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1.
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
2.
A simple, high frequency, and reproducible method for plant regeneration through direct organogenesis from cotyledonary leaf
explants of Jatropha curcas was developed using Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ) or 6-benzyl
aminopurine (BAP). Medium containing TDZ has greater influence on regeneration as compared to BAP. The induced shoot buds
were transferred to MS medium containing 10 μM kinetin (Kn), 4.5 μM BAP, and 5.5 μM α-naphthaleneacetic acid (NAA) for shoot
proliferation. The proliferated shoots could be elongated on MS medium supplemented with different concentrations and combinations
of BAP, indole-3-acetic acid (IAA), NAA, and indole-3-butyric acid (IBA). MS medium with 2.25 μM BAP and 8.5 μM IAA was found
to be the best combination for shoot elongation. However, significant differences in plant regeneration and shoot elongation
were observed among the genotypes studied. Rooting was achieved when the basal cut end of elongated shoots were dipped in
half strength MS liquid medium containing different concentrations and combinations of IBA, IAA, and NAA for 4 days, followed
by transfer to growth regulators free half strength MS medium supplemented 0.25 mg l−1 activated charcoal. Elongated shoot treated with 15 μM IBA, 5.7 μM IAA, and 11 μM NAA resulted in highest percent rooting.
The rooted plants could be established in soil with more than 90% survival rate. The method developed may be useful in improvement
of J. curcas through genetic modification. 相似文献
3.
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
4.
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
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.
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
7.
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
8.
Xueping Shi Xigang Dai Guofeng Liu Junwei Zhang Guogui Ning Manzhu Bao 《In vitro cellular & developmental biology. Plant》2010,46(2):117-125
An efficient protocol for secondary somatic embryogenesis in camphor tree is reported. Secondary somatic embryos (SSEs), initially
obtained from the primary embryos of a nascent embryogenic culture in 2002, were proliferated and maintained for more than
4 yr via cyclic secondary somatic embryogenesis. Throughout this period, the embryo populations retained a high level of competence
for plant regeneration. SSEs were produced on the surfaces of the cotyledons and radicular ends of maternal somatic embryos
(MSEs). Histological observations of the various stages of secondary embryo development revealed four typical stages, namely,
globular, heart-shaped, torpedo, and cotyledonary. The process of secondary embryogenesis continued in a cyclic way, with
each newly formed embryo producing a subsequent generation of secondary embryos. In order to progress developmentally beyond
proliferation cycles, cotyledonary embryos from one of embryogenic lines (L14) were cultured on Murashige and Skoog (MS) medium
with 0.1–3.0 mg l−1 abscisic acid (ABA) or 0.05–1.0 mg l−1 thidiazuron (TDZ) in darkness for 2 mo to achieve maturation. Matured embryos were then transferred to MS-based germination
medium containing either 0.1 mg l−1 TDZ, 0.2 mg l−1 indole-3-butyric acid (IBA), and 0.5 mg l−1 6-benzylaminopurine (BA) or 0.1 mg l−1 TDZ and 0.2 mg l−1 IBA and were cultured in light for germination. Over 50% of embryos matured in the presence of 0.5 mg l−1 ABA were able to germinate with shoots and poor root system. Frequencies of embryos germinating normal shoots among different
genotypes did not change significantly. A total of 93% of the shoots from the germinated embryos converted to plantlets on
half strength MS medium with 0.5 mg l−1 IBA by 3 wk. Plantlets acclimatized successfully to ex vitro conditions and developed as field-grown plants with normal appearance. 相似文献
9.
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
10.
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
11.
N. R. Nayak P. K. Chand S. P. Rath S. N. Patnaik 《In vitro cellular & developmental biology. Plant》1998,34(3):185-188
Summary An efficient procedure is outlined forin vitro regeneration of an epiphytic orchid,Cymbidium aloifolium (L.) Sw. using rhizomes developed from seeds. Murashige and Skoog's (1962) medium (MS) containing indole-3-acetic acid (IAA),
indole-3-butyric acid (IBA), or 1-naphthaleneacetic acid (NAA) stimulated growth and proliferation of rhizomes with NAA being
most effective at 5.0 mg.l−1 (27.0 μM). Shoot bud differentiation was induced in the apical portions of the rhizomes on MS medium containing kinetin (Kn) or N6-benzyladenine (BA). The highest frequency of shoot regeneration (91.5%) and the maximum number of shoot buds formed (3.5
shoots/rhizome) were recorded with BA at 1.0 mg.l−1 (4.4 μM). NAA (0.1 mg.l−1, 0.54 μM), whenever added to the medium in conjunction with BA (1.0 mg.l−1, 4.4 μM), slightly enhanced the frequency of shoot bud regeneration (92.6%) and the number of shoot buds formed (5.2 shoots/rhizome).
Moreover, an NAA-BA combination induced rooting in regenerated shoots thereby producing complete plantlets in one step. Shoots
developed on cytokinin-supplemented medium were rooted on MS containing NAA at 1.0 mg.l−1 (5.4 μM). Regenerated plantlets were acclimated and eventually established in a garden. 相似文献
12.
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
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.
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
15.
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
16.
Salema Valencio Francis Sunil Kumar Senapati Gyana Ranjan Rout 《In vitro cellular & developmental biology. Plant》2007,43(2):140-143
An efficient protocol was developed for in vitro clonal propagation of Curculigo orchioides Gaertn. through apical meristem culture. Multiple shoots were induced from apical meristems grown on Murashige and Skoog
(MS) basal medium supplemented with 1.5 mg l−1 6-benzyladenine (BA), 100 mg l−1 adenine sulfate (Ads) and 3% sucrose. Inclusion of indole-3-butyric acid (IBA) or indole-3-acetic acid (IAA) in the culture
medium improved the formation of multiple shoots. The highest frequency of multiplication was obtained on MS medium supplemented
with 1.5 mg l−1 BA, 100 mg l−1 Ads, 0.25 mg l−1 IBA and 3% sucrose. Rooting was achieved upon transferring the micro-shoots to half-strength MS medium containing 0.25 mg
l−1 IBA and 2% sucrose. Micropropagtated plantlets were hardened in the greenhouse and successfully established in soil. 相似文献
17.
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
18.
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
19.
Buhara Yucesan Arzu Ucar Turker Ekrem Gurel 《Plant Cell, Tissue and Organ Culture》2007,91(3):243-250
A very efficient and rapid regeneration system via multiple shoot formation was developed for Cichorium intybus L. when leaf explants excised from sterile seedlings were cultured on medium supplemented with different concentrations and
combinations of various plant growth regulators. In a comparison of leaf lamina and petiole explants, lamina explants produced
over three times more shoots than petiole explants, with a mean of 7.5 shoots compared to 2.4. Of the combinations of KIN/IAA,
KIN/NAA, BAP/IAA, or BAP/NAA, 0.5 mg l−1 KIN combined with 0.3 mg l−1 IAA was the most effective, producing a mean of 19.7 shoots per lamina explant while the control treatment involving no plant
growth regulators produced no shoots at all. When either cytokinin was used alone, BAP was found nearly twice more successful
than KIN. However, the most effective treatment of all was the combination of 0.01 mg l−1 TDZ and 1.0 mg l−1 IAA, producing as many as 35.8 shoots per lamina explant. This rate of shoot regeneration is remarkably higher than those
previously reported for C. intybus, most likely due to the highly inductive effect of TDZ, which was tested for the first time in this species. Rooting of the
shoots was readily achieved on medium containing different concentrations of IAA or IBA. IAA was more effective than IBA and
resulted in the highest frequency of shoots that rooted (100%) and mean number of roots per shoot (4.2) when used at 0.5 mg l−1. Hardening off process resulted in a production of more than 80% healthy plantlets. 相似文献
20.
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