 1 microM 2,4-D and 10 microM Kn. These calli differentiated shoots when transferred to a medium containing BAP; 5 microM BAP was optimum for young calli (75% cultures differentiated shoots), but older calli showed the best regeneration with 7.5 microM BAP. Shoots elongated at a lower concentration of BAP-0.5 microM. These shoots were multiplied by forced axillary branching and rooted in vitro. The plants were subsequently established in soil. Of the plants that regenerated from anther callus 60% were haploid, 20% were diploid and 20% were aneuploid. 相似文献
7.
Anita R. Sehrawat Sanjogta Uppal J. B. Chowdhury 《Journal of plant biochemistry and biotechnology.》2002,11(2):105-108
Different explants of Rauwolfia serpentina were tested for their capacity and root differentiation. MS sedium containing 2.0 mg I-1 BAP or 1.5 mg I-1 BAP with 0.5 mg I-* NAA gave the best shoot proliferation from shoot apices (84.12%). Multiple shoots subcultured on the same media gave higher number of shoots (6–9). Callus formed at the cut ends of explants produced shoots when subcultured on the above media. Rooting was achieved after transfer of shoots either to hormone free or half strength or full strength MS medium with different concentrations of IAA (0.5,1.0 mg I-1 and IBA (0.5,1.0 mg I-1).The complete regenerated plantlets have been successfully transferred to earthen pots in green house. Maximum root differentiation from leaves, stem and nodal cuttings derived calli was obtained on MS medium supplemented with NAA (2.0 mg I-1) and BAP (1.5 mg I-1). Identification of reserpine and other alkaloids from differentiated roots holds an interesting alternative for controlled production of alkaloids from this endangered, overexploited medicinal plant species. 相似文献
8.
爪哇三七组织培养植株再生 总被引:1,自引:0,他引:1
本文报道了爪哇三七通过组织培养再生植株的方法。爪哇三七的叶片,叶柄或茎段外植体分别接种于6种分化培养基上,均难以直接分化。在附加1mgL~(12),4-D和0.1mgL~(-1)KT的B_5培养基上的叶片形成无色疏松的愈伤组织转接于MS附加1mgL~(-1)BAP和0.1mg L~(-1)NAA培养基上,5天后开始形成绿色球胚状结构,继而形成不定芽或丛生芽。这些不定芽或丛生芽在锈根培养基中迅速长成根系发达的完整植株。此外,本试验比较了不同浓度及不同组合的激素对爪哇三七外植体的脱分化、再分化的影响。并讨论了此项工作对柑桔裂皮病类病毒(简称CEV)的复制及致病机理研究的意义。 相似文献
9.
Liangtao Tian Yanni Ke Shirui Gan Yongqin Chen Yong Chen Zhifan Yang Xiaoguang Wang 《Plant Growth Regulation》2012,68(2):319-324
Sapium sebiferum is a potential bioenergy plant that can be cultivated under various soil, water and climate conditions for both oil-rich seeds and woody biomass. An efficient protocol for regenerating triploid plants of S. sebiferum was established using mature endosperms as explants. Green and compact calli were induced from endosperms within 30 days on Murashige and Skoog medium (MS) containing 1.0–5.0 mg/l 6-benzylaminopurine (BAP) or in combination with 0.2 mg/l α-naphthalene acetic acid (NAA). Within 45 days after endosperm-derived calli were cut into pieces and cultured on media supplemented with 1.0–2.0 mg/l BAP alone or plus 0.1 mg/l NAA, more than 60 % of the callus explants initiated adventitious buds. The buds elongated into shoots after transfer onto a MS medium containing 0.1 mg/l BAP and 1.0 g/l activated charcoal. Approximate 80 % of shoots rooted on a MS medium amended with 1.0 g/l activated charcoal and 1.0 or 2.0 mg/l indole-3-butyric acid within 30 days. The triploidy of the endosperm-derived plantlets was confirmed by flow cytometric analysis, and the triploid plants grew normally after transplantation. 相似文献
10.
Hypocotyls of Albizia odoratissima cultured on shoot induction medium (MS medium with 7.5 μM BAP and 0.5 μM NAA) showed adventitious shoot organogenesis under
light with 16 h photoperiod. Similar cultures under total darkness produced non-morphogenic calli. The changes in the specific
peroxidase and catalase activity, total protein content and acidic isoperoxidase pattern were compared between the culture
showing shoot organogenesis and culture producing non-morphogenic calli. It was found that in vitro shoot bud differentiation
is accompanied by an increase of the specific activities of peroxidase and catalase in culture kept under light. In parallel
with the above changes the total protein content reached to the maximum level and also a new isoperoxidase (P10) expressed
on the 21st day in cultures kept under light. Conversely, culture producing non-morphogenic calli underwent a reverse change
in specific peroxidase activity. This change in antioxidant enzyme activities corresponds to the histological observation
of shoot bud differentiation in cultures kept under light. 相似文献
11.
To maintain embryogenic cell lines ofPimpinella brachycarpa, we suspension-cultured friable and rapidly growing yellowish calli in an MS liquid medium containing 0.2 ~ 2,4-D and 0.5pM BAP. Efficient somatic embryogenesis was achieved when selected cells were then transferred to an MS medium (0.2% gelrite)
that contained 0.2gM 2,4-D, 0.5 uM BAP, and 10.0 laM TDZ (thidiazuron). These cells were cultured at 27°C under continuous illumination (21.5
I~E m-2 s-l). Embryogenic calli expanded about four-fold, and developed into pale yellow calli. Somatic embryogenesis was initiated only
from glossy and nodular-type calli. After two more weeks of culture, globular embryos appeared on the surface of calli grown
in the MS medium that contained 10.0 /aM TDZ only, or in combination with 0.5 gM NAA. Experimenting with 2,4-D, an auxin,
to promote embryogenic calli resulted in excessive browning and death. We overcame this problem by growing glossy embryogenic
and nodular calli on media that contained 10.0 gM TDZ. Calli that were not treated with TDZ turned dark brown and were not
viable. Up to 74% of the calli showed somatic embryos when the medium was supplemented with 10.0 uM TDZ and 0.5 uM NAA. Embryos
from these TDZ-induced, somatic embryogenic calli grew efficiently, forming multiple shoots and developing into normal plants.
Therefore, efficient differentiation of suspension-cultured cell clusters into embryogenic calli, along with treatment of
subsequent somatic embryos by TDZ, suggests that TDZ probably helps in establishing the optimum cytokinin-auxin ratio required
for induction and expression of somatic embryogenesis. 相似文献
12.
P. B. Kirti S. R. Bhat V. Dinesh Kumar S. Prakash V. L. Chopra 《Journal of plant biochemistry and biotechnology.》2001,10(1):49-51
We report here a simple protocol for regenerating plants from leaf protoplasts of vegetable Brassicas, viz., cabbage, cauliflower and broccoli. Protoplasts from in vitro grown leaf material were cultured in Kao’s medium with a supplementation of 2,4-D, NAA, BAP and glucose, initially in dark for 3d and subsequently in light. Dilution of protoplast cultures was effected on the 7th, 10th and 13th day of culture initiation with Kao’s medium supplemented with sucrose, and reduced 2,4-13 content; NAA was omitted. Micro-colonies were plated on a K3 medium having 2,4-D, BAP and sucrose gelled with agarose. Transfer of calli to another K3 medium with zeatin regenerated shoots from cauliflower protoplast derived calli, whereas a medium with kinetin and zeatin supported shoot regeneration in cabbage and broccoli. Shoot regeneration occurred within 6-6 weeks of culture initiation. Shoots were easily rooted on MS medium without growth regulators. 相似文献
13.
甘薯叶柄原生质体有效植株再生 总被引:3,自引:0,他引:3
将甘薯(Ipomoea batatas (L.)Lam.)‘元气’和‘白星’(‘White Star’)的叶柄原生质体培养在含有0.05 mg·L-1 2,4-D和0.5 mg·L-1 KT的改良MS液体培养基中,3~4 d后细胞开始分裂。培养8~9周后,将直径达1~2 mm的愈伤组织转移到添加0.05~0.2 mg· L-1 2,4-D和0~0.5 mg·L-1 KT或添加0.5~2.0 mg ·L-1 NAA和1.0~3.0 mg·L-1 BAP 的MS固体增殖培养基上使其增殖。转移3~5周后,将愈伤组织再转移到MS基本培养基或转移到添加2.0~3.0 mg·L-1 BAP的MS培养基上。当进一步转移到MS基本培养基上后,从愈伤组织或从愈伤组织形成的不定根上再生出植株。‘元气’植株再生率高达60.0 %,White Star高达43.4%。 相似文献
14.
Morphogenetic responses of cultured cells of cambial origin of a mature tree — Dalbergia sissoo Roxb
Regeneration of plantlets was achieved from cell suspension derived calli of cambial origin from mature elite trees of Dalbergia sissoo. Callus proliferation occurred on the cambial tissue pieces cultured on MS medium supplemented with 2, 4-dichlorophenoxyacetic acid (2.0 mg/1) and benzylaminopurine (0.1 mg/l). Suspension cultures were obtained by transferring and agitating callus lumps in liquid medium composed as above. Aggregates of about 30 cells were plated on semi solid medium, which developed into calli. Shoot bud differentiation was observed in the calli transferred to medium devoid of auxin but containing 0.5–2.0 mg/1 benzylaminopurine. The isolated microshoots were rooted on modified MS medium containing low organic salts and auxins.Abbreviations BAP
6-benzylaminopurine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- IAA
indole-3-acetic acid
- IPA
indole-3-propionic acid
- KN
kinetin
- MS
Murashige and Skoog (1962)
- NAA
1-naphthaleneacetic acid
- IBA
indole-3-butyric acid 相似文献
15.
Summary Frieble calli, obtained from petioles ofGeranium roberlianum were used for the production of cell suspension cultures in liquid MS modified medium supplemented with BAP and NAA. Casamino acids were shown to be necessary for suspension cultures establishment With a 15.9×104 cell. ml–1 concentration a td=38.2h was achieved. 相似文献
16.
A effective protocol for complete plant regeneration via somatic embryogenesis has been developed for Ocimum basilicum L. Callus was initiated from leaf explant of young plant on supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) 1.0 mg l(-1), 3% sucrose and 0.9% agar. The calli showed differentiation of globular structure embryos when transferred to MS medium containing 2,4-D 0.5 mg l(-1) and BAP 1.0 mg l(-1). The maximum globular structure embryos were further enlarged and produced somatic embryos in MS basal medium supplemented with BAP 1.0 mg l(-1)+NAA 1.0 mg l(-1) + KN 0.5 mg l(-1). Continued formation of globular embryo and germination of embryos occurred in this medium. Complete plantlets were transferred onto specially made plastic cup containing soilrite followed by their transfer to the garden soil. Survival rate of the plantlets under ex vitro condition was 80%. 相似文献
17.
María del Pilar Nicasio-Torres Mariana Meckes-Fischer Lucía Aguilar-Santamaría María Luisa Garduño-Ramírez Víctor Manuel Chávez-Ávila Francisco Cruz-Sosa 《Acta Physiologiae Plantarum》2012,34(1):307-316
The antidiabetic properties of Cecropia obtusifolia are attributed to chlorogenic acid (CGA) and isoorientin (ISO) phenolic compounds; both compounds possess hypoglycemic, hypolipidemic,
and antioxidant properties. As a potential strategy for an adequate supply of authentic plant raw material, the aim of this
study was to establish in vitro conditions for the development of cell suspension cultures that produce these bioactive compounds.
Callus cultures of leaf explants from acclimatized tree and in vitro plantlets were set up using different auxin levels; treatments
with 2,4-dichlorophenoxyacetic acid (2,4-D) and α-naphthalene acetic acid (NAA) to 8.92 μM with 6-benzylaminopurine (BAP)
at 2.22 μM stimulate highest callus production. Seedling cotyledon, hypocotyl, leaf, and stem explants developed calli bearing
roots with 2,4-D. With NAA, hypocotyl, cotyledon, and leaf explants developed morphogenic calli; 75% of stem explants formed
calli, and the remaining calli developed shoots. Determined CGA concentrations in calli were similar to those detected in
the leaves of wild trees, and ISO was not produced. Cell suspension cultures were established from leaf explants friable calli
with 8.92 μM 2,4-D in combination with 2.22 μM BAP, employing 4 and 5% inocula in fresh weight; CGA levels were maintained
and ISO was produced only at the end of logarithmic growth. On diminishing nitrate content in Murashige and Skoog (MS) medium
to 8.0 mM, maximum cell biomasses diminished, CGA production is increased and twice with 16.0 and, instead of CGA production
is tripled and quadrupled with 16.0 and 8.0 mM nitrates, respectively, and ISO synthesis was induced earlier and for a longer
time period, increasing its levels at the end of culture. Two compounds with ultraviolet spectra similar to those of caffeic
and ferulic acids were formed. Our results offer a protocol of cell suspension cultures for C. obtusifolia bioactive production and hypoglycemic property conservation. 相似文献
18.
The in vitro studies with Cardiospermum halicacabum indicated that the different explants, i.e cotyledon, hypocotyl, cotyledonary node, leaf, internode and node had the potential to produce calli on Murashige and Skoog (MS) medium supplemented with benzylaminopurine (BAP) and napthalene acetic acid (NAA). Calli of different explant origin showed variable growth responses on different BAP concentrations. The shoots were favourably formed from the calli of leaf and cotyledon explants. The maximum number of shoots were produced from calli subcultured on MS + BAP (17.8 µM). The roots were initiated on growth regulator free MS medium. 相似文献
19.
In vitro regeneration of plantlets and multiplication of Sesbania bispinosa (Jacq.) W.F. Wight plants from cultured callus tissue were demonstrated. Callus was established from both cotyledons and mature leaflets on Murashige and Skoog (MS) basal medium supplemented with BAP (0.5 mg/l) and 2,4-D (2 mg/l). Callus mediated shoot bud differentiation was studied under defined nutritional, hormonal and cultural conditions. Various concentrations of BAP or kinetin (Kn) with coconut milk (CM) in MS media induced different levels of shoot bud differentiation as well as multiplication. Multiple shoot bud differentiation occurred in most of the primary calli. The best medium for shoot bud differentiation from cotyledon derived callus, contained BAP (2 mg/l) and 15% CM (V/V). More efficient shoot bud organogenesis was recorded with BAP than Kn. Supplementation with CM in MS media accelerated shoot bud organogenesis in differentiating callus tissue. Rooting of differentiated shoots was achieved by a three step culture procedure involving (a) MS solid medium containing IBA (2 mg/l), (b) growth regulator free half strength MS medium with 1% charcoal, and (c) half strength MS liquid medium free of vitamins, growth regulators and charcoal.Abbreviations IAA
indoleacetic acid
- IBA
indole-3-butyric acid
- NAA
naphthaleneacetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- BAP
6-benzylaminopurine
- Kn
kinetin
- CM
coconut milk
- MS
Murashige and Skoog's medium
- SBI
shoot bud inducing medium 相似文献
20.
Hee-Ju Yu Soo Kyung Oh Man-Ho Oh Dong-Woog Choi Young Myung Kwon Sang-Gu Kim 《Plant cell reports》1997,16(5):261-266
We previously studied the production of shikonin derivatives by cell lines ofLithospermum erythrorhizon. As a result, we have obtained a cell line LE 87, which exhibited high cell growth and high shikonin production. In the present study, the effects of auxins (2,4-D, IAA, picloram, and NAA) and cytokinins (BAP and kinetin) on organogenesis and somatic embryogenesis in this shikonin-producing cell line were investigated. The highest organogenic and embryogenic efficiency was obtained on MS medium supplemented with 10 µM NAA and 0.3 µM kinetin. Subcultured calli showed different morphogenic frequencies depending on the NAA and kinetin concentration. Morphologically normal plants have been regenerated via mostly organogenesis. Shoots subsequently produced roots on plant growth regulator-free MS medium and developed into plantlets. In most cases, a few thin roots were formed at the bases of the shoots after four weeks on the rooting medium. More than fifty green plantlets were transplanted to soil in pots and developed into phenotypically normal plants 8 weeks after being transferred to soil. The regenerated plants grew to maturity, flowered, and set seeds by only artificial pollination.Abbreviations BAP
6-benzylaminopurine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- IAA
indole-3-acetic acid
- NAA
-naphthalene-acetic acid
- MS
Murashige and Skoog (1962) medium
Communicated by S. Gleddie 相似文献