Somatic embryogenesis from in vitro anther culture of apomictic buffel grass genotypes and analysis of regenerated plants using flow cytometry

Abstract Taraxacum belorussicum Val. N. Tikhom, a poorly known and obligatory apomictic species, is an attractive plant material for studying the embryological, genetic and molecular mechanisms of apomixis. This work aims to obtain an efficient protocol for Taraxacum belorussicum regeneration. Four types of explants (cotyledons, hypocotyls, meristems and roots) that were taken from 2-weeks-old seedlings were used for in vitro cultures, and a fast and efficient protocol of T. belorussicum regeneration was obtained. Various ½ MS-based media containing IAA (5.71 µM), TDZ (4.54 µM) and PSK (100 nM) were chosen to assess the morphogenetic abilities of selected T. belorussicum explants. Studies on the role of PSK were done in three independent experiments, where the most significant factors were always light and darkness. All explants produced callus by the third day of culture and adventitious shoots after 7 days, although in an asynchronous indirect manner, and with different intensities for all explant types. The most preferred medium culture for hypocotyl, cotyledon and meristem explants was ½ MS + TDZ, and ½ MS + IAA + TDZ + PSK for roots which were the only explant sensitive to PSK. A short darkness pretreatment (8 days) in PSK medium was found suitable to enhance organogenesis. Secondary organogenesis was observed for regenerated plants on meristem explants from the ½ MS + IAA + TDZ + PSK medium. A weak somatic embryogenesis was observed for hypocotyl and cotyledon explants from ½ MS + IAA + TDZ and ½ MS + IAA + TDZ + PSK media. Histological and scanning electron microscope images (SEM) of T. belorussicum confirmed indirect organogenesis and somatic embryogenesis. Plant material treated with aniline blue solution revealed the presence of callose in the cell walls of cotyledon and hypocotyl explants. The presence of extracellular matrix (ECM) and heterogenic structure of callus was also verified by scanning electron microscopy and light microscopy, confirming the high morphogenetic ability of T. belorussicum.

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Morphoagronomic characterization of Sorghum cultivar ‘CIAP 132R-05’ obtained by somatic embryogenesis in the conversion phase using different substrates

Emirates Journal of Food and Agriculture ◽

10.9755/ejfa.2019.v31.i4.1930 ◽

2019 ◽

pp. 248

Author(s):

Silvio de J. Martínez Medina ◽

Raúl Barbón Rodríguez, Rafael Gómez-Kosky, Novisel Veitía Rodríguez ◽

María Esther González Vega ◽

Orlando Saucedo Castillo ◽

Eduardo Fidel Héctor Ardisana ◽

...

Keyword(s):

Somatic Embryogenesis ◽

Plant Height ◽

Somatic Embryos ◽

Morphological Characters ◽

Morphological Characterization ◽

Stem Diameter ◽

Limb Length ◽

Regenerated Plants

Sorghum [Sorghum bicolor (L.) Moench] is an important crop after wheat, corn, rice and barley. The aim of this research was to study morphological characterization of the plant population of grain sorghum cultivar CIAP 132R-05 regenerated via somatic embryogenesis under semi controlled conditions in green house. Plants regenerated from somatic embryos were compared to plants grown from botanical seeds. Both populations of plants were characterized morphologically, taking in account quantitative traits (plant height (cm), stem diameter, number of active leaves per plant, limb length and width, number of active roots per plant, length of the roots (cm), fresh weight (gMF), foliar area (dm2), number of shoots per plant) and qualitative traits (albino or variegated plants, color of the leaves). Quantitative characteristics of both populations corresponded with those listed in the National Register of Commercial Varieties of Cuba. However; the population of plants derived from somatic embryos showed significantly higher values for plant height, stem diameter, limb length and width, which may be associated with the physiological rejuvenation produced by the effect of in vitro culture. This allowed to determine the phenotypic stability of the regenerated plants via somatic embryogenesis, by assessing morphological characters in field conditions. The results can be applied to the in vitro propagation of elite plants selected as a product of conventional breeding programs and obtained through the use of different biotechnological methods.

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Histology of organogenesis and somatic embryogenesis in excised root cultures of an endangered species Tylophora indica (Asclepiadaceae)

Australian Journal of Botany ◽

10.1071/bt09220 ◽

2010 ◽

Vol 58 (3) ◽

pp. 198 ◽

Cited By ~ 19

Author(s):

Aastha Sahai ◽

Anwar Shahzad ◽

Shiwali Sharma

Keyword(s):

Somatic Embryogenesis ◽

Induction Medium ◽

Root Induction ◽

Ms Medium ◽

Tylophora Indica ◽

Regenerated Plants ◽

Maturation Medium ◽

Half Strength ◽

One Year

This paper reports an efficient regeneration protocol through parallel organogenic and embryogenic pathways from green root segments (GRSs) of Tylophora indica (Burm.f) Merrill. GRSs explants from one year old in vitro cultures were cultured on Murashige and Skoog (MS) medium containing various cytokinins. Five µmol/L of 6-benzyladenine (BA) was most responsive for organogenesis in 1.5 cm long GRSs. Repeated subculture on medium containing both BA (5 µmol/L) and 1-naphthleneacetic acid (NAA) (0.1 µmol/L) promoted multiplication and proliferation of direct shoot buds (46.80 ± 0.96) and callus mediated somatic embryogenesis (18.07 ± 0.33). Germinated embryos isolated from callus were transferred onto maturation medium consisting of half-strength MS medium either devoid of plant growth regulators (PGRs) or with various concentrations of gibberellic acid (GA). Microshoots were excised during subculture and transferred onto root induction medium, thus ensuring a continuous supply of germplasm. Morphogenic variations were noticed in types of roots induced on various auxins. Regenerated plantlets and emblings hardened best on vermiculite with a survival rate of 90% and 70% respectively. However, the emblings were healthier in comparison to the regenerated plants. Histological analysis showed the origin and development of organogenesis.

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Screening of Brazilian soybean genotypes with high potential for somatic embryogenesis and plant regeneration

Pesquisa Agropecuária Brasileira ◽

10.1590/s0100-204x2010000700012 ◽

2010 ◽

Vol 45 (7) ◽

pp. 715-720 ◽

Cited By ~ 4

Author(s):

Annette Droste ◽

Anelise Machado da Silva ◽

Iliane Freitas de Souza ◽

Beatriz Wiebke-Strohm ◽

Lauro Bücker-Neto ◽

...

Keyword(s):

Somatic Embryogenesis ◽

Plant Regeneration ◽

Good Alternative ◽

High Potential ◽

Embryo Induction ◽

Regenerated Plants ◽

Maturation Medium ◽

Secondary Embryo ◽

Soybean Genotypes

The aim of this work was to identify Brazilian soybean (Glycine max) genotypes with potential to respond to in vitro culture stimuli for primary somatic embryo induction, secondary embryo proliferation and plant regeneration. Differences among eight tested cultivars were observed at each stage. Two cultivars, IAS-5 and BRSMG 68 Vencedora, were selected for the evaluation of the capacity for embryo differentiation and plant regeneration. These cultivars had high embryo induction frequencies, repetitive embryogenic proliferation, and low precocious embryo germination in the initial experiment. The effect of abscisic acid (ABA) and charcoal addition on plant regeneration was investigated. The addition of ABA to proliferation medium and of ABA and activated charcoal to maturation medium increased embryo differentiation rates, which resulted in a higher number of regenerated plants. The BRSMG 68 Vencedora cultivar was found to have a high potential for embryo induction, embryo proliferation and plant regeneration. The potential of this cultivar for somatic embryogenesis was similar to that observed for cultivar IAS-5, which is currently used for soybean transformation in Brazil. BRSMG 68 Vencedora may be a good alternative genotype for soybean genetic engineering via somatic embryogenesis protocols.

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Gametic and somatic embryogenesis through in vitro anther culture of different Citrus genotypes

Plant Biosystems - An International Journal Dealing with all Aspects of Plant Biology ◽

10.1080/11263504.2014.987847 ◽

2015 ◽

Vol 150 (2) ◽

pp. 304-312 ◽

Cited By ~ 10

Author(s):

J. C. Cardoso ◽

A. M. Abdelgalel ◽

B. Chiancone ◽

R. R. Latado ◽

O. Lain ◽

...

Keyword(s):

Somatic Embryogenesis ◽

Anther Culture

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Detection of somaclonal variants in somatic embryogenesis-regenerated plants of Vitis vinifera by flow cytometry and microsatellite markers

Plant Cell Tissue and Organ Culture (PCTOC) ◽

10.1007/s11240-010-9753-1 ◽

2010 ◽

Vol 103 (1) ◽

pp. 49-59 ◽

Cited By ~ 33

Author(s):

María Jesús Prado ◽

Eleazar Rodriguez ◽

Laura Rey ◽

María Victoria González ◽

Conceição Santos ◽

...

Keyword(s):

Flow Cytometry ◽

Somatic Embryogenesis ◽

Vitis Vinifera ◽

Microsatellite Markers ◽

Somaclonal Variants ◽

Regenerated Plants

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Effect of Plant Growth Regulators on Somatic Embryogenesis and Plantlet Development of Turkey Berry (Solanum torvum SW)

European Journal of Medicinal Plants ◽

10.9734/ejmp/2021/v32i730400 ◽

2021 ◽

pp. 1-8

Author(s):

Ghan Singh Maloth ◽

Rajinikanth Marka ◽

Rama Swamy Nanna

Keyword(s):

Somatic Embryogenesis ◽

Plant Regeneration ◽

Somatic Embryos ◽

Leaf Explants ◽

Direct Somatic Embryogenesis ◽

Ms Medium ◽

Solanum Torvum ◽

Plantlet Formation ◽

Regenerated Plants

In the present study it was reported on direct somatic embryogenesis and plant regeneration from cotyledon and leaf explants of Turkey berry/pea egg plant (Solanum torvum SW), a medicinally important plant. Somatic embryogenesis has several advantages over other routes of in vitro plant regeneration. Somatic embryogenesis was induced directly from cotyledon and leaf explants on MS medium fortified with BAP (0.5 mg/L)+NAA (0.5-6.0 mg/L). High percentage of somatic embryogenesis (90%), maximum number of somatic embryos formation (62±0.18) along with high percentage (76%) conversion of somatic embryos into bipolar embryos was observed on cotyledon explants in 0.5 mg/L BAP+2.5 mg/L NAA. At the same concentration of BAP (0.5 mg/L)+NAA (2.5 mg/L) also resulted on the maximum percentage of somatic embryogenesis (92%), the highest number of somatic embryos formation (88±0.15) and the highest percentage (76%) of somatic embryos conversion into bipolar embryos in leaf explants. A mixture of globular, heart and torpedo-shaped embryos were germinated on MS medium supplemented with 0.5 mg/L IAA+1.0-4.0 mg/L BAP. Maximum germination frequency (75±0.14) of somatic embryos and plantlet formation was found in 0.5 mg/L IAA+2.0 mg/L BAP, but they didn’t germinate on ½ MSO and MSO media. The survival rate of regenerated plants after field transfer was recorded to be 75%. These regenerated plants were found morphologically similar to donor plants. The present protocol can be used for conservation of the species and also for genetic transformation experiments in S. torvum.

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Time of In Vitro Anther Culture May Moderate Action of Copper and Silver Ions that Affect the Relationship between DNA Methylation Change and the Yield of Barley Green Regenerants

Plants ◽

10.3390/plants9091064 ◽

2020 ◽

Vol 9 (9) ◽

pp. 1064

Author(s):

Piotr T. Bednarek ◽

Renata Orłowska

Keyword(s):

Anther Culture ◽

De Novo ◽

Silver Ions ◽

Dna Demethylation ◽

Induction Medium ◽

Green Plants ◽

Ion Concentrations ◽

Regenerated Plants ◽

Short Time

Plant anther culture allows for the regeneration of uniform and homozygous double haploids. However, off-type regenerants may appear as a result of so-called tissue culture-induced variation (TCIV). In addition, the presence of Cu2+ and Ag+ ions in the culture medium might influence the number of green plants. The regenerants were obtained via anther cultures of barley under varying Cu2+ and Ag+ ion concentrations in the induction medium during distinct time conditions. DArTseqMet markers were evaluated based on regenerants and donor plants and delivering data on DNA demethylation (DM) and de novo methylation (DNM) and changes in methylation (Delta). The number of green regenerated plants per 100 anthers (GPs) was evaluated. The Cu2+ and Ag+ ion concentrations moderated relationships between Delta and the number of green plants conditional on time of tissue cultures. Depending on the ions, moderated moderation is valid within the different time of anther culture. When the highest concentration of copper is analyzed, plant regeneration is possible under short ‘Time’ (21 days) of anther culture wherein Delta is negative or under elongated Time when Delta is positive. Under 21 days of culture, the highest concentration of silver ions and when Delta is negative, some regenerants could be evaluated. However, under high Ag+ concentration when Time of culture is long and Delta positive, the highest number of green plants could be obtained.

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Genetics of anther culture capability traits and its association with seed yield traits in sesame

Indian Journal of Genetics and Plant Breeding (The) ◽

10.31742/ijgpb.80.4.7 ◽

2020 ◽

Vol 80 (04) ◽

Author(s):

Manasi Dash ◽

Smrutirekha Naik ◽

Banshidhar Pradhan

Keyword(s):

Somatic Embryogenesis ◽

Anther Culture ◽

Seed Yield ◽

Callus Induction ◽

Combining Ability ◽

Hybrid Breeding ◽

Direct Selection ◽

Early Screening ◽

Induction Frequency

Successful development of heterotic hybrids in the autogamous sesame crop depends largely on the general combining ability of the parents as well as the specific combining ability of the hybrids produced from them. Homogeneity of Vr -Wr values and non-significance of regression coefficient (b) in a 6 parents half diallal cross were found in respect of capsule length, seeds per capsule, plant height, callus induction frequency, days to callus induction and response to somatic embryogenesis indicating validity of Hayman’s assumptions. The estimates of D, H1 and H2 components indicated predominance of non-additive gene action in the inheritance of these six traits, and the ADD estimates indicated overdominance for all these characters. The estimates of F and KD/KR indicated presence of more of dominant alleles in the parental population except for response to somatic embryogenesis. Strong positive association was recorded between seed yield and in vitro traits viz., callus induction frequency, callus fresh weight and somatic embryogenesis. This indicates that anther culture technique can assist in early screening for combining ability of parents as well as direct selection for heterotic crosses at an earlier stage of hybrid breeding programme.

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