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1.
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
2.
Root segments from seedlings of Panax ginseng produced adventitious roots directly when cultured on 1/2 MS solid medium lacking NH4NO3 and containing 3.0 mg l−1 IBA. Using this adventitious root formation, we developed rapid and efficient transgenic root formation directly from adventitious
root segments in P. ginseng. Root segments were co-cultivated with Agrobacterium tumefaciens (GV3101) caring β-glucuronidase (GUS) gene. Putative transgenic adventitious roots were formed directly from root segments on medium with 400 mg l−1 cefotaxime and 50 mg l−1 kanamycin. Kanamycin resistant adventitious roots were selected and proliferated as individual lines by subculturing on medium
with 300 mg l−1 cefotaxime and 50 mg l−1 kanamycin at two weeks subculture interval. Frequency of transient and stable expression of GUS gene was enhanced by acetosyringon (50 mg l−1) treatment. Integration of transgene into the plants was confirmed by the X-gluc reaction, PCR and Southern analysis. Production
of transgenic plants was achieved via somatic embryogenesis from the embryogenic callus derived from independent lines of
adventitious roots. The protocol for rapid induction of transgenic adventitious roots directly from adventitious roots can
be applied for a new Agrobacterium tumefaciens-mediated genetic transformation protocol in P. ginseng. 相似文献
3.
Su-Juan Zhao Zhong-Chun Zhang Xiang Gao Gulsum Tohsun Bao-Sheng Qiu 《Plant Cell, Tissue and Organ Culture》2009,99(1):9-16
An efficient micropropagation system for mining ecotype Sedum alfredii Hance, a newly identified Zn/Cd hyperaccumulator, was developed. Frequency of callus induction reached up to 70% from leaves
incubated on Murashige and Skoog (MS) medium supplemented with 1.0 mg l−1 2,4-dichlorophenoxy acetic acid (2,4-D) and 0.5 mg l−1 6-benzyladenine (BA), and 83% from internodal stem segments grown on MS medium with 0.1 mg l−1 2,4-D and 0.1 mg l−1 BA. Callus proliferated rapidly on MS medium containing 0.2 mg l−1 2,4-D and 0.05 mg l−1 thidiazuron. The highest number of adventitious buds per callus (17.3) and frequency of shoot regeneration (93%) were obtained
when calli were grown on MS medium supplemented with 2.0 mg l−1 BA and 0.3 mg l−1 α-naphthalene acetic acid (NAA). Elongation of shoots was achieved when these were incubated on MS medium containing 3.0 mg l−1 gibberellic acid. Induction of roots was highest (21.4 roots per shoot) when shoots were transferred to MS medium containing
2.0 mg l−1 indole 3-butyric acid rather than either indole 3-acetic acid or NAA. When these in vitro plants were acclimatized and transferred
to the greenhouse, and grown in hydroponic solutions containing 200 μM cadmium (Cd), they exhibited high efficiency of Cd
transport, from roots to shoots, and hyperaccumulation of Cd. 相似文献
4.
Guohua Ma Jaime A. Teixeira da Silva Guojiang Wu 《Journal of Plant Growth Regulation》2011,30(1):114-116
The induction of adventitious buds from apical shoot explants of Euphorbia tirucalli was studied. On average, 10.5 adventitious buds were efficiently induced in a ring on the segment from one apical explant
on MS (Murashige and Skoog) medium supplemented with 0.5 mg l−1 thidiazuron and 0.5 mg l−1 benzylaminopurine. The adventitious buds could develop into adventitious shoots during subsequent cultures on hormone-free
MS medium. For rooting, shoot clumps were cultured on half-strength MS medium containing 0.2 mg l−1 α-naphthaleneacetic acid or indole-3-butyric acid. All the rooted plants survived establishment in soil within 2 months. 相似文献
5.
Praveen Nagella Hosakatte Niranjana Murthy 《Plant Cell, Tissue and Organ Culture》2011,104(1):119-124
Withania somnifera is an important medicinal plant that contains withanolides and withaferins, both bioactive compounds. We have tested the
effects of macroelements and nitrogen source in W. somnifera cell suspension cultures with the aim of optimizing the production of biomass and withanolide A. The effects of the macroelements
NH4NO3, KNO3, CaCl2, MgSO4 and KH2PO4 at concentrations of 0.0, 0.5, 1.0, 1.5 and 2.0× strength and of the nitrogen source [NH4
+/NO3
− (mM/mM) ratio of: 0.00/18.80, 7.19/18.80, 14.38/18.80, 21.57/18.80, 28.75/18.80, 14.38/0.00, 14.38/9.40, 14.38/18.80, 14.38/28.20,
and 14.38/37.60 (mM)] in Murashige and Skoog medium were tested for biomass and withanolide A production. The highest accumulation
of biomass [147.81 g l−1 fresh weight (FW) and 14.02 g l−1 (dry weight (DW)] was recorded in the medium containing a 0.5× concentration of NH4NO3, and the highest production of withanolide A content was recorded in the medium with 2.0× KNO3 (4.36 mg g−1 DW). The NH4
+/NO3
− ratio also influenced cell growth and withanolide A production, with both parameters being larger when the NO3
− concentration was higher than that of NH4
+. Maximum biomass growth (110.45 g l−1 FW and 9.29 g l−1 DW) was achieved at an NH4
+/NO3
− ratio of 7.19/18.80, while withanolide A production was greatest (3.96 mg g−1 DW) when the NH4
+/NO3
− ratio was 14.38/37.60 mM. 相似文献
6.
Weimei Jiang Luxi Chen Qi Pan Yingxiong Qiu Yingying Shen Chengxin Fu 《Acta Physiologiae Plantarum》2012,34(2):631-639
Dysosma versipellis (Hance) M. Cheng is an endangered plant due to overharvesting for the extraction of podophyllotoxin. Thus, the in vitro technique
is valuable for the propagation of this species. When the explants of rhizome buds were cultured on Murashige and Skoog’s
(MS) medium with 6-benzyladenine (BA) (1.0 mg l−1), gibberellic acid (GA3) (0.5 mg l−1) and zeatin (Zea) (0.5 mg l−1), multiple buds were regenerated directly on the explants without callusing within 6 weeks. Callus was induced from the leaf
segment cultures on MS basal medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) (0.5 mg l−1) and BA (0.2 mg l−1) within 4 weeks. The adventitious buds were differentiated when the calli were subcultured on MS medium supplemented with
BA (1.0 mg l−1) and thidiazuron (TDZ) (0.2 mg l−1) within 6 weeks. The adventitious buds obtained from callus and the rhizome-buds rooted with a frequency of 100% on half
strength MS medium fortified with indole-3-butyric acid (IBA) 0.5 mg l−1 and activated charcoal (AC) 0.5 g l−1 for 4 weeks. The rooted shoots were successfully transplanted from a mixture of vermiculite:soil (1:1 v/v) to the field with
a survival rate of 85%. Podophyllotoxin production in calli, cultured rhizomes, rhizomes of transplanting plants from the
garden and rhizomes in the wild field was confirmed by high-performance liquid chromatography (HPLC) analysis. Our results
suggest that calli, cultured rhizomes and rhizomes of transplanting plants would be the potential sources of podophyllotoxin. 相似文献
7.
Morinda citrifolia adventitious roots were cultured in shake flasks using Murashige and Skoog medium with different types and concentrations
of auxin and cytokinin. Root (fresh weight and dry weight) accumulation was enhanced at 5 mg l−1 indole butyric acid (IBA) and at 7 and 9 mg l−1 naphthalene acetic acid (NAA). On the other hand, 9 mg l−1 NAA decreased the anthraquinone, phenolic and flavonoid contents more severely than 9 mg l−1 IBA. When adventitious roots were treated with kinetin (0.1, 0.3 and 0.5 mg l−1) and thidiazuron (TDZ; 0.1, 0.3 and 0.5 mg l−1) in combination with 5 mg l−1 IBA, fresh weight and dry weight decreased but secondary metabolite content increased. The secondary metabolite content (including
1,1-diphenyl-2-picrylhydrazyl activity) increased more in TDZ-treated than in kinetin-treated roots. Antioxidative enzymes
such as catalase (CAT) and guaiacol peroxidase (G-POD), which play important roles in plant defense, also increased. A strong
decrease in ascorbate peroxidase activity resulted in a high accumulation of hydrogen peroxide. This indicates that adventitious
roots can grow under stress conditions with induced CAT and G-POD activities and higher accumulations of secondary metabolites.
These results suggest that 5 mg l−1 IBA supplementation is useful for growth and secondary metabolite production in adventitious roots of M. citrifolia. 相似文献
8.
The present study describes the potential of in vitro grown adventitious roots of Hypericum perforatum L. commonly known as St. John’s wort at low nutrient and auxin levels in the liquid medium for micropropagation. Roots were
regenerated from shoot-derived callus on MS medium containing 4.0 mg l−1 Indole-3 acetic acid (IAA). IAA and Indole-3 butyric acid (IBA) were equally effective for the induction of roots from shoot
cultures. Half strength MS medium containing 1.0 mg l−1 IAA was most found suitable for culturing roots in liquid medium. A total biomass of 4.13 ± 0.67 g comprising 226 ± 34.4
shoots and shoot buds along with roots was obtained per culture starting with 200 mg roots inoculum. Pretreatment with kinetin
(2.0 mg l−1) enhanced the shoot multiplication. Shoots proliferated profusely from excised roots in static liquid medium supported with
glass bead matrix. Growtek™ vessel was found suitable and cost effective system for high throughput plantlet production. In vitro grown roots regardless
of their source of origin were an excellent and easy to handle source of explant for aseptic production of plantlets without
loosing the morphogenetic potential over the generations. The plants exhibited 84–99% similarity among themselves through
RAPD. The in vitro shoots produced can either be multiplied or rooted perpetually, and alternatively they can also be explored
for the in vitro production of hypericin and hyperforin. 相似文献
9.
Kusampudi Shilpa Chinnasamy Selvakkumar Arun Kumar Senthil Baddireddi Subhadra Lakshmi 《Plant Cell, Tissue and Organ Culture》2010,101(1):105-109
Young leaf explants of Ocimum sanctum L. incubated on solidified Murashige and Skoog (MS) medium supplemented with 2 mg l−1 1-naphthaleneacetic acid (NAA) and 0.2 mg l−1 kinetin (Kn) developed rhizogenic callus. When these were subcultured onto MS medium supplemented with 1.5 mg l−1 2, 4-dichlorophenoxyacetic acid (2, 4-D) and 0.5 mg l−1 NAA, friable rhizogenic callus was observed. Upon transfer of this friable callus onto liquid MS medium containing 4 mg l−1 NAA and 1.3 mg l−1 6-benzyladnine (BA) under continuous agitation at 90 rpm and 16 h photoperiod, roots with an optimum dry weight of 1,460 mg l−1 were obtained. An ethyl acetate extract of these roots exhibited 1, 1–diphenyl-2-picrylhydrazyl (DPPH) radical scavenging
activity. 相似文献
10.
Mashitha Pise Jaishree Rudra Sunita Bundale Deovrat Begde Nandita Nashikkar Avinash Upadhyay 《In vitro cellular & developmental biology. Plant》2012,48(1):85-91
Asparagus racemosus is an important monocot medicinal plant that is in great demand for its steroidal saponins called shatavarins. This study
was initiated to optimize the conditions for production of shatavarins in cell cultures of A. racemosus in a modified Murashige and Skoog (MS) medium supplemented with six different combinations of growth regulators. Biomass
accumulation was correlated with saponin production over a 30-d culture cycle. Biomass and saponin accumulation patterns were
dependent on combinations of growth regulators and the pH of the medium. Maximum levels of saponin and biomass accumulation
were recorded on day 25 of the culture cycle within a pH range of 3.4 to 5.6. Total saponin produced by the in vitro cultures was 20-fold higher than amounts produced by cultivated plants. Saponin accumulation was not a biomass-associated
phenomenon; cultures which showed the highest biomass accumulation were not the highest saponin accumulators. Maximum biomass
(28.30 ± 0.29 g l−1) and maximum levels of shatavarin IV(11.48 ± 0.61 mg g−1) accumulation was found using a medium containing 2.0 mg l−1 2,4-D, 2 g l−1 casein hydrolysate and 0.005% pectinase. The highest levels of sarsapogenin, secreted and intracellular (4.02 ± 0.09 mg g−1), accumulated using a medium containing 1.0 mg l−1 NAA, 1.0 mg l−1 2,4-D, 0.5 mg l−1 BAP, 2 g l−1 casein hydrolysate and 0.005% pectinase, after 25 d. Shatavarins were secreted into the medium and can be isolated easily
for further purification. 相似文献
11.
A. Śliwińska O. Olszowska M. Furmanowa A. Nosov 《In vitro cellular & developmental biology. Plant》2008,44(2):69-77
Efficient plant regeneration through somatic embryogenesis was achieved in Polyscias filicifolia. Embryogenic calluses were induced on Murashige and Skoog (MS) basal medium supplemented with 0.5 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.0 mg l−1 benzylaminopurine (BAP; type I callus) and on MS medium with 2.0 mg l−1 2,4-D and 0.01 mg l−1 kinetin (type II callus) from leaf explants of a 2-yr-old plant. Primary somatic embryos (PSEs) developed after four passages
of suspension culture established from embryogenic callus when cultured in liquid half-strength MS medium (1/2 MS) without
growth regulators. PSEs in the cotyledonary stage were multiplied by adventitious embryogenesis. Single secondary somatic
embryos (SSEs) or their clusters developed at the base of PSE hypocotyls and regenerated into plantlets in a one-step process
on plant growth regulator-free 1/2 MS medium. Low sucrose concentration of 15 g l−1 promoted development of normal SSEs. All SSEs regenerated into single, well-rooted plantlets on a Nitsch and Nitsch medium
supplemented with 0.5 mg l−1 kinetin, 0.1 mg l−1 indole-3-butyric acid, and 10 mg l−1 adenine sulfate. Subsequent two subculture cycles on the same medium were necessary to obtain plantlets sufficiency developed
to allow successful transfer to the soil. Rooted plantlets were established in a peat mixture with 90% survival, with the
plants showing normal morphological characteristics. 相似文献
12.
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
13.
Xi-Hua Cui Hosakatte Niranjana Murthy Chun-Hua Wu Kee-Yoeup Paek 《Plant Cell, Tissue and Organ Culture》2010,103(1):7-14
In this study, we investigated the influence of initial sucrose concentration on the accumulation of biomass, phenols, flavonoids,
chlorogenic acid, and hypericin in adventitious root cultures of Hypericum perforatum L. Cultures were initiated in shake flasks by using half-strength Murashige and Skoog (MS) medium, 1.0 mg l−1 indolebutyric acid (IBA), 0.1 m g l−1 kinetin, and different concentrations 0, 1, 3, 5, 7, or 9% in w/v) of sucrose and were maintained in darkness. The medium
supplemented with 3% (w/v) sucrose resulted in the optimum biomass accumulation, but higher sucrose concentrations (5, 7,
and 9%) inhibited biomass accumulation due to the relatively higher osmotic pressure. However, the amount of total phenols,
flavonoids, chlorogenic acid, and total hypericin was increased with the roots grown in the medium supplemented with 5, 7,
and 9% (w/v) sucrose. The antioxidant potential of methanolic extract [1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic
acid; ABTS) radical scavenging activities] of H. perforatum adventitious roots was also assessed and correlated with the metabolite accumulation. Cultures maintained with higher initial
sucrose concentration (5, 7, and 9% w/v) showed increased accumulation of phenols, flavonoids, chlorogenic acid, and total
hypericin, and this might be due to the osmotic stress at elevated sucrose concentrations. To verify the effect of osmotic
stress on lipid peroxidation, the levels of hydrogen peroxide (H2O2), malondialdehyde (MDA), and proline were determined in the adventitious roots and the results revealed a marked increase
in the concentrations of these compounds. These results suggest that optimal adventitious root biomass could be achieved in
the MS medium with 3% (w/v) sucrose and increased sucrose concentration resulted in osmotic stress and, in turn, induces the
accumulation of secondary metabolites. 相似文献
14.
Xi-Hua Cui Hosakatte Niranjana Murthy Chun-Hua Wu Kee-Yoeup Paek 《In vitro cellular & developmental biology. Plant》2010,46(5):437-444
The present study investigated the effect of nitrogen source (NH4+; NO3−) at different concentrations on the accumulation of biomass and secondary metabolites in adventitious root cultures of Hypericum perforatum L. Cultures were initiated in shake flasks by using half-strength Murashige and Skoog (MS) medium with B5 vitamins, 1.0 mg l−1 indole-3-butyric acid, 0.1 mg l−1 kinetin, 3% (w/v) sucrose, and different ratios of ammonium and nitrate (0:30, 5:25, 10:20, 15:15, 20:10, 25:5, and 30:0 mM, using NH4Cl and KNO3). The cultures were maintained in darkness. The medium supplemented with 5:25 (mM) NH4+/NO3− resulted in the optimum accumulation of biomass and total phenols and flavonoids. The antioxidant potential of a methanolic
extract, measured as the 1, 1-diphenyl-2-picrylhydrazyl and 2, 2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) radical
scavenging activities, of H. perforatum adventitious roots showed that antioxidant activity was high from root extracts that were grown on higher concentrations
of NO3− nitrogen (15, 20, and 25 mM). Further, assessment of hydrogen peroxide (H2O2) and malondialdehyde content of the root extracts revealed that cultures supplemented with higher levels of NO3− nitrogen (15–30 mM) were under oxidative stress, which boosted the levels of secondary metabolites in the adventitious roots.
These results suggest that optimal adventitious root biomass could be achieved with the supplementation of cultures with 5:25
ratios of MS nitrogen sources. 相似文献
15.
N. Irvani M. Solouki M. Omidi A. R. Zare S. Shahnazi 《Plant Cell, Tissue and Organ Culture》2010,100(3):293-299
Dorema ammoniacum D. Don. (Apiaceae), a native medicinal plant in Iran, is classified as a vulnerable species. Root, hypocotyl, and cotyledon
segments were cultured on Murashige and Skoog (MS) (1962) medium supplemented with either 2,4-dichlorophenyoxyacetic acid
(2,4-D) or naphathalene acetic acid (NAA), at 0–2 mg l−1, alone or in combination with either benzyladenine (BA) or kinetin (KN), at 0–2 mg l−1 for callus induction. The best response (100%) was observed from root segments on MS medium containing 1 mg l−1 NAA and 2 mg l−1 BA. The calli derived from various explants were subcultured on MS medium supplemented with BA (1–4 mg l−1) alone or in combination with NAA or indole-3-butyric acid (IBA), at 0.2 or 0.5 mg l−1 for shoot induction. Calli derived from hypocotyl segments showed significantly higher frequency of plantlet regeneration
and number of plantlets than the calli derived from root and cotyledon segments. Therefore, MS medium supplemented with 2 mg l−1 BA and 0.2 mg l−1 IBA produced the highest frequency of shoot regeneration (87.3%) in hypocotyl-derived callus. The optimal medium for rooting
contained 2.5 mg l−1 IBA on which 87.03% of the regenerated shoots developed roots with an average number of 5.2 roots per shoots within 30 days.
These plantlets were hardened and transferred to the soil. The described method can be successfully employed for the large-scale
multiplication and conservation of germplasm this plant. 相似文献
16.
Guang-Zhe Lin Xiao-Mei Zhao Soon-Kwan Hong Yu-Ji Lian 《Plant Cell, Tissue and Organ Culture》2011,106(1):93-103
We have developed a system for the in vitro regeneration of pasqueflowers (Pulsatilla koreana Nakai). The system was based on somatic embryogenesis and shoot organogenesis. Over a growth period of 6 weeks, multiple
shoots were initiated from leaf, petiole, and pedicel explants on Murashige and Skoog (MS) medium containing 0.5 mg l−1 indole-3-acetic acid (IAA) and zeatin (Zn), kinetin (Kin), or 6-benzyladenine (BA). We achieved 100% of adventitious shoot
induced when petiole and pedicel explants were cultured on MS, 0.5–2.0 mg l−1 Zn, and 0.5 mg l−1 IAA. Somatic embryos developed from the explants and generated shoots on MS medium containing 0.25 mg l−1 Zn and 0.5 mg l−1 IAA. Globular and heart-shaped stages of somatic embryos were observed. Histological studies have revealed the stages of
development of somatic embryos. For propagation and growth, the regenerated shoots from organogenic or embryogenic calluses
were transferred to MS medium containing either (1) 1.5 mg l−1 Zn and 0.05 mg l−1 IAA or (2) 1.0 mg l−1 BA and 0.05 mg l−1 IAA. After the length of the shoots reached 3 cm, the shoots initiated by organogenesis as well as those initiated by somatic
embryogenesis were transferred to the root induction medium. After 2 months of culture in half-strength MS with 1.5 mg l−1 α-naphthalene acetic acid (NAA), the rooting ratio was 93%. Finally, the rooted plantlets were acclimatized in a mixture
of mountain soil and perlite. 相似文献
17.
A simple protocol for direct shoot organogenesis and plant regeneration in Lessertia frutescens using hypocotyl and cotyledon segments is reported. l-canavanine content in the derived shoots is also quantified. Media containing different concentrations and combinations of
the cytokinins kinetin (K) and benzyladenine (BA) were tested for shoot induction potential. The best shoot regeneration rate
(83%) was obtained from hypocotyl segments cultured in Murashige and Skoog (MS) medium supplemented with 1 mg l−1 K; these hypocotyls also produced the largest number of shoots per explant (3.5) and the longest shoots per explant (13.3
mm). The best shoot regeneration rate (46%) using cotyledons as explant material was obtained in MS medium supplemented with
1 mg l−1 K and 1 mg l−1 BA or with 5 mg l−1 K and 0.5 mg l−1 BA. The highest number of cotyledon-derived shoots (1.5) was obtained in MS medium containing 2 mg l−1 K and 0.5 mg l−1 BA, and the longest cotyledon-derived shoots (6.1 mm) were obtained in MS medium containing 1 mg l−1 K and 0.5 mg l−1 BA. Shoots derived from hypocotyls cultured on media containing 1 mg l−1 K contained the highest quantity of l-canavanine (1.42 mg g−1) relative to the control (0.52 mg g−1). Shoots derived from cotyledons cultured on media containing 2 mg l−1 K contained the highest quantity of l-canavanine (2.07 mg g−1) compared to the control. Scanning electron microscopy revealed that shoots regenerated directly from the wounded epidermal
tissue, although callus formation was observed in most cultures. Young shoot clusters proliferated into healthy adventitious
shoots that were subsequently transferred directly onto rooting medium (MS medium containing 4 mg l−1 indole-3-butyric acid), eliminating the need for an additional multiplication or elongation phase. The in vitro plants were
successfully acclimatized in a growth chamber, achieving an 85% survival rate. 相似文献
18.
An efficient regeneration protocol for genetic transformation was developed from the leaves of an 11-year-old Phtinia × fraseri “Red Robin” tree. A high frequency of adventitious buds (88.63 ± 1.38%) and the highest maximum mean number of adventitious
buds per explant (4.65 ± 0.48) were obtained in light conditions on Murashige–Skoog (MS) medium containing 2 mg l−1 benzyladenine (BA) and 0.2 mg l−1 α-Naphthaleneacetic acid (NAA). After preculturing for 6 days, over 95% of the shoots successfully rooted on 1/2 MS medium
supplemented with 0.3 mg l−1 indole-3-butyric acid (IBA) within 3 weeks. For genetic transformation, two crucial parameters (30 mg l−1 kanamycin for selection and 30-min suspension time) were optimized. Taken together, a reliable transformation system with
an efficiency of more than 5% was established. This genetic transformation protocol can be utilized for further genetic manipulation
of the Photinia tree. 相似文献
19.
Qi Zhao Chuanfang Wu Wenguo Wang Shu Yuan Jinku Bao Fang Chen 《Plant Cell, Tissue and Organ Culture》2009,99(3):269-275
Polygonatum cyrtonema Hua. lectins (PCLs) were extracted from plantlets regenerated from rhizome explants of P. cyrtonema. Rhizome explants demonstrated a high frequency of callus induction (72.5%) and adventitious shoots differentiation (83.7%)
on Murashige Skoog (MS) medium supplemented with 2.0 mg l−1 2,4-dichlorophenoxyacetic acid and 1.0 mg l−1 6-benzyladenine. The adventitious shoots could root readily on 1/2 MS medium + 0.5 mg l−1 α-naphthaleneacetic acid and regenerate plantlets with a survival rate of 75.0%. Regenerated rhizomes were freeze-dried,
macerated and prepared for total RNAs and proteins extraction. The PCL gene was cloned and its expression level was measured by RT-PCR. Western blot using a lectin-specific antibody revealed a
similar amount in regenerated rhizomes compared to wild rhizomes, Furthermore, lectin derived from regenerated rhizomes retained
its ability to haemagglutinate rabbit blood cells. 相似文献
20.
Kailash Choudhary M. Singh M. S. Rathore N. S. Shekhawat 《Plant biotechnology reports》2009,3(3):205-211
An efficient in vitro regeneration protocol for moth bean [Vigna aconitifolia (Jacq.) Marechal] via somatic embryogenesis has been developed. Embryogenic callus cultures were established from the cotyledonary
node as explant on semi-solid Murashige and Skoog (MS) medium supplemented with 0.75 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.5 mg l−1 6-benzylaminopurine (BA) and with various additives (50 mg l−1 ascorbic acid and 25 mg l−1 each of adenine sulphate, citric acid and l-arginine). Numerous somatic embryos differentiated on MS basal nutrient medium supplemented with 0.25 mg l−1 2,4-D and 0.5 mg l−1 of kinetin (Kin). Sustained cell division resulted in the formation of cell aggregates, which progressed to the globular-
and heart-shaped somatic embryos and then, if they differentiated properly, to the torpedo shape and cotyledonary stages.
The transfer of embryos onto fresh MS basal medium containing 0.2 mg l−1 BA and 2.0 mg l−1 gibberellic acid enabled the embryos to achieve complete maturation and germination. More than 80% of somatic embryos were
converted into true-to-type fertile plants. In vitro-regenerated plantlets with well-developed roots were successfully hardened
in a greenhouse and established in soil. 相似文献