scholarly journals In vitro culture of Nonea pulla DC. seeds.

Author(s):  
E.F. Semenova ◽  
◽  
K.V. Vedernikova ◽  
E.Yu. Schetneva ◽  
◽  
...  

Nonea pulla DC. is a promising perennial medicinal plant growing in the Crimea. Controlled in vitro cultivation of nonea seeds allows improving the up-to-date techniques of seedlings preparation. The conducted experiments confirmed the low germinating capacity of seeds (5–9 %). To increase this parameter and to speed up the introduction process, we investigated the Nonea pulla in vitro culture. The initial phases of germination were expectedly observed during seeds cultivation. The seed swelling, rupture of pericarp and seed hull, release of germ with cotyledons, dehiscence of cotyledons were detected. Moreover, in some cases, no subsequent development was observed. However, normal germs formed in 60% of cases. Seeds also sprouted without the prior cold stratification. For the following growth, plants required a relatively simple culture medium. The maximum development conditions were reached after 1.0–1.5 months of in vitro cultivation (26±2 °С, illuminance of 2000–3000 lux, 16-hour photoperiod).

2018 ◽  
Vol 22 ◽  
pp. 257-261 ◽  
Author(s):  
S. I. Kovtun ◽  
A. B. Zyuzyun ◽  
O. V. Shcherbak ◽  
P. A. Trotskiy

Aim. Investigated effect nanomaterial of highly dispersed ultra fine silica (UFS) by carbohydrate – sucrose (UFS/sucrose) on the effectiveness of meiotic maturation cows oocytes in vitro. Methods. The fresh and frozen – thawed cow oocyte-cumulus complex (OCC) was divided into four groups: three experimental in which the cultivation was carried out in a medium containing of 0, 1; 0, 01 and 0,001 % UFS/sucrose and control without adding nanomaterial. Results. It was concluded that the addition of UFS/sucrose in 0.001 % concentration is effective for elevation level of oocytes maturation and provides reception of 76,8% oocytes that induced the metaphase II of meiosis. Adding carbohydrate – sucrose (UFS/sucrose) in 0.001 % concentration to the culture medium frozen – thawed cow generative cells, make positive effect on in vitro fertilization and provide embryos quantity enhancement to 33.3 %. Conclusions. Addition of UFS/sucrose in 0.001 % concentration to the culture medium have increase effect and promote level in vitro maturation of cows oocytes rising to 76.8 %. Usage of UFS/sucrose in 0.001 % concentration as part of in vitro culture medium for cows oocyte-cumulus complex conduce rising quantity of cattle embryos to 33.3 % after in vitro fecundation frozen – thawed and maturation oocytes. Keywords: oocytes, in vitro culture, embryos, nanomaterial, ultra fine silica (UFS).


2017 ◽  
Vol 52 (11) ◽  
pp. 1123-1126 ◽  
Author(s):  
Filipe Almendagna Rodrigues ◽  
Renata Alves Lara Silva Rezende ◽  
Moacir Pasqual ◽  
Maria Teresa Gomes Lopes

Abstract: The objective of this work was to evaluate the effects of the solidifying agents agar and phytagel and of activated charcoal on the in vitro cultivation of two maná cubiu (Solanum sessiliflorum) varieties: Thaís and Santa Luzia. The phytotechnical characteristics analyzed included number of leaves, number of roots, shoot and root length, and fresh matter of shoot and root. Regardless of the variety, phytagel was superior to agar as a culture medium. A greater number of leaves and longer shoots were observed in the Santa Luzia variety, in the absence of charcoal. The Thaís variety showed longer shoots and roots in the presence of charcoal.


2018 ◽  
Vol 24 (1/2) ◽  
pp. 46-54
Author(s):  
Joseila Maldaner ◽  
Raíssa Schwalbert ◽  
Gerusa Pauli Kist Steffen ◽  
Cleber Witt Saldanha ◽  
Rejane Flores ◽  
...  

Desmodium incanum is a perennial species, native in grassland of Rio Grande do Sul, Brazil and popularly known as "pega-pega". Due its good characteristics as forage, the degradation of the natural fields became an alarming reality. The conservation of these species requires the knowledge of the nutritional requirements and the abiotic conditions that the species is adapted. So, besides in vitro culture as germplasm conservation, it is suggested practices such as acclimatization of plants to increase the survival percentage in ex vitro environment. The plant nutritional status in the period prior to the acclimatization may influences the results of survival and development during this process. To this end, it was tested the influence of nutrition of the seedling during in vitro cultivation on the D. incanum acclimatization through variations in the salt composition of the culture medium. Our results showed that the culture medium which provided the greatest survival rate and better plant growth was MS medium with half the standard concentration of salts. On the other hand, we observed that the iron halving, in culture medium, during in vitro culture was highly detrimental to the acclimatization process of D. incanum. So we could confirm the importance of nutritional status on in vitro culture for acclimatization stage.


2021 ◽  
Vol 2 (2) ◽  
pp. 538-553
Author(s):  
Natacha Coelho ◽  
Alexandra Filipe ◽  
Bruno Medronho ◽  
Solange Magalhães ◽  
Carla Vitorino ◽  
...  

In vitro culture is an important biotechnological tool in plant research and an appropriate culture media is a key for a successful plant development under in vitro conditions. The use of natural compounds to improve culture media has been growing and biopolymers are interesting alternatives to synthetic compounds due to their low toxicity, biodegradability, renewability, and availability. In the present study, different culture media containing one biopolymer (chitosan, gum arabic) or a biopolymer derivative [hydroxyethyl cellulose (HEC), carboxymethyl cellulose (CMC)], at 100 or 1000 mg L−1, were tested regarding their influence on the growth and physiological responses of Thymus lotocephalus in vitro culture. Cellulose-based biopolymers (HEC and CMC) and gum arabic were used for the first time in plant culture media. The results showed that CMC at 100 mg L−1 significantly improved shoot elongation while chitosan, at the highest concentration, was detrimental to T. lotocephalus. Concerning only the evaluated physiological parameters, all tested biopolymers and biopolymer derivatives are safe to plants as there was no evidence of stress-induced changes on T. lotocephalus. The rheological and microstructural features of the culture media were assessed to understand how the biopolymers and biopolymer derivatives added to the culture medium could influence shoot growth. As expected, all media presented a gel-like behaviour with minor differences in the complex viscosity at the beginning of the culture period. Most media showed increased viscosity overtime. The surface area increased with the addition of biopolymers and biopolymer derivatives to the culture media and the average pore size was considerably lower for CMC at 100 mg L−1. The smaller pores of this medium might be related to a more efficient nutrients and water uptake by T. lotocephalus shoots, leading to a significant improvement in shoot elongation. In short, this study demonstrated that the different types of biopolymers and biopolymer derivatives added to culture medium can modify their microstructure and at the right concentrations, are harmless to T. lotocephalus shoots growing in vitro, and that CMC improves shoot length.


2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
F Du ◽  
R Li ◽  
Q Zhang ◽  
W Wang

Abstract Study question what is the source, prevalence, and influence of microbial contamination on in vitro fertilization (IVF) and embryo transfer (ET) cycles? Summary answer Microbial contamination mainly occurs on Day 2, most caused by Escherichia coli carried with semen. ICSI could prevent contamination effectively and get good clinical outcomes. What is known already Microbial contamination occurs in IVF-ET system occasionally, which is hard to stop happening. The IVF culture system and laboratory environment, the patients’ follicular fluid and semen are not absolutely sterile, while the antibiotics in culture medium isn’t effective for all microbe types, and the artificial operations may bring in microbes. Generally, microbial contamination leads to degradation of embryos, reduction the number of embryos available, and infection of female reproductive tract, which would increase the cost of patients’ time, money, and bring psychological damages. A better understanding of embryo contamination in IVF culture system is of added value. Study design, size, duration A total of 29583 IVF-ET cycles were enrolled in this prospective observational study, from January 2010 to December 2020, included 70 microbial contamination cycles discovered in Day1-Day3 (D1-D3) of in vitro culture. Follicular fluid and semen saved on oocyte retrieval day, and culture medium contaminated were examined and identified for microorganisms at each contamination cycle. Participants/materials, setting, methods Compared the contamination rate of different insemination methods (IVF/ICSI/IVF+ICSI), different in vitro culture days (D1-D3), and different samples examination (follicular fluid, semen, culture medium) respectively, identified the source of microorganism types, compared the IVF culture outcomes and clinical outcomes between total contamination group (TC group, 42 cases) and partial contamination group (PC group, 28 cases). Main results and the role of chance A total of 70 microbial contamination cases occurred in 29583 oocyte retrieving cycles (0.24%), and it was observed only in IVF embryos but never in ICSI (Intracytoplasmic sperm injection) embryos. 38 contamination cases occurred on D2 with a highest ratio (54.3%) compared to D1 (32.9%) and D3(12.9%); Compared with follicular fluid, semen was the main cause inducing contamination from D1 to D3, and Escherichia coli in semen and culture medium, Enterococcus faecalis in follicular fluid proved to be the most common sources. Compared with TC group, the PC group showed a lower rate of No-available embryos (21.4% vs 81.0%) and a higher rate of blastocyst formation (41.2% vs 28.6%), In addition, the clinical pregnancy rate of PC group was higher than that of TC group in both fresh and frozen-thawed embryo transfer cycles (31.3% vs 16.7%, 38.5% vs 0.0%). Limitations, reasons for caution Further study is still necessary to better understand the sources that induce microbial contamination embryos, and more efficient methods are required to remove the microbes on these contaminated embryos so as better develop and manage a sterile micro-environment for successful embryo growth. Wider implications of the findings: The differential embryonic microbe types associated to different IVF culture and clinical outcomes in patients undergoing IVF-ET might have profound implications for understanding the microbial sources and making a better management of IVF culture system. Trial registration number Not applicable


Development ◽  
1974 ◽  
Vol 32 (2) ◽  
pp. 515-532
Author(s):  
A. Colman

RNA synthesis can be maintained in large oocytes of Xenopus laevis during periods of in vitro culture of at least 10 days. A simple salt medium, modified Barth's solution, is found to be as effective a culture medium for these oocytes as several other complex media. The newly synthesized RNA is characterized electrophoretically and shown to consist predominantly of ribosomal RNA precursor, 28S and 18S ribosomal RNA, and 4S RNA. The distribution of this RNA within the oocyte is detected autoradiographically, where it is found to be greatly concentrated over the nucleoli. No qualitative alterations in either of these parameters are found during culture, within the limits of sensitivity of the assay procedures.


1997 ◽  
Vol 35 (9) ◽  
pp. 2430-2431 ◽  
Author(s):  
M E Limoncu ◽  
I C Balcioğlu ◽  
K Yereli ◽  
Y Ozbel ◽  
A Ozbilgin

2020 ◽  
Vol 26 (1) ◽  
pp. 18-34 ◽  
Author(s):  
Elizangela Rodrigues Santos ◽  
João Paulo Rodrigues Martins ◽  
Luiz Carlos de Almeida Rodrigues ◽  
Andreia Barcelos Passos Lima Gontijo ◽  
Antelmo Ralph Falqueto

Abstract When propagated in vitro, explants receive all the nutrients needed for their growth, including carbohydrates, from the culture medium. However, it is not well understood how the type and concentration of carbohydrates can affect the functioning of the photosynthetic apparatus (particularly photosystem II) of these plants. The aim was to assess the morphophysiological responses of Billbergia zebrina plants in function of sources and concentrations of carbohydrates during in vitro culture. Side shoots of plants previously established in vitro were individualized and transferred to a culture medium containing fructose, glucose or sucrose in four concentrations (0, 15, 30 or 45 g L−1). After growth for 55 days, the chlorophyll a fluorescence transient, leaf anatomy and growth were analyzed. The concentration and type of carbohydrate employed during in vitro culture did not decrease the photosynthetic apparatus performance. However, concentrations above 30 g L−1 led to anatomical modifications, revealing some degree of stress suffered by the plants. When grown in concentrations of 15 and 30 g L−1, irrespective of the carbohydrate used, the plants presented greater stomatal density. The supplementation of the culture medium with monosaccharides caused alterations in the development of the xylem vessels, such as increased number and diameter, allowing adjustment to the microenvironmental conditions. The in vitro conditions influenced the photosynthetic and anatomical responses of plants. The concentration interval from 15 to 30 g L−1 sucrose had a better effect by not causing large changes in the performance of the photosynthetic apparatus and anatomy of plants.


Author(s):  
Yu. Khoma ◽  
L. Khudolieieva ◽  
N. Kutsokon

Soil salinization is an important abiotic factor negatively affecting plant growth, development and productivity. Fast-growing poplar and willow trees are important plants for bioenergy production demonstrating varying degrees of adaptation to different habitats. The study of salt resistance in different clones of poplars and willows will reveal genotypes that can be planted in saline soils for producing biomass for the bioenergy industry. Therefore, the aim of the study was to investigate the effects of salt stress on poplar plants of clone 'INRA 353-38' (Populus tremula × P. tremuloides) and willow clone 'Zhytomyrska – 1' (Salix sp.) under in vitro culture. For this purpose the plants were cultivated on MS nutrient medium with the addition of sodium chloride in concentrations 25 mM, 50 mM and 100 mM. The control plants were grown on the sodium chloridefree medium. The plant status (with a 4-score scale), the intensity of their growth (by shoot length) and rooting capacity (by the number of roots) were assessed on the 10th and the 30th day of cultivation. The results obtained indicate a high level of sensitivity to sodium chloride of both studied clones under in vitro cultivation. But the willow 'Zhytomyrska – 1' had a higher sensitivity to salt stress comparing to hybrid polar 'ІNRA 353-38' since growth parameters of willow were significantly decreased even under the concentration of sodium chloride 50 mM, and in the case of short term influence (10 days) of the highest concentration of sodium chloride (100 mM) all willow plants terminated their growth and quickly died. The growth parameters of hybrid poplar were declined within a month, mainly under the highest concentration of sodium chloride, but even under such conditions some part of the shoots were able to survive.


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