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1.
Nisar Ahmad Hina Fazal Bilal Haider Abbasi Muhammad Rashid Tariq Mahmood Nighat Fatima 《Plant Cell, Tissue and Organ Culture》2010,102(1):129-134
The organogenic potential and antioxidant potential (1, 1-diphenyl-2-picrylhydrazyl-scavenging activity) of the medicinal
plant Piper nigrum L. (black pepper) were investigated. Callus induction and shoot regeneration were induced from leaf explants of potted plants
cultured on MS medium supplemented with different plant growth regulators. The best callogenic response was observed on explants
cultured for 30 days on MS medium supplemented with either 0.5 or 1.5 mg l−1 6-benzyladenine (BA) + 1.0 mg l−1 α-naphthaleneacetic acid. Subsequent transfer of the callogenic explants onto MS medium supplemented with 1.5 mg l−1 BA + 1.0 mg l−1 gibberellic acid (GA3) achieved 85% shoot organogenesis after 30 days of culture. The maximum number (7.2) of shoots/explant was recorded for explants
cultured in MS medium supplemented with 1.0 mg l−1 BA. Following the transfer of shoots to an elongation medium, the longest shoots (5.4 cm) were observed on MS medium supplemented
with 1.0 mg l−1 BA + 1.0 mg l−1 GA3. The elongated shoots were rooted on MS medium supplemented with different concentrations of indole butyric acid. An assay
of the antioxidant potential of the in vitro-grown tissues revealed that the antioxidant activity of the regenerated shoots
was significantly higher than that of callus and the regenerated plantlets. 相似文献
2.
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
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.
Pueraria tuberosa, a medicinally important leguminous plant, yielding various isoflavanones including puerarin, is threatened, thus requiring
conservation. In this study, fresh shoot sprouts of P. tuberosa, produced by tubers, were used as explants for in vitro micropropagation. Surface-sterilized nodal shoots were incubated
on Murashige and Skoog (MS) medium supplemented with 8.88 μM benzyladenine (BA), 50 mg l−1 ascorbic acid, and 25 mg l−1 of each of citric acid and adenine sulphate. Cut ends of nodal stem segments rapidly turned brown, and cultures failed to
establish. When 100 mg l−1 ascorbic acid (ABA) and 25.0 mg l−1 polyvinyl pyrrolidone (PVP) were added to the medium, explants remained healthy, and cultures were established. Bud-breaking
of nodal stem explants resulted in multiple shoot formation. Shoots proliferated (35–40 shoots per culture vessel) on MS medium
as described above, but supplemented with 4.44 μM BA and 0.57 μM indole acetic acid (IAA) and additives. After 4–5 passages,
proliferating shoots exhibited tip-browning and decline in growth and multiplication. However, when shoots were transferred
to fresh shoot proliferation medium supplemented with 2.32 μM kinetin (Kn), sustained growth and high rate of shoot proliferation
(50–60 shoots per culture vessel) was observed. Shoots rooted when transferred to medium consisting of half- strength MS medium
with 9.84 μM indole butyric acid (IBA) and 0.02% activated charcoal. Alternatively, individual shoots were pulsed with 984.0 μM
IBA and transferred to glass bottles containing sterile and moistened soilrite. These shoots rooted ex-vitro and were acclimatized
in the greenhouse. Plants were then analyzed for puerarin content using HPLC, and leaves showed maximum accumulation of purerarin. 相似文献
5.
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
6.
Bilal Haider Abbasi Murad Khan Bin Guo Saleem Ahmed Bokhari Mir Ajab Khan 《Plant Cell, Tissue and Organ Culture》2011,105(3):337-344
The regeneration potential and antioxidative enzyme activities of economically important Brassica rapa var. turnip were evaluated. Calli were induced from leaf explants of seed-derived plantlets on Murashige and Skoog (MS) medium incorporated
with different concentrations of various plant growth regulators (PGRs). The highest leaf explant response (83%) was recorded
for 2.0 mg l−1 benzyladenine (BA) and 1.0 mg l−1 α-naphthaleneacetic acid (NAA). Subsequent subculturing of callus after 3 weeks of culture, on medium with similar compositions
of PGRs, induced shoot organogenesis. The highest shoot induction response (83%) was recorded for 5.0 mg l−1 BA after 5 weeks of transfer. However, 7.8 shoots/explant were recorded for 2.0 mg l−1 BA. The transferring of shoots to elongation medium resulted in 5.1-cm-long shoots on 10 mg l−1 of gibberellic acid (GA3). Rooted plantlets were obtained on MS medium containing different concentrations of indole butyric acid (IBA). The determination
of activities of antioxidative enzymes (superoxide dismutase [SOD], ascorbate peroxidase [APX], catalase [CAT], glutathione
peroxidase [GPX], and peroxidase [POD]) revealed involvement of these enzymes in callus formation and differentiation. All
of the activities were interlinked with each other and played significant roles in the scavenging of toxic free radicals.
This study will help in the advancement of a regeneration protocol for B. rapa var. turnip and the understanding of the functions of antioxidative enzymes in plant differentiation. 相似文献
7.
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
8.
A. V. Raghu Kuzhiyumparambil Unnikrishnan S. P. Geetha Gerald Martin Indira Balachandran 《In vitro cellular & developmental biology. Plant》2011,47(4):506-515
Embelia ribes, an important vulnerable medicinal liana, was regenerated through organogenesis and embryogenesis using leaf explants. Leaf
explants produced organogenic calluses on MS medium supplemented with 1.0 mg l−1 2,4-dichlorophenoxy acetic acid (2,4-D) and 0.5 mg l−1 6-benzylaminopurine. Shoot regeneration was obtained from organogenic calluses on MS medium containing different concentrations
of thidiazuron (TDZ) and indole-3-acetic acid (IAA). The frequency of shoot bud organogenesis was highest (23.9 shoots/explant)
in MS medium containing 0.5 mg l−1 TDZ and 0.1 mg l−1 IAA. The best result for induction of embryogenic callus was noticed in the combination of 2.0 mg l−1 TDZ and 0.5 mg l−1 2,4-D. This callus, maintained in the same medium, showed the highest differentiation of embryos (56.5%) after 6 wk of culture.
Embryos were transferred to MS medium supplemented with different concentrations of TDZ, and this facilitated conversion of
embryos into plants. After 6 wk of subculture, MS medium with 0.05 mg l−1 TDZ favored the highest percentage (52.2%) embryo conversion. As per the present protocol, 52.2% of the embryos underwent
conversion, and a mean number of 29.5 shoots per culture was obtained. Shoots developed from both types of calluses were rooted
on half-strength MS basal medium supplemented with 1.0 mg l−1 indole-3-butyric acid. HPLC-UV assay demonstrated the highest embelin content (5.33% w/w) in the embryogenic callus cultures. Embelin was isolated from embryogenic callus and was identified using IR and 1H NMR studies. 相似文献
9.
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
10.
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
11.
The influence of the basal medium and different plant growth regulators on micropropagation of nodal explants from mature
trees of lemon cultivars was investigated. Although the basal medium did not affect any of the variables, explants on DKW
medium were greener. Several combinations of 6-benzyladenine (BA) and gibberellic acid (GA) were used to optimise the proliferation
phase. The number of shoots was dependent on the BA and GA concentrations and the best results were obtained with 2 mg l−1 BA and 1 or 2 mg l−1 GA. Explants length was shorter with the higher BA concentrations and, in all genotypes, shoot length was greater with 2 mg l−1 GA. The best results for productivity (number of shoots × the average shoot length) were obtained with 2 mg l−1 BA and 2 mg l−1 GA, although explants with chlorosis and narrow leaves were observed. The presence of BA and GA in the proliferation medium
was essential for the explant multiplication but GA had a greater influence. The transfer of in vitro shoots to rooting media,
containing different concentrations of indole butyric acid (IBA) and indole acetic acid (IAA) produced complete plantlets.
Lemon shoots rooted well in all rooting combinations. The highest rooting percentages were obtained on media containing 3 mg l−1 IBA alone or IBA in combination with 1 mg l−1 IAA and on these media the highest numbers of roots were produced. The average root length was affected significantly by
the IBA and IAA concentrations. Root length was greater when only 3 mg l−1 IBA was used, and in this rooting medium explants had a better appearance, with greener and larger leaves. The success during
the acclimatisation was close to 100% and the plantlets exhibited normal growth in soil under greenhouse conditions. 相似文献
12.
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
13.
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
14.
An efficient in vitro propagation is described for Spondias mangifera Willd., a medicinally important tree, using nodal explants obtained from 4-week-old seedlings. The frequency of shoot regeneration
from seedling node was affected by various concentrations of BAP and successive transfer of mother explant. MS (Murashige
and Skoog, Physiol Plant 15:473–497, 1962) medium supplemented with 1.0 mg l−1 of 6-benzylaminopurine (BAP) was optimal for shoot multiplication. Upon this medium, highest number of shoots (about 10.6)
per explants was obtained after fourth subculture of mother explants. Half-strength MS medium containing IAA (1.0 mg l−1) was most effective for rooting of shoots. Regenerated plantlets were successfully acclimatized and transferred into soil
with 80–90% survival rate. The regenerated plants were morphologically uniform and exhibited similar growth characteristics
and vegetative morphology to the mother plants. This is the first report on micropropagation of S. mangifera, which can be applied for further genetic transformation assays and pharmaceutical purposes. 相似文献
15.
Kulkarni Anjali A. Thengane S.R. Krishnamurthy K.V. 《Plant Cell, Tissue and Organ Culture》2000,62(3):203-209
In vitro studies were initiated with Withania somnifera (L.) Dun. for rapid micropropagation of selected chemotypes using nodes, internodes, hypocotyls and embryo explants. Direct
regeneration of shoot buds was observed in MS basal medium supplemented with various concentrations of either benzyladenine
(BA) or thidiazouron (TDZ) depending on the explant. Nodal explants formed multiple shoots both from pre-existing and de novo buds on Murashige and Skoog's medium (MS) containing 0.1–5.0 mg l−1 BA and a ring of de novo shoot buds on MS medium containing 0.2 and 0.3 mg l−1 TDZ. Internodal explants formed shoot buds on MS with 1.0 and 5.0 mg l−1 BA while the hypocotyl explants gave rise to multiple shoots only on MS with 0.5 mg l−1 BA. Isolated embryos gave rise to many shoot buds on MS with 0.2 and 0.3 mg l−1 TDZ. The shoot buds elongated and rooted either on MS medium with 0.01 mg l−1 BA or on half strength MS medium lacking growth regulators, which depended upon the growth regulator used in the shoot bud
induction medium. Except for the embryo-derived plantlets, all other plantlets could be acclimatized with 100% success.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
16.
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
17.
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
18.
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
19.
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
20.
Guirong Qiao Jing Zhou Jing Jiang Yuehua Sun Luanyin Pan Honggai Song Jingmin Jiang Renying Zhuo Xiaojuan Wang Zongxiu Sun 《Plant Cell, Tissue and Organ Culture》2010,102(2):163-170
Malaxis acuminata is a terrestrial orchid that grows in shady areas of semi-evergreen to shrubby forests. It is highly valued for its medicinal
properties as dried pseudo-bulbs are important ingredients of several Ayurvedic preparations. In this study, adventitious
shoot buds were induced from internodal explants of M. acuminata grown on Murashige and Skoog (MS) medium supplemented with different concentrations of 6-benzyladenine (BA), kinetin (Kn),
and thidiazuron (TDZ). Of the three cytokinins used, TDZ at 3 mg l−1 induced the highest frequency (82%) of organogenic explants. However, all responding explants produced only a single adventitious
shoot irrespective of the type and concentration of the cytokinin. Adding 0.5 mg l−1 α naphthaleneacetic acid (NAA) to the medium enhanced adventitious shoot formation. In the presence of 3 mg l−1 TDZ and 0.5 mg l−1 NAA, frequency of organogenesis was 96% with a mean number of 6.1 shoots per explant. Prolonged culture or subculture on
the same medium did not promote further shoot production. However, transfer of these cultures to MS medium supplemented with
3 mg l−1 TDZ and 0.5 mg l−1 NAA and various concentrations of different polyamines (PAs), including spermine, spermidine, and putrescine, significantly
increased mean shoot number per explant. The highest frequency of shoot induction (100%) and mean shoot number per explant
(14.6) was observed on MS medium with 3 mg l−1 TDZ, 0.5 mg l−1 NAA, and 0.4 mM spermidine. Regenerated shoots were excised and subcultured on an elongation medium consisting of MS medium
with 3 mg l−1 BA. Moreover, the highest frequency of rooting (96%) and mean number of roots per shoot (3.3) was observed on MS medium with
4 mg l−1 indole-3-butyric acid (IBA) and 1.5 mg l−1 activated charcoal (AC). Almost 90% of rooted shoots were successfully acclimatized and established ex vitro. 相似文献