scholarly journals Obtaining calli and regenerated plants in anther cultures of pea

2014 ◽  
Vol 50 (No. 2) ◽  
pp. 123-129 ◽  
Author(s):  
S. Bobkov
Keyword(s):  
Developmental Stages ◽  
Callus Formation ◽  
Naphthaleneacetic Acid ◽  
Embryogenic Calli ◽  
Nutrient Media ◽  
Shoot Morphogenesis ◽  
Regenerated Plants ◽  
Wide Range ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Anther Cultures

Pea (Pisum sativum L.) is a species for which there is no efficient method for the recovery of haploid plants yet. This research investigated the influence of various genotypes, nutrient media, and stress treatments on callus formation, embryogenesis and plant regeneration in anther cultures of pea. A wide range of pea genotypes and nutrient media was studied. Morphogenic calli were initiated on media supplemented with &alpha;-naphthaleneacetic acid (NAA), 6-benzyladenine (BA), and 2,4-dichlorophenoxyacetic acid (2,4-D) without application of stress treatments. Embryogenic calli and embryos were regenerated on media with low sucrose content in the presence of 2,4-D or indole-3-butyric acid (IBA) after cold stress (4&deg;C) of isolated buds, alone or in combination with in&nbsp;vitro treatment of isolated anthers at higher temperatures (35&ndash;38&deg;C). The efficiency of regeneration via shoot morphogenesis on different nutrient media and the peculiarities of regeneration from embryogenic calli were investigated. Green embryogenic calli initiated on 2,4-D were able to develop through shoot morphogenesis on a medium supplemented with BA and NAA. This process led to regeneration of hypertrophic embryos at various developmental stages. The origin of regenerated plants (i.e. from microspores or somatic anther cells) was estimated using marker alleles determining morphological traits. Almost all R<sub>0</sub> regenerants derived from morphogenic calli originated from anther somatic cells.

Selectivity of auxin for induction and growth of callus from excised embryo of spring and winter wheat

1984 ◽  
Vol 62 (7) ◽  
pp. 1393-1397 ◽  
Author(s):  
M. D. Zhou ◽  
T. T. Lee
Keyword(s):  
Winter Wheat ◽  
Chinese Spring ◽  
Shoot Growth ◽  
Callus Formation ◽  
Triticum Aestivum L ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Methoxybenzoic Acid

The callus-promoting activity of most commonly known as well as some rarely tested auxins was compared with that of 2,4-dichlorophenoxyacetic acid (2,4-D) for in vitro culture of the excised embryo of spring and winter wheat (Triticum aestivum L.), cv. Chinese Spring and cv. Fredrick. Different auxins in a concentration range from 1 to 50 μM showed widely different activities. Also the two wheat cultivars responded differently to the auxins. When rapid callus formation with limited root growth was used as the basis for comparison, 2-(2-methyl-4-chlorophenoxy)propionic acid (2-MCPP), α-naphthaleneacetic acid, 3,6-dichloro-2-methoxybenzoic acid (dicamba), 4-amino-3,5,6,trichloropicolinic acid (picloram), γ-(2,4-dichlorophenoxy)butyric acid, 2,4,5-trichlorophenoxyacetic acid, and 2,4,5-trichlorophenoxypropionic acid, in the order of effectiveness, were superior to 2,4,-D for callus induction from the embryo of 'Chinese Spring,' although the concentration required was higher than that of 2,4-D. For the winter wheat 'Fredrick,' however, only picloram, dicamba, and 2-MCPP performed as well as 2,4-D. All auxins tested promoted shoot growth; 2-methyl-4-chlorophenoxypropionic acid was most effective for 'Chinese Spring,' whereas picloram was most effective for 'Fredrick.'

scholarly journals Immature embryo is the potential source for in vitro plant regeneration in Jatropha curcas

2014 ◽  
Vol 20 ◽  
pp. 125-134
Author(s):  
MR Islam ◽  
MA Bari
Keyword(s):  
Developmental Stages ◽  
Callus Formation ◽  
Immature Embryo ◽  
Basal Medium ◽  
Coconut Water ◽  
Immature Embryos ◽  
Plantlet Formation ◽  
Potential Source ◽  
Wide Range

Context: Jatropha belongs the spurge family Euphorbiaceae. Special interest mounting for its biodiesel which has created enthusiasm in cultivation of the species for oil extraction. Objectives: The study was conducted to develop the protocol for tissue and callus culture in Bangladeshi Jatropha curcus plant particularly to identify the most suitable explants for its wide scale micropropagation. Materials and Methods: Immature embryos taken from four developmental stages of fruits were cultured on growth regulator free MS liquid medium. After fifteen days of germination, elongated hypocotyls and two cotyledonary leaves were used as explants. Results: Embryo derived seedlings acted as the potential source of explants both for callus and plantlets. The immature embryo of size 0.87cm produced highest callus formation (83.33%) on MS medium supplemented with lower concentration of 2, 4-D (0.5 mg/l) and coconut water 2% (v/v). Immature embryos grown on MS basal medium supplemented with 2,4-D (0.2 mg/l, 0.5 mg/l and 1.0 mg/l) alone or in combination with coconut water 2% (v/v) exhibited a wide range of callus induction percentage (26-100%) for hypocotyls and (20 - 40%) for cotyledonary leaves. Conclusion: The age of immature embryo and addition of growth adjuvants and growth additive to the culture medium played the role in promoting better callus and plantlet formation. DOI: http://dx.doi.org/10.3329/jbs.v20i0.17726 J. bio-sci.  20:  125-134, 2012

Embryo induction and plant regeneration of Callerya speciosa (Fabaceae) through anther culture

2017 ◽  
Vol 65 (1) ◽  
pp. 80 ◽  
Author(s):  
Bilan Huang ◽  
Li Xu ◽  
Kelie Li ◽  
Yunlu Fu ◽  
Zhiying Li
Keyword(s):  
Anther Culture ◽  
Culture Medium ◽  
Basal Medium ◽  
Dichlorophenoxyacetic Acid ◽  
Anther Wall ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Anther Cultures

An in vitro protocol for Callerya speciosa (Champ.) Schot regeneration through embryogenesis was developed using the anthers as the explants. The late uninucleate stage of the microspore was optimal for the anther culture of C. speciosa. Embryonic callus was induced on a MS basal medium supplemented with 4.4 µM 6-benzylaminopurine (BA) and 9.04 µM 2,4-dichlorophenoxyacetic acid (2,4-D). Embryos were obtained on MS medium supplemented with 2.2 µM BA and 0.5 µM naphthaleneacetic acid (NAA). The highest percentage (16.7%) of embryos was achieved using the culture medium MS + 0.25 µM NAA + 1.1 µM BA. The highest percentage of embryos that developed into plants was 18.3%. However, haploid plants were not observed, which may have been due to the collection of the calli from the anther wall. The results presented here demonstrate the establishment of a highly efficient and rapid system for regenerating C. speciosa using anther cultures.

scholarly journals Somatic embryogenesis from leaf transverse thin cell layer derived-callus of Vietnamese ginseng (Panax vietnamensis Ha et Grushv.)

2016 ◽  
Vol 14 (1) ◽  
pp. 63-73
Author(s):  
Vu Thi Hien ◽  
Nguyen Phuc Huy ◽  
Bui Van The Vinh ◽  
Hoang Xuan Chien ◽  
Hoang Thanh Tung ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Cell Layer ◽  
Culture Media ◽  
Callus Formation ◽  
Basal Medium ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Transverse Thin Cell Layer ◽  
Cell Layers ◽  
2,4 Dichlorophenoxyacetic Acid

No report on plant regeneration via somatic embryogenesis of P. vietnamensis has been previously published. In the present study, somatic embryogenesis via callus formation from cultures of leaf transverse thin cell layers (tTCLs) of Vietnamese ginseng (Panax vietnamensis Ha et Grushv.) was investigated. α-naphthaleneacetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (BA) and thidiazuron (TDZ) were added separately and in combination into the culture media. Explant necrosis or low callogenesis rates were observed when 1-mm wide leaf tTCLs were cultured on media with TDZ, BA, 2,4-D or NAA. On the other hand, calli were successfully induced from the tTCL explants cultured on medium supplemented with either 2,4-D and BA or 2,4-D and TDZ. Callogenesis was observed under both light and dark conditions. The highest callogenesis rate (100%) was obtained on Murashige and Skoog (MS) basal medium supplemented with 1.0 mg l-1 2,4-D in combination with 0.1 mg l-1 TDZ in darkness after eight weeks of culture. White calli were cut into small pieces (1.0 x 1.0 cm dimension) and placed on MS media containing 1.0 mg l-1 2,4-D, 0.5 mg l-1 NAA and TDZ at various concentrations (0.01; 0.1; 0.2; and 0.5 mg l-1), and the best callus proliferation was recorded on medium containing 1.0 mg l-1 2,4-D and 0.2 mg l-1 TDZ. Somatic embryogenesis, with a success rate of 53.3% and 35 embryos per explant, was achieved when calli were subcultured onto MS medium supplemented with 1.0 mg l-1 2,4-D, 0.5 mg l-1 NAA and 0.2 mg l-1 TDZ.

scholarly journals Regeneration of Ornamental Cherry (Prunus) Taxa from Mature Stored Seed

HortScience ◽  
2000 ◽  
Vol 35 (4) ◽  
pp. 745-748 ◽  
Author(s):  
Karen E. Hokanson ◽  
Margaret R. Pooler
Keyword(s):  
Callus Formation ◽  
Adventitious Shoot ◽  
Shoot Formation ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Adventitious Shoot Regeneration ◽  
Potential System ◽  
Regeneration In Vitro ◽  
2,4 Dichlorophenoxyacetic Acid

Callus formation and adventitious shoot regeneration in vitro from mature stored seed were evaluated in eight ornamental cherry (Prunus) taxa: P. campanulata Maxim., P. maackii Rupr., P. sargentii Rehd., P. serrula Franch., P. serrulata Lindl., P. subhirtella Miq., P. virginiana L., and P. yedoensis Matsum. Several portions of the embryo (cotyledons and hypocotyl sections) and nine combinations of growth regulators (BA, 2,4-D, IBA, NAA, and TDZ) were compared. Effects of embryo portions and growth regulator treatments were generally small within taxa, but shoot formation differed among taxa. About 20% to 50% of the embryos from P. virginiana and P. serrula and ≈5% to 30% of those from P. maackii produced shoots. The other taxa generally did not produce shoots. Regeneration from mature stored seed in the responsive taxa represents a potential system for genetic transformation. Chemical names used: 6-benzyladenine (BA); 2,4-dichlorophenoxyacetic acid (2,4-D); indole-3-butyric acid (IBA); α-naphthaleneacetic acid (NAA); thidiazuron (TDZ).

Callus formation and shoot bud differentiation in anther culture of Solanum surattense

1979 ◽  
Vol 57 (22) ◽  
pp. 2524-2527 ◽  
Author(s):  
S. Sinha ◽  
R. P. Roy ◽  
K. K. Jha
Keyword(s):  
Anther Culture ◽  
Callus Formation ◽  
Low Frequency ◽  
Basal Medium ◽  
Pollen Grains ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Cytological Examination ◽  
Shoot Bud ◽  
2,4 Dichlorophenoxyacetic Acid

In anther culture of Solatium surattense, the Murashige and Skoog's medium supplemented with 2,4-dichlorophenoxyacetic acid (2.2 mg/L), indoleacetic acid or naphthaleneacetic acid (1.9 mg/L), and kinetin (2.2 mg/L) served as “callus-producing medium.” Histological and cytological observations indicated that the callus originated from the pollen grains. Synergistic action of kinetin (5.0 mg/L) and coconut milk (15%) in basal medium was able to induce differentiation of shoot buds either from the anthers directly or from the callus. Directly differentiating buds were formed by whole shoot bud morphogenesis of pollen. They were produced at a low frequency and showed presence of well-developed radicular and plumular regions. But the buds originating from callus lacked radicular ends. Root initiation in such buds was achieved by transferring them to basal medium. Cytological examination of the androgenic plantlets revealed a chromosomal series ranging from the haploid to the hexaploid with a few aneuploids.

scholarly journals Somatic embryogenesis and plant regeneration from callus cultures of Cleome rosea Vahl

2010 ◽  
Vol 53 (3) ◽  
pp. 679-686 ◽  
Author(s):  
Claudia Simões ◽  
Norma Albarello ◽  
Cátia Henriques Callado ◽  
Tatiana Carvalho de Castro ◽  
Elisabeth Mansur
Keyword(s):  
Somatic Embryogenesis ◽  
Callus Formation ◽  
Callus Cultures ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Stem Explants ◽  
Ms Medium ◽  
Fold Reduction ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Embryogenic Callus Formation

This paper describes a protocol for the efficient vegetative propagation of Cleome rosea by somatic embryogenesis. Leaf and stem explants from nursery-grown seedlings of C. rosea were cultivated on Murashige and Skoog (MS) medium supplemented with indole-3-acetic acid (IAA), a -naphthaleneacetic acid (NAA), 4-amino-3,5,6-trichloropicolinic acid (picloram) or 2,4-dichlorophenoxyacetic acid (2,4-D). Nodular calli were produced from both explant types in the presence of 4.5 and 9.0 µM 2,4-D. Embryo development and maturation were achieved when calli from stem explants were transferred to media containing a ten-fold reduction of 2,4-D concentration initially used (0.45 and 0.90 µM). Leaf-derived calli did not form embryos with the same treatments. The highest frequency of embryogenic callus formation (85%) and number of embryo per callus (13.45 ± 2.8) were achieved during the first subculture on medium supplemented with 0.90 µM 2,4-D. Embryo conversion into plantlets was achieved following transfer to growth regulator-free MS medium solidified with 2 g.L-1 phytagel. An acclimatization rate of 53% was found three months after transfer to ex vitro conditions and the recovered plants presented a normal phenotypic aspect.

scholarly journals Somatic Embryogenesis and Plant Regeneration from Immature Persimmon (Diospyros kaki Thunb.) Embryos

HortScience ◽  
2008 ◽  
Vol 43 (1) ◽  
pp. 211-214 ◽  
Author(s):  
Akira Sugiura ◽  
Yoshiko Matsuda-Habu ◽  
Mei Gao ◽  
Tomoya Esumi ◽  
Ryutaro Tao
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Cultured Cells ◽  
Callus Formation ◽  
Formation Rate ◽  
Adventitious Bud ◽  
Diospyros Kaki ◽  
Embryogenic Calli ◽  
Globular Embryos ◽  
2,4 Dichlorophenoxyacetic Acid

In persimmon, plant regeneration from cultured cells usually takes place through adventitious bud formation. If somatic embryogenesis were possible, the efficiency of mass propagation and genetic engineering would be greatly improved. We attempted to induce somatic embryogenesis from immature embryos and plant regeneration from the induced embryos. Hypocotyls and cotyledons from immature ‘Fuyu’ and ‘Jiro’ seeds were cultured in the dark in Murashige and Skoog medium solidified with gellan gum and supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzyladenine (BA) at various concentrations. Callus formation started at ≈2 weeks of culture, and the callus formation rate was highest at 3 or 10 μm combinations of 2,4-D and BA. The initially formed calli gradually became brown or black from which white embryogenic calli (EC) appeared secondarily. After ≈8 weeks of culture, globular embryos were formed from these EC, and the formation proceeded until 20 weeks of culture. Formation of globular embryos was higher with ‘Fuyu’ than ‘Jiro’, especially with hypocotyls. When EC with globular embryos were transferred to fresh medium with no plant growth regulators, ≈70% developed to the torpedo-type embryo stage in 6 weeks. The torpedo-type embryos thus formed were germinated and rooted in agar medium with or without zeatin in several weeks without entering dormancy. After germination and rooting, the plantlets were transferred to the same medium and acclimatized for another 4 weeks. As the embryos germinated and rooted simultaneously, the plantlets were easy to grow in pots without transplanting shock. This is the first report on plant regeneration through somatic embryogenesis of persimmon.

scholarly journals Direct Shoot Regeneration from Vaccinium pahalae (Ohelo) and V. myrtillus (Bilberry) Leaf Explants

HortScience ◽  
1996 ◽  
Vol 31 (7) ◽  
pp. 1225-1228 ◽  
Author(s):  
Rida A. Shibli ◽  
M.A.L. Smith
Keyword(s):  
Callus Formation ◽  
Leaf Explants ◽  
Shoot Proliferation ◽  
Direct Organogenesis ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Direct Shoot Regeneration ◽  
The Media ◽  
2,4 Dichlorophenoxyacetic Acid

Ohelo (V. pahalae Skottsb.) and bilberry (V. myrtillus L.) shoots were regenerated via direct organogenesis from whole leaves and leaf sections and also from hypocotyl explants of bilberry. Explants preincubated for 1 to 2 weeks in darkness yielded ≈75% regeneration frequencies and the highest number of regenerating shoots/explant on TDZ-supplemented media (0.9 to 2.7 μm). When 2iP or zeatin were substituted as the cytokinin source, frequencies of regeneration and shoot productivity were significantly lower. Explants held under constant illumination (no dark pretreatment) had significantly lower regeneration frequencies in all tested cytokinin-supplemented media. 2,4-D stimulated callus formation, but did not support regeneration from vegetative explants. Cells from callus and suspension cultures did not exhibit regeneration in any of the media that supported organogenesis from leaves. Regenerants were successfully micropropagated, although callus formation caused by zeatin and high 2iP levels interfered with shoot proliferation. Zeatin induced hyperhydricity in shoots from both species, but more severely in ohelo. Ex vitro rooting after treatment with 4.9 μm IBA or 5.4 μm NAA was 95% and 60% successful for bilberry and ohelo, respectively, and plants were readily acclimatized after an interval in a fog chamber. Bilberry microshoots also rooted in vitro in the absence of growth regulator treatment. Chemical names used: 1H-indole-3-butanoic acid (IBA); N-(3-methyl-2-butenyl)-1-H-purine-6-amine (2iP); 6-furfurylaminopurine (kinetin); 1-naphthaleneacetic acid (NAA); thidiazuron=1-phenyl-3-(1,2,3-thiadiazio-5-yl)urea (TDZ); 2,4-dichlorophenoxyacetic acid (2,4-D); 6-(4-hydroxy-3-methylbut-2-enylamino) purine (zeatin).

scholarly journals Somatic Embryogenesis and Plant Regeneration from Muscadine Grape Leaf Explants

HortScience ◽  
1993 ◽  
Vol 28 (1) ◽  
pp. 53-55 ◽  
Author(s):  
Carol Robacker
Keyword(s):  
Somatic Embryogenesis ◽  
Growth Regulators ◽  
Leaf Explants ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Vitis Rotundifolia ◽  
Regenerated Plants ◽  
Immature Leaf ◽  
Grape Leaf ◽  
2,4 Dichlorophenoxyacetic Acid

Immature leaf laminae and petioles of `Regale' and `Fry' muscadine grapes (Vitis rotundifolia Michx.) were cultured on Nitsch and Nitsch (NN) medium supplemented with 9.0 μm 2,4-D and 4.4 μm BA, and gelled with agar. Callus and original explant tissues were transferred to NN medium containing 10.7 μm NAA and 0.9 μm BA to proliferate embryogenic callus, which, when transferred to NN medium without growth regulators, yielded globular embryos. The embryos matured and germinated after being subcultured to fresh medium without growth regulators. Somatic embryogenesis incidence was greater from petioles than laminae: 90% of `Regale' and 50% of `Fry' petioles formed embryos, compared with 14% and 2% of laminae, respectively. Culturing germinated somatic embryos on NN medium with 1 μm BA enhanced shoot growth. Regenerated plants flowered and appeared morphologically normal. Chemical names used: N-(phenylmethyl)- 1H -purin-6-amine (BA); 2,4-dichlorophenoxyacetic acid (2,4-D); α- naphthaleneacetic acid (NAA).

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