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1.
Li-Hua Zhu Xiao-Qin Wu Hong-Ye Qu Jing Ji Jian-ren Ye 《Plant Cell, Tissue and Organ Culture》2010,102(1):121-128
A protocol was developed for the micropropagation of Pinus massoniana and mycorrhiza formation on rooted microshoots. Seedling explants were first cultured on Gresshoff and Doy (GD) medium supplemented
with 6-benzyladenine (BA) alone or in combination with α-napthaleneacetic acid (NAA) to stimulate the formation of intercotyledonary
axillary buds. The frequency of axillary bud induction was up to 97% on medium supplemented with 4.0 mg l−1 BA and 0. 2 mg l−1 NAA, and the average number of buds per explant reached up to 5.5 on medium with 4.0 mg l−1 BA and 0.1 mg l−1 NAA. Axillary buds elongated rapidly after being transferred to half-strength GD medium containing activated charcoal (0.1%
w/v). Shoot proliferation was achieved by cutting elongated shoots into stem segments and subculturing on GD medium containing
2 mg l−1 BA and 0.2 mg l−1 NAA. Root primordia were induced in 82% of shoots when transferred to half-strength GD medium containing 0.2 mg l−1 NAA. Root elongation was achieved in a hormone-free GD agar medium or a perlite substrate. Rooted plantlets were inoculated
with the mycelium of ectomycorrhizal fungus Pisolithus tinctorius and the formation of ectomycorrhiza-like structures was achieved in vitro. 相似文献
2.
Teodora Janković Branka Vinterhalter Dijana Krstić-Milošević Radomirka Nikolić Dragan Vinterhalter Slobodan Milosavljević 《Acta Physiologiae Plantarum》2011,33(4):1515-1520
Shoot cultures of Gentianella bulgarica established from seedling epicotyls were maintained on MS medium supplemented with BA 0.2 mg l−1 + NAA 0.1 mg l−1. Cultures were prone to precocious flowering requiring the use of small shoot buds for multiplication purposes. The contents
of three xanthone compounds identified as DGL, BGL, and DMB, in different plant material were determined by HPLC. The analysis
revealed that the production of xanthones was affected by different concentrations of BA in medium. Shoot cultures grown at
higher BA concentrations contained more DGL than material grown in nature. The concentrations of other two xanthones were
lower in shoot cultures than in plants from nature. The radical scavenging activity of plant extracts and xanthones was investigated
by DPPH test. Samples from plants grown in nature showed the highest activity (IC50 = 0.26 mg ml−1), while the extracts of shoot cultures grown in media with higher concentrations of BA showed moderate activities (IC50 from 1.6 to 4.4 mg ml−1). 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.
Junli Wang Jue Wang Kun Liu Xuan Xiao Weizhen Gong Yuan Lu Mingfei Liu Dongting Xu 《In vitro cellular & developmental biology. Plant》2010,46(5):445-450
An efficient micropropagation system for Hylotelephium tatarinowii (Maxim.) H. Ohba, a rare medicinal plant, has been developed. Callus induced from leaf explants placed onto Murashige and
Skoog (MS) medium with supplementation of plant growth regulators. When the concentration of 2,4-dicholorophenoxy acetic acid
was as high as 2.0 mg l−1 in combination with 0.5 mg l−1 6-benzylaminopurine (6-BAP), the callus induction rate reached 92.1%. Adventitious shoots were observed on callus exposed
to 1.0 mg l−1 6-BAP, with 81.5% frequency of shoot regeneration after 30 d. Flower buds appeared after subculture. Regenerated shoots could
flower normally in vitro. Up to 100% of the regenerated shoots formed complete plantlets on half-strength MS medium without any growth regulator, with
an average of 5.9 roots per shoot explant. Quantitative analysis of flavonoids and rutin showed that the phytochemical profile
of callus and regenerated plants was similar to that of wild plants. 相似文献
6.
Shelake Rahul Mahadev Angappan Kathithachalam Murugan Marimuthu 《In vitro cellular & developmental biology. Plant》2011,47(5):611-617
In vitro propagation has played a key role for obtaining large numbers of virus free, homogenous plants, and for breeding of plantains
and bananas (Musa spp.). Explant sources utilized for banana micropropagation include suckers, shoot tips, and floral buds. The present study
employed male floral meristems as explant material for micropropagation of hill banana ecotypes (AAB) ‘Virupakshi’ and ‘Sirumalai.’
Immature male floral buds were collected from healthy plants from hill banana growing areas. Exposure of explants to ethyl
alcohol (70%, v/v) for 30 s, then mercuric chloride (0.1%, w/v) for 30 s, followed by three independent rinses of 5 min each in autoclaved, double-distilled water satisfactorily reduced
the contamination. Male floral bud explants were cultured on Murashige and Skoog (MS) basal medium supplemented with different
combinations of 6-benzylaminopurine (BAP), coconut water, naphthaleneacetic acid, gibberellic acid, and additional supplements.
MS medium supplemented with 5 mg l−1 BAP and coconut water (15%) was the most efficient media for shoot initiation and multiple shoot formation (15 shoots from
a single part of a floral bud). The best response for shoot elongation was obtained using the combination of basal MS, 5 mg l−1 BAP, 1 mg l−1 naphthaleneacetic acid and 1.5 mg l−1 gibberellic acid. Regenerated shoots were rooted in basal MS medium within 15–20 d. The rooted plantlets were transferred
to a soil mixture and maintained at a temperature of 25 ± 2°C for 10 d and then at room temperature (30–32°C) for 2 wk, before
transferring to a greenhouse. The regenerated plantlets showed 100% survival. 相似文献
7.
Soha E. Mostafa Nabila S. Karam Rida A. Shibli Feras Q. Alali 《Plant Cell, Tissue and Organ Culture》2010,103(1):111-121
A protocol for micropropagation of Arbutus andrachne from seeds was developed. Results indicated that none of the seeds cultured on Murashige and Skoog (MS) medium, with or without
plant growth regulators (PGRs), germinated. Seeds soaked in 250 mg l−1 gibberellic acid (GA3) at 4°C for 3 days, then cultured on water-agar medium containing 2.0 mg l−1 GA3 exhibited 80–100% germination and developed into usable seedlings. Shoot proliferation was tested on MS or B5 medium containing
different concentrations of cytokinin. No shoot proliferation was observed on PGR-free medium. Proliferation was more successful
on MS than on B5 medium. On both media, the most successful proliferation was obtained using zeatin as a cytokinin type. Rooting
was tested on MS medium containing different concentrations of auxin. Rooting failed on PGR-free medium and on medium containing
indole-3-acetic acid (IAA), 0.25 or 0.5 mg l−1 indole-3-butyric acid (IBA), or 0.25, 0.5 or 2.0 mg l−1 α-naphthaleneacetic acid (NAA). Rooting (40%) was most successful with 1.0 mg l−1 NAA. Rooted plantlets exhibited 80% survival in all mixtures of peatmoss and perlite, and acclimatized plants were successfully
grown in the greenhouse. Quantitative analysis of arbutin performed on in vivo and in vitro leaves using high-performance
liquid chromatography (HPLC) revealed that in vivo leaves contained higher arbutin content (0.3–0.81% w/w) than in vitro leaves
(0.09% w/w). The highest yield of arbutin in vivo was detected in leaves collected in August, and the lowest yield in leaves
collected in December. 相似文献
8.
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
9.
María del Pilar Nicasio-Torres Juan Carlos Erazo-Gómez Francisco Cruz-Sosa 《Acta Physiologiae Plantarum》2009,31(5):905-914
Two Cecropia species (Cecropia obtusifolia and C. peltata), known as guarumbo, are employed in Mexican traditional medicine to treat diabetes mellitus; the leaves of both species contain phenolic bioactive
compounds such as chlorogenic acid (CA) and isoorientine (ISO), which have been attributed with hypoglycemic, hypolipidemic,
and antioxidant properties. An in vitro propagation protocol was developed from existing apical bud meristem from C. obtusifolia seedlings; the shoot generation was induced on Murashige and Skoog (MS) medium supplemented with varying concentrations of
6-benzylaminopurine and kinetin (Kn) combined with either α-naphtalene acetic acid (NAA) or indole-3-acetic acid (IAA) auxins.
Best morphogenetic response was developed with Kn 26.64 μM combined with either NAA or IAA 0.57 μM, respectively; likewise,
C. peltata-seedling apical buds were subjected to these best selected treatments. Cecropia obtusifolia and C. peltata shoots were rooted in growth regulator-free half-strength MS medium, and regenerated whole plants were adapted successfully
under greenhouse conditions and field. Leaves from both Cecropia-micropropagated plants produced the phenolic compounds CA and ISO, with highest concentrations in leaves from 18-month C. obtusifolia (12.28 ± 7.06 mg g−1 dry leaves of CA and 8.30 ± 2.70 mg g−1 dry leaves of ISO) growth in the field. Our results offer a protocol of apical-bud use for multiplication and curative-property
conservation of the two previously mentioned important Mexican medicinal plants. 相似文献
10.
The halophyte Leymus chinensis (Trin.) is a perennial rhizome grass (tribe Gramineae) that is widely distributed in China, Mongolia and Siberia, where it
is produced as a forage product. In this report, we establish a highly reproducible plant regeneration system through somatic
embryogenesis. Two explants, mature seeds and leaf base segments were used; these parts displayed different responses to combinations
of growth factors that affect embryogenic callus induction, callus type optimization and plant regeneration. The highest callus
induction frequency was obtained on Murashige and Skoog (MS) medium supplemented with 2.0 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D) in the presence of 5.0 mg l−1
l-glutamic acid. The inclusion of 5.0 mg l−1
l-glutamic acid was found to significantly promote primary callus induction, embryogenic callus formation and callus status
improvement. Subculturing on maintenance medium for 1–2 months before plant regeneration was found to be essential for the
optimization of callus type and the maturation of embryogenic callus. Callus relative water content and growth rate were simultaneously
investigated during callus maintenance, and found to possibly be related to callus type. Shoots were differentiated from the
embryogenic callus on the optimal medium with MS salts containing 0.2–0.5 mg l−1 α-naphthalene acetic acid (NAA), 2.0 mg l−1 kinetin (Kn) and 2.0 g l−1 casamino acids in 71.0 and 69.2% of wild-type (WT) and Jisheng No.1 (JS) plants, respectively. Plant regeneration was variable
depending on NAA levels, and the addition of casamino acids stimulated the maturation of embryogenic callus and plant regeneration.
Transferring callus with shoots onto half-strength MS medium resulted in rooting within 1 week. The growth of regenerated
plants was also surveyed in the field. This is the first report of plant regeneration through somatic embryogenesis from mature
seeds and leaf base segments of L. chinensis. 相似文献
11.
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
12.
Saba Ambreen Memon Xilin Hou Bo Zhu Joseph N. Wolukau 《Acta Physiologiae Plantarum》2009,31(6):1191-1196
The objective of the present work was selection of cultivar and suitable medium for regenerating shoots from leaf segments
of non-heading Chinese cabbage. We evaluated six types of supplemented media with 2.0, 5.0 and 10.0 mg l−1 6-BA; 1.0 and 2.0 mg l−1 TDZ; 0.1, 0.3, 0.5, 0.8 and 1.0 mg l−1NAA; 3.0, 5.0 and 7.5 mg l−1AgNO3; 0.01 mg l−1 2–4, D and 4.0 mg l−1 KT for shoot regeneration and six cultivars “Sanchidaye”, “Liuchuandasuomian”, “Qingyou 4”, “Liangbaiye”, “AiKang 5” and
“Hanxiao F3”, furthermore for root formation three types of supplemented media with 0.2, 0.3, 0.5 mg l−1 NAA, and for survival rate two types of base media: turf + vermiculite + manure (1:2:0.2) and soil + vermiculite (1:2). Culturing
leaf segments on MS medium supplemented with 2 mg l−1 TDZ; 0.5 mg l−1 NAA and 7.5 mg l−1 AgNO3 gave the highest number of shoots per leaf segment (66) while roots were best formed on the medium supplemented with 0.2 mg l−1 NAA. Survival rate was highest (61.6%) in the turf: vermiculite: manure (1:2:0.2) medium. The highest percentage of responding
leaf segments, number of shoots per leaf segment, rooting percentage and survival rate were observed in “Liuchuandasuomian”.
The plantlets were transferred to the soil and grown into mature plants in pots. These results could be used for preliminary
selections of cultivars to transfer disease resistance (Bt) gene through agrobacterium in non-heading Chinese cabbage. 相似文献
13.
Bilal Haider Abbasi Mubarak Ali Khan Tariq Mahmood Mushtaq Ahmad Muhammad Fayyaz Chaudhary Mir Ajab Khan 《Plant Cell, Tissue and Organ Culture》2010,101(3):371-376
The morphogenic potential and free-radical scavenging activity of the medicinal plant, Silybum marianum L. (milk thistle) were investigated. Callus development and shoot organogenesis were induced from leaf explants of wild-grown
plants incubated on media supplemented with different plant growth regulators (PGRs). The highest frequency of callus induction
was observed on explants incubated on Murashige and Skoog (MS) medium supplemented with 5.0 mg l−1 6-benzyladenine (BA) after 20 days of culture. Subsequent transfer of callogenic explants onto MS medium supplemented with
2.0 mg l−1 gibberellic acid (GA3) and 1.0 mg l−1 α-naphthaleneacetic acid (NAA) resulted in 25.5 ± 2.0 shoots per culture flask after 30 days following culture. Moreover,
when shoots were transferred to an elongation medium, the longest shoots were observed on MS medium supplemented with 0.5 mg l−1 BA and 1.0 mg l−1 NAA, and these shoots were rooted on a PGR-free MS basal medium. Assay of antioxidant activity of in vitro and in vivo grown
tissues revealed that significantly higher antioxidant activity was observed in callus than all other regenerated tissues
and wild-grown plants. 相似文献
14.
The role of different growth regulators in callus induction, shoot regeneration, floral induction and chlorophyll content
of the obligatory parasitic plant Cuscuta
reflexa has been studied. Callus development was excellent from the nodal part of the shoot explants in modified Murashige and Skoog
(MMS) media supplemented with 2 mg L−1 benzyl adenine (MMS1c). Supplementation of 2 mg L−1 naphthalene acetic acid (NAA) along with MMS1c (MMS2c) was responsible for estimable shoot induction and development in callus.
2,4-Dichloro acetic acid (2,4-D) played a crucial role in the floral induction of C. reflexa in vitro. MMS supplemented with 2 mg L−1 NAA and 2 mg L−1 2,4-D (MMS3b) supported floral induction after shooting in vitro. MMS supplemented with 3 mg L−1 2,4-D (MMS4a) rapidly induced flower directly from the stem explants without showing any elongation of shoot. MMS1c along
with MMS3b (MMS5a) showed callus proliferation followed by shoot elongation and floral induction. In vitro MMS5a grown plants
show a sharp increase in the chlorophyll contents. Cytokinin treatment further increases the chlorophyll level of the plant. 相似文献
15.
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
16.
Trifolium alexandrinum L. (Egyptian clover) is one of the most important forage crops in the world. Its regeneration in tissue culture has been
described in a few reports but the efficiency, accurate time scales and applicability to various genotypes of the described
procedures are uncertain. Therefore their suitability for genetic transformation is unclear. In this study, were report new
fast procedures for regeneration of Egyptian clover that are applicable to the regeneration of various genotypes (Mescawi-ahaly,
Sakha3 and Sakha4). Shoots were regenerated from intact and wounded cotyledons as well as hypocotyls of Mescawi-ahaly on naphthaleneacetic
acid/benzyladenine (NAA/BA) and naphthaleneacetic acid/thidiazuron (NAA/TDZ) media. The highest shoot regeneration frequencies
were obtained from intact cotyledons on NAA/BA (0.05 mg l−1 NAA combined with 2.0 mg l−1 BA) and NAA/TDZ (0.05 mg l−1 NAA combined with 1.0 mg l−1 TDZ) media (66.2 and 43.1% respectively) compared to 18.4 and 10.1% for wounded cotyledons on NAA/BA and NAA/TDZ respectively.
21.0% shoot regeneration frequency was observed for hypocotyls on NAA/BA (2.0 mg l−1 NAA combined with 0.5 mg l−1 BA) medium but no regeneration was obtained on NAA/TDZ medium. Rooting of the regenerated shoots was induced on indole butyric
acid (IBA: 0.24 mg l−1) or NAA (2.0 mg l−1) media where IBA medium supported significantly higher frequencies of rooting as well as survival of the whole plantlets
after transfer to soil. However, the rooting and survival frequencies also depended on the type of explant and the medium
used for shoot regeneration. The two cultivars Sakha3 and Sakha4 were regenerated using the culture conditions optimized for
Mescawi-ahaly with comparable efficiencies, indicating that the described procedure is not genotype dependent. The time scale
of whole plantlet regeneration ranged from 7.5 weeks for intact and wounded cotyledons to 10 weeks for hypocotyl explants. 相似文献
17.
Athip Sakunphueak Pharkphoom Panichayupakaranant 《Plant Cell, Tissue and Organ Culture》2010,102(1):9-15
The effects of type of explant (leaves and roots), donor plants, and plant growth regulators on naphthoquinone (NQ) production
of Impatiens balsamina L. root cultures were evaluated. The root cultures were initiated in liquid Gamborg’s B5 medium supplemented with 0.1 mg l−1 α-naphthaleneacetic acid (NAA), 0.1 mg l−1 kinetin (Kn) and 1.0 mg l−1 6-benzyladenine (BA). The present investigation indicated that the root cultures established from the leaf explants produced
higher total NQ content [1.01 ± 0.046 mg/g dry weight (DW)] than those established from the root explants (0.62 ± 0.023 mg/g
DW). The leaf explants of four I. balsamina strains including white flower plant (IbW), pink flower plant (IbP), violet flower plant (IbV) and red flower plant (IbR)
were used to establish the root cultures. Based on HPLC analysis, IbP strain produced the highest total NQ content (3.39 ± 0.072 mg/g
DW), while IbR strain produced the lowest one (1.45 ± 0.055 mg/g DW). The root cultures established from the IbP explant were
capable of producing higher content of total NQs (2.76 ± 0.093 mg/g DW) than those established from the other strains. The
results suggest that the tissue cultures initiated from the high-yielding donor plants should be capable of producing higher
content of secondary compounds than those initiated from low-yielding donor plants. In addition, plant growth regulator manipulation
exhibited that a combination of 0.1 mg l−1 NAA, 1.0 mg l−1 Kn and 2.0 mg l−1 BA is capable of increasing NQ production (2.97 ± 0.072 mg/g DW) in I. balsamina root cultures. 相似文献
18.
Tatjana Vujović Đurđina Ružić Radosav Cerović Gordana Šurlan Momirović 《Plant Growth Regulation》2010,61(3):265-275
Adventitious shoot regeneration from leaves of blackberry cultivar Čačanska Bestrna was examined using 30 different combinations
of treatment. Young, fully expanded leaves taken from in vitro proliferating shoots were cultured on Murashige and Skoog (MS)
medium containing either N
6-benzylaminopurine (BAP) (2.0 mg l−1) or thidiazuron (TDZ) (1.0 and 2.0 mg l−1) alone, or either of them combined with indol-3-butyric acid (IBA), α-naphthalene acetic acid (NAA) and 2,4-dichlorophenoxyacetic
acid (2,4-D) at different concentrations (0.1, 1.0 and 2.0 mg l−1). Indirect shoot formation was observed in 12 different treatments, though the efficacy varied greatly among types and concentrations
of plant growth regulators. TDZ at 1.0 mg l−1, applied either alone or combined with IBA, was significantly more effective than BAP in inducing shoot regeneration. The
highest regeneration rate (41.66%) was obtained on medium containing TDZ alone. Cytological, flow cytometry and isozyme analyses
were used for screening of genetic stability/instability in regenerants. Cytological study, based on chromosome counts in
root tip meristems, and flow cytometry analysis indicated that adventitious shoots of Čačanska Bestrna are tetraploid with
2n = 4x = 28 as well as those derived from axillary buds. However, a study conducted on the peroxidase patterns of the different
blackberry regenerating lines showed differences between some lines and control plants (in vivo plants and micropropagated
plants). These differences were visible with 3,3′,5,5′-tetramethylbenzidine (TMBZ) as hydrogenous donor for peroxidase detection. 相似文献
19.
Saikat Gantait Nirmal Mandal Somnath Bhattacharyya Prakash Kanti Das 《In vitro cellular & developmental biology. Plant》2010,46(6):537-548
A novel, efficient, and simple protocol was developed on in vitro mass propagation and acclimatization of Gerbera jamesonii Bolus cv. Sciella, an ornamental plant with attractive flowers. Shoot tip was used as the primary explant for in vitro establishment in which Murashige and Skoog (MS) medium supplemented with a low level of NAA (0.5 mg l−1) and BAP (1.5 mg l−1) promoted earliest axillary bud initiation within 5 d in 91.6% of the inoculants. Five axillary buds were initiated from
a single explant within 13 d after inoculation. A very high rate of shoot multiplication (14 shoots per inoculated axillary
bud) and proliferation was achieved when MS medium was fortified with a relatively higher level of BAP (2 mg l−1) and 60 mg l−1 ADS within 27 d of multiple shoot culture. A maximum number of well-developed roots per plant was observed in MS medium with
0.5 mg l−1 IAA in the next 26 d. In the easy low-cost acclimatization process of 20 d, a combination of sand, soil, cow urine, and tea
leaves extract (1:1:1:1; v/v) ensured 95% survival rate. Sixty-one well-acclimatized plants were obtained from a single shoot tip within 86 d. The sustained
multiple shoot culture for 15 mo paved the way toward the conservation of genetic resources as well as beneficial economics.
The clonal fidelity study of micropropagated and sustained cultured clones using ISSR primers ensured the continuous supply
of quality propagules retaining genetic uniformity. The in vitro-generated plants performed better over conventionally propagated plants in the field condition. 相似文献
20.
Ehsan Ullah Khan Xing-Zheng Fu Ji-Hong Liu 《Plant Cell, Tissue and Organ Culture》2012,109(2):383-390
In this study, attempts were made to develop a protocol for regeneration of transgenic plants via Agrobacterium tumefaciens-mediated transformation of leaf segments from ‘Valencia’ sweet orange (Citrus sinensis L. Osbeck) using gfp (green fluorescence protein) as a vital marker. Sensitivity of the leaf segments regeneration to kanamycin was evaluated,
which showed that 50 mg l−1 was the best among the tested concentrations. In addition, factors affecting the frequency of transient gfp expression were optimized, including leaf age, Agrobacterium concentration, infection time, and co-cultivation period. Adventitious shoots regenerated on medium containing Murashige
and Tucker basal medium plus 0.1 mg l−1 α-naphthaleneacetic acid (NAA), 0.5 mg l−1 6-benzyladenine (BA) and 0.5 mg l−1 kinetin (KT). The leaf segments from 3-month-old in vitro seedlings, Agrobacterium concentration at OD600 of 0.6, 10-min immersion, and co-cultivation for 3 days yielded the highest frequency of transient gfp expression, shoots regeneration response and transformation efficiency. By applying these optimized parameters we recovered
independent transformed plants at the transformation efficiency of 23.33% on selection medium (MT salts augmented with 0.5 mg l−1 BA, 0.5 mg l−1 KT, 0.1 mg l−1 NAA, 50 mg l−1 kanamycin and 250 mg l−1 cefotaxime). Expression of gfp in the leaf segments and regenerated shoots was confirmed using fluorescence microscope. Polymerase chain reaction (PCR)
analysis using gfp and nptII gene-specific primers further confirmed the integration of the transgene in the independent transgenic plants. The transformation
methodology described here may pave the way for generating transgenic plants using leaf segments as explants. 相似文献