scholarly journals Optimized somatic embryogenesis and plant regeneration in elite Argentinian sugarcane (Saccharum spp.) cultivars

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Valentina Di Pauli ◽  
Paola Daniela Fontana ◽  
Dalia Marcela Lewi ◽  
Arturo Felipe ◽  
Luis Ernesto Erazzú
Keyword(s):  
Tissue Culture ◽  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Culture Conditions ◽  
Light Conditions ◽  
Saccharum Spp ◽  
Regenerated Plants ◽  
Embryogenic Calluses ◽  
Indirect Somatic Embryogenesis ◽  
Good Number

Abstract Background Biotechnological breeding of elite sugarcane cultivars is currently limited because of the difficulty of regenerating plants by tissue culture. Here, we report that commercially elite sugarcane genotypes, which are adapted to Argentinian agro-ecological conditions, are capable of being regenerated via indirect somatic embryogenesis. Leaf rolls of five elite genotypes were cultured following two callus induction protocols using different concentrations of 2,4-D as the growth regulator. Embryogenic calluses were regenerated under light conditions. Regenerated plants were subsequently acclimatized in the greenhouse under two acclimatization procedures before being transplanted to the field. Results Four of the five genotypes were able to form somatic embryos following the two induction protocols. The variables related to embryogenic callus production were influenced by the interaction between genotype and culture conditions. For plant regeneration, the embryogenic calluses were further cultured on an IBA-supplemented medium, where we observed a high genotype dependence. Calluses from the four cultivars regenerated a good number of plants. With the procedures described here, we obtained more than 90% of well-acclimatized plants both in the greenhouse and in the field. Conclusions This protocol provides a simple way to regenerate sugarcane plants through indirect somatic embryogenesis. Also, the results confirm that tissue culture ability is highly genotype-dependent in sugarcane. Our findings suggest that these elite cultivars could be good candidates for biotechnological breeding.

scholarly journals Barley somatic embryogenesis-an attempt to modify variation induced in tissue culture

2021 ◽  
Vol 28 (1) ◽  
Author(s):  
Renata Orłowska
Keyword(s):  
Tissue Culture ◽  
Somatic Embryogenesis ◽  
Taguchi Method ◽  
Culture Conditions ◽  
Dna Demethylation ◽  
Silver Ion ◽  
Donor Plant ◽  
In Vitro Tissue Culture ◽  
Induced Variation

Abstract Background Somatic embryogenesis is a phenomenon carried out in an environment that generates abiotic stress. Thus, regenerants may differ from the source of explants at the morphological, genetic, and epigenetic levels. The DNA changes may be the outcome of induction media ingredients (i.e., copper and silver ions) and their concentrations and time of in vitro cultures. Results This study optimised the level of copper and silver ion concentration in culture media parallel with the induction medium longevity step towards obtaining barley regenerants via somatic embryogenesis with a minimum or maximum level of tissue culture-induced differences between the donor plant and its regenerants. The optimisation process is based on tissue culture-induced variation evaluated via the metAFLP approach for regenerants derived under varying in vitro tissue culture conditions and exploited by the Taguchi method. In the optimisation and verification experiments, various copper and silver ion concentrations and the different number of days differentiated the tested trials concerning the tissue culture-induced variation level, DNA demethylation, and de novo methylation, including symmetric (CG, CHG) and asymmetric (CHH) DNA sequence contexts. Verification of optimised conditions towards obtaining regenerants with minimum and maximum variability compared to donor plants proved useful. The main changes that discriminate optimised conditions belonged to DNA demethylation events with particular stress on CHG context. Conclusions The combination of tissue culture-induced variation evaluated for eight experimental trials and implementation of the Taguchi method allowed the optimisation of the in vitro tissue culture conditions towards the minimum and maximum differences between a source of tissue explants (donor plant) and its regenerants from somatic embryos. The tissue culture-induced variation characteristic is mostly affected by demethylation with preferences towards CHG sequence context.

Optimisation of Tissue Culture Conditions for Transformation Studies Using Immature Embryos of Australian Barley Cultivars

1995 ◽  
Vol 43 (5) ◽  
pp. 499 ◽  
Author(s):  
AM Walmsley ◽  
RJ Henry ◽  
RG Birch
Keyword(s):  
Tissue Culture ◽  
Plant Regeneration ◽  
Culture Medium ◽  
Immature Embryo ◽  
Culture Conditions ◽  
Medium Composition ◽  
Immature Embryos ◽  
Systematic Testing ◽  
Barley Cultivars ◽  
Key Variables

Eight Australian barley cultivars were tested for efficiency of embryonic callus initiation and plant regeneration, from immature embryo explants in tissue culture. Optimisation of tissue culture conditions was performed for cultivars Bandulla, Clipper, Schooner and Tallon in an attempt to increase regeneration frequencies to levels suitable for genetic engineering of barley. Variables tested were 2,4-D concentration, salt composition, carbon source and immature embryo explant. Optimal culture medium composition varied between cultivars. Shoot regeneration rates from culture of isolated scutellar tissues were low for all four cultivars. Halved, immature embryos produced most shoots for cultivars Clipper, Schooner and Tallon, whereas Bandulla performed best with entire immature embryo explants. Clipper (a malting barley) and Bandulla (a feed barley) are suggested as model Australian cultivars for transformation studies. Immature embryos of Bandulla produced an average of 5.3 shoots and Clipper 10.1 shoots per embryo under optimal conditions. Our results show that rates of somatic embryo and plant regeneration sufficient for use in transformation studies can be achieved for diverse Australian Barley cultivars, through systematic testing of a range of key variables including explant type and medium composition.

scholarly journals Factors influencing somatic embryogenesis and regeneration ability in somatic tissue culture of spring and winter rye

2008 ◽  
Vol 13 (4) ◽  
pp. 363 ◽  
Author(s):  
R. MA ◽  
S. PULLI
Keyword(s):  
Tissue Culture ◽  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Developmental Stage ◽  
Embryogenic Callus ◽  
Immature Embryo ◽  
Green Plant ◽  
Somatic Tissue ◽  
Winter Rye ◽  
Effi Ciency

Rye is an important crop in Northern and Eastern Europe. However, the application of various biotechnologies in rye breeding has been limited duo to its recalcitrant in tissue culture. In order to improve somatic tissue effi ciency, key factors affecting somatic embryogenesis and reproducible green plant regeneration of rye (Secale cereale L.) were evaluated and optimised. In this study, a total 27 rye genotypes including 10 spring and 17 winter genotypes were involved in the investigation. Genotype, culture medium, sugar, gel agent and auxin infl uenced somatic embryogenesis of immature embryo signifi cantly. One-two weeks cold pretreatment of young embryo enhanced somatic embryogenesis and green plant regeneration. In culture of immature embryos, infl orescences and leaf segments of the seedlings, explants signifi cantly infl uenced the culture effi ciency. Highest embryogenic callus yield resulted from rye immature embryo as explant compared to young infl orescence and leaf segment of seedling. Developmental stage of embryo played an important role in somatic embryogenesis. Late spherical coleoptile stage (embryo size 0.5–1mm in length) was optimal developmental stage of immature embryo for culture. Morphogenetic potential of embryogenic callus decreased with an increasing number of subcultures, and this ability could be maintained in vitro for a maximum of 8 months of culturing.;

scholarly journals Callus Induction and Plant Regeneration Efficiency According to Tissue Culture Conditions in Teff grass (Eragrostis)

2013 ◽  
Vol 19 (1) ◽  
pp. 55-62
Author(s):  
Ki-Won Lee ◽  
Jin Young Moon ◽  
Hyung Soo Park ◽  
Gi Jun Choi ◽  
Ki-Yong Kim ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Plant Regeneration ◽  
Callus Induction ◽  
Culture Conditions ◽  
Regeneration Efficiency

scholarly journals Plant regeneration via somatic embryogenesis from immature leaves in Tetrapleura tetraptera (Schum. & Thonn.) Taub.

2011 ◽  
Vol 63 (4) ◽  
pp. 1135-1145 ◽  
Author(s):  
J.T. Opabode ◽  
O.A. Akinyemiju ◽  
O.O. Ayeni
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Leaf Explants ◽  
Basal Medium ◽  
Shoot Elongation ◽  
Leaf Petiole ◽  
Immature Leaf ◽  
Embryogenic Calluses ◽  
Tetrapleura Tetraptera ◽  
Secondary Embryos

Plant regeneration via somatic embryogenesis was assessed using immature leaf, petiole and apical meristem explants in Tetrapleura tetraptera. Somatic embryos were induced in the immature leaf using MS basal medium supplemented with 2,4-D and matured on MS basal medium containing BAP. Medium supplemented with 12 mg/l 2,4-D had the highest (43.1%) percentage of embryogenic calluses from immature leaf explants. Conversion of embryogenic callus to mature primary somatic embryo occurred in the medium that contained 1.2 mg/l BAP. Development of secondary embryogenic calluses to matured secondary embryos was highest (98.0%) in the medium with 0.4 mg/l BAP, while the highest average number of mature secondary embryos (6.0) was obtained in the same medium. Medium supplemented with 1.0 mg/l BAP and 0.5 mg/l IBA had the highest (38.7%) percentage of explants with shootbuds. The highest (18.1%) percentage of shoot elongation was obtained in medium with 1.0 mg/l BAP and 20 mg/l IBA. Shootbuds survived and produced roots on medium free of plant growth regulators. Shoots obtained on medium supplemented with 1.0 mg/l BAP and 20 mg/g IBA recorded the highest number of roots per plantlet (7.5) with no apparent morphological abnormality.

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