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1.
Meiru Li Hongqing Li Xiaoying Hu Xiaoping Pan Guojiang Wu 《Plant Cell, Tissue and Organ Culture》2010,102(3):321-327
Zoysia tenuifolia Willd. ex Trin. is one of the most popularly cultivated turfgrass. This is the first report of successful plant regeneration
and genetic transformation protocols for Z. tenuifolia using Agrobacterium tumefaciens. Initial calli was induced from stem nodes incubated on a Murashige and Skoog (1962) (MS) medium supplemented with 2 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 1 mg l−1 6-benzyladenine (BA), with a frequency of 53%. Compact calli were selected and subcultured monthly on the fresh medium. Sixty-nine
percent of the calli could be induced to regenerate plantlets when the calli incubated on a MS medium supplemented with 0.2 mg l−1 BA under darkness. For genetic transformation, calli were incubated with A. tumefaciens strain EHA105 harboring the binary vector pCAMBIA 1301 which contains the hpt gene as a selectable marker for hygromycin resistance and an intron-containing β-glucuronidase gene (gus-int) as a reporter gene. Following co-cultivation, about 12% of the callus explants produced hygromycin resistant calli on
MS medium supplemented with 2 mg l−1 2,4-D, 1 mg l−1 BA, 50 mg l−1 hygromycin, 500 mg l−1 cefotaxime after 8 weeks. Shoots were regenerated following transfer of the resistant calli to shoot induction medium containing
0.2 mg l−1 BA, 50 mg l−1 hygromycin, and 250 mg l−1 cefotaxime, and about 46% of the resistant calli differentiated into shoots. Finally, all the resistant shoots were rooted
on 1/2 MS media supplemented with 50 mg l−1 hygromycin, 250 mg l−1 cefotaxime. The transgenic nature of the transformants was demonstrated by the detection of β-glucuronidase activity in the primary transformants and by PCR and Southern hybridization analysis. About 5% of the total
inoculated callus explants produced transgenic plants after approximately 5 months. The procedure described will be useful
for both, the introduction of desired genes into Z. tenuifolia and the molecular analysis of gene function. 相似文献
2.
Aline Vieira Santos Maria de Fátima Arrigoni-Blank Arie Fitzgerald Blank Leandro Eugênio Cardamone Diniz Roberta Miranda Pereira Fernandes 《Plant Cell, Tissue and Organ Culture》2011,107(1):35-43
Patchouli is an aromatic shrub of commercial interest because its essential oil is rich in patchoulol. This study aimed to
evaluate the effect of growth regulators on callus production, analyze the essential oil production in calli and evaluate
metabolic differences between callus, in vitro grown-plantlets and greenhouse-grown plants in three different accessions of
patchouli. Calli were induced from leaf explants on media supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) in combination
with 6-benzyladenine (BA). The largest size calli from different accessions were obtained in the presence of the two plant
growth regulators (PGRs). For accession POG014, presence of 0.022 mg l−1 2,4-D plus 0.022 mg l−1 BA were optimum. For accession POG021, presence of 0.110 mg l−1 2,4-D plus 0.022 mg l−1 of BA induced the largest callus, whereas for accession POG002, 0.022 mg l−1 2,4-D and 0.225 mg l−1 BA, as well as 0.11 mg l−1 2,4-D and 0.022 mg l−1 BA promoted the development of largest callus. Among all accessions, peroxidase activity was highest in organogenic calli
of accession POG014, whereas, polyphenol oxidase activity was highest in in vitro-grown plantlets of accession POG021. Biochemical
variables differed significantly among the treatments, with the exception of total sugar levels. The highest concentrations
of total sugars were observed in the calli and in vitro-grown plantlets of POG014 and POG021. Essential oils were not detected
in callus tissues. 相似文献
3.
Meiru Li Hongqing Li Xiaoying Hu Xiaoping Pan Guojiang Wu 《Plant Cell, Tissue and Organ Culture》2011,106(3):363-371
Saussurea involucrata is a valuable traditional Chinese medicinal herb. This is the first report of a successful genetic transformation protocol
for S. involucrata using Agrobacterium tumefaciens. Leaf explants were incubated with A. tumefaciens strain EHA105 harboring the binary vector pCAMBIA 1301, which contains the hpt gene as a selectable marker for hygromycin resistance and an intron-containing β-glucuronidase gene as a reporter gene. Following
co-cultivation, about 23.7% of the explants produced hygromycin-resistant calli on MS basal medium (Murashige and Skoog in
Physiol Plant 15: 473–497, 1962) supplemented with 1 mg l−1 benzyladenine (BA), 0.1 mg l−1 α-naphthaleneacetic acid (NAA), 0.1 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D), 20 mg l−1 hygromycin, and 500 mg l−1 cefotaxime. Shoots were regenerated following transfer of the resistant calli to shoot induction medium containing 1.5 mg l−1 BA, 0.1 mg l−1 NAA, 0.25 mg l−1 gibberellic acid (GA3), 20 mg l−1 hygromycin, and 250 mg l−1 cefotaxime, and about 67.5% of the resistant calli differentiated into shoots. Finally, 80% of the hygromycin-resistant shoots
rooted on MS media supplemented with 0.2 mg l−1 NAA, 20 mg l−1 hygromycin, and 250 mg l−1 cefotaxime. The transgenic nature of the transformants was demonstrated by detection of β-glucuronidase activity in the primary
transformants and by Southern blot hybridization analysis. About 16% of the total inoculated leaf explants produced transgenic
plants after approximately 5 months. Using this optimized transformation system, a rice ortholog of the Arabidopsis FLOWERING LOCUS T gene, Hd3a, was transferred into S. involucrata. Introduction of this gene caused an early-flowering phenotype in S. involucrata. 相似文献
4.
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
5.
Bishnoi U.S. Jain R.K. Gupta K.R. Chowdhury V.K. Chowdhury J.B. 《Plant Cell, Tissue and Organ Culture》2000,61(2):153-159
An improved procedure has been developed for high frequency androgenesis in indica × Basmati rice hybrids using a liquid culture
medium. Anthers from fourteen genotypes comprising of indica × Basmati rice F1 hybrids, F2 plants and the parental rice cultivars, were floated in liquid RZM, N6M, and Heh5M media. Anther culture frequencies (percentage
of anthers forming calluses) in most of the genotypes were significantly higher in RZM medium (16–75%) compared to those obtained
in N6M (7–29%) and Heh5M (7–41%) media. Agarose (1.0% w/v)-solidified MSR1 medium containing 3.0% (w/v) maltose, 1 mg l−1 kinetin, 1 mg l−1 6-benzyladenine (BA) and 0.5 mg l−1α-naphthalene acetic acid (NAA) induced green shoot regeneration at high frequencies compared to the medium (MSR2) lacking
BA. In all the genotypes, microspore calluses initiated in RZM medium regenerated green shoots with over tenfold higher frequencies
compared to the calluses initiated in other two media. High plant regeneration frequencies (up to 270 green plants/1000 anthers)
were obtained from microspore-derived calluses of some of the F1 hybrids (Gobind × Basmati 370, Gobind × Taraori Basmati) and F2 plants (Gobind × Basmati 370, Gobind × Taraori Basmati, HKR86-3 × Taraori Basmati) as compared to their actual parents. Cytological
analysis of the root tips of the progeny seedlings of the microspore-derived plants revealed haploids at a frequency of about
50%; 22% of the microspore- derived plants had > 5% spikelet fertility and were diploid. Use of RZM liquid and MSR1 media,
respectively for anther culture and plant regeneration resulted in several fold increase in the recovery of green plants from
recalcitrant indica × Basmati rice F1 hybrids/F2 plants which were comparable to those reported for japonica rice varieties/hybrids leading to the improved feasibility of
using doubled haploids in genetic, breeding and mapping research with indica rice.
This revised version was published online in June 2006 with corrections to the Cover Date. 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.
This study demonstrates the morphogenic potential of pulvinus, an important organ situated at the base of the petiole or rachis
of leguminous plants. Plant regeneration via pulvinus-derived calli of Caesalpinia bonduc has been achieved. Organogenic calli have been derived from the explant 45 days after culture on Murashige and Skoog (MS)
medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) alone or in combination with 6-benzylaminopurine (BA). Optimum
callus induction (100%) occurred when the pulvini were cultured on MS medium fortified with 6 mg l−1 2,4-D and 1 mg l−1 BA. The highest shoot induction was obtained when the calli were transferred to MS medium supplemented with 5 mg l−1 BA and 1 mg l−1 indole-3-acetic acid (IAA). On this medium, 87% cultures responded with an average number of 4.2 shoots per culture. The
maximum root induction from the regenerated shoots was observed on half strength MS medium containing 6 mg l−1 indole-3-butyric acid (IBA). Here 100% shoots rooted with a mean number of 6.3 roots per shoot. The regenerated plantlets
were acclimatized and subsequently showed normal growth. This efficient protocol will be helpful for propagating elite clones
on a mass scale and could be utilized for genetic transformation study. 相似文献
8.
Morteza Shabannejad Mamaghani Mohammad Hassan Assareh Mansoor Omidi Mohammad Matinizadeh Abbas Ghamari-Zare Shokofeh Shahrzad Massih Forootan 《Plant Growth Regulation》2009,59(3):199-205
The green twigs of 1-year-old Eucalyptus microtheca F. Muell seedlings were cultured on modified MS medium, supplemented with α-naphthalene acetic acid (NAA) and kinetin (Kin)
hormones at 12 different concentrations. After 4 weeks, the combination of 1 mg l−1 NAA + 1 mg l−1 Kin induced the highest number of axillary shoots. Meanwhile, embryogenic calli were observed in media containing 4 mg l−1 NAA + 0.5 mg l−1 Kin, without any regeneration. The hormone treatments were followed by subculturing the twigs in different levels of thidiazuron
(TDZ). The combination of 1 mg l−1 NAA + 1 mg l−1 Kin together with 0.01 mg l−1 TDZ resulted in an increase of direct shoot, while higher amounts of TDZ led to adventitious shoot induction. Somatic embryogenesis
was observed in the treatment containing 0.01 mg l−1 TDZ + 4 mg l−1 NAA + 0.5 mg l−1Kin. The peroxidase (POD) band patterns in regenerated plantlets were investigated in order to determine the effect of different
levels of TDZ on loci synthesis. A dimer locus, a tetramer locus and two epigenetic bands (a new band for NAA + Kin and the
other for TDZ) were observed in the POD profiles. In case of low (0.01 mg l−1 and 0.1 mg l−1) levels of TDZ, one heterozygote allele was disappeared from dimer locus, while at higher TDZ levels, the dimer locus lost
its stability and tetramer locus showed a high activity. Thus, POD allele patterns seems to be a feasible marker for different
types of regeneration. 相似文献
9.
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
10.
This work presents a rapid and reliable micropropagation method for a Lycaste hybrid using a field-grown axillary bud culture system. Intact buds (2–4 mm in length) were excised from a mature pseudobulb
and were cultured in half-strength MS basal medium, which was supplemented with 0.5 mg l−1 benzyladenine (BA), 1.0 mg l−1 thidiazuron (TDZ) and 2% (w/v) sucrose. After 2 months, the calli exhibited vigorous growth and eventually turned green,
forming protocorm-like bodies (PLBs) originating in the surface of each callus. The results of this work reveal that the combination
of 0.5 mg l−1 BA and 1.0 mg l−1 TDZ treatments was highly effective in indirectly multiplying shoots from callus-PLB mixed explants, which yielded up to
400 shoots in the fourth time subcultures (within 24 weeks). Histological observations showed the apical meristem of adventitious
bud is based on a longitudinal section of a callus sample. Histological and scanning electronic microscopy also indicated
that PLBs derived from calli could be regarded as organogenesis but not somatic embryogenesis. Shoots with a length of around
2–3 cm generated in vitro were excised and cultured in MS medium supplemented with 0.5 mg l−1 IBA exhibited the best rooting response (78.3%), and an average of 1.8 roots per explant was produced within 4 weeks. 相似文献
11.
In vitro regeneration protocol for Anethum graveolens (Apiaceae) was developed using leaf explants. MS basal medium used in experiments was augmented with various hormones for
caulogenic and rhizogenic response. The optimum callus induction (100%) was obtained by leaf explants on MS media fortified
with BA (0.5 mg l−1) singly and in combination with NAA (0.1 and 0.2 mg l−1). BA at 0.5 mg l−1, KN at 1.0 mg l−1 and NAA at 0.1 mg l−1 induced highest number of multiple shoots (10.0 ± 0.25) per explant and they also showed in vitro flowering within 3 weeks
of culture. Influence of adenine sulfate on regeneration frequency of callus was evaluated. The highest frequency of rooting
(100%) with 6.0 ± 0.25 roots per explants was obtained in one-fourth strength MS medium supplemented with 1/4 MS + IBA 0.5 mg l−1 within 4 weeks of transfer to the rooting medium. In vitro flowering (35%) was obtained with MS fortified with BA alone and
also in combination with KN and NAA (5.3 ± 0.42 flowers per explants). In vitro flowering response was tested with different
carbohydrates (fructose, glucose, mannose and sorbitol) and optimized. Hardening was successfully attained under controlled
conditions inside the plant tissue culture room. The proposed method could effectively be applied for the conservation and
clonal propagation to meet the pharmaceutical demands of this medicinally important species. 相似文献
12.
A. Othmani C. Bayoudh N. Drira M. Marrakchi M. Trifi 《Plant Cell, Tissue and Organ Culture》2009,97(1):71-79
Plant regeneration through somatic embryogenesis from young leaf explants (5–10 mm long) adjacent to the apex of 5–6 year
old offshoots of Tunisian date palm (Phœnix dactylifera L.), cultivar Boufeggous was successfully achieved. Factors affecting embryogenic callus initiation, including plant growth
regulators and explant size, were investigated. The highest induction frequencies of embryogenic calli occurred after 6–7 months
on MS medium supplemented with 10 mg l−1 2,4-D and 0.3 mg l−1 activated charcoal. The subculture of these calli onto maintenance medium resulted in the formation of proembryos. Fine chopping
and partial desiccation (6 and 12 h) of embryogenic calli with proembryos prior to transfer to MS medium supplemented with
1 mg l−1 ABA stimulated the rapid maturation of somatic embryos. Maturated somatic embryo yield per 0.5 g FW of embryogenic callus
was 51 embryos with an average maturation time of 55 days. This was increased to 422 with finely chopped callus, and 124 and
306 embryos following 6 and 12 h desiccation treatments, respectively. The average time to maturation for these 3 treatments
was 35, 43 and 38 days, respectively. Subsequent substitution of ABA in MS medium with 1 mg l−1 NAA resulted in the germination and conversion of 81% of the somatic embryos into plantlets with normal roots and shoots.
The growth of regenerated somatic plants was also monitored in the field. 相似文献
13.
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
14.
The effect of increasing concentration of polycyclic aromatic hydrocarbon (PAH) fluoranthene (FLT; 0.1, 1 and 5 mg l−1) on the growth, ethylene production and anatomy of stems of 21-day-old pea plants cultivated in vitro in MS medium, with
or without FLT, enriched with 0.1 mg l−1 indole-3-acetic acid (IAA) or with combination of 0.1 mg l−1 IAA + 0.1 mg l−1 N6-benzyladenine (BA) were investigated. The low concentration of 0.1 mg l−1 FLT, in both IAA- and IAA + BA-treated plants, significantly stimulated the growth of pea callus, while higher concentrations
1 mg l−1 and especially 5 mg l−1 FLT significantly inhibited it. Pea shoots were significantly influenced only after application of 5 mg l−1 FLT in IAA treatment. Significantly increased production of ethylene was found in IAA + BA treatments in all concentrations
of FLT, whereas in IAA treatments in 1 and 5 mg l−1 FLT. The lysigenous aerenchyma formation in the cortex of pea stems significantly increased in all FLT treatments and its
highest proportion was found in plants exposed to 1 mg l−1 FLT. 相似文献
15.
Plant regeneration was achieved through direct and indirect somatic embryogenesis in Eucalyptus camaldulensis. Callus was induced from mature zygotic embryos and from cotyledon explants collected from 10, 15, 25, and 30-day-old seedlings
cultured on Murashige and Skoog (MS) basal medium supplemented with different concentrations of naphthaleneacetic acid (NAA).
Maximum callus induction from mature zygotic embryos was obtained on MS basal medium containing 1 mg l−1 NAA. The frequency of callus development varied based on the age of the cotyledon explants 10-day-old explants giving highest
percentage on MS basal medium supplemented with 1 mg l−1 NAA. Callus obtained from mature zygotic embryos gave highest frequency of somatic embryogenesis on MS basal medium containing
0.5 mg l−1 benzyladenine (BA) and 0.1 mg l−1 NAA. Separate age wise culture of the calli, obtained from cotyledons of different ages cultured separately, revealed high
somatic embryogenic potential on callus from 10-day-old cotyledons. Direct somatic embryogenesis too was obtained from hypocotyl
explants without an intervening callus phase on MS basal medium containing 0.5 mg l−1 BA. The effects of abscisic acid (ABA), sucrose, and different strengths of MS medium on somatic embryo maturation and germination
were also investigated. Number of mature somatic embryos increased with lower concentrations (0–1 mg l−1) of ABA while no significant differences were observed at higher concentrations (2–5 mg l−1) of ABA. Compared to basal medium containing lower concentrations of sucrose (1%), the MS medium supplemented with higher
levels of sucrose (4%) showed significantly lower frequency of mature somatic embryos. Basal medium without any dilution gave
the highest number of immature embryos. However, the number of mature embryos was high at higher medium dilutions. 相似文献
16.
Yantree Devi Sankar-Thomas Katja Saare-Surminski Reinhard Lieberei 《Plant Cell, Tissue and Organ Culture》2008,95(2):163-173
The present study describes a protocol for plant regeneration via somatic embryogenesis in temporary immersion system (TIS)
for Camptotheca acuminata. Somatic embryos were induced by culturing hypocotyl segments from 14-day-old in vitro grown C. acuminata seedlings in TIS. Hypocotyl segments were placed in culture vessels modified with a mechanical device to support the fixation
of explants. Cultures were maintained under a 16 h photoperiod with a light intensity of 60 μmol m−2 s−1 PPF at 25 ± 1°C. After 16 weeks of incubation embryogenic calli were formed above the edge of the mechanical device in the
basal Murashige and Skoog (MS) medium containing 35 g l−1 sucrose and without hormonal supplementation. For plantlet regeneration, somatic embryos at cotyledonary stage were cultured
in three different concentrations of 6-benzylamino-purine (0.5, 1.0 and 1.5 mg l−1 BAP) and in plant growth regulator (PGR) free medium. In general, 0.5 mg l−1 BAP was found to be the most effective concentration for growth and development of Camptotheca embryos in TIS. Conversion of somatic embryos into plantlets was also successfully achieved on sterile substrates moistened
with 0.5 mg l−1 BAP. Plantlets derived from cotyledonary embryos were rooted in vitro with 0.5 mg l−1 indole-3-butyric acid (IBA) before transfer to ex vitro conditions. 相似文献
17.
Jatropha curcas L. (Physic nut) is a commercially important non-edible oil seed crop known for its use as an alternate source of biodiesel.
In order to investigate the morphogenic potential of immature embryo, explants from four developmental stages were cultured
on medium supplemented with combinations of auxins and cytokinins. It was found that the size of embryo is critical for the
establishment of callus. Immature embryos (1.1–1.5 cm) obtained from the fruits 6 weeks after pollination showed a good response
of morphogenic callus induction (85.7%) and subsequent plant regeneration (70%) with the maximum number of plantlets (4.7/explant)
on Murashige and Skoog’s (MS) medium supplemented with IBA (0.5 mg l−1) and BA (1.0 mg l−1). The above medium when supplemented with growth adjuvants such as 100 mg l−1 casein hydrolysate + 200 mg l−1
l-glutamine + 8.0 mg l−1 CuSO4 resulted in an even higher frequency of callus induction (100%). Plant regeneration (90%) with the maximum number of plantlets
(10/explant) was achieved on MS medium supplemented with 500 mg l−1 polyvinyl pyrrolidone + 30 mg l−1 citric acid + 1 mg l−1 BA + 0.5 mg l−1 Kn + 0.25 mg l−1 IBA. It was observed that plantlet regeneration could occur either through organogenesis of morphogenic callus or via multiplication
of pre-existing meristem in immature embryos. The age of immature embryos and addition of a combination of growth adjuvants
to the culture medium appear to be critical for obtaining high regeneration rates. Well-developed shoots rooted on half-strength
MS medium supplemented with 0.5 mg l−1 IBA and 342 mg l−1 trehalose. The rooted plants after acclimatization were successfully transferred to the field in different agro-climatic
zones in India. This protocol has been successfully evaluated on five elite lines of J. curcas. 相似文献
18.
The cell cultures of Cayratia trifolia (Vitaceae) a tropical lianas, were maintained in Murashige and Skoog’s medium containing 0.25 mg l−1 naphthalene acetic acid, 0.2 mg l−1 kinetin and 250 mg l−1 casein hydrolysate. Cell suspension cultures of C. trifolia accumulate stilbenes (piceid, resveratrol, viniferin, ampelopsin) which on addition of 0.1–0.5 mg l−1 morphactin in the medium containing naphthalene acetic acid and kinetin declined. Morphactin or 2 isopentenyl adenine alone
at 0.1 mg l−1 concentration enhanced stilbenes which on combination markedly enhanced the yield to ~5 mg l−1 at 15th day. 相似文献
19.
Ethrel treatment enhanced isoflavonoids accumulation in cell suspension cultures of Pueraria tuberosa, a woody legume 总被引:1,自引:0,他引:1
The cell cultures of Pueraria tuberosa, a perennial leguminous lianas, were maintained in modified MS medium (KNO3 475 mg l−1, thiamine 1 mg l−1, biotin 1 mg l−1, calcium pantothenate 1 mg l−1) containing 0.1 mg l−1 2,4,5-trichloroacetic acid and 0.1 mg l−1 kinetin. Isoflavonoids (puerarin, genistin, daidzein, genistein) accumulation in cell suspension cultures was increased by
14-fold to ~12 mg l−1 after 48 h of adding 100 μM ethrel. Ethrel inhibitors (silver nitrate and silver thiosulfate) completely inhibited this effect
in the presence of ethrel and isoflavonoids were not detected in the spent medium. The increase was dose dependent and can
be explored to trigger high yield of isoflavonoids production. 相似文献
20.
In vitro regeneration and morphogenesis studies in common bean 总被引:1,自引:0,他引:1
Kingdom Kwapata Robab Sabzikar Mariam B. Sticklen James D. Kelly 《Plant Cell, Tissue and Organ Culture》2010,100(1):97-105
An efficient protocol for high frequency in vitro regeneration of multiple shoots and somatic embryos from the embryonic axis
of common bean (Phaseolus vulgaris) was developed. Ten common bean cultivars representing a wide range of diversity among current commercial market classes
were used for in vitro regeneration evaluation in our study. These cultivars were tested on 63 different media formulations
consisting of combinations of cytokinins, namely benzyladenine (BA) and thidiazuron (TDZ) at concentration levels of 0.0,
1.0, 2.5, 5.0 and 10.0 mg l−1 and auxin, namely naphthalene acetic acid (NAA) and indole-3-acetic acid (IAA) at concentration levels of 0.0, 0.05, 0.1
and 1.0 mg l−1. P. vulgaris cv. Olathe pinto bean performed the best producing over 20 multiple shoots per explant while cv. Condor black bean was the
poorest with nine multiple shoots per explant. The optimum media for regeneration of multiple shoots was 4.4 mg l−1 Murashige and Skoog (MS) containing 2.5 mg l−1 BA and 0.1 mg l−1 IAA supplemented with 30 mg l−1 silver nitrate. Adventitious shoots and somatic embryos were regenerated on 4.4 mg l−1 MS medium containing 1 mg l−1 TDZ and 0.05 mg l−1 NAA supplemented with 30 mg l−1 silver nitrate or activated charcoal. Efficient and effective rooting of plantlets was achieved by dipping the cut end base
of in vitro regenerated shoots in 1.0 mg l−1 indole-3-butyric acid (IBA) solution and culturing on media containing 4.4 mg l−1 MS supplemented by 0.1 mg l−1 IAA, NAA or IBA. 相似文献