Ovule and Embryo Culture of Arachis hypogaea and Interspecific Hybrids1

Abstract Interspecific hybridization in Arachis is difficult between species within sectional groups and nearly impossible among more distantly related species. Embryos usually abort early in the reproductive cycle; thus in vitro techniques are necessary to recover many desirable hybrid combinations in the genus. The objectives of this investigation were to develop techniques whereby mature plants could be recovered from otherwise aborting embryos. First, ovule culture was performed using eight genotypes, three levels of kinetin, and the two basal media Murashige and Skoog (MS) and N6. Two-tenths mg/L kinetin in media resulted in 24% of the ovules swelling to a size of 3-4 mm which could be used for excising embryos. Embryo culture was next performed on five genotypes. The transfer series (I) 0.2 mg/L kinetin for 21 days, (2) 0.5 mg/L 6-benzylamino-purine (BAP) for 14 days and, (3) MS without growth regulators resulted in 34.6% of ovules producing plants across genotypes; other transfer series either resulted in a lower percentage of plant recovery and/or tissues of some genotypes which did not survive to maturity. The BAP medium induced shoot growth, while root growth was induced on the MS without growth regulator medium. Approximately 90% of embryos transferred to a mist system after 7-9 weeks in vitro survived transplanting to soil. Two interspecific hybrids were recovered from incompatible hexaploid x diploid crosses, but only after roots were induced using a MS basal medium with 4 mg/L 1-naphthaleneacetic acid:2 mg/L indole-3-butyric acid in a fourth tissue transfer. The experiments illustrated the feasibility of rescuing embryos of A. hypogaea and interspecific peanut hybrids. The process is slow and will be most applicable to wide crosses which cannot be obtained by more conventional methods.

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SOMACLONAL VARIATION IN CULTURED IN VITRO TOBACCO PLANTS FROM THE HYBRID NICOTIANA GOSSEY DOMIN. X N. TABACUM L.

Israel Journal of Plant Sciences ◽

10.1080/07929978.1998.10676705 ◽

1998 ◽

Vol 46 (1) ◽

pp. 35-40 ◽

Cited By ~ 2

Author(s):

Violeta Nikova ◽

Maria Palakarcheva ◽

Roumiana Pundeva ◽

Dobrinka Krusteva

Keyword(s):

Somaclonal Variation ◽

Pollen Viability ◽

Basal Medium ◽

Leaf Size ◽

Culture Method ◽

Stem Length ◽

Naphthaleneacetic Acid ◽

In Vitro Techniques ◽

Plant Forms

Incompatibility between the speciesNicotiana gosseyDomin. andN. tabacumL., resulting in complete sterility of F1hybrid, was overcome using in vitro techniques. Explants of stem parenchyma were put in culture and were regularly subcultured on Murashige and Skoog basal medium, supplemented with different compounds: α-naphthaleneacetic acid (2 mg L−1) and kinetin (0.5 mg L−1) for callus induction, kinetin (2 mg L−1) and indule-3-acetic acid (0.5 mg L−1) for organ formation, and ferulic acid (2 mg L−1) for rooting. The regenerants obtained showed significant morphological and cytological variability. They differed from each other as well as from the initial material in stem length, leaf size, and flower morphology. Most of the plants examined were mixoploids. A great number of aberrations were established during the meiotic division of the regenerants. Pollen viability varied from 0 to 92%. The results of our investigations showed that the tissue culture method could be successfully applied not only for overcoming species incompatibility but also for inducing somaclonal variation and creation of a large variety of plant forms.

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

Canadian Journal of Botany ◽

10.1139/b84-189 ◽

1984 ◽

Vol 62 (7) ◽

pp. 1393-1397 ◽

Cited By ~ 6

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.'

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In vitro organogenesis and plant regeneration of Passiflora xishuangbannaensis, a species with extremely small populations

10.21203/rs.3.rs-360926/v1 ◽

2021 ◽

Author(s):

Yuan-yuan Meng ◽

Shi-jie Song ◽

Sven Landrein

Keyword(s):

Plant Regeneration ◽

Plant Growth ◽

Plant Growth Regulators ◽

Growth Regulators ◽

Basal Medium ◽

Ex Situ ◽

Naphthaleneacetic Acid ◽

South American ◽

South American Species

Abstract Passiflora xishuangbannaensis (Passifloraceae) is endemic to a few sites of Mengyang nature reserve in Yunnan, Xishuangbanna and less than 40 individuals have been recorded. Nine Passiflora species are endemic to Yunnan with most species occurring in South America, making P. xishuangbannaensis highly significant and emblematic to the conservation work in the region. This study is designed to provide the first protocol for in vitro organogenesis and plant regeneration for ex situ conservation and reintroduction for an Asian Passiflora species. Using internodes, petioles and tendrils we optimize calli formation and root elongation using several plant growth regulators, individually or in combination. We also assess the genetic stability of regenerated cells. The maximum callus induction and shoot bud differentiation were both achieved on half Murashige and Skoog basal medium supplemented with 4.44 µM 6-Benzylaminopurine and 1.08 µM 1-Naphthaleneacetic acid. The best rooting was achieved from 30 days old, regenerated shoots on half Murashige and Skoog basal medium supplemented with 1.08 µM 1-Naphthaleneacetic acid. Micropropagated plants were subjected to inter simple sequence repeat markers analyses. Collectively, 86 bands were generated from 6 primers of which 12 bands were polymorphic, showing genetic variation between the regenerated plantlets and the original plant. Response to plant growth regulators was more specific than most other studies using South American species, which could be explained by the morphological and physiological differences between South American and Asian Passiflora species

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Genetic transformation of Stella De Oro daylily by particle bombardment

Canadian Journal of Plant Science ◽

10.4141/p03-025 ◽

2003 ◽

Vol 83 (4) ◽

pp. 873-876 ◽

Cited By ~ 3

Author(s):

A. N. Aziz ◽

R. J. Sauvé ◽

S. Zhou

Keyword(s):

Southern Blotting ◽

Basal Medium ◽

Callus Cultures ◽

Naphthaleneacetic Acid ◽

Ms Medium ◽

Chain Reaction ◽

Polymerase Chain ◽

Semi Solid ◽

Independent Transformation

Daylily (Hemerocallis sp. ‘Stella de Oro’) callus cultures initiated from ovules were bombarded with gold particles coated with plasmid harboring Basta® resistance gene. Resulting putative transgenic calli were selected after 3 wk on semi-solid Murashige and Skoog’s (MS) basal medium supplemented with 10 mg L-1 1-naphthaleneacetic acid, 2 mg L-1 6-benzylaminopurine and 3 mg L-1 phosphinothricin (PPT). Surviving calli regenerated shoots after 2 mo on semi-solid MS medium supplemented with 2 mg L-1 thiadiazuron and 1 mg L-1 PPT. Polymerase chain reaction and Southern blotting were used to confirm independent transformation events. Key words: Basta® resistance, in vitro, Hemerocallis

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In vitro study of Tinospora cordifolia (Willd.) Miers (Menispermaceae)

Botanica Orientalis Journal of Plant Science ◽

10.3126/botor.v6i0.2918 ◽

2010 ◽

Vol 6 ◽

pp. 103-105 ◽

Cited By ~ 5

Author(s):

Aditi Singh ◽

Saroj K Sah ◽

Aunji Pradhan ◽

Sabari Rajbahak ◽

Niran Maharajan

Keyword(s):

Medicinal Plant ◽

Callus Induction ◽

Shoot Growth ◽

Tinospora Cordifolia ◽

Nodal Segments ◽

Naphthaleneacetic Acid ◽

Nodal Segment ◽

Root Induction ◽

Ms Medium

In vitro study was carried out in an important medicinal plant Tinospora cordifolia (Willd.) Miers belonging to the family: Menispermaceae. Vegetative parts such as stem, leaf and nodal explants were excised from an elite in vivo grown mature plant and thereafter cultured on Murashige-Skoog (MS) medium supplemented with different hormonal concentrations for callus induction and organogenesis. Callus formation occurred from nodal segments, leaf and inter-node explants when planted on different combinations of hormones. Tinospora cordifolia showed response for in vitro shoot growth from the nodal segment. The best shoot growth was observed on MS medium supplemented with kinetin (1.5 mg/l). Similarly, the best result for root induction was obtained on MS medium supplemented with 6-benzylaminopurine (1.0 mg/l) and naphthaleneacetic acid (2.5 mg/l). Key-words: callus induction; explants; medicinal plant; MS medium; tissue culture.DOI: 10.3126/botor.v6i0.2918 Botanica Orientalis - Journal of Plant Science (2009) 6: 103-105

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In vitro propagation of Crambe maritima

Canadian Journal of Botany ◽

10.1139/b87-010 ◽

1987 ◽

Vol 65 (1) ◽

pp. 72-75 ◽

Cited By ~ 2

Author(s):

J. Y. Peron ◽

E. Regnier

Keyword(s):

In Vitro Propagation ◽

Indoleacetic Acid ◽

Adventitious Shoots ◽

Basal Medium ◽

Naphthaleneacetic Acid ◽

Petiole Explants ◽

Acclimatization Period

A method for rapid micropropagation of sea kale (Crambe maritima L.) was developed. Petiole explants placed in vitro on a medium containing 0.5 mg/L indoleacetic acid (IAA), 6.0 mg/L kinetin, and 1.5 mg/L benzylaminopurine developed callus within 15 days and shoots within 28 days. Nearly four adventitious shoots could be developed within 3 weeks by placing the initial shoot on media without IAA. To develop roots, the shoots were then transferred to the basal medium containing 0.1 to 1.0 mg/L indolbutyric or α-naphthaleneacetic acid. Rooted plantlets were obtained within 2 or 3 weeks. After an acclimatization period of 6 weeks in a greenhouse in unsterilized medium, the plantlets could be set outdoors.

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120 EFFECT OF INDIVIDUAL CULTURE SYSTEM USING WOW OR CELL-TAK ON DEVELOPMENT OF IN VITRO-PRODUCED BOVINE EMBRYOS

Reproduction Fertility and Development ◽

10.1071/rdv21n1ab120 ◽

2009 ◽

Vol 21 (1) ◽

pp. 160

Author(s):

S. Matoba ◽

P. Lonergan

Keyword(s):

Embryo Culture ◽

Culture System ◽

Basal Medium ◽

Bovine Embryos ◽

Paracrine Effects ◽

Individual Culture ◽

Cell Adhesive ◽

Developmental Rates ◽

One Way Anova

The culture of embryos individually in vitro is generally associated with poorer developmental rates. However, the ability to do this successfully would greatly facilitate studies where identification of individual embryos, or the embryos from a particular donor, is necessary. The objective of this study was to examine the effect of culture system on the development of individual IVP bovine embryos. Presumptive zygotes (n = 1301, 6 replicates), produced by IVM/IVF, were used. The aim of Experiment 1 was to compare development of bovine embryos in SOF or CR1aa supplemented with 5% FCS. Zygotes were cultured in droplets under oil as follows: (i) 20/25 μL, (ii) 20/100 μL or (iii) 20/100 μL individually in the Well of the Well (WOW) system (Vajta et al. 2000 Mol. Reprod. Dev. 55, 254–264). Twenty WOW were prepared in a 100 μL droplet of medium under oil using a sterile rod. The aim of Experiment 2 was to compare development of embryos cultured in groups but individually identifiable on the cell adhesive Cell-Tak (Stokes et al. 2005 Dev. Biol. 284, 62–71) or in the WOW system. Zygotes were cultured as follows: (i) 20/20 μL, (ii) 20/20 μL with Cell-Tak, (iii) 20/100 μL with Cell-Tak or (iv) 20/100 μL in WOW. A drop of Cell-Tak (1 μL/20 μL medium) was placed on the base of the dish, dried for 20 min, washed with sterile water and dried completely. Once dried, the area was covered with 10 μL of FCS-free medium and groups of 20 zygotes were placed on the Cell-Tak in a 5 × 4 grid formation a maximum of 160 μm apart. Then, an additional 10 μL or 90 μL medium supplemented with FCS was added to give a final volume of 20 or 100 μL. Cleavage and blastocyst rates were assessed on Day 2 and Days 7–9, respectively. Data (means ± SE) were analyzed by one way ANOVA. In Experiment 1, there were no differences between SOF and CR1aa with respect to culture of embryos individually in WOW (P > 0.05); therefore, SOF was used as the basal medium for Experiment 2. There were no differences (P > 0.05) between the cleavage and blastocyst rate among drop sizes and individual culture systems; individual culture, irrespective of the system used (Cell-Tak or WOW), resulted in the similar developmental rates to the control. In conclusion, individual embryo culture offers the opportunity to study embryo development in a more powerful manner. Furthermore, the use of the cell adhesive Cell-Tak may be more practical because it removes the potential variability associated with well dimensions in the WOW system and may improve any potential paracrine effects during embryo culture. Further studies are required to establish the viability of such embryos after transfer. Table 1.Effect of individual culture system on development of IVP bovine embryos Supported by Science Foundation Ireland.

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Total Polyphenol Content, in vitro Antifungal and Antioxidant Activities of Callus Cultures from Inula crithmoides

Natural Product Communications ◽

10.1177/1934578x1300801122 ◽

2013 ◽

Vol 8 (11) ◽

pp. 1934578X1300801 ◽

Cited By ~ 2

Author(s):

Anahi Bucchini ◽

Laura Giamperi ◽

Donata Ricci

Keyword(s):

Methanol Extract ◽

Antioxidant Activities ◽

Leaf Explants ◽

Basal Medium ◽

Alternaria Solani ◽

Antimicrobial Activities ◽

Callus Cultures ◽

Naphthaleneacetic Acid ◽

Stable Free Radical

This is the first report on the antioxidant and antifungal activities of callus cultures from Inula crithmoides L. (Asteraceae). Callus cultures were initiated from leaf sections, on initial culture MS basal medium supplemented with various concentrations of 2,4-D (2,4-dichlorophenoxyacetic acid), NAA (1-naphthaleneacetic acid) and IBA (indole-3-butyric acid) and a 72% survival was achieved. Significant differences between the various auxins used as phytohormones on callus growth were found. Maximum callusing was noticed on the leaf explants grown on MS basal medium supplemented with 1 mgL–1 2,4-D. Subsequently the antioxidant and antimicrobial activities of the methanol extract from calli were investigated. Antioxidant studies suggested that the methanol extracts of dark-grown and light-grown callus were able to reduce the stable free radical 2,2-diphenyl-1-picrilhydrazyl (DPPH). In the inhibition against lipid peroxidation, extracts of dark-grown callus showed the strongest effect with IC50 values better than those of the standards. The methanol extract of callus cultures had significant antifungal activity only against two of the fungi tested: Alternaria solani and Phytophthora cryptogea. Against all the other tested fungi, the I. crithmoides calli extracts showed fungistatic activity.

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Embryo Rescue in Wide Crosses in Arachis. 3. In vitro Culture of Peg Tips of A. hypogaea Selfs and Interspecific Hybrids1

Peanut Science ◽

10.3146/i0095-3679-14-2-5 ◽

1987 ◽

Vol 14 (2) ◽

pp. 70-74 ◽

Cited By ~ 3

Author(s):

J. P. Moss ◽

H. T. Stalker

Keyword(s):

Gibberellic Acid ◽

In Vitro Culture ◽

Successful Treatment ◽

Interspecific Hybrids ◽

Embryo Rescue ◽

Wide Crosses ◽

Reproductive Tissues ◽

Globular Stage ◽

Callus Production

Abstract Embryo rescue in wide crosses in Arachis has only been achieved from culturing ovules excised from well developed pods, or from immature pods derived from flowers treated with gibberellic acid (GA) and which had embryos large enough to dissect without injury. The objective of this study was to determine whether reproductive tissues could grow in vitro without the need to dissect them from the peg tip and to determine the effects of GA application on flowers at the time of pollination, age of peg when cultured, and the presence of the peg meristem on reproductive growth, callus production, and peg elongation in vitro. Pegs elongated in culture only when the peg meristem was not removed. Ovules enlarged and grew out of the surrounding peg tissue in 3.8 to 32.8% of the cultures. Significantly more ovules grew when the peg meristem was removed (p < 0.01) and when 10- and 20- day-old pegs were cultured (p < 0.05). Overall, the most successful treatment for growth of ovules was treating flowers with GA at pollination and culturing without the peg meristem 10 days after pollination when 25.0 and 32.8% of all hybrid and selfed ovules, respectively, grew. Embryo growth was observed in an average of 8.4 and 17.6% of embryo sacs in hybrid and self peg tips, respectively, with several embryos reaching the globular stage after 21 days in vitro. This illustrates the potential for culturing young reproductive tissues of Arachis to recover interspecific hybrids.

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