Spore germination and the life cycle of Frankia in vitro

1989 ◽  
Vol 35 (8) ◽  
pp. 801-806 ◽  
Author(s):  
S. S. Tzean ◽  
John G. Torrey

Bacterial spores of Frankia produced in defined culture media were collected by filtration after washing in amounts approximating 106 spores/mL. Frankia strains UFGCeI5 from Casuarina equisetifolia and UFGCgI1 from C. glauca showed spontaneous release of spores in culture; strains HFPCcI3 from C. cunninghamiana and HFPAllI1 from Allocasuarina lehmanniana showed low spore release in culture unless homogenized. Spore germination was tested on plates of agar nutrient media under different physical and chemical environments. Strain CeI5 showed about 15% germination within 2 days in a defined (BAP) medium with an optimum pH of 6.0–6.8 at 28–35 °C. Under these conditions, strain CcI3 germinated less than 0.5%. In a series of trials with increasingly complex media, strain CeI5 showed 75% spore germination in 3 days at 28 °C and pH 6.7 in the most complex medium tested. Additions of specific single organic compounds to BAP medium caused either strong inhibition or slight stimulation of spore germination. Frankia strains that showed spontaneous spore release germinated better than strains that did not release. Spore germination in Frankia strains is markedly influenced by their strain origin and by the physical and chemical environment in which they are placed.Key words: Casuarina, Frankia, life cycle, spore germination.

2020 ◽  
Vol 21 (11) ◽  
Author(s):  
Yupi ISNAINI ◽  
Titien Ngatinem Praptosuwiryo

Abstract. Isnaini Y, Praptosuwiryo TNg. 2020. In vitro spore germination and early gametophyte development of Cibotium barometz (L.) J. Sm. in different media. Biodiversitas 21: 5373-5381. Cibotium barometz (L.) J. Sm. is known as the golden chicken fern and included in Appendix II of CITES. It is an important export commodity for traditional and modern medicine. Globally, populations of this species are under significant pressure due to overexploitation in the wild. In vitro culture is one of the technologies used for ex-situ propagation and conservation of rare and endangered ferns and lycophytes. This study’s objectives were: (i) to observe in vitro spore germination and early gametophyte development of C. barometz, and (ii) to determine the best culture medium for rapid spore germination and early development of the gametophytes. The sterilized spores were sown in half-strength Murashige & Skoog (½MS) basal medium supplemented with combinations of 6-Benzylaminopurine (BAP) and α-Naphthalene acetic acid (NAA). A factorial combination of four BAP concentrations (0, 2, 4, and 6 mg L-1) with four concentrations of NAA (0; 0.01; 0.03 and 0.05 mg L-1) created 16 treatments replicated in a Completely Randomized Design. Spore germination of C. barometz was observed to be Vittaria-type, and its prothallial development was Drynaria-type. Spore germination started 7-14 days after sowing. Young heart-shape gametophytes consisting of 110-240 cells were formed in 45-61 days after sowing. The two best spore culture media for rapid spore germination and development of C. barometz gametophytes were ½ MS with or without 2 mg L-1 BAP.


2014 ◽  
Vol 2014 ◽  
pp. 1-7 ◽  
Author(s):  
Melekber Sulusoglu ◽  
Aysun Cavusoglu

Pollen quality is important for growers and breeders. This study was carried out to determinein vitropollen viability and pollen germination in seven genotypes of cherry laurel (Prunus laurocerasusL.). Two pollen viability tests, TTC (2,3,5-triphenyl tetrazolium chloride) and IKI (iodine potassium iodide), were used. Pollen traits of genotypes were studied using anin vitromedium containing 0%, 5%, 10%, 15%, and 20% sucrose to determine the best sucrose concentrations for germination. In the second step, the germinated pollen was counted 1, 4, 6, 10, 12, 24, and 48 hours later until there was no further germination. The viability rates were different according to genotypes and tests used. The IKI and TTC staining tests and pollen germination had low correlation (r2= 0.0614 andr2= 0.0015, resp.). Painted pollen rate was higher and pollen was well-stained with IKI test and pollen viability estimated with TTC staining test was better than that estimated with the IKI staining test. 15% sucrose gave the best germination rates in most of the genotypes. Pollen germination rates were recorded periodically from one hour to 48 hours in 15% sucrose and the results showed that pollen germination rates increased after 6 hours of being placed in culture media.


Zygote ◽  
2014 ◽  
Vol 23 (3) ◽  
pp. 327-335 ◽  
Author(s):  
Hruda Nanda Malik ◽  
Dinesh Kumar Singhal ◽  
Shrabani Saugandhika ◽  
Amit Dubey ◽  
Ayan Mukherjee ◽  
...  

SummaryThe present study was carried out to investigate the effects of different activation methods and culture media on the in vitro development of parthenogenetic goat blastocysts. Calcium (Ca2+) ionophore, ethanol or a combination of the two, used as activating reagents, and embryo development medium (EDM), modified Charles Rosenkrans (mCR2a) medium and research vitro cleave (RVCL) medium were used to evaluate the developmental competence of goat blastocysts. Quantitative expression of apoptosis, stress and developmental competence-related genes were analysed in different stages of embryos. In RVCL medium, the cleavage rate of Ca2+ ionophore-treated oocytes (79.61 ± 0.86) was significantly (P < 0.05) higher than in ethanol (74.90 ± 1.51) or in the combination of both Ca2+ ionophore and ethanol. In mCR2a or EDM, hatched blastocyst production rate of Ca2+ ionophore-treated oocytes (8.33 ± 1.44) was significantly higher than in ethanol (6.46 ± 0.11) or in the combined treatment (6.70 ± 0.24). In ethanol, the cleavage, blastocyst and hatched blastocyst production rates in RVCL medium (74.90 ± 1.51, 18.30 ± 1.52 and 8.24 ± 0.15, respectively) were significantly higher than in EDM (67.81 ± 3.21, 14.59 ± 0.27 and 5.59 ± 0.42) or mCR2a medium (65.09 ± 1.57, 15.36 ± 0.52 and 6.46 ± 0.11). The expression of BAX, Oct-4 and GlUT1 transcripts increased gradually from 2-cell stage to blastocyst-stage embryos, whereas the transcript levels of Bcl-2 and MnSOD were significantly lower in blastocysts. In addition, different activation methods and culture media had little effect on the pattern of variation and relative abundance of the above genes in different stages of parthenogenetic activated goat embryos. In conclusion, Ca2+ ionophore as the activating agent, and RVCL as the culture medium are better than other tested options for development of parthenogenetic activated goat blastocysts.


1999 ◽  
Vol 4 (2) ◽  
pp. 19
Author(s):  
K.P. Akhtar ◽  
M. Asif ◽  
M.A. Khan ◽  
M.J. Jaskani ◽  
I.A. Khan

Mango malformation occurs in most mango growing regions of the world. Floral and vegetative malformation have been reported. There is general agreement that the fungal pathogen Fusarium moniliforme var. subglutinans or Fusarium subglutinans is the causal agent. Healthy and malformed samples of both floral and vegetative tissues were collected from different varieties of mango grown in several locations to verify the association of F.moniliforme with mango malformation disease in Pakistan. The fungus was isolated and cultured. Frequency of fungal association with the disease ranged between 90- 94%, There was less recovery of fungus from asymptomatic tissue (12- 15%). There was no difference among the commercial mango varieties in the level of susceptibility to this disease. However, seedling germplasm and land races showing resistance to mango malformation were identified. The in vitro growth characters of the fungus were determined on different culture media, at varying temperatures, light and pH conditions. Mycelial growth on potato dextrose agar was better than nine other media tested. At pH 7.00, the ideal temperature for growth was between 25-30° C. Normally, the malformation is not controlled by fungicide application. The in vitro sensitivity of fungus to six fungicides at three concentrations was determined to seek potential means of chemical control.


1999 ◽  
Vol 1999 ◽  
pp. 62-62
Author(s):  
N.C. Farrar ◽  
M.E. Staines ◽  
G.J. McCallum ◽  
P. Haggarty ◽  
J.J. Robinson ◽  
...  

Serum, which is routinely included in many embryo culture media, can decrease the viability of bovine and ovine embryos produced in cultures employing synthetic oviduct fluid (SOF; Kuran et al., 1999) and represents a possible route for transmission of disease. Alternative approaches include the use of chemically defined culture media but results from studies which avoid sera and its derivatives (e.g., albumin) are generally less favourable due to a lack of knowledge regarding the embryo's response to specific nutrients, most notably fatty acids. As a preliminary step towards investigating fatty acid influences on bovine embryo developmentin vitro, the present study examined the effect of adding palmitic acid (C16:0) to SOF plus bovine serum albumin (BSA) on the performance of this semi-defined culture medium and contrasted it with embryo production in SOF supplemented with serum.


1992 ◽  
Vol 37 (1) ◽  
pp. 127-146 ◽  
Author(s):  
Barry D. Bavister ◽  
Teresa A. Rose-Hellekant ◽  
Tanu Pinyopummintr

2021 ◽  
Vol 10 (3) ◽  
pp. e5210312994
Author(s):  
Paula Fernanda de Azevedo ◽  
Ana Carolina de Almeida ◽  
Rodrigo Domiciano Marques ◽  
Christiane Luciana da Costa ◽  
Anderson Roberto Benedetti ◽  
...  

Cassava root rot causes significant production losses. Difficulties of management, along with the lack of chemical fungicides officially registered by the Ministry of Agriculture, Livestock and Supply (MAPA), require alternative control methods. This study investigated the in vitro antagonistic activity of Trichoderma harzianum as well as a biological fertilizer MICROGEO® on Fusarium solani. The phytophatogenic strains of F. solani, called F1 and F2 were isolated from rotted cassava tubers and T. harzianum, strain ESALQ 1306, from a biological fungicide. Continuous liquid composting of bovine ruminal content, water and MICROGEO® produced the biological fertilizer. Dual culture method was used at the bioassay with T. harzianum. Sterilized (St) and unsterilized (USt) biological fertilizer were tested in different concentrations (% v/v) diluted in the culture media. Colony diameters were measured daily in order to establish the mycelial growth velocity index, inhibition percentage, aside from the sporulation rate and spore germination percentage. The mycelial growth of F. solani isolates was interrupted after hyphae encounter with T. harzianum, due to the occurrence of mycoparasitism, but without influence on the sporulation rate. Sterilized biological fertilizer induced no biocontrol, whereas the unsterilized product (concentration 2.5%) inhibited approximately 64% and 85% of the mycelial growth of isolates F1 and F2, respectively. Moreover, spore germination declined with increasing concentration. In conclusion, T. harzianum and the unsterilized biofertilizer showed in vitro antagonistic activity on F. solani.


2019 ◽  
Vol 70 (11) ◽  
pp. 4000-4002

Phomopsis incarcerata, known as the pathogen which caused the dieback of Rosa branches was detected in many orchards in Romania. Our investigations have approached a series of bio-ecological aspects of this pathogen: isolation, purification and obtaining the pathogen; determination and identification of the pathogen; establishing in vitro parameters of fungal development (nitrogen source). The isolate used in this study was obtained from Rosa spp. branches and was cultivated on three culture media: potato dextrose agar (PDA), malt extract agar and water agar which included five amino-acids: cysteine, glycine, beta-alanine, leucine and tryptophan. Leucine and glycine were favorable for the dynamics of the fungus.When the fungus grew on water agar, the sporulation was completely inhibited. The optimum pH values for the growth and creation of the Phomopsis incarcerata are in the range of 4.4-7, so weak to neutral acid. Keywords: Phomopsis, nitrogen sources, Rosa, pathogen


2012 ◽  
Vol 36 (2) ◽  
pp. 199-203
Author(s):  
Imad M. AL-Maeeni

The present study included 98 bovine ovary collected from AL-Shulla slaughter house immediately after the animal was slaughtered. Ovaries were preserved in physiological saline at 37 Co and transport to the laboratory within 3-4 hrs. Diameters of the follicles were measured and divided into large (6-10 mm) and small (1-5 mm) follicles. Cumulus-oocyte complexes (COCs) were aspirated from large and small follicles using a 18 G needle connected to a 10 ml syringe COCs surrounded by compact and thick cumulus cell were cultured in an integrated culture media RPMI-1640 , than were divided in to two groups , the first group (control media) using integrated culture media only , the second group alcoholic extract of Tribulus terrestris (Tt) plant was added to the culture media RPMI-1640 once a concentration of 50μg/ml , and the other a concentration of 25μ/ml . The results of the current study demonstrated superiority of alcoholic extract of 50μg/ml in the percentage of oocytes maturation which optended or drawn from large and small follicles compared with a concentration of 25μg/ml of the alcoholic extract it was observable the effect of adding alcoholic extract of T. t. plant in two concentrations of 25μg/ml and or 50μg/ml on the percentage of mature oocytes compared to control media (culture media RPMI-1640) .The present results indicated that the oocytes retrieved from larger follicles is better than small follicles in maturing oocytes maturation. In conclusion, alcoholic concentration of 50μg/ml preference extracted first and then the concentration of 25μg/ml T. t. plant extract that was added to culture media RPMI-1640 , superiority in percentage of the mature oocytes compared to control culture media RPMI-1640 , also the large follicle were better than small follicles in terms of equality suitable for mature oocytes.


2018 ◽  
Vol 8 (3) ◽  
pp. 20170037 ◽  
Author(s):  
Jekaterina Kazantseva ◽  
Irina Hussainova ◽  
Roman Ivanov ◽  
Toomas Neuman ◽  
Michael Gasik

A challenge in regenerative medicine is governed by the need to have control over the fate of stem cells that is regulated by the physical and chemical microenvironment in vitro and in vivo . The differentiation of the stem cells into specific lineages is commonly guided by use of specific culture media. For the first time, we demonstrate that human mesenchymal stem cells are capable of turning spontaneously towards neurogenic lineage when seeded on graphene-augmented, highly anisotropic ceramic nanofibres without special differentiation media, contrary to commonly thought requirement of ‘soft’ substrates for the same purpose. Furthermore, pro-inflammatory gene expression is simultaneously suppressed, and expression of factors promoting focal adhesion and monocytes taxis is upregulated. This opens new possibilities of using local topo-mechanical cues of the ‘graphenized’ scaffold surfaces to guide stem cell proliferation and differentiation, which can be used in studies of neurological diseases and cell therapy.


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