scholarly journals Optimized Protocol for In Vitro Pollen Germination in Yam (Dioscorea spp.)

Plants ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 795
Author(s):  
Jean M. Mondo ◽  
Paterne A. Agre ◽  
Robert Asiedu ◽  
Malachy O. Akoroda ◽  
Asrat Asfaw
Keyword(s):  
Pollen Germination ◽  
Incubation Temperature ◽  
Optimal Growth ◽  
Germination Percentage ◽  
Medium Composition ◽  
Assessment Protocol ◽  
Critical Problems ◽  
Growing Conditions

Yam (Dioscorea spp.) plants are mostly dioecious and sometimes monoecious. Low, irregular, and asynchronous flowering of the genotypes are critical problems in yam breeding. Selecting suitable pollen parents and preserving yam pollen for future use are potential means of controlling these constraints and optimizing hybridization practice in yam breeding programs. However, implementing such procedures requires a robust protocol for pollen collection and viability testing to monitor pollen quality in the field and in storage. This study, therefore, aimed at optimizing the pollen germination assessment protocol for yam. The standard medium composition was stepwisely modified, the optimal growth condition was tested, and in vivo predictions were made. This study showed that the differences in yam pollen germination percentage are primarily linked to the genotype and growing conditions (i.e., medium viscosity, incubation temperature, and time to use) rather than the medium composition. The inclusion of polyethylene glycol (PEG) in the culture medium caused 67–75% inhibition of germination in D. alata. Although the in vivo fertilization was dependent on female parents, the in vitro germination test predicted the percentage fruit set at 25.2–79.7% and 26.4–59.7% accuracy for D. rotundata and D. alata genotypes, respectively. This study provides a reliable in vitro yam pollen germination protocol to support pollen management and preservation efforts in yam breeding.

scholarly journals Influence of Light Conditions and Medium Composition on Morphophysiological Characteristics of Stevia rebaudiana Bertoni In Vitro and In Vivo

Horticulturae ◽  
2021 ◽  
Vol 7 (7) ◽  
pp. 195
Author(s):  
Alla A. Shulgina ◽  
Elena A. Kalashnikova ◽  
Ivan G. Tarakanov ◽  
Rima N. Kirakosyan ◽  
Mikhail Yu. Cherednichenko ◽  
...  
Keyword(s):  
Culture Media ◽  
Stevia Rebaudiana ◽  
Adventitious Shoot ◽  
Optimal Growth ◽  
Medium Composition ◽  
Light Spectra ◽  
Stevia Rebaudiana Bertoni ◽  
Multiplication Coefficient

We investigated the influence of different conditions (light composition and plant growth regulators (PGRs) in culture media) on the morphophysiological parameters of Stevia rebaudiana Bertoni in vitro and in vivo. Both PGRs and the light spectra applied were found to significantly affect plant morphogenesis. During the micropropagation stage of S. rebaudiana, optimal growth, with a multiplication coefficient of 15, was obtained in an MS culture medium containing 2,4-epibrassinolide (Epin) and indole-3-acetic acid (IAA) at concentrations of 0.1 and 0.5 mg L−1, respectively. During the rooting stage, we found that the addition of 0.5 mg L−1 hydroxycinnamic acid (Zircon) to the MS medium led to an optimal root formation frequency of 85% and resulted in the formation of strong plants with well-developed leaf blades. Cultivation on media containing 0.1 mg L−1 Epin and 0.5 mg L−1 IAA and receiving coherent light irradiation on a weekly basis resulted in a 100% increase in the multiplication coefficient, better adventitious shoot growth, and a 33% increase in the number of leaves. S. rebaudiana microshoots, cultured on MS media containing 1.0 mg L−1 6-benzylaminopurine (BAP) and 0.5 mg L−1 IAA with red monochrome light treatments, increased the multiplication coefficient by 30% compared with controls (white light, media without PGRs).

scholarly journals Effect of Boron and Calcium on in Vitro and in Vivo Lowbush Blueberry Pollen Germination

HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 784B-784
Author(s):  
Youzhi Chen ◽  
John M. Smagula
Keyword(s):  
Pollen Germination ◽  
Pollen Grains ◽  
Germination Percentage ◽  
Control Treatment ◽  
Water Control ◽  
Flower Bud ◽  
Lowbush Blueberry ◽  
To Receive

Ten clones of lowbush blueberry (Vaccinium angustifolium) having low leaf boron (B) concentrations (<20 ppm) were selected to receive fall foliar B (400 ppm), Ca (4000 ppm), B (400 ppm) + Ca (4000 ppm), or water (control). B concentration was raised in stem and bud tissue 3 months after application, but Ca concentration was unaffected. Two randomly selected 5-inch sod plugs from treatment plots within each clone were transported to cold storage at 2.7C for 1000 h to satisfy flower bud dormancy, then to a growth chamber at 24C to blossom. Pollen from plants receiving B had lower in vitro germination rates on 5% agar with 12% lactose after 20 h compared to control and Ca treatments. For in vivo germination, 10 blossoms were randomly selected on sod plugs of each treatment plot to receive 15 control-treatment pollen grains, which were allowed to germinate for 3 days. With the aid of fluorescence microscopy, a higher pollen germination percentage was observed in blossoms of plants receiving B, Ca, and B + Ca. B and Ca may have more influence on the ability of the stigma to stimulate pollen germination than on the germinability of pollen grains themselves.

Differentiation of mesenchymal stem cells from humans and animals into insulin-producing cells: An overview in vitro induction forms

2020 ◽  
Vol 16 ◽  
Author(s):  
Bruna O. S. Câmara ◽  
Bruno M. Bertassoli ◽  
Natália M. Ocarino ◽  
Rogéria Serakides
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Culture Medium ◽  
Adipogenic Differentiation ◽  
Medium Composition ◽  
Cell Therapies ◽  
Insulin Producing Cells ◽  
Msc Differentiation

The use of stem cells in cell therapies has shown promising results in the treatment of several diseases, including diabetes mellitus, in both humans and animals. Mesenchymal stem cells (MSCs) can be isolated from various locations, including bone marrow, adipose tissues, synovia, muscles, dental pulp, umbilical cords, and the placenta. In vitro, by manipulating the composition of the culture medium or transfection, MSCs can differentiate into several cell lineages, including insulin-producing cells (IPCs). Unlike osteogenic, chondrogenic, and adipogenic differentiation, for which the culture medium and time are similar between studies, studies involving the induction of MSC differentiation in IPCs differ greatly. This divergence is usually evident in relation to the differentiation technique used, the composition of the culture medium, the cultivation time, which can vary from a few hours to several months, and the number of steps to complete differentiation. However, although there is no “gold standard” differentiation medium composition, most prominent studies mention the use of nicotinamide, exedin-4, ß-mercaptoethanol, fibroblast growth factor b (FGFb), and glucose in the culture medium to promote the differentiation of MSCs into IPCs. Therefore, the purpose of this review is to investigate the stages of MSC differentiation into IPCs both in vivo and in vitro, as well as address differentiation techniques and molecular actions and mechanisms by which some substances, such as nicotinamide, exedin-4, ßmercaptoethanol, FGFb, and glucose, participate in the differentiation process.

scholarly journals An optimised GAS-pharyngeal cell biofilm model

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Heema K. N. Vyas ◽  
Jason D. McArthur ◽  
Martina L. Sanderson-Smith
Keyword(s):  
Crystal Violet ◽  
Biofilm Formation ◽  
Cell Model ◽  
Matrix Material ◽  
Three Dimensional ◽  
Growth Period ◽  
Optimal Growth ◽  
Abiotic Surfaces

AbstractGroup A Streptococcus (GAS) causes 700 million infections and accounts for half a million deaths per year. Biofilm formation has been implicated in both pharyngeal and dermal GAS infections. In vitro, plate-based assays have shown that several GAS M-types form biofilms, and multiple GAS virulence factors have been linked to biofilm formation. Although the contributions of these plate-based studies have been valuable, most have failed to mimic the host environment, with many studies utilising abiotic surfaces. GAS is a human specific pathogen, and colonisation and subsequent biofilm formation is likely facilitated by distinct interactions with host tissue surfaces. As such, a host cell-GAS model has been optimised to support and grow GAS biofilms of a variety of GAS M-types. Improvements and adjustments to the crystal violet biofilm biomass assay have also been tailored to reproducibly detect delicate GAS biofilms. We propose 72 h as an optimal growth period for yielding detectable biofilm biomass. GAS biofilms formed are robust and durable, and can be reproducibly assessed via staining/washing intensive assays such as crystal violet with the aid of methanol fixation prior to staining. Lastly, SEM imaging of GAS biofilms formed by this model revealed GAS cocci chains arranged into three-dimensional aggregated structures with EPS matrix material. Taken together, we outline an efficacious GAS biofilm pharyngeal cell model that can support long-term GAS biofilm formation, with biofilms formed closely resembling those seen in vivo.

In Vitro Germination of Eucalyptus Pollen: Response to Variation in Boric Acid and Sucrose

1989 ◽  
Vol 37 (5) ◽  
pp. 429 ◽  
Author(s):  
BM Potts ◽  
JB Marsden-Smedley
Keyword(s):  
Pollen Tube ◽  
Boric Acid ◽  
Pollen Tube Growth ◽  
Pollen Germination ◽  
Sucrose Concentration ◽  
Response Surfaces ◽  
Tube Growth ◽  
Pollen Competition

The effect of boric acid (0-450 ppm) and sucrose (0-40%) on pollen germination and pollen tube growth in Eucalyptus globulus, E. morrisbyi, E. ovata and E. tirnigera was examined in vitro. Over the con- centrations tested, sucrose had by far the largest effect upon both pollen germination and tube lengths. The optimum sucrose concentration for pollen germination (30%) and pollen tube growth (20%) differed markedly with very little (<lo%) germination occurring in the absence of sucrose. The interaction of sucrose and boric acid was significant. However, in general both pollen germination and pollen tube growth were increased by the addition of up to 100 ppm boric acid, but above this level the response plateauxed. The four species differed significantly in their pattern of response to both boric acid and sucrose and the predicted optima derived from analysis of response surfaces differed between species. The predicted sucrose concentration for optimal germination and growth of E. urnigera pollen was consistently less than the other species and in terms of the optimal level of boric acid for pollen tube growth species can be ranked in the order E. globulus > E. ovata > E. morrisbyi = E. urnigera. Pollen germination and tube growth of all four species on a medium comprising 20% sucrose and 200 ppm boric acid would not differ significantly from the observed maximum response of each species and this could suffice as a generalised medium. However, if only percentage germination is to be assessed 30% sucrose would be preferable. It is argued that subtle interspecific differences in optimal in vitro con- ditions for pollen germination and pollen tube growth are likely to reflect differences in pollen physiology which in vivo may have important implications for the success of hybridisation where pollen competition occurs.

scholarly journals Foliar Application of Boron to Almond Trees Affects Pollen Quality

2000 ◽  
Vol 125 (2) ◽  
pp. 265-270 ◽  
Author(s):  
A.M.S. Nyomora ◽  
P.H. Brown ◽  
K. Pinney ◽  
V.S. Polito
Keyword(s):  
Pollen Germination ◽  
Foliar Application ◽  
Culture Media ◽  
Pollen Viability ◽  
Prunus Dulcis ◽  
Pollen Tubes ◽  
Tube Growth ◽  
Mature Trees

The effect of boron (B) on in vivo and in vitro development of almond [Prunus dulcis (Mill.) D.A. Webb (syn. P. amygdalus Batsch)] pollen and pollen tubes and the resultant effect on fruit set was studied in mature trees. The cultivars Mono (pistil donor) and Butte (pollinizer) in an orchard with low soil B in Fresno, California were sprayed with B at 0, 0.8, 1.7, or 2.5 kg·ha-1 during Fall 1993. Pollen viability as indicated by the fluorescein diacetate method (FDA) was >85% and was not affected by field-applied B, however, in vivo pollen germination and tube growth were enhanced by foliar-applied B. More effect of applied B on in vivo growth appeared as pollen tubes progressed toward the ovary. For in vitro germination, foliar-applied B reduced bursting of tubes, and addition of B to the culture media significantly increased pollen germination and pollen tube growth.

Viability and in Vitro Germination of Johnsongrass (Sorghum Halepense) Pollen

2007 ◽  
Vol 21 (1) ◽  
pp. 23-29 ◽  
Author(s):  
Ian C. Burke ◽  
John W. Wilcut ◽  
Nina S. Allen
Keyword(s):  
Suspension Culture ◽  
Boric Acid ◽  
Pollen Germination ◽  
Pollen Viability ◽  
Calcium Nitrate ◽  
Medium Composition ◽  
In Vitro Tests ◽  
In Vitro Germination ◽  
Agar Culture

A high proportion of viable pollen grains must germinate to study the physiology of pollen growth to reduce the confounding effects of environmental influences on pollen germination. The objectives of this study were to evaluate the nuclear state and develop a suitable medium and culture method for in vitro germination of johnsongrass pollen. Johnsongrass pollen was trinucleate, and in vitro tests for pollen viability using Alexander's stain and a fluorochromatic reaction method (FCR) indicated johnsongrass pollen was viable (92.6 to 98.4%). A factorial treatment arrangement of four concentrations of sucrose, two concentrations of boric acid, and two concentrations of calcium nitrate were used to determine the optimum pollen-germination medium composition in suspension culture, agar culture, and cellophane membrane culture. Germination was highest in a suspension culture with a medium containing 0.3 M sucrose, 2.4 mM boric acid, and 3 mM calcium nitrate. Pollen germination using this medium was 78.9% when anthers were harvested just before anthesis.

scholarly journals Molecular Mechanisms of Pollen-Pistil Interactions in Interspecific Crossing Barriers in the Tomato Family

2000 ◽  
Author(s):  
Yoram Eyal ◽  
Sheila McCormick
Keyword(s):  
Pollen Germination ◽  
Molecular Mechanisms ◽  
Functional Characterization ◽  
Dominant Negative ◽  
Peptide Sequence ◽  
Peptide Ligand ◽  
Hybrid Breakdown ◽  
Degenerate Primers

During the evolutionary process of speciation in plants, naturally occurring barriers to reproduction have developed that affect the transfer of genes within and between related species. These barriers can occur at several different levels beginning with pollination-barriers and ending with hybrid-breakdown. The interaction between pollen and pistils presents one of the major barriers to intra- and inter-specific crosses and is the focus of this research project. Our long-term goal in this research proposal was defined to resolve questions on recognition and communication during pollen-pistil interactions in the extended tomato family. In this context, this work was initiated and planned to study the potential involvement of tomato pollen-specific receptor-like kinases (RLK's) in the interaction between pollen and pistils. By special permission from BARD the objectives of this research were extended to include studies on pollen-pistil interactions and pollination barriers in horticultural crops with an emphasis on citrus. Functional characterization of 2 pollen-specific RLK's from tomato was carried out. The data shows that both encode functional kinases that were active as recombinant proteins. One of the kinases was shown to accumulate mainly after pollen germination and to be phosphorylated in-vitro in pollen membranes as well as in-vivo. The presence of style extract resulted in dephosphorylation of the RLK, although no species specificity was observed. This data implies a role for at least one RLK in pollination events following pollen germination. However, a transgenic plant analysis of the RLK's comprising overexpression, dominant-negative and anti-sense constructs failed to provide answers on their role in pollination. While genetic effects on some of the plants were observed in both the Israeli and American labs, no clear functional answers were obtained. An alternative approach to addressing function was pursued by screening for an artificial ligand for the receptor domain using a peptide phage display library. An enriched peptide sequence was obtained and will be used to design a peptide-ligand to be tested for its effect o pollen germination and tube growth. Self-incompatibility (SI) in citrus was studied on 3 varieties of pummelo. SI was observed using fluorescence microscopy in each of the 3 varieties and compatibility relations between varieties was determined. An initial screen for an S-RNase SI mechanism yielded only a cDNA homologous to the group of S-like RNases, suggesting that SI results from an as yet unknown mechanism. 2D gel electrophoresis was applied to compare pollen and style profiles of different compatibility groups. A "polymorphic" protein band from style extracts was observed, isolated and micro-sequenced. Degenerate primers designed based on the peptide sequence date will be used to isolate the relevant genes i order to study their potential involvement in SI. A study on SI in the apple cultivar Top red was initiated. SI was found, as previously shown, to be complete thus requiring a compatible pollinator variety. A new S-RNase allele was discovered fro Top red styles and was found to be highly homologous to pear S-RNases, suggesting that evolution of these genes pre-dated speciation into apples and pears but not to other Rosaceae species. The new allele provides molecular-genetic tools to determine potential pollinators for the variety Top red as well as a tool to break-down SI in this important variety.

scholarly journals Establishment of Excised Date Palm (Phoenix dactylifera L.) Roots in Vitro

HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 875D-875
Author(s):  
Yousef I. Dlaigan ◽  
A.E. Said ◽  
M.A. El-Hamady
Keyword(s):  
Date Palm ◽  
Incubation Temperature ◽  
Culture Media ◽  
Root Culture ◽  
Optimal Growth ◽  
Phoenix Dactylifera ◽  
Tween 20 ◽  
Distilled Water ◽  
Root Tips

Several trials were conducted with the objective of obtaining an explant for the establishment of date palm root culture in vitro. These trials included disinfecting and germinating seeds of three cultivars on several autoclaved culture media, the influence of incubation temperature on different germination parameters, and the quality of roots before excision and after culture in nutrient media. Three culture media were used: distilled water only; minimal organics that consisted of MS salts, 3% sucrose, modified White's organics, 0.01% inositol, and 0.15% activated charcoal; and 1/2 MS salts mixture, 3% sucrose, and 1/2 modified White's organics. All three media were solidified with 0.7% agar. The seeds were incubated at 25 or 35C for germination. The study revealed the difficulty of seed disinfection. We immersed seeds in 20% to 40% Clorox, with two to four drops of Tween-20, for 30 to 60 minutes and then rinsed them four to five times in deionized distilled water before culturing. The minimal organics medium supported optimal growth of excised roots, and incubation at 35C significantly improved germination. The use of 10-mm-long root tips as explants for culture initiation gave the best growth and elongation. In addition, the growth and elongation of excised root tips increased significantly as the distance from it to the apex of the cotyledonary sheath increased.

scholarly journals Investigations of suitable pollinator for 0900 Ziraat sweet cherry cv.: pollen performance tests, germination tests, germination procedures, in vitro and in vivo pollinations

2008 ◽  
Vol 34 (No. 2) ◽  
pp. 47-53 ◽  
Author(s):  
F. Tosun ◽  
F. Koyuncu
Keyword(s):  
Pollen Tube ◽  
Pollen Germination ◽  
Sweet Cherry ◽  
Fruit Set ◽  
Pollen Viability ◽  
Performance Tests ◽  
Pollen Performance ◽  
Growth Experiments

The objective of this study was to determine suitable cultivars to be used as pollinators for 0900 Ziraat. 0900 Ziraat was used as a female cultivar; Bigarreu Gaucher, Bing, Noble, Starks Gold, Stella, Van, and Vista were used for pollination in the experiments. Starks Gold had the highest values in terms of anther number, average number of pollens per anther, number of pollen per flower and the morphological homogeneity. The pollen viability rates showed significant differences according to stain tests. <i>In vitro</i> pollen germination in 0.5% agar + 15% sucrose + 5 ppm boric acid medium increased with increasing incubation period, and the highest germination was obtained after 48 hours for all cultivars. In orchard trials parallel to pollen tube growth experiments in the laboratory, 0900 Ziraat &times; Starks Gold combination gave the best fruit set results.

Sign in / Sign up

Export Citation Format

Share Document