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1.
Jing Li Yang Bo Zhao Eun Soo Seong Myong Jo Kim Won Hee Kang Na Young Kim Chang Yeon Yu Cheng Hao Li 《Plant biotechnology reports》2010,4(4):261-267
We describe culture conditions for a high-efficiency in vitro regeneration system of Papaver
nudicaule through somatic embryogenesis and secondary somatic embryogenesis. The embryogenic callus induction rate was highest when
petiole explants were cultured on Murashige and Skoog (MS) medium containing 1.0 mg l−1 α-naphthaleneacetic acid (NAA) and 0.1 mg l−1 6-benzyladenine (BA) (36.7%). When transferred to plant growth regulator (PGR)-free medium, 430 somatic embryos formed asynchronously
from 90 mg of embryogenic callus in each 100-ml flask. Early-stage somatic embryos were transferred to MS medium containing
1.0 mg l−1 BA and 1.0 mg l−1 NAA to germinate at high frequency (97.6%). One-third-strength MS medium with 1.0% sucrose and 1.0 mg l−1 GA3 had the highest frequency of plantlet conversion from somatic embryos (91.2%). Over 90% of regenerated plantlets were successfully
acclimated in the greenhouse. Secondary somatic embryos were frequently induced directly when the excised hypocotyls of the
primary somatic embryos were cultured on MS medium without PGRs. Sucrose concentration significantly affected the induction
of secondary embryos. The highest induction rate (89.5) and number of secondary somatic embryos per explant (9.3) were obtained
by 1% sucrose. Most secondary embryos (87.2–94.3%) developed into the cotyledonary stage on induction medium. All cotyledonary
secondary embryos were converted into plantlets both in liquid and on semisolid 1/3-strength MS medium with 1.0% sucrose. 相似文献
2.
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
3.
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
4.
Ai Hua Chen Jing Li Yang Yu Da Niu Chuan Ping Yang Gui Feng Liu Chang Yeon Yu Cheng Hao Li 《Plant Cell, Tissue and Organ Culture》2010,102(3):357-364
We developed a new protocol for highly efficient somatic embryogenesis and plantlet conversion of Schisandra chinensis. Friable embryogenic callus was induced from cotyledonary leaves and hypocotyls of germinated zygotic embryos on Murashige
and Skoog (MS) agar medium containing 2,4-dichlorophenoxyacetic acid (2,4-D). Preculture of zygotic embryos on 2,4-D-containing
medium increased embryogenic callus induction efficiency. The highest embryogenic callus induction frequency of 56.7% was
obtained from shoot apical meristem-containing hypocotyl explants from 1-week-old germinated embryos on MS medium containing
4.0 mg l−1 2,4-D. Embryogenic callus proliferation, somatic embryo (SE) formation, and subsequent plantlet conversion occurred under
optimal culture conditions. The effects of MS medium strength, sucrose, gibberellic acid (GA3), and 6-benzyladenine (BA) on SE formation and plantlet conversion were evaluated. Low MS medium strength (1/4 to 1/2) was
necessary for SE formation, and the optimal sucrose concentration was 2.0%. Supplementing medium with GA3 negatively impacted SE formation and subsequent development. BA significantly increased the number of SEs and the plantlet
conversion capacity. One-third-strength MS medium with 1.0% sucrose and 0.5 mg l−1 BA produced the highest number of SEs (309 embryos from 9 mg embryogenic callus) and the highest frequency of plantlet conversion
from germinated SEs (52.6%). When transplanted to soil, 90% of the regenerated plants developed into normal plants. 相似文献
5.
Effects of post-harvest application of two plant growth regulators viz., gibberellic acid (GA3) and benzyl adenine (BA) with sucrose in the vase solution on cell membrane stability and vase life of gladiolus were investigated.
The vase solution treatment combinations of GA3 and BA with sucrose significantly increased the membrane stability index and enhanced the vase life as compared to the sucrose
alone treatments or the controls. Vase solution treatment of GA3 (50 mg l−1), followed by BA (50 mg l−1) with sucrose (50 g l−1) significantly increased solution uptake, fresh weight and dry weight of cut spikes. The same treatments also enhanced the
concentration of reducing and non-reducing sugars in gladioli petals 4 days after treatment (DAT). Cut spikes in vase solution
enriched with 50 mg l−1 GA3 + 50 g l−1 sucrose showed higher antioxidative enzyme activities of superoxide dismutase (SOD) and glutathione reductase (GR), lower
lipoxygenase (LOX) activity and lipid peroxidation (measured as TBARS). Petal membrane stability index was also highest in
cut spikes 6 DAT with 50 mg l−1 GA3 + 50 g l−1 sucrose vase solution. Treatment of gladiolus cut spikes with 50 mg l−1 GA3 + 50 g l−1 sucrose vase solution showed two fold increase in vase life and improved flower quality with a higher number of open flower
per spike at any one time. These results suggest that post-harvest application of GA3 (50 mg l−1) with sucrose (50 g l−1) maintains higher spike fresh and dry weight, improves anti-oxidative defence, stabilizes membrane integrity leading to a
delay in petal cell death. 相似文献
6.
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
7.
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
8.
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
9.
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
10.
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
11.
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
12.
Bo Long Alex X. Niemiera Zhi-ying Cheng Chun-lin Long 《Plant Cell, Tissue and Organ Culture》2010,101(2):151-162
The effects of seed maturity, media type, carbon source, and organic nutrient additives on seed germination, protocorm development,
and plant growth of Paphiopedilum villosum var. densissimum Z. J. Liu et S. C. Chen were investigated. Micropropagation frequency was enhanced through the use of 200-day-old seed, Knudson
C (KC) medium, and the presence of both glucose and coconut milk in the medium. The effects of various plant growth regulators
on the frequency of shoot organogenesis in four Paphiopedilum species were also investigated. Explants of P. villosum var. densissimum and P. insigne (Lindl.) Stein incubated in the presence of 5 mg l−1 6-benzyladenine (BA) with 0.5 mg l−1 α-naphthalene acetic acid (NAA) and 0.2 mg l−1 BA with 0.1 mg l−1 NAA, respectively, showed a twofold increase in the frequency of shoot organogenesis. For explants of P. bellatulum (Rchb. f.) Stein and P. armeniacum S. C. Chen et F. Y. Liu, the combination of 5.5 mg l−1 BA with 0.5 mg l−1 NAA and 4 mg l−1 BA with 0.1 mg l−1 NAA, respectively, resulted in the highest frequencies of shoot organogenesis. 相似文献
13.
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
14.
Elena Palomo-Ríos Araceli Barceló-Muñoz José A. Mercado Fernando Pliego-Alfaro 《Plant Cell, Tissue and Organ Culture》2012,109(2):201-211
Key factors influencing the efficiency of transformation of embryogenic cultures, induced from immature zygotic embryos, of
avocado cv. ‘Duke 7’ were evaluated. Initially, the sensitivity of somatic embryos to the antibiotics kanamycin, used for
selection, carbenicillin, cefotaxime and timentin, all used for elimination of Agrobacterium cells, were evaluated. Isolated globular somatic embryos were more sensitive to kanamycin than embryogenic masses, and 25 mg l−1 kanamycin completely restricted callus proliferation. Cefotaxime at 500 mg l−1 partially inhibited proliferation of embryogenic cultures, while both carbenicillin and timentin did not affect callus growth.
For genetic transformation, somatic embryos were infected with A. tumefaciens containing the pBINUbiGUSint plasmid. After 2 days, the embryos were transferred to selection medium supplemented with 50 mg l−1 kanamycin and 250 mg l−1 timentin for 2 months. Then, kanamycin level was increased to 100 mg l−1 for two additional months. The A. tumefaciens strain AGL1 yielded higher transformation rates, 6%, than EHA105 or LBA4404, 1.2%. The percentage of kanamycin resistant
calli obtained was significantly influenced by the embryogenic line used as source of explants. Genetic transformation was
confirmed by PCR and Southern blot analysis. A significant improvement in the germination rate was obtained when transgenic
embryos were cultured in liquid MS medium with 4.44 μM BA and 2.89 μM GA3 for 3 days in a roller drum and later transferred to the same medium gelled with 7 g l−1 agar. Plants from five independent transgenic lines were acclimated and grown in the greenhouse, being phenotipically similar
to control plants. 相似文献
15.
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
16.
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
17.
Iyyakkannu Sivanesan Mi Young Lim Byoung Ryong Jeong 《Plant Cell, Tissue and Organ Culture》2011,107(2):365-369
A simple and efficient protocol was developed for somatic embryogenesis from leaf and petiole explants of Campanula punctata Lam. var. rubriflora Makino. Somatic embryos (SE) were obtained with greater frequency from petiole explants than from leaf explants when cultured
on Murashige and Skoog (MS) medium supplemented with 2.0 mg L−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.0 mg L−1 6-benzyladenine (BA). On this medium, a mean number of 19.5 and 31.2 SE were developed per leaf and petiole explants, respectively.
Embryos were induced both light and dark conditions but culturing the explants 2 weeks in the dark followed by 3 weeks under
light resulted in high frequency of embryo formation. Globular embryos germinated best on MS medium supplemented with 0.3%
(w/v) activated charcoal (AC) and 1.0 mg L−1 GA3. The germinated plantlets grew further on MS medium containing 0.3% AC. Plantlets were successfully acclimatized in the greenhouse
with 94% survival rate. This is the first report on induction of somatic embryogenesis in this genus and also has implications
for genetic transformation, and mass clonal propagation. 相似文献
18.
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
19.
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
20.
The influence of increasing concentrations (0.1, 1.0 and 5.0 mg l−1) of fluoranthene (FLT) on growth, endogenous abscisic acid (ABA) level and primary photosynthetic processes in 21-day-old
pea plants (Pisum sativum L.) in vitro was investigated. Murashige and Skoog’s (MS) medium, with or without FLT, was enriched with indole-3-acetic
acid (IAA; 0.1 mg l−1) or a combination of IAA (0.1 mg l−1) plus N6-benzyladenine (BA; 0.1 mg l−1). The level of endogenous ABA significantly increased with increasing FLT concentrations in the presence of both IAA and
IAA plus BA. An increased level of endogenous ABA was observed in plants treated with IAA alone. The growth of shoot, callus
and the content of photosynthetic pigments (chlorophyll a and b, carotenoids), in both IAA- and IAA plus BA-treated plants, were significantly stimulated by FLT at its lowest concentration
(0.1 mg l−1) assayed in this study. However, FLT at higher concentrations (1.0 and 5.0 mg l−1) significantly inhibited all these parameters. Chlorophyll fluorescence imaging showed that FLT only at the highest concentration
(5.0 mg l−1) in the presence of IAA (0.1 mg l−1) significantly increased F0, but decreased FV/FM and ΦII. 相似文献