scholarly journals In vitro pollen culture and the regeneration of Brassica campestris L. plants

1995 ◽  
Vol 4 (5-6) ◽  
pp. 513-518
Author(s):  
Yang-Dong Guo ◽  
Seppo Pulli
Keyword(s):  
Culture Medium ◽  
Brassica Campestris ◽  
Higher Plants ◽  
Oilseed Crop ◽  
Pollen Culture ◽  
Culture Techniques ◽  
Liquid Culture Medium ◽  
Embryo Yield ◽  
Colchicine Solution

Brassica campestris (Brassica rapa L. ssp. oleifera) is an important oilseed crop, particularly in Finland. Pollen culture techniques for haploid production have been developed, but B. campestris is relatively recalcitrant in pollen culture. Twenty eight genotypes of B. campestris were included in this study. The donor plants were grown in the greenhouse and transferred to the growth cabinet before bolting. Buds (2-4 mm long) were selected, macerated in B 5 medium, then NLN liquid culture medium was added. The microspores were incubated in the dark at 32°C for 72 h, then at 25°C for a further three weeks. Nineteen genotypes produced microspore-derived embryos. The highest yield was more than 300 embryos per 100 buds. Activated charcoal (150 mg/L) promoted embryogenesis, pollen development was faster and the embryo yield was higher. Plants were regenerated after transferring embryos to a solid B 5 medium. Colchicine solution was used to double the chromosome complements. About 100 regenerate plants have been obtained in our laboratory, and these haploids will be useful for the oilcrop breeding.

Fate of Safening Agent L-2-Oxothiazolidine-4-Carboxylic Acid in Sorghum (Sorghum bicolor)

Weed Science ◽  
1990 ◽  
Vol 38 (3) ◽  
pp. 201-205 ◽  
Author(s):  
James L. Hilton ◽  
Parthasarathy Pillai ◽  
Helen A. Norman
Keyword(s):  
Carboxylic Acid ◽  
Sorghum Bicolor ◽  
Culture Medium ◽  
In Vitro Assay ◽  
Thiol Compounds ◽  
Vitro Assay ◽  
Herbicide Safener ◽  
Liquid Culture Medium ◽  
Germinating Seeds

The herbicide safener OTC (L-2-oxothiazolidine-4-carboxylic acid) increased the amount of reduced thiol compounds in sorghum [Sorghum bicolor(L.) Moench. ‘DK 427′] seedlings. When seedlings were grown in liquid culture medium containing35S-OTC, the compound was metabolized to radiolabeled cysteine and glutathione. The addition of tridiphane [2-(3,5-dichlorophenyl)-2-(2,2,2-trichloroethyl)oxirane] increased conversion of35S-OTC to cysteine and resulted in the formation of one additional35S-labeled compound. When35S-glutathione was injected into germinating seeds it was converted to35S-cysteine and both thiols were subsequently found in roots and shoots. Seeds injected with35S-OTC both translocated the compound to developing roots and shoots and metabolized35S-OTC to cysteine and glutathione. Excised roots and shoots also metabolized35S-OTC to the thiols. In an in vitro assay the enzyme 5-oxoprolinase converted OTC to cysteine.

232 ADDITION OF VERY LOW AMOUNTS OF SERUM (ESTRUS COW SERUM) IMPROVES IN VITRO EMBRYO PRODUCTION IN DAIRY CATTLE

2015 ◽  
Vol 27 (1) ◽  
pp. 205 ◽  
Author(s):  
E. Mullaart ◽  
F. Dotinga ◽  
C. Ponsart ◽  
H. Knijn ◽  
J. Schouten
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Calf Serum ◽  
High Serum ◽  
In Vitro Production ◽  
Embryo Production ◽  
Chi Square ◽  
Embryo Yield ◽  
Vitro Production

Improving the efficiency of the in vitro production (IVP) process is very important because it results in more embryos to be used in breeding programs or as commercial service. At CRV, a culture medium consisting of SOF with amino acids and BSA is used. In the past, richer culture media were used with 10% fetal calf serum combined with BRL cell co-culture. Although the efficiency of the IVP process of these media was good, these rather high serum concentrations were quite often related to large offspring syndrome (LOS). The switch to a culture system without serum resulted in a significant reduction in LOS but also in a reduction of embryo yield. The aim of the present study was to investigate the effect of adding low amounts of serum to the culture medium on efficiency of embryo production. Immature cumulus-oocyte complexes (COC) were recovered from ovaries 6 to 8 h upon slaughter. The COC were matured in vitro in TCM199/FCS/LH/FSH supplemented with cysteamine (0.1 mM). Subsequently, matured oocytes were fertilised with frozen-thawed gradient-separated semen and further cultured for 7 days in SOFaaBSA. The SOF medium contained either 0 (control), 0.1, 0.5, or 1.0% oestrus cow serum (ECS). Embryos development was scored at Day 7. Three replicates were performed and results were analysed by chi-square analyses. The results clearly show that adding ECS significantly improved embryo production (Table 1). Interestingly, already very low amounts (0.1%) of serum gave a significant increase in embryo percentage. In conclusion, addition of very low amounts of ECS (0.1%) is beneficial for embryo production, resulting in significantly higher embryo production (from 19 to 27%). In a subsequent field trial with OPU-derived embryos, the effect of addition of 0.1% ECS on birth weight (LOS) of the calves has to be investigated. Table 1.Percentage of blastocysts at Day 7 after culture in SOF medium with different amounts of serum

scholarly journals In vitro establishment of Bambusa oldhamii Munro from field-grown matrices and molecular identification of endophytic bactéria

2019 ◽  
Vol 49 ◽  
Author(s):  
Ana Paula de Azevedo Pasqualini ◽  
Gabriela Xavier Schneider ◽  
Hugo Pacheco de Freitas Fraga ◽  
Luiz Antonio Biasi ◽  
Marguerite Quoirin
Keyword(s):  
Molecular Identification ◽  
Culture Medium ◽  
Bacterial Isolate ◽  
Fungal Growth ◽  
Endophytic Bacterium ◽  
Liquid Culture Medium ◽  
Rdna Sequencing ◽  
In Vitro Establishment ◽  
Bambusa Oldhamii

ABSTRACT In plant micropropagation, the establishment stage is difficult, due to the presence of microorganisms in tissues from field-grown matrices, especially for bamboo. This study aimed to establish an efficient asepsis protocol for Bambusa oldhamii explants from field plants, as well as to carry out the molecular identification of a possible endophytic bacterial isolate. The explants were exposed to 70 % alcohol, 1 % sodium hypochlorite (NaOCl), 0.1 % mercuric chloride (HgCl2), thiophanate-methyl (Cercobin®) and chlorhexidine digluconate (2 % Riohex®) in different combinations, and introduced into Murashige and Skoog culture medium (solid or liquid), supplemented or not with 4 mL L-1 of Plant Preservative Mixture (PPMTM), totaling seven treatments. The asepsis and immersion of the explants in the liquid culture medium containing 4 mL L-1 of PPMTM visually inhibited the bacterial and fungal growth, allowed the development of shoots with a mean length of 2.2 cm and posterior subcultures, being the best treatment used for the in vitro establishment of B. oldhamii. The molecular identification of an endophytic bacterium performed by 16S rDNA sequencing allowed to identify the bacterial isolate as Ralstonia sp., with 100 % of similarity, and the phylogenetic analysis grouped it with Ralstonia pickettii. In addition, the bacterial isolate showed to be sensitive to 4 mL L-1 of PPMTM by the minimum inhibitory concentration test.

scholarly journals Effects of a commercial biocide, kasugamycin and consistency of the culture medium on the in vitro establishment of bamboo1

2019 ◽  
Vol 49 ◽  
Author(s):  
Daniela Werner Ribeiro dos Santos ◽  
Théo Piucco Rocker ◽  
Thiago Sanches Ornellas ◽  
Miguel Pedro Guerra
Keyword(s):  
Liquid Culture ◽  
Culture Medium ◽  
Liquid Culture Medium ◽  
Bambusa Vulgaris ◽  
Phyllostachys Bambusoides ◽  
In Vitro Establishment ◽  
Dendrocalamus Asper

ABSTRACT The contamination by microorganisms and oxidation of explants in the in vitro establishment of bamboo are recurrent problems for its micropropagation. In the present study, effects of the biocide Plant Preservative Mixture (PPM™), the antibiotic kasugamycin and the consistency of the culture medium were evaluated in the in vitro establishment of Bambusa vulgaris,Phyllostachys bambusoides and Dendrocalamus asper. The presence of PPM™ in the culture medium had a significant effect using 2 mL L-1 or 4 mL L-1 concentrations, as well as in the liquid culture medium, increasing the plants established in the autumn. Kasugamycin promoted variable responses for all the three species, depending on the season. There was interaction among the factors, demonstrating that higher rates of viable plants can be obtained by combining different strategies to reduce the oxidation and contamination. For the in vitro establishment of B. vulgaris,P. bambusoides and D. asper, it is recommended to add 2 mL L-1 or 4 mL L-1 of PPM™ to the liquid culture medium.

scholarly journals Influence of genotype on protein and oil concentration of soybean seeds

2010 ◽  
Vol 53 (4) ◽  
pp. 793-799 ◽  
Author(s):  
Esmael Lopes dos Santos ◽  
Antonio Eduardo Pípolo ◽  
Ricardo Tadeu de Faria ◽  
Cássio Egidio Cavenaghi Prete
Keyword(s):  
Protein Concentration ◽  
Culture Medium ◽  
Seed Oil ◽  
Soybean Seeds ◽  
Glutamine Concentration ◽  
Liquid Culture Medium ◽  
Immature Seeds ◽  
Oil Concentration

Aiming at evaluating genotype influence on the concentration of protein and oil, immature seeds of cultivars CD 202 and CD 206 were removed from the mother-plant, in the stage R5, and were grown in vitro, in a liquid culture medium which contained 20, 40 and 60 mM of glutamine, during eight days. Afterwards, the concentrations of oil and protein were compared to the contents of the seeds cultivated in vivo. With a higher availability of glutamine for the seed, there was an increase of protein content. The genotypes were statistically different as far as the protein concentration was concerned,which confirmed that the genotype had influence on the concentration of protein in the seed. Oil and protein concentrations were inversely related when a variation of glutamine concentration occurred.

scholarly journals Early studies on the effect of peptide growth factor phytosulfokine-α on Brassica oleracea var. capitata L. protoplasts

2017 ◽  
Vol 86 (3) ◽  
Author(s):  
Agnieszka Kiełkowska ◽  
Adela Adamus
Keyword(s):  
Brassica Oleracea ◽  
Shoot Regeneration ◽  
Liquid Culture ◽  
Culture Medium ◽  
Breeding Lines ◽  
Liquid Culture Medium ◽  
Peptide Growth Factor ◽  
The Media ◽  
Cells Cultured

<span>Phytosulfokines (PSK) are peptidyl growth factors with the potential of inducing cell proliferation. We examined the effect of supplementation of liquid culture medium with 0.1 µM phytosulfokine-α (PSK-α) on protoplast viability and division frequencies in seven accessions of <em>Brassica oleracea</em> var. <em>capitata</em> L., including cultivars and breeding lines. Protoplasts were isolated from leaves and hypocotyls of in vitro grown plants and immobilized in calcium-alginate layers. Cabbage protoplast-derived cells cultured in medium supplemented with 0.1 µM of PSK-α had higher viability and division frequencies compared to cells cultured in PSK-α-free control medium. The effect of PSK-α was more pronounced in low-responding accessions (‘Sława z Gołębiewa’, ‘Ramkila F1’, LM, and LM98); however, in two cultivars with very low response (‘Badger Shipper’ and ‘Oregon 123’), although the division frequencies in the media supplemented with PSK-α were increased over the control, the differences were not significant. Obtained callus colonies were subjected to regeneration. PSK-α supplemented into the liquid culture medium had an indirect effect on shoot regeneration by inducing sustained cell divisions leading to an increase in shoot regeneration in Sława z Gołębiewa and both breeding lines.</span>

CORRELATION BETWEEN THE EFFECTS OF HORMONES ON THE SYNTHESIS OF DNA IN EXPLANTS FROM INDUCED RAT MAMMARY TUMOURS AND THE GROWTH OF THE TUMOURS

1974 ◽  
Vol 62 (2) ◽  
pp. 225-240 ◽  
Author(s):  
D. LEWIS ◽  
R. C. HALLOWES
Keyword(s):  
Organ Culture ◽  
Dna Synthesis ◽  
Culture Medium ◽  
Mammary Glands ◽  
Sprague Dawley ◽  
Culture Techniques ◽  
Mammary Tumours ◽  
Sprague Dawley Rats

SUMMARY Explants from 32 mammary tumours induced in Sprague—Dawley rats by 9,10-dimethyl-1,2-benzanthracene (DMBA) were maintained in organ culture for up to 48 h. Insulin, corticosterone, prolactin, growth hormone and oestradiol were added to the culture medium in various combinations and their effects on the DNA synthesis of the explants was studied. DNA synthesis was stimulated by insulin in explants from 30 out of the 32 tumours examined and this group of 30 responsive tumours could be further subdivided. Explants from 16 tumours showed a greater rate of DNA synthesis in medium containing insulin plus corticosterone plus prolactin than in medium containing insulin alone and this higher rate was decreased by oestradiol; this group is referred to as 'prolactin-responsive'. Explants from the remaining 14 tumours did not show a greater rate of DNA synthesis in medium that contained insulin plus corticosterone plus prolactin than in medium containing insulin alone and neither rate was decreased by oestradiol; this group is referred to as 'insulin-responsive'. Explants from two tumours were not stimulated by insulin and these tumours are referred to as 'non-responsive'. After oophorectomy or administration of ergocryptine to tumour-bearing rats, the prolactin-responsive tumours regressed whereas the non-responsive tumours continued to grow. Explants taken from prolactin-responsive tumours 2 weeks after either oophorectomy or administration of ergocryptine were still prolactin-responsive but those taken from insulin-responsive tumours 2 weeks after the same treatment were now also prolactin-responsive. The non-responsive tumours remained non-responsive. The effects of hormones on the DNA synthesis in vitro of explants from growing DMBA-induced tumours were thus different from those on explants of mammary glands from virgin or pregnant Sprague—Dawley rats. It was concluded that it was possible to predict by organ culture techniques the response in vivo of growing mammary tumours to oophorectomy and ergocryptine administration.

scholarly journals MODULATION OF CULTURE MEDIUM ON THE EX SITU CONSERVATION OF Neoregelia mucugensis Leme (BROMELIACEAE)1

2021 ◽  
Vol 34 (4) ◽  
pp. 763-771
Author(s):  
ANDRESSA PRISCILA PIANCÓ SANTOS LIMA ◽  
FERNANDA DE JESUS OLIVEIRA BASTOS ◽  
ALONE LIMABRITO ◽  
GILÊNIO BORGES FERNANDES ◽  
JOSÉ RANIERE FERREIRA DE SANTANA
Keyword(s):  
Culture Medium ◽  
Ex Situ Conservation ◽  
Slow Growth ◽  
Genetic Material ◽  
Natural Populations ◽  
Ex Situ ◽  
Chapada Diamantina ◽  
Culture Techniques ◽  
Plant Collections

ABSTRACT Bromeliads are the target of predatory extractivism and consequently many species are included in the red list of threatened species, such as those belonging to the genus Neoregelia. Although Neoregelia mucugensis has not been evaluated for the degree of threat, its exploitation is exclusively extractive and its occurrence in Chapada Diamantina-BA is subject to the action of fires that affect the region annually. In this context, applying techniques aimed at protecting this genetic resource is fundamental for both the maintenance of its natural populations and the ex situ conservation of this genetic material. Plant tissue culture techniques have been successfully applied for the conservation of several bromeliad species. One of the methods used is slow growth, which consists in reducing plant metabolism and consequently decelerating its growth, which allows the maintenance of in vitro plant collections without the need for subculture. In this context, the objective of this study was to test the reduction of salts in the culture medium and the addition of osmoregulators on the induction of slow growth of N. mucugensis. Plants were subjected to treatments composed of different concentrations of MS medium and mannitol for a period of 12 months, when then analyses were conducted to evaluate growth, chlorophyll content and regeneration capacity of shoots in vitro. It was found that the treatment containing MS ½ and 7.8 g.L-1 of mannitol is indicated for in vitro conservation of N. mucugensis with maintenance of the regenerative capacity of its tissues.

scholarly journals The Effects of Heat Shock Protein 70 Addition in the Culture Medium on the Development and Quality of In Vitro Produced Heat Shocked Bovine Embryos

Animals ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 3347
Author(s):  
Konstantina Stamperna ◽  
Themistoklis Giannoulis ◽  
Eleni Dovolou ◽  
Maria Kalemkeridou ◽  
Ioannis Nanas ◽  
...  
Keyword(s):  
Gene Expression ◽  
Culture Medium ◽  
Heat Exposure ◽  
Developmental Competence ◽  
Culture Period ◽  
Negative Effects ◽  
Altered Expression ◽  
Embryo Yield ◽  
Bovine Oocytes

The aims of the present study were to examine the effects of HSP70 addition in the in vitro culture medium of day 3 embryos on their developmental competence and quality. Bovine oocytes (n = 1442) were in vitro matured, inseminated and cultured for the first two days according to standardized methods. The presumptive zygotes were randomly allocated in three experimental groups: Control, C (embryos cultured at 39 °C throughout the culture period), group C41 (temperature was raised to 41 °C from the 48th to 72nd h post insemination (p.i.) and then it returned at 39 °C for the remaining culture period), and group H41 (the temperature modification was the same as in C41 and during heat exposure, HSP70 was added in the culture medium). Cleavage and embryo yield were assessed 48 h p.i. and on days 7, 8, 9, respectively and gene expression in day 7 blastocysts was assessed by RT-PCR. Blastocyst yield was the highest in group C39; and higher in group H41 compared to group C41. From the gene expression analyses, altered expression of 11 genes was detected among groups. The analysis of the orchestrated patterns of gene expression differed between groups. The results of this study confirm the devastating effects of heat stress on embryo development and provide evidence that HSP70 addition at the critical stages can partly counterbalance, without neutralizing, the negative effects of the heat insult on embryos, acting mainly through mechanisms related to energy deployment.

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