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1.
When cotyledonary explants, excised from in vitro germinated seedlings, of pomegranate (Punica granatum L.) were incubated on solid Murashige and Skoog (1962) medium supplemented with 21 μM naptheleneacetic acid (NAA) and 9 μM 6-benzyladenine (BA), 80% of explants developed callus.
A high frequency of shoot organogensis was obtained when explants were incubated on MS medium supplemented with 8 μM BA, 6 μM
NAA, and 6 μM giberrellic acid (GA3). However, adding 24 μM silver nitrate (AgNO3) to this medium markedly enhanced shoot regeneration frequency (63%) and mean number of shoots per explant (11.26) and length
of shoots (2.22 cm). Highest frequency of in vitro rooting, mean number of roots/shoot (4.32), and mean root length (2.71 cm)
were obtained when regenerated shoots were transferred to half-strength MS medium supplemented with 0.02% activated charcoal.
Well-rooted plantlets were acclimatized, and then transferred to soil medium. Moreover, when zygotic embryos of P. granatum, excised from seeds collected at 16 weeks following full bloom, were incubated on MS medium containing 30 g l−1 sucrose, 15% coconut water, 21 μM NAA, and 9 μM BA, they developed the highest frequency of embryogenic callus, clumps with
globular embryos, and mean number of both globular and heart-shaped embryos per callus clump. Subjecting zygotic embryo explants
to six-week dark incubation period was essential for embryogenic callus induction, and these were subsequently transferred
to 16 h photoperiod for further growth and development of somatic embryos. Germination of somatic embryos was observed when
these were transferred to MS medium was supplemented with 60 g l−1 sucrose. 相似文献
2.
Plant regeneration through somatic embryogenesis was achieved using immature zygotic embryos (ZE) of Sorbus pohuashanensis as explants. Over 50% of immature ZEs from immature seed collected at 30 days after pollination produced direct somatic embryos
(SEs) on Murashige and Skoog (MS) medium supplemented with 0–0.44 μM 6-benzyladenine (BA) in combination with 5.73 μM naphthaleneacetic
acid (NAA) or with 0.91–2.26 μM 2,4-dichlorophenoxyacetic acid (2,4-D) alone. Fourteen to 23 SEs per explant were regenerated
on MS medium supplemented with BA 0.44 μM in combination with NAA 5.73 μM. SE formation decreased when sucrose concentrations
were higher than 40 g L−1. Repetitive embryogenesis occurred following culture on solid MS medium containing 12 μM abscisic acid, 75 g L−1 polyethylene glycol, and 20 g L−1 sucrose at 25 ± 1°C under a 16-h photoperiod with a light intensity of 40 μmol m−2 s−1. Over 40% of the mature SEs germinated on solid MS medium under light condition described previously. Up to 40% of the regenerated
plantlets were successfully acclimatized under greenhouse conditions. Plantlets derived from SEs grew vigorously with similar
morphology as those germinated from ZEs. Histological studies of explants at various developmental stages of somatic embryogenesis
revealed that SEs passed through globular, heart, torpedo, and mature stages. Similar to ZE suspensors, similar structures
of SE degenerated in later stages of embryo development. ZE and SE are a effective means of regenerating tissue culture plantlets
for S. pohuashanesis. 相似文献
3.
4.
Dong-Mei Kong John E. Preece Hai-Long Shen 《Plant Cell, Tissue and Organ Culture》2012,108(3):485-492
Somatic embryogenesis was obtained from immature cotyledon explants that were cultured on half-strength Murashige and Skoog
(MS) salts and vitamins with 5.4 μM naphthaleneacetic acid (NAA) and 0.2 μM thidiazuron (TDZ) plus a 4 × 4 factorial combination
of 0, 9.8, 24.6, or 49.2 μM indole-3-butyric acid (IBA) and 0, 8.9, 22.2, or 44.4 μM 6-benzyladenine (BA). The addition of
44.4 μM BA improved the percentage of cotyledon explants that produced somatic embryos to >20%, if 9.8 or 24.6 μM IBA was
also present. Somatic embryogenesis was >30% when seeds were harvested on 31 July or 15 August. The addition of 50 or 70 g l−1 sucrose enhanced embryogenesis. Histological examination showed that somatic embryos originated from epidermis cells of zygotic
embryos. A peak germination rate (69%) was attained when somatic embryos were desiccated for 10 min before they produced green
cotyledons and elongating shoot tips. Of the germinated embryos from this desiccation treatment, 65.6% also grew roots and
therefore converted into plants. Chilling somatic embryos at 4°C for 15 days resulted in the highest germination rate (69.4%),
which was significantly higher than those without chilling treatment (27.6%). However <10% of the chilled germinated embryos
formed roots and grew into plants. Plantlets from somatic embryos were transplanted into a 2 vermiculite: 1 sphagnum peat
medium, where they had a survival rate of 80.8%, and had no morphological abnormalities. 相似文献
5.
Simões-Gurgel Claudia Cordeiro Lívia da Silva de Castro Tatiana Carvalho Callado Cátia Henriques Albarello Norma Mansur Elisabeth 《Plant Cell, Tissue and Organ Culture》2011,106(3):537-545
The effects of different levels of Murashige and Skoog (MS) basal medium, 2,4-dichlorophenoxyacetic acid (2,4-D), and sucrose
on anthocyanin production and biomass accumulation of cell suspension cultures of Cleome rosea were investigated. Cultures were established in liquid MS medium containing 30 g l−1 sucrose and supplemented with 0.90 μM 2,4-D. Proliferating cell suspension cultures achieved the highest growth capacity,
a fourfold increase in biomass accumulation, following subculture at the exponential growth phase, 14–18 days of culture.
Moreover, the presence of 2,4-D was essential for anthocyanin production and biomass accumulation. On the other hand, increasing
levels of sucrose above 30 g l−1 resulted in a drastic reduction in biomass accumulation. Anthocyanin production was highest in cell suspension cultures grown
on half-strength MS medium (1/2 MS), 30 g l−1 sucrose, and 0.45 μM 2,4-D. These cell suspension cultures were mainly composed of small aggregates of spherical cells with
similar morphology observed in anthocyanin-producing and non-producing cultures. Moreover, microscopic analysis of anthocyanin-producing
cultures showed the presence of mixtures of non-pigmented, low-pigmented, and high-pigmented cells. 相似文献
6.
Abhinav Grover Jayashankar S. Yadav Ranjita Biswas Choppakatla S. S. Pavan Punita Mishra Virendra S. Bisaria Durai Sundar 《Plant Cell, Tissue and Organ Culture》2012,108(2):323-331
Cell suspension cultures of Camellia sinensis were established in 250 ml shake flasks. Flasks contained 50 ml liquid medium of either Murashige and Skoog (MS), N/5 MS
or Heller medium containing different levels of 6-benzyladenine (BA) (0.05–2 mg l−1), 2,4-dichlorophenoxyacetic acid (2,4-D) (1–10 mg l−1), and sucrose (10–50 g l−1). Moreover, the pH of the medium was varied from 5.2–6.2. In addition, cultures were subjected to light irradiation as well
as to complete darkness. Following optimization of aroma and terpenoid extraction methods, cell cultures were analyzed for
the volatile compounds using GC/MS. A total of 43 compounds were identified using the micro SDE apparatus. Among the major
monoterpenoids obtained were α-terpineol and nerol. Moreover, other high aroma-value compounds, including 2-ethyl hexanol,
benzyl alcohol, benzene acetaldehyde, nonanal and phenylethylalcohol were also detected. The highest levels of these compounds
were obtained from cell suspension cultures grown in MS medium containing 5 mg l−1 2,4-D, 1 mg l−1 BA and 30 g l−1 sucrose at pH of 5.8 with incubation in complete darkness. 相似文献
7.
Toshinari Godo Miho Komori Emi Nakaoki Tomohisa Yukawa Kazumitsu Miyoshi 《In vitro cellular & developmental biology. Plant》2010,46(3):323-328
The effects of culture conditions on the asymbiotic germination of mature seeds of Calanthe tricarinata Lindl., an endangered terrestrial cool-climate orchid, were examined. Specifically, conditions such as illumination, temperature,
and the addition of plant growth regulators to the medium were studied. Mature seeds were harvested from plants that had been
collected in Toyama Prefecture, Japan, and maintained at the Botanic Gardens of Toyama. Solidified “New Dogashima” medium
was used as the basal medium, and it was supplemented with 6-benzyladenopurine (BA) or α-naphthalene acetic acid (NAA). White
light at 40 μmol m−2 s−1, with a 16-h photoperiod, inhibited the germination of seeds by 53–80%, as compared to dark controls in genotypes examined.
The optimal temperature for the germination of seeds in darkness was 20°C and the germination frequency reached 60%, whereas
it was only 28% at 25°C. While both NAA and BA stimulated germination, BA was more effective than NAA. After storage for 18 mo
at 5°C, seeds incubated on medium that contained 0.2 mg l−1 BA germinated at a frequency of 36%, which was twice that of seeds grown without any plant growth regulators. The frequency
of subsequent germination decreased during storage of seeds at 5°C for approximately 2 yr, dropping from 61% to 13%. The protocorms
obtained in this study were developed to plantlets readily after transferring to fresh 1/2 MS medium without any plant growth
regulators. They were successfully acclimatized in green house after two to three subcultures in vitro. The significant role of a reproducible protocol for the germination of mature seeds is discussed in terms of the ex situ conservation of endangered orchid species. 相似文献
8.
Yohana de Oliveira Fernanda Pinto André Luís Lopes da Silva Ivan Guedes Luiz Antonio Biasi Marguerite Quoirin 《In vitro cellular & developmental biology. Plant》2010,46(2):192-197
Melaleuca alternifolia is cultivated for the production of an essential oil useful in the cosmetic and pharmaceutical industries. Despite the economic
importance of this species, there is little knowledge about its in vitro propagation. The aim of this study was to establish an efficient protocol for micropropagation of M. alternifolia. With the goal of in vitro multiplication by axillary shoot proliferation, both solid and liquid MS and WPM media were tested with supplementation with
BA at 0, 0.55, 1.11, 2.22, 3.33, and 4.44 μM. The best result for shoot multiplication was obtained when either 0.55 μM BA
was added into solid MS medium or 1.11 μM BA was added into liquid MS medium, with 5.6 and 11.8 shoots per explant generated,
respectively. On solid or liquid WPM medium supplemented with 0.55 μM BA, the proliferation rates were 5.5 and 4.7, respectively.
Three auxins (NAA, IAA, and IBA) were tested at 0.53 and 2.64 μM during the rooting stage. Several sucrose concentrations
(15, 30, and 45 g L−1) were compared to a sucrose-free medium. Rooting performances on four culture media were then compared: MS, half-strength
MS (MS/2), MS + activated charcoal (AC), and MS/2 + AC. The results showed that auxin addition to culture medium is not necessary
for in vitro rooting. Rooted microcuttings from different culture media were acclimatized in a greenhouse, and the survival percentage was evaluated.
All shoots cultured in an auxin-free MS medium supplemented with sucrose (30 g L−1) produced roots, and all plants survived during acclimatization. Activated charcoal added in rooting medium reduced rooting
rates. 相似文献
9.
Cheng-Hao Li Bao-Guang Liu Tae-Dong Kim Heung-Kyu Moon Yong-Eui Choi 《Plant biotechnology reports》2008,2(4):259-265
Picea koraiensis, called Korean spruce, is an evergreen tree and found mostly in northeast Asia. In this study, plant regeneration via somatic
embryogenesis from open-pollinated immature zygotic embryos of nine genotypes of elite trees was established. Immature zygotic
embryos were cultured onto RJW medium modified from 505 medium with 21.48 μM NAA, 2.22 μM BA, and 2.32 μM KT. The average
frequency for all nine genotypes was 74.2%. Embryogenic calluses of the nine genotypes of elite trees were subcultured on
RJW basal medium containing 8.06 μM NAA, 1.11 μM BA, and 1.16 μM kinetin. The calluses of three lines, 3#, 9#, and 2#, were actively proliferated but others were not. Somatic embryogenesis was induced from the embryogenic callus in genotypes
of 3#, 9#, and 2# on RJW medium with ABA and 60 g l−1 sucrose. Cotyledonary somatic embryos were subjected to a drying process. The drying of embryos by uncapping the culture
bottle for 5 days on a clean bench resulted in a high frequency of germination of somatic embryos (87% in RJW medium). However,
plantlet conversion from germinated embryos was greatly reduced and the optimal medium for plant conversion was 1/2 WPM or
1/2 BMI medium. In conclusion, we have, for the first time, established a plant regeneration system via somatic embryogenesis
in the Korean spruce, which can be applied for rapid micropropagation of elite trees. 相似文献
10.
T. Zhang 《In vitro cellular & developmental biology. Plant》2007,43(2):91-94
This study investigated the factors affecting in vitro flowering of Perilla frutescens. The shoots regenerated from cotyledonary and hypocotyl explants cultured on Murashige and Skoog (MS) medium supplemented
with benzyladenine (BA) and indole-3-acetic acid, each at 0.5 mg l−1, were excised and transferred to MS medium containing 30 g l−1 of sucrose, 8.25 g l−1 of ammonium nitrate, and 1.0 mg l−1 of BA. After 40 d of culture, 86.2% of shoots flowered and most of which self-fertilized in vitro and produced mature fruits with viable seeds. These seeds were germinated and plants were grown to maturity and flowered
in soil under greenhouse conditions. The in vitro flowering system reported in this study may facilitate rapid breeding of P. frutescens and offers a model system for studying the physiological mechanism of flowering. 相似文献
11.
The genus Dierama comprises plants with a potential to be developed as ornamentals. D. erectum seeds were decontaminated and germinated on 1/10th strength Murashige and Skoog (Physiol Plant 15:473–497, 1962) (MS) media
without plant growth regulators or sucrose. In an experiment investigating the effects of 6-benzyladenine (BA), meta-Topolin (mT), kinetin (KIN) and zeatin (Z) with or without α-naphthaleneacetic acid (NAA), the highest shoot number per hypocotyl (4.20 ± 0.51)
was obtained from MS medium supplemented with 1.0 μM Z after 8 weeks. This was followed by a combination of 2.0 μM KIN and
2.0 μM NAA with 3.67 ± 0.81 shoots per explant. BA treatments produced 3.20 ± 0.22 shoots per hypocotyl explant when 2.0 μM
was combined with 1.0 μM NAA, while mT gave 3.09 ± 0.99 shoots per explant when 2.0 μM mT was combined with 2.0 μM NAA. Adventitious shoot regeneration was optimised when shoots were grown under a 16-h photoperiod
at 100 μmol m−2 s−1 on MS medium supplemented with 1.0 μM BA. This resulted in an average of 12.73 ± 1.03 shoots per hypocotyl explant. Various
concentrations of ancymidol, activated charcoal and sucrose did not promote in vitro corm formation of this species. Plants
rooted successfully after 8 weeks on MS medium supplemented with 1.0 μM indole-3-butyric acid (IBA) and had an average root
number of 2.73 ± 0.40. After 2 months of acclimatisation, plants had formed corms. The largest corms (of diameter 0.45 ± 0.03 cm)
were produced in plants pre-treated with 0.5 μM IBA. The highest plant survival percentage of 73% was also associated with
this treatment. 相似文献
12.
D. P. Barik S. K. Naik A. Mudgal P. K. Chand 《In vitro cellular & developmental biology. Plant》2007,43(2):144-148
This study describes a reproducible protocol for rapid mass propagation of a multipurpose legume, Clitoria ternatea L., using cotyledonary node explants derived from axenic seedlings. Multiple shoots were induced in Murashige and Skoog (MS)
medium supplemented with N
6-benzyladenine (BA), zeatin riboside, or thidiazuron. N
6-Benzyladenine at 1.0 mg l−1 (4.44 μM) was most effective for shoot proliferation. Multiple shoots were also induced in nodal segments of in vitro-raised shoots grown on MS medium containing 1.0 mg l−1 (4.44 μM) of BA. Rooting was best induced in shoots grown on half-strength MS medium with 0.25 mg l−1 (1.42 μM) of indole-3-butyric acid. Plants were acclimatized in vermicompost and established in soil where they flowered
and formed mature seeds. 相似文献
13.
Zita Demeter Gyula Surányi V. Attila Molnár Gábor Sramkó Dániel Beyer Zoltán Kónya Gábor Vasas Márta M-Hamvas Csaba Máthé 《Plant Cell, Tissue and Organ Culture》2010,100(3):349-353
Crocus heuffelianus belongs to the C. vernus (Iridaceae) species aggregate. In the Carpathian Basin and particularly in Hungary it is considered an endangered species. Therefore
our aim was to establish a tissue culture system with potential of germplasm preservation of this taxon. For in vitro culture
experiments, shoot primordia from corms were the most suitable. We induced an embryogenic callus line from those explants
on basal Murashige-Skoog (MS) medium supplemented with Gamborg’s vitamins, 2% (w/v) sucrose, 10 mg l−1 (53.7 μM) α-naphthaleneacetic acid (NAA) and 1 mg l−1 (4.44 μM) 6-benzyladenine (BA). Globular stage embryos developed on this medium and several culture conditions were used
in an attempt to obtain mature embryos and plant regeneration. Firstly a decrease of auxin/cytokinin concentration and ratio,
then secondly a decrease in the strength of culture medium and the concentration of carbon source was used, which was effective
in embryogenesis and the production of plants. Regeneration medium used in the second step was fourfold diluted MS medium
and Gamborg’s vitamins supplemented with 1% (w/v) sucrose, 0.05 mg l−1 (0.26 μM) NAA and 0.5 mg l−1 (2.22 μM) BA, with a 14/10 h photoperiod. Under these conditions we could detect all the stages of somatic embryo development
characteristic for Iridaceae. This is the first report demonstrating the production of stable tissue culture of C. heuffelianus with potential use in germplasm preservation via plant regeneration. This study could also contribute to a better understanding
of somatic embryogenesis in the Crocus genus. 相似文献
14.
A reproducible protocol for clonal propagation of Spilanthes acmella has been established. Routinely, the cultures were established in spring (January–April) season because of the highest aseptic
culture establishment and high frequency shoot proliferation. Incorporation of 5 μM N6-benzyladenine (BA) to Murashige and Skoog (MS) basal medium showed 100% bud-break and promoted multiple shoot proliferation
in cultures. Interestingly, a higher concentration of BA (7–15 μM) promoted stunted shoots with pale leaves while a lower
concentration (1–3 μM) resulted in shoots with long internodes and excessive adventitious root proliferation from all over
their surface. For recurrent shoot multiplication, single node segments from in vitro-developed shoots were excised and cultured
on MS + BA (5 μM) medium where 20.3-fold shoot multiplication was achieved every 5 weeks. Finally, these shoots were successfully
rooted on half-strength MS medium (major salts reduced to half-strength) with 50 g l−1 sucrose, with a frequency of 100%. Transplantation survival of micropropagated plants was 88.9%. Additionally, accumulation
of scopoletin, a phytoalexin, was revealed for the first time in the uninfected leaves of Spilanthes. Further, the quantitative estimation by HPLC with a fluorescence detector showed that the amounts of scopoletin content
(0.10 μg g−1 DW) in the leaves of micropropagated plants are comparable to those of field-grown mother plants. The study thus signifies
the effectiveness of in vitro methodology for true-to-type plant regeneration of Spilanthes and their later utility for biosynthesis and constant production of scopoletin throughout the year. 相似文献
15.
In this work, the effect of different inducing factors on trans-resveratrol extracellular production in Monastrell grapevine suspension cultured cells is evaluated. A detailed analysis
provides the optimal concentrations of cyclodextrins, methyljasmonate and UV irradiation dosage, optimal cell density, elicitation
time and sucrose content in the culture media. The results indicate that trans-resveratrol production decreases as the initial cell density increases for a constant elicitor concentration in Monastrell
suspension cultured cells treated with cyclodextrins individually or in combination with methyljasmonate; the decrease observed
in cell cultures elicited with cyclodextrins alone is far more drastic than those observed in the combined treatment. trans-Resveratrol extracellular production observed by the joint use of cyclodextrins and methyljasmonate (1,447.8 ± 60.4 μmol
trans-resveratrol g−1 dry weight) is lower when these chemical compounds are combined with UV light short exposure (669.9 ± 45.2 μmol trans-resveratrol g−1 dry weight). Likewise, trans-resveratrol production is dependent on levels of sucrose in the elicitation medium with the maximal levels observed with
20 g l−1 sucrose and the joint action of cyclodextrins and 100 μM methyljasmonate. The sucrose concentration did not seem to limit
the process although it affects significantly the specific productivity since the lowest sucrose concentration is 10 g l−1, the highest productivity is reached (100.7 ± 5.8 μmol trans-resveratrol g−1 dry weight g−1 sucrose) using cyclodextrins and 25 μM methyljasmonate. 相似文献
16.
Muthu Thiruvengadam K. T. Rekha Chang-Hsien Yang Narayanasamypillai Jayabalan Ill-Min Chung 《Plant biotechnology reports》2010,4(4):321-328
An efficient protocol for in vitro organogenesis was achieved from callus-derived immature and mature leaf explants of Momordica charantia, a very important vegetable and medicinal plant. Calluses were induced from immature leaf explants excised from in vitro
(15-day-old seedlings) mature leaf explants of vivo plants (45 days old). The explants were grown on Murashige and Skoog (MS)
medium with Gamborg (B5) vitamins containing 30 g l−1 sucrose, 2.2 g l−1 Gelrite, and 7.7 μM naphthalene acetic acid (NAA) with 2.2 μM thidiazuron (TDZ). Regeneration of adventitious shoots from
callus (30–40 shoots per explant) was achieved on MS medium containing 5.5 μM TDZ, 2.2 μM NAA, and 3.3 μM silver nitrate (AgNO3). The shoots (1.0 cm length) were excised from callus and elongated in MS medium fortified with 3.5 μM gibberellic acid (GA3). The elongated shoots were rooted in MS medium supplemented with 4.0 μM indole 3-butyric acid (IBA). Rooted plants were
acclimatized in the greenhouse and subsequently established in soil with a survival rate of 90%. This protocol yielded an
average of 40 plants per leaf explant with a culture period of 98 days. 相似文献
17.
Federico A. Gutiérrez-Miceli Lourdes Arias Nicolás Juarez-Rodríguez Miguel Abud-Archila Aldo Amaro-Reyes Luc Dendooven 《In vitro cellular & developmental biology. Plant》2010,46(1):57-63
This paper reports on the optimum concentrations of naphthalene acetic acid (NAA) and 6-benzyladenine (BA) to stimulate callus
growth and NAA; kinetin and silver nitrate (AgNO3) for callus redifferentiation in Dianthus caryophyllus L. Meristems were excised and placed in MS medium with 30 g l−1 sucrose and 9.0 μM 2,4-d. Callus clusters were transferred to MS medium containing NAA (0, 1.7, 3.3, and 5.0 μM) and BA (0, 1.7, 3.3, and 5.0 μM)
for proliferation and to MS medium with 30 g l−1 sucrose, 2.5 g l−1 phytagel, kinetin (0, 33, and 66 μM); NAA (0, 7.95, and 15.9 μM) and AgNO3 (0, 23.54 and 47.08 μM) for shoot and root induction. Treatments were applied according to a Box–Behnken design. After callus
growth and redifferentiation, plants were incubated in the greenhouse at 18 ± 2°C for 4 wk and at 20–26°C for 4 wk. Finally,
plants were changed to near-commercial greenhouse conditions with different day (30–35°C) and night (16–24°C) temperatures.
Results showed better callus growth at higher NAA concentrations. A maximum callus weight was found with 5.0 μM NAA but without
BA. A maximum of 78% calluses with shoots was obtained with 15.9 μM NAA, 47.08 μM AgNO3, and 0.74 μM kinetin and 58% with roots with 15.7 μM NAA and 47.08 μM AgNO3, but without kinetin. The shoots obtained showed little hyperhydricity. Vigorous plants were obtained after gradual acclimatization
with an 80% survival rate under nursery conditions. 相似文献
18.
Tissue culture and plant regeneration of blue grama grass, Bouteloua gracilis (H.B.K.) Lag. Ex Steud
Gerardo Armando Aguado-Santacruz José Luis Cabrera-Ponce Víctor Olalde-Portugal M. A. Rosario Sánchez-González Judith Márouez-Guzmán Luis Herrera-Estrella 《In vitro cellular & developmental biology. Plant》2001,37(2):182-189
Summary As a first step towards applying biotechnology to blue grama, Bouteloua gracilis (H. B. K.) Lag. ex Steud., we have developed a regenerable tissue culture system for this grass. Shoot apices were isolated
from 3-d-old seedlings and cultured in 15 different growth regulator formulations combining 2,4-dichlorophenoxyacetic acid
(2,4-D), Picloram (4-amino-3, 5,6-trichloropicolinic acid), N6-benzyladenine (BA) or adenine (6-aminopurine). The highest induction of organogenic callus was obtained with formulations
containing 1 mg l−1 (4.52 μM) 2,4-D plus 0.5 mg l−1 (2.22 μM) BA. and 2 mg l−1 (8.88 μM) BA plus 1 mg l−1 (4.14 μM) Picloram with or without 40 mg l−1 (296.08 μM) adenine. Lower frequencies of induction were obtained for embryogenic as compared to organogenic callus. The most efficient
treatments for induction of embryogenic callus contained 2 mg l−1 (9.05 μM) 2,4-D combined with 0.25 (1.11 μM) or 0.50 mg l−1 (2.22 μM) BA, or 1 mg l−1 (4.52 μM) 2,4-D with 0.50 mg l−1 (2.22 μM) BA. Regeneration was achieved in hormonefree Murashige anmd Skoog (MS) medium, half-strength MS medium or MS medium plus
1 mg l−1 (1.44 μM) gibberellic acid. The number of plantlets regenerated per 500 mg callus fresh weight on MS medium ranged from 9 for 2 mg
l−1 (9.05 μM) 2,4-D to 62.2 for induction medium containing 2 mg l−1 (8,28 μM) Picloram, 1 mg l−1 (4.44 μM) BA and 40 mg l−1 (296.08 μM) adenine. Regnerated plants grown in soil under greenhouse conditions reached maturity and produced seeds. 相似文献
19.
Smita Srivastava Ashok Kumar Srivastava 《In vitro cellular & developmental biology. Plant》2012,48(1):73-84
Azadirachtin, a well-known biopesticide, is a secondary metabolite extracted from the seeds of Azadirachta indica. In the present study, azadirachtin was produced in hairy roots of A. indica, generated by Agrobacterium rhizogenes-mediated transformation of leaf explants. Liquid cultures of A. indica hairy roots were developed with a liquid-to-flask volume ratio of 0.15. The kinetics of growth and azadirachtin production
were established in a basal plant growth medium containing MS medium major and minor salts, Gamborg’s medium vitamins, and
30 g l−1 sucrose. The highest azadirachtin accumulation in the hairy roots (up to 3.3 mg g−1) and azadirachtin production (∼44 mg l−1) was obtained on Day 25 of the growth cycle, with a biomass production of 13.3 g l−1 dry weight. To enhance the production of azadirachtin, a Plackett–Burman experimental design protocol was used to identify
key medium nutrients and concentrations to support high root biomass production and azadirachtin accumulation in hairy roots.
The optimal nutrients and concentrations were as follows: 40 g l−1 sucrose, 0.19 g l−1 potassium dihydrogen phosphate, 3.1 g l−1 potassium nitrate, and 0.41 g l−1 magnesium sulfate. Concentrations were determined by a central composite design protocol and verified in shake-flask cultivation.
The optimized medium composition yielded a root biomass production of 14.2 g l−1 and azadirachtin accumulation of 5.2 mg g−1, which was equivalent to an overall azadirachtin production of 73.84 mg l−1, 68% more than that obtained under non-optimized conditions. 相似文献
20.
Summary
Dendrobium candidum Wall. Ex Lindl. is an important species used in the formulation of Shih-hu, a Chinese traditional medicine. An efficient
protocol for in vitro propagation of D. candidum using the axenic nodal segments of the shoots, originated from the in vitro germinated seedlings, was developed. The seeds from 120-d-old capsules after pollination were first germinated on half-strength
Murashige and Skoog (MS) basal medium supplemented with 30 g l−1 sucrose. After 4 mo., the seedlings were subcultured on a similar medium supplemented with 1 ml l−1 HYPONeX, 80 g l−1 potato homogenate and 2 g l−1 activated charcoal for further growth. Axenic nodal segments excised from 9-mo.-old seedlings were cultured on the medium
in the presence of 2 mg l−1 benzyladenine (BA) and 0.1 mg l−1 naphthaleneacetic acid (NAA). After 75 d, 73.2% of the explants gave rise to buds/shoots. The elongated shoots were rooted
on the medium containing 0.2 mg l−1 NAA and the plantlets were successfully acclimatized in soil. 相似文献