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1.
Payam Pour Mohammadi Ahmad Moieni Asa Ebrahimi Farzad Javidfar 《Plant Cell, Tissue and Organ Culture》2012,108(2):251-256
An efficient method for producing doubled haploid plants of oilseed rape (Brassica
napus L.) was established using in vitro colchicine treatment of haploid embryos. Haploid embryos in the cotyledonary stage were
treated with one of four colchicine concentrations (125, 250, 500 and 1,000 mg/L); for one of three treatment durations (12,
24 and 36 h) at one of the two temperatures (8 and 25°C) and were compared to control embryos (without colchicine treatment).
The number of chromosomes, seed recovery, size and density of leaf stomata, and pollen grain size from regenerated plants
were determined. No doubled haploid plants were regenerated from control embryos; however, the doubled haploid plants were
regenerated from colchicine-treated embryos. A high doubling efficiency, 64.29 and 66.66% of regenerated plants, was obtained
from 250 mg/L colchicine treatment for 24 h and 500 mg/L colchicine treatment for 36 h, respectively, at 8°C. Following 500 mg/L
colchicine treatment for 36 h, a few plants regenerated (9 plants). At the higher colchicine concentration (1,000 mg/L), no
plant regenerated. These results indicate that the colchicine treatment of embryos derived from microspores can induce efficient
chromosome doubling for the production of doubled haploid lines of oilseed rape. 相似文献
2.
In vitro induction of autotetraploids from diploid yellow passion fruit mediated by colchicine and oryzalin 总被引:2,自引:0,他引:2
M. M. Rêgo E. R. Rêgo C. H. Bruckner F. L. Finger W. C. Otoni 《Plant Cell, Tissue and Organ Culture》2011,107(3):451-459
In vitro chromosome doubling from hypocotyl segments of yellow passion fruit (Passiflora edulis Sims.) was carried out in the presence of either colchicine (0, 25, 250 and 1,250 μM) or oryzalin (0, 5, 15 and 30 μM). Murashige
and Skoog (in Physiol Plant 15:473–497, 1962)(MS)-based regeneration medium containing 250 or 1,250 μM colchicine markedly affected explant development leading to browning
and death of the hypocotyl segments. Oryzalin has similar effect to colchicine in inducing polyploidy. In vitro regenerated
autotetraploid plants induced by 25 μM colchicine or 15 μM oryzalin were further acclimatized and cultivated in hydroponics
system in greenhouse. Autotetraploids plants were more vigorous than the control diploids. The chromosome number of diploid
plants was 2n = 2x = 18, whereas that found on autotetraploid plants were 2n = 4x = 36. The stomata sizes of the autotetraploids were significantly larger than those on the diploid counterparts, while the
frequency of stomata was significantly reduced. Similarly, the chloroplast number of guard cells of autotetraploid plants
increased significantly. Two albino plants (4%) were generated in medium with 25 μM colchicine, indicating phytotoxic effects.
These plants are being grown to full maturity in order to test their potential to use in a breeding program. 相似文献
3.
Tetraploid plants were produced from leaf explants of diploid Populus pseudo-simonii by treating the leaves with colchicine. Leaf explants were cultured on MS basal medium containing 1.78 μM BA and 1.08 μM
NAA for 0, 6 and 12 days, and then transferred to the same MS liquid medium with colchicine at concentrations of 25, 50 and
75 μM for 1, 2 and 3 days. The highest efficiency of tetraploid induction was 14.6% by treating leaf explants that were pre-cultured
for 6 days and then cultured in liquid MS with 50 μM colchicine for 3 days. Flow cytometric analysis was used to screen the
tetraploids out from the regenerated plants and chromosome number counting was employed to confirm the polyploidy level. Size
and frequency of leaf stomata between diploid and tetraploid plants were demonstrated to have significant differences. 相似文献
4.
Iva Viehmannová Eloy Fernández Cusimamani Miroslav Bechyne Miroslava Vyvadilová Marie Greplová 《Plant Cell, Tissue and Organ Culture》2009,97(1):21-25
In vitro chromosome doubling was induced in octoploid (2n = 58) yacon using oryzalin and colchicine as mitotic spindle inhibitors. Nodal segments of in vitro cultured plants, 5–15 mm
long, were exposed to 20, 25, or 30 μM oryzalin and 1, 3, or 5 mM colchicine for 24 or 48 h. The resulting ploidy level was
determined by chromosome counting and flow cytometry. Out of 240 nodal segments, 3.33% hexadecaploid (2n = 116) plants were regenerated after the application of oryzalin. The greatest proportions of hexadecaploid plants (1.6%)
were obtained after 48 h of 25 μM oryzalin treatment. With the colchicine treatment, only 0.42% hexadecaploid plants were
detected and their survival rate was significantly lower in comparison with the oryzalin treatment. In hexadecaploid yacon,
significantly higher levels of saccharides were detected (FOS 13.9 g/100 g FM, fructose 4.6 g/100 g FM and glucose 2.1 g/100 g FM)
compared to the octoploid control (FOS 5.3 g/100 g FM, fructose 2.9 g/100 g FM and glucose 1.0 g/100 g FM). These results
indicate that in vitro treatment of nodal segments with oryzalin solution could be an effective procedure for chromosome doubling
and the polyploidy breeding can help to increase the FOS content in the tuberous roots. 相似文献
5.
Recovery of doubled haploid (DH) progeny from haploid melon plants for use in breeding programs requires efficient chromosome
doubling procedures. We describe improved procedures for recovery of fruits and viable seeds from parthenogenetic melon plants.
Plant regeneration from nodal explants treated with 500 mg/L colchicine for 12 h was increased from 40 to 88% by transferring
the treated explants to medium supplemented with a combination of growth regulators [5 μM IAA; 5 μM BA; 1 μM ABA; 30 μM AgNO3). Prolonged exposure (2–7 days) to colchicine inhibited regeneration from nodal explants but had less effect on shoot tip
explants. Many colchicine-treated plantlets flowered in vitro, allowing early assessment of their male fertility. Production
of stained pollen in plants from nodal explants was highest after 0.5–2 days of colchicine treatment and on plants from shoot
tips after 1–2 days. In vitro pollen counts correlated well with counts from greenhouse grown plants and with fruit set. The
fruit set rate for colchicine-treated plants with a high pollen number was 47%. Appropriate colchicine treatment and culture
of nodal explants as well as tip explants can substantially increase the number of fertile plants and DH lines recovered from
parthenogenetic melons. 相似文献
6.
Jin-Hu Wu A. Ross Ferguson Brian G. Murray 《Plant Cell, Tissue and Organ Culture》2011,106(3):503-511
Tetraploid plants were induced by colchicine treatment of in vitro leaf petiole segment cultures of five diploid Actinidia chinensis Planch. genotypes, including the commercially important, yellow-fleshed cultivar ‘Hort16A’, three female selections with red-fleshed fruit and
one male pollinizer. Petiole segments were incubated on a shoot regeneration medium for a period of 4 weeks, and subsequently
microshoots were treated with 0.05 or 0.1% colchicine. About one-third of the regenerated shoots were tetraploid following
0.05% colchicine treatment, more than with 0.1% colchicine treatment. Similar rates of tetraploid induction were achieved
with all the genotypes tested. The efficiency of induction of polyploidy depended on the interaction between the types of
in vitro culture chosen and the concentration of colchicine used. There are no previous reports of colchicine being used so
successfully to induce polyploidy in Actinidia. 相似文献
7.
Cryopreservation of shoot-tips by droplet vitrification applicable to all taro (Colocasia esculenta var. esculenta) accessions 总被引:1,自引:0,他引:1
Rajnesh Sant Bart Panis Mary Taylor Anand Tyagi 《Plant Cell, Tissue and Organ Culture》2008,92(1):107-111
The application of the droplet vitrification cryopreservation technique to taro accessions from a range of Asia Pacific countries
is presented. The optimum protocol involves excision of about 0.8 mm shoot-tips from in vitro plants, 20–40 min PVS2 exposure
at 0°C followed by rapid plunge into liquid nitrogen. Thawing was done at room temperature (25°C) and shoot-tips inoculated
on MS medium with 0.1 M sucrose regenerated into plantlets 4–6 weeks later. This new droplet vitrification protocol improved
the mean post-thaw regeneration rates to 73–100% from 21–30% obtained with the previous cryo-vial vitrification protocol. 相似文献
8.
Liliana Hernandez-Vazquez Mercedes Bonfill Elisabeth Moyano Rosa M. Cusido Arturo Navarro-Oca?a Javier Palazon 《Biotechnology letters》2010,32(2):315-319
Plant cell cultures of Centella asiatica produce small quantities of centellosides: madecassosid > asiaticosid > madecassic acid > asiatic acid. To obtain a more
efficient production system of these bioactive triterpenoid compounds, we developed a process where the substrate, α-amyrin,
was converted into centellosides by cell suspensions of C. asiatica. When α-amyrin in acetone was added at 0.01 mg/ml−1 to the culture medium, together with the permeabilizing agent DMSO, after 7 days nearly 50% had penetrated the plant cells,
of which almost 84% was transformed into centellosides. The system therefore efficiently converts α-amyrin into centellosides,
thus opening a new possibility for the production of these compounds. 相似文献
9.
Pathirana R Frew T Hedderley D Timmerman-Vaughan G Morgan E 《Plant cell reports》2011,30(6):1055-1065
Protocols were developed for the generation of haploid or doubled haploid plants from developing microspores and ovules of
Gentiana triflora. Plant regeneration was achieved using flower buds harvested at the mid to late uninucleate stages of microspore development
and then treated at 4°C for 48 h prior to culture. Anthers and ovaries were cultured on modified Nitsch and Nitsch medium
supplemented with a combination of naphthoxyacetic acid and benzylaminopurine. The explants either regenerated new plantlets
directly or produced callus that regenerated into plantlets upon transfer to basal media supplemented with benzylaminopurine.
Among seven genotypes of different ploidy levels used, 0–32.6% of cultured ovary pieces and 0–18.4% of cultured anthers regenerated
plants, with all the genotypes responding either through ovary or anther culture. Flow cytometry confirmed that 98% of regenerated
plants were either diploid or haploid. Diploid regenerants were shown to be gamete-derived by observing parental band loss
using RAPD markers. Haploid plants were propagated on a proliferation medium and then treated with oryzalin for 4 weeks before
transfer back to proliferation medium. Most of the resulting plants were diploids. Over 150 independently derived diploidised
haploid plants have been deflasked. The protocol has been successfully used to regenerate plants from developing gametes of
seven different diploid, triploid and tetraploid G. triflora genotypes. 相似文献
10.
Rocío Elizabeth Escobar-Guzmán Fernando Hernández-Godínez Octavio Martínez de la Vega Neftalí Ochoa-Alejo 《Plant Cell, Tissue and Organ Culture》2009,96(2):181-189
Eight cultivars and two accessions of Physalis ixocarpa Brot. were tested for their capacity to regenerate embryos and plants from anther cultures. Anthers were pretreated at 4°C
for 2 days and then at 35°C for 8 days in the dark while cultured on MS medium supplemented with 0.045 μM 2,4-D + 0.03 mg l−1 vitamin B12 (MS1) or with 2.26 μM 2,4-D + 0.1 mg l−1 vitamin B12 (MS3). Anther incubation proceeded under a 16 h photoperiod at 25 ± 2°C. Embryo formation occurred after 6 weeks of incubation
in these conditions. Androgenetic responses were cultivar- and culture medium-dependent, with the greatest embryo yields recorded
for cv. Chapingo (36.3%) on MS1 medium, and with wild-type 2 (21.8%) on MS3. Further development of regenerated embryos was
promoted on MS medium supplemented with 0.54 μM NAA, 8.88 μM BA and 50 mg l−1 casein hydrolysate. The regenerated plants were cultured on half-strength mineral salts MS medium with 2.85 μM IAA to enhance
root formation. Rooted plantlets were transferred to pots and acclimatized to the greenhouse. Ploidy analysis of regenerated
plants using flow cytometry revealed 72% diploids, 15% haploids and 7% triploids. AFLP analysis of regenerated plants from
anthers of a single parental plant showed different polymorphic patterns indicating their gametophytic origin. 相似文献
11.
12.
Cryopreservation of in vitro-grown shoot-tips of hop (Humulus lupulus L.) using encapsulation/dehydration 总被引:4,自引:0,他引:4
A cryopreservation procedure using encapsulation/dehydration was established for shoot-tips obtained from in vitro-grown shoots of hop. After dissection, shoot-tips were encapsulated in medium with alginate and 0.5 M sucrose. Optimal conditions consisted of preculture for 2 days in solid medium with 0.75 M sucrose, or in increasing sucrose concentrations, desiccation for 4 h with silicagel in a flow cabinet (16% water content)
followed by rapid freezing and slow thawing. Shoot recovery after freezing 60 min in liquid nitrogen was around 80%. No phenotypical
changes were observed in the recovered plants from cryopreserved shoot-tips growing in the field.
Received: 20 April 1997 / Revised: 20 February 1998 / Accepted: 1 Dezember 1998 相似文献
13.
14.
Ravinder Kaur Grewal Monika Lulsdorf Janine Croser Sergio Ochatt Albert Vandenberg Thomas D. Warkentin 《Plant cell reports》2009,28(8):1289-1299
This is the first report on the production of double-haploid chickpea embryos and regenerated plants through anther culture
using Canadian cultivar CDC Xena (kabuli) and Australian cultivar Sonali (desi). Maximum anther induction rates were 69% for
Sonali and 63% for CDC Xena. Under optimal conditions, embryo formation occurred within 15–20 days of culture initiation with
2.3 embryos produced per anther for CDC Xena and 2.0 embryos per anther for Sonali. For anther induction, the following stress
treatments were used: (1) flower clusters were treated at 4°C for 4 days, (2) anthers were subjected to electric shock treatment
of three exponentially decaying pulses of 50–400 V with 25 μF capacitance and 25 Ω resistance, (3) anthers were centrifuged
at 168–1,509g for 2–15 min, and finally (4) anthers were cultured for 4 days in high-osmotic pressure (563 mmol) liquid medium. Anthers
were then transferred to a solid embryo development medium and, 15–20 days later, embryo development was observed concomitant
with a small amount of callus growth of 0.1–3 mm. Anther-derived embryos were regenerated on plant regeneration medium. Electroporation
treatment of anthers enhanced root formation, which is often a major hurdle in legume regeneration protocols. Cytological
studies using DAPI staining showed a wide range of ploidy levels from haploid to tetraploid in 10–30-day-old calli. Flow cytometric
analysis of calli, embryos and regenerated plants showed haploid profiles and/or spontaneous doubling of the chromosomes during
early regeneration stages. 相似文献
15.
A protocol for in vitro induction of crape myrtle tetraploids using nodes from in vitro-grown shoots (2n = 48) was established. Nodal buds were excised from in vitro-grown shoots, maintained on proliferation medium containing
Murashige and Skoog medium supplemented with 4.44 μM 6-benzyladenine , 0.54 μM α-naphthaleneacetic acid, and treated with
a range of concentrations of colchicine under three different conditions. Nodal bud explants treated in liquid proliferation
medium supplemented with either 15 or 20 mM colchicine for 24 h turned necrotic and died; whereas, those cultured on solid
proliferation medium supplemented with either 125 or 250 μM colchicine for 30 days survived, but no tetraploid plants were
obtained. However, when explants were cultured in liquid proliferation medium containing 250, 500 or 750 μM colchicine for
10 days, tetraploid plants (2n = 96) were obtained. Incubation of explants in medium containing 750 μM colchicine promoted the highest frequency of survival
(40%) of explants and of recovered tetraploids (60%). Morphological and anatomical characteristics of leaves, including leaf
index, stomata size and number, stomata index (length/width), and number of chloroplasts in guard cells correlated with ploidy
of crape myrtle plants. The number of chloroplasts in guard cells of stomata was a stable and reliable marker in discriminating
plants of different ploidy levels. Chromosome counts and flow cytometry confirmed these findings. 相似文献
16.
Yong Duck Kim Ji Yun Min Won Jung Kim Young Min Kang Hyun Shik Moon Cheul Ho Lee D. Theertha Prasad Myung Suk Choi 《In vitro cellular & developmental biology. Plant》2008,44(3):203-208
An efficient in vitro plant regeneration system was established from callus culture of Scopolia parviflora. Callus was induced from adventitious roots on B5 medium with 0.45–9.04 μM 2,4-dichlorophenoxyacetic acid (2,4-D). In vitro plantlet regeneration was achieved on B5 medium supplemented with 44.38 μM benzyladenine (BA), 3% sucrose, and 0.38% gelrite.
Plantlets were transplanted to artificial soil and grown to maturity successfully in a greenhouse. The tropane alkaloid contents
in regenerated plants were analyzed using high-performance liquid chromatography (HPLC), and were found to be higher than
those of adventitious roots, native growing plants, and acclimated plants. Regenerated plants from organogenic callus cultures
produced a greater amount of tropane alkaloids. 相似文献
17.
18.
Jaindra Nath Tripathi Abubaker Muwonge Leena Tripathi 《In vitro cellular & developmental biology. Plant》2012,48(2):216-224
In banana and plantain research, it is essential to establish embryogenic cell suspensions together with a highly efficient
regeneration and transformation system. This article describes the development of an embryogenic cell suspension (ECS), regeneration,
and transformation for plantain cv. “Gonja manjaya”. ECS was established using highly proliferative multiple buds. The frequency
of embryogenic friable callus formation was 56.8% of the cultured explants. Friable embryogenic calli with many translucent
proembryos were transferred to liquid medium and homogenous cell suspensions were established within 3–4 mo. Approximately
25,000 to 30,000 plants per 1.0 ml of settled cell volume were regenerated in approximately 13–14 mo. ECSs were transformed
using Agrobacterium strain EHA 105 harboring the binary vector pBI121. About 50–60 transgenic plants per 0.5 ml settled cell volume were regenerated
on selective medium containing 100 mg l−1 kanamycin. Histochemical GUS assays using different tissues of putatively transformed plants demonstrated stable expression
of uidA gene. The presence and integration of the uidA gene were confirmed by PCR and Southern blot analysis, respectively. This is the first report showing establishment of embryogenic
cell suspension cultures and Agrobacterium-mediated transformation of an important plantain cultivar, “Gonja manjaya”. This study shows the huge potential for genetic
transformation of plantains for disease or pest resistance, as well as tolerance to abiotic factors such as drought stress
using this robust regeneration and transformation protocol. 相似文献
19.
Efficient production of doubled haploid plants through colchicine treatment of anther-derived maize callus 总被引:3,自引:0,他引:3
Y. Wan J. F. Petolino J. M. Widholm 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1989,77(6):889-892
Summary A chromosome doubling technique, involving colchicine treatment of an embryogenic, haploid callus line of maize (Zea mays L., derived through anther culture), was evaluated. Two colchicine levels (0.025% and 0.05%) and three treatment durations (24, 48, and 72 h) were used and compared to untreated controls. Chromosome counts and seed recovery from regenerated plants were determined. No doubled haploid plants were regenerated from calli without colchicine treatment. After treatment with colchicine for 24 h, the callus tissue regenerated about 50% doubled haploid plants. All of the plants regenerated from the calli treated with colchicine for 72 h were doubled haploids, except for a few tetraploid plants. No significant difference in chromosome doubling was observed between the two colchicine levels. Most of the doubled haploid plants produced viable pollen and a total of 107 of 136 doubled haploid plants produced from 1 to 256 seeds. Less extensive studies with two other genotypes gave similar results. These results demonstrate that colchicine treatment of haploid callus tissue can be a very effective and relatively easy method of obtaining a high frequency of doubled haploid plants through anther culture. 相似文献
20.
A. Othmani C. Bayoudh N. Drira M. Marrakchi M. Trifi 《Plant Cell, Tissue and Organ Culture》2009,97(1):71-79
Plant regeneration through somatic embryogenesis from young leaf explants (5–10 mm long) adjacent to the apex of 5–6 year
old offshoots of Tunisian date palm (Phœnix dactylifera L.), cultivar Boufeggous was successfully achieved. Factors affecting embryogenic callus initiation, including plant growth
regulators and explant size, were investigated. The highest induction frequencies of embryogenic calli occurred after 6–7 months
on MS medium supplemented with 10 mg l−1 2,4-D and 0.3 mg l−1 activated charcoal. The subculture of these calli onto maintenance medium resulted in the formation of proembryos. Fine chopping
and partial desiccation (6 and 12 h) of embryogenic calli with proembryos prior to transfer to MS medium supplemented with
1 mg l−1 ABA stimulated the rapid maturation of somatic embryos. Maturated somatic embryo yield per 0.5 g FW of embryogenic callus
was 51 embryos with an average maturation time of 55 days. This was increased to 422 with finely chopped callus, and 124 and
306 embryos following 6 and 12 h desiccation treatments, respectively. The average time to maturation for these 3 treatments
was 35, 43 and 38 days, respectively. Subsequent substitution of ABA in MS medium with 1 mg l−1 NAA resulted in the germination and conversion of 81% of the somatic embryos into plantlets with normal roots and shoots.
The growth of regenerated somatic plants was also monitored in the field. 相似文献