scholarly journals Somatic Embryogenesis in Vitis for Genome Editing: Optimization of Protocols for Recalcitrant Genotypes

Horticulturae ◽  
2021 ◽  
Vol 7 (11) ◽  
pp. 511
Author(s):  
Lucia Rosaria Forleo ◽  
Margherita D’Amico ◽  
Teodora Basile ◽  
Antonio Domenico Marsico ◽  
Maria Francesca Cardone ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Citric Acid ◽  
Fold Increase ◽  
Chlorine Concentration ◽  
Growth Media ◽  
Embryogenic Calluses ◽  
Grape Varieties ◽  
Qualitative Characteristics ◽  
Breeding Techniques ◽  
Embryogenic Response

New Plant Breeding Techniques (NPBTs) protocols have been developed to produce new grape varieties with improved quantitative and qualitative characteristics. Reliable transformation protocols for grapes are based on the generation/induction of embryogenic callus cells that are then transformed. Varieties such as Italia have proven to be very recalcitrant to regeneration via somatic embryogenesis. In this work, the development of a protocol for improved production of embryogenic calluses is described. Two sterilization protocols were tested: (a) a lower active chlorine concentration for a longer time (LS); and (b) a higher chlorine concentration for a shorter time (HS), in combination with the absence or presence of citric acid in the growing substrate in the first growth media. The embryogenic calluses formation in Chardonnay, a cv. with a high embryogenic response, was significantly higher in presence of citric acid in the initial growing substrate regardless of the sterilization protocol. In Aglianico, a cv. with a lower embryogenic response, no significant differences were observed. Instead, in a recalcitrant cv. as Italia, we obtained a 13-fold increase in embryogenic calluses formation performing sterilization of flowers with the HS protocol compared to LS.

scholarly journals Factors affecting in vitro cultivation of grape (Vitis vinifera L.): a review

2020 ◽  
Vol 10 (1) ◽  
pp. 1-5 ◽  
Author(s):  
Fikadu Kumsa
Keyword(s):  
Vitis Vinifera ◽  
Review Paper ◽  
Vitis Vinifera L ◽  
Growth Media ◽  
In Vitro Cultivation ◽  
Factors Affecting ◽  
Current Review ◽  
Research Gap ◽  
Grape Varieties

Grape (Vitis vinifera L.) is globally cultivated as commercial fruit crop usually used for fruit purpose or industrial product. The objective of the current review is to review and identify the research gap on the effect of different growth media and vitrification on shooting and rooting performance of grape. Factors affecting rooting of grape cuttings can be internal or external factors. Currently, grapevines are very sensitive to disease in the conventional method of propagation. Even if tissue culture is recommended for healthy propagation of the grape varieties, still factors affecting the growth of the plant verifications were reported. This, review paper progressively revised for the existing factors and possible solutions during in vitro propagation of grapevines. Int. J. Agril. Res. Innov. Tech. 10(1): 1-5, June 2020

The role of chitinases and glucanases in somatic embryogenesis of black pine and hybrid firs

2013 ◽  
Vol 8 (12) ◽  
pp. 1172-1182 ◽  
Author(s):  
Lenka Fráterová ◽  
Terézia Salaj ◽  
Ildikó Matušíková ◽  
Ján Salaj
Keyword(s):  
Somatic Embryogenesis ◽  
Culture Media ◽  
Extracellular Fluid ◽  
Abies Alba ◽  
Pinus Nigra ◽  
Callus Cultures ◽  
Growth Media ◽  
Large Variability ◽  
Embryogenic Cell ◽  
Embryogenic Potential

AbstractGlucanase and chitinase enzymes play an important role in different plant processes including defense against pathogens and morphogenesis. Moreover, their role in the processes of somatic embryogenesis has been demonstrated. It has been suggested, that the presence of this type of proteins might be a marker for embryogenic potential of callus cultures. In this work we screened for the presence of glucanases and chitinases in liquid growth media of a set of conifer embryogenic cell lines in order to find correlation with their embryogenic potential. We have found that none of the 12 chitinase isoforms detected in culture media of Pinus nigra Arn. or the nine chitinases detected in media with Abies alba × A. cephalonica and Abies alba × A. numidica embryogenic tissues could be linked to their embryogenic capacity. Similarly, none of the six glucanase isoforms detected in the extracellular fluid of Pinus nigra Arn. cultures can be assigned as a marker of embryogenic potential. Thus, our data indicate the large variability and doubtless importance of glucanases and chitinases for cell growth and development of somatic embryos, however, do not support the premise that they are markers of embryogenesis.

Transcriptional profiling of genes involved in embryogenic, non-embryogenic calluses and somatic embryogenesis of Valencia sweet orange by SSH-based microarray

Planta ◽  
2012 ◽  
Vol 236 (4) ◽  
pp. 1107-1124 ◽  
Author(s):  
Xiao-Xia Ge ◽  
Li-Jun Chai ◽  
Zheng Liu ◽  
Xiao-Meng Wu ◽  
Xiu-Xin Deng ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Sweet Orange ◽  
Transcriptional Profiling ◽  
Embryogenic Calluses

scholarly journals Effect of grape variety (Vitis vinifera L.) and grape pomace fermentation conditions on some volatile compounds of the produced grape pomace distillate

OENO One ◽  
2004 ◽  
Vol 38 (4) ◽  
pp. 225 ◽  
Author(s):  
Maria Gerogiannaki-Christopoulou ◽  
Nikolaos V. Kyriakidis ◽  
Panagiotis E. Athanasopoulos
Keyword(s):  
Citric Acid ◽  
Vitis Vinifera ◽  
Volatile Compounds ◽  
Internal Standard ◽  
Grape Pomace ◽  
Grape Variety ◽  
Ionization Detector ◽  
Flame Ionization ◽  
Grape Varieties

<p style="text-align: justify;">Agrape pomace distillate was produced from 4 Greek white grape varieties (<em>Vitis vinifera</em> L.). Pomace was fermented with and without addition of citric acid, acting as an antibacterial agent, during fermentation. Fermented grape pomace was distilled in a traditional copper distillation apparatus.. Five major volatile compounds, including methanol, were measured. Pentanol-3 was used as an internal standard. Flame ionization detector (FID) coupled to capillary gas chromatography was used for determination of five volatile distillate components. The addition of citric acid resulted in the reduction of methanol content by about 15%. All the other components studied did not affect in any appreciable degree.</p>

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.

Influence of auxins on somatic embryogenesis and alkaloid accumulation in Leucojum aestivum callus

2013 ◽  
Vol 8 (6) ◽  
pp. 591-599 ◽  
Author(s):  
Agata Ptak ◽  
Anna Tahchy ◽  
Edyta Skrzypek ◽  
Tomasz Wójtowicz ◽  
Dominique Laurain-Mattar
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryo ◽  
Symptomatic Treatment ◽  
Dichlorophenoxyacetic Acid ◽  
Leucojum Aestivum ◽  
Anisic Acid ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Embryogenic Response

AbstractIn vitro cultures of Leucojum aestivum are considered as an alternative for the production of galanthamine, which is used for the symptomatic treatment of Alzheimer’s disease. We studied the effects of auxins 2,4-dichlorophenoxyacetic acid (2,4-D), 4-amino-3,5,6-trichloropicolinic acid (picloram), 3,6-dichloro-o-anisic acid (dicamba) at concentrations of 25 and 50 µM on the induction of embryogenic callus and its capacity to induce somatic embryogenesis and alkaloid accumulation. The embryogenic response of the explants was from 30% for 25 µM of dicamba to 100% for picloram (for both 25 and 50 µM). 2,4-D (50 µM) stimulated greater callus proliferation and somatic embryo induction as compared to the other auxins. Polyethylene glycol (PEG) stimulated somatic embryo maturation. Callus grown on media containing 50 µM of auxins produced fewer phenolic compounds as compared with callus grown on media containing 25 µM of auxins. GC-MS analyses showed seven alkaloids in the in vivo bulbs and two to four in callus culture. Galanthamine was detected in callus cultivated with 2,4-D (25, 50 µM), picloram (25 µM), and dicamba (50 µM). Other alkaloids, trisphaeridine, tazettine, and 11-hydroxyvittatine were accumulated only in callus growing on medium with picloram (50 µM).

scholarly journals SEPARATION OF HUMAN LYMPHOID CELLS INTO G1, S, AND G2 CELL CYCLE POPULATIONS BY USE OF A VELOCITY SEDIMENTATION TECHNIQUE

1973 ◽  
Vol 137 (2) ◽  
pp. 343-358 ◽  
Author(s):  
Lloyd K. Everson ◽  
Donald N. Buell ◽  
G. Nicholas Rogentine
Keyword(s):  
Cell Cycle ◽  
Surface Membrane ◽  
Fold Increase ◽  
Growth Patterns ◽  
Cell Cycle Phase ◽  
Lymphoid Cells ◽  
Velocity Sedimentation ◽  
Cycle Phase ◽  
Growth Media ◽  
Human Lymphoid Cell

Human lymphoid tissue culture cells can be separated according to cell size and corresponding cell cycle phase with a velocity sedimentation centrifugation method employing a continuous 5–20% wt/wt Ficoll gradient. A 7-fold increase in streaming limit was achieved by placing a buffer zone of isosmolar 5% Ficoll on top of the gradient before application of the cell load. The various pooled populations of cells from upper, middle, and lower areas of the gradient were characterized using autoradiographic, TCA-precipitable 3H]thymidine incorporation, and Fuelgen microspectrophotometric methods. The upper range of the gradient contains cells in the G1 cell cycle phase; the lower range, cells in the G2 phase; cells found in the middle of the gradient belong largely to the S phase of the cell cycle. These gradient-separated cell pools contained relatively little contamination with cells from other phases of the cell cycle and, when explanted from the gradient into fresh growth media, showed growth patterns characteristic of synchronized cell populations. This system of cell separation provides a useful tool for investigating the relationship of the cell cycle to surface membrane and metabolic characteristics in human lymphoid cell culture systems.

scholarly journals Efficient anaerobic consumption of D-xylose by E. coli BL21(DE3) via xylR adaptive mutation

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Jung Min Heo ◽  
Hyun Ju Kim ◽  
Sang Jun Lee
Keyword(s):  
Consumption Rate ◽  
Xylose Isomerase ◽  
Fold Increase ◽  
Adaptive Mutation ◽  
Growth Media ◽  
Missense Mutations ◽  
Xylose Consumption ◽  
E Coli ◽  
Fermentation Technology ◽  
Xylose Transporter

Abstract Background Microorganisms can prioritize the uptake of different sugars depending on their metabolic needs and preferences. When both D-glucose and D-xylose are present in growth media, E. coli cells typically consume D-glucose first and then D-xylose. Similarly, when E. coli BL21(DE3) is provided with both D-glucose and D-xylose under anaerobic conditions, glucose is consumed first, whereas D-xylose is consumed very slowly. Results When BL21(DE3) was adaptively evolved via subculture, the consumption rate of D-xylose increased gradually. Strains JH001 and JH019, whose D-xylose consumption rate was faster, were isolated after subculture. Genome analysis of the JH001 and JH019 strains revealed that C91A (Q31K) and C740T (A247V) missense mutations in the xylR gene (which encodes the XylR transcriptional activator), respectively, controlled the expression of the xyl operon. RT-qPCR analyses demonstrated that the XylR mutation caused a 10.9-fold and 3.5-fold increase in the expression of the xylA (xylose isomerase) and xylF (xylose transporter) genes, respectively, in the adaptively evolved JH001 and JH019 strains. A C91A adaptive mutation was introduced into a new BL21(DE3) background via single-base genome editing, resulting in immediate and efficient D-xylose consumption. Conclusions Anaerobically-adapted BL21(DE3) cells were obtained through short-term adaptive evolution and xylR mutations responsible for faster D-xylose consumption were identified, which may aid in the improvement of microbial fermentation technology.

NUTRITIONAL AND METABOLIC STUDIES OF DEBARYOMYCES HANSENII

1962 ◽  
Vol 8 (2) ◽  
pp. 213-220 ◽  
Author(s):  
Emanuel Merdinger ◽  
Salem Shair
Keyword(s):  
Citric Acid ◽  
Sodium Acetate ◽  
Debaryomyces Hansenii ◽  
Chloride Concentration ◽  
Normal Growth ◽  
Nitrogen Sources ◽  
Logarithmic Phase ◽  
Growth Media ◽  
Metabolic Products ◽  
Esterified Sterols

The growth of strain NRRL Y-1448 of Debaryomyces hansenii was studied by using various growth media. A sodium chloride concentration of 2% to 3% and a pH of 5.5 were optimal for growth. No growth occurred below pH 4 and above pH 8. Either thiamine or biotin was required for normal growth. Growth occurred in media containing asparagine plus sodium acetate or asparagine plus citric acid but not citric acid plus acetate. Urea or ammonium sulphate were satisfactory nitrogen sources. The logarithmic phase of growth was reached twice as fast in media containing fructose as in media containing glucose. The metabolic products of growth were alcohol, acetic acid, pyruvic acid, and both free and esterified sterols.

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