Bourgeonnement in vitro a partir d'épiderme séparé de feuille de Bryophyllum Daigremontianum (Crassulacées)

1976 ◽  
Vol 54 (9) ◽  
pp. 852-867 ◽  
Author(s):  
C. Bigot

The lower epidermis from leaves of Bryophyllum Daigremontianum Berger has been mechanically separated and cultured in liquid or solid medium; cytokinin and auxin in combination have been necessary to obtain a lamellar callus, with the Murashige and Skoog's macroelements being more efficient than White's, Knop's, or Lin and Staba's. Numerous buds have been initiated after 3 or 4 weeks, especially with naphthaleneacetic acid (1 mg/litre) in combination with 6-benzylaminopurine (0.2 mg/litre). The best results for bud formation have been obtained with the epidermis of the first and second well developed leaves. The buds, rooted in vitro, developed into normal plants. The histological study has showed that the callus was originated only from the banal epidermal cells. The stomatic cells have not been activated and died. The other tissues of the leaf have never produced any bud in the same culture conditions; then, as for other species, the epidermis has a greater ability to initiate a budding programme.

2015 ◽  
Vol 60 (3) ◽  
pp. 1226-1233 ◽  
Author(s):  
Petros Ioannou ◽  
Aggeliki Andrianaki ◽  
Tonia Akoumianaki ◽  
Irene Kyrmizi ◽  
Nathaniel Albert ◽  
...  

The modestin vitroactivity of echinocandins againstAspergillusimplies that host-related factors augment the action of these antifungal agentsin vivo. We found that, in contrast to the other antifungal agents (voriconazole, amphotericin B) tested, caspofungin exhibited a profound increase in activity against variousAspergillusspecies under conditions of cell culture growth, as evidenced by a ≥4-fold decrease in minimum effective concentrations (MECs) (P= 0. 0005). Importantly, the enhanced activity of caspofungin againstAspergillusspp. under cell culture conditions was strictly dependent on serum albumin and was not observed with the other two echinocandins, micafungin and anidulafungin. Of interest, fluorescently labeled albumin bound preferentially on the surface of germinatingAspergillushyphae, and this interaction was further enhanced upon treatment with caspofungin. In addition, supplementation of cell culture medium with albumin resulted in a significant, 5-fold increase in association of fluorescently labeled caspofungin withAspergillushyphae (P< 0.0001). Collectively, we found a novel synergistic interaction between albumin and caspofungin, with albumin acting as a potential carrier molecule to facilitate antifungal drug delivery toAspergillushyphae.


2018 ◽  
Vol 46 (1) ◽  
pp. 52-57 ◽  
Author(s):  
Alina TREJGELL ◽  
Monika KAMIŃSKA ◽  
Karolina LISOWSKA ◽  
Andrzej TRETYN

This is the first communication of micropropagation system for Inula germanica using seedling explants germinated in vitro. The development of this system gives the possibility of future reintroduction of I. germanica providing a way to stabilize or re-establish its population. Shoot tips and fragments of cotyledons, hypocotyls and roots were isolated from ten-day-old seedlings. Explants were put on MS medium containing 1.0 mg l-1 benzylaminopurine and 0.1 mg l-1 naphthaleneacetic acid and cultured under continuous white fluorescent light (45 μmol.m-2.s-1) at 26 ± 1 °C. The highest percentage of shoot organogenesis (83.3%) was recorded for hypocotyl, while the highest average number of shoots per explant (12.0) was recorded for shoot tips. In subsequent subcultures, multiplication rate decreased to 3.0-4.9 shoots per explant. Less than 19% shoots were able to root on the solid medium without auxins. The highest rooting efficiency (69.3%) was recorded for solid medium supplemented with indolebutyric acid, but growth of roots was inhibited. The percentage of rooted shoots (62.2%) and number of roots per shoot (2.4 per shoot) into the liquid medium were comparable to medium with 0.1 mg·l-1 indolebutyric acid. showing a positive impact on the process of acclimatization. The regenerated plants were able to flowering in the first year after acclimatization. Developed micropropagation system for I. germanica is efficient and can be a useful tool for the active protection of this species.


2013 ◽  
Vol 58 (No. 10) ◽  
pp. 443-452 ◽  
Author(s):  
B. Heidari ◽  
A. Shirazi ◽  
M.-M. Naderi ◽  
M.-M. Akhondi ◽  
H. Hassanpour ◽  
...  

Considering the advent of mesenchymal stem cells (MSCs) as a new source of somatic cells in embryo co-culture system, the current study was aimed to compare in vitro embryo development using embryonic MSCs monolayer with embryonic fibroblast cells (EFCs), oviductal epithelial cells (OECs), and cell-free culture system. The IVM/IVF presumptive sheep zygotes were randomly cultured in different culture conditions as follows: (1) SOFaaBSA medium for the whole culture period (SOF, n = 371), (2) SOFaaBSA medium for the first 3 days followed by co-culturing with MSCs for the next 5 days (SOF-MSCs, n = 120), (3) co-culturing with MSCs for the first 3 days followed by culture in SOFaaBSA medium for the next 5 days (MSCs-SOF, n = 133), (4) co-culturing with MSCs for the whole culture period (MSCs, n = 212), (5) SOFaaBSA medium for the first 3 days followed by co-culturing with EFCs for the next 5 days (SOF-EFCs, n = 132), (6) co-culturing with EFCs for the first 3 days followed by culture in SOFaaBSA medium for the next 5 days (EFCs-SOF, n = 165), (7) co-culturing with EFCs for the whole culture period (EFCs, n = 236), and (8) co-culturing with OECs for the whole culture period (OECs, n = 255). One-Way ANOVA by multiple pairwise comparisons using Tukey&rsquo;s test was performed. Co-culturing in MSCs group had no superiority over EFCs and OECs groups. Though, when co-culturing with MSCs and EFCs was limited to the first 3 days of culture, the embryo development indices were improved compared to the other co-cultured groups. Considering both the hatching rate and total cell number, the application of MSCs for the first 3 days of culture (MSCs-SOF) was superior to the other co-culture and SOF groups. &nbsp;


2014 ◽  
Vol 22 (1) ◽  
pp. 49-61 ◽  
Author(s):  
Rossen S. Sokolov ◽  
Bistra Y. Atanassova ◽  
Elena T. Iakimova

AbstractThe objective of this study was to assess the regeneration response of in vitro cultured Magnolia × soulangeana ‘Alexandrina’ and Magnolia liliiflora ‘Nigra’ to nutrient medium composition. In the primary culture (initiated from dormant axillary buds) combinations of Murashige and Skoog (MS) basal salts with 6-benzylaminopurine and α-naphthaleneacetic acid were tested. The primary explants of cv. ‘Alexandrina’ expressed higher regeneration rate than cv. ‘Nigra’. For both species, the regen eration was most strongly potentiated at addition of 0.25 mg dm−3 of the cytokinin alone. The auxin exerted undesir–able effects. Several basal salts media were applied in proliferation stage and their physiological effects were evaluated in reference to traditionally used MS. At culturing on Chée & Pool C2d Vitis Medium (VM) that is for the first time introduced to magnolia and on MS, M. liliiflora formed more but less elongated shoots than M. soulangeana. However, on VM, substantial increase (25-30%) of the number of axillary shoots and leaves, shoot length and fresh and dry weights over MS was established for both species. This suggested VM as promising composition of nutrients in multiplication stage. Microshoots obtained on MS, VM, Rugini Olive Medium and DKW Juglans Medium were successfully rooted in vitro and subsequently established ex vitro. The findings expand the information on magnolia response to culture conditions and contribute to elaboration of innovative elements of protocols for establishing tissue cultures with high regeneration capacity.


1970 ◽  
Vol 65 (3) ◽  
pp. 466-476 ◽  
Author(s):  
Richard R. Gala

ABSTRACT Factors influencing prolactin production and anterior pituitary (AP) tissue survival in vitro were investigated. Media 199, NCTC-109, MB752/1 and Trowells-T8 were compared for their ability to support the rat AP in vitro and found to be similar, although Trowells-T8 appeared to be less well suited for this tissue. Culturing AP in Hanks balanced salt solution (BSS) resulted in a lower prolactin production and tissue survival than in Medium 199; nevertheless, there was a tenfold net synthesis of prolactin. Streptomycin and penicillin at a level of 200 units each/ml and nystatin at a level of 50 units/ml were found to control the growth of microorganisms without any deleterious effects on the AP. Amphotercine B at a level of 25 μg/ml, on the other hand, was incompatible with AP tissue survival and function. When pituitary fragments weighed 2.8 mg or more, prolactin production and tissue survival were decreased relative to AP fragments which weighed 1.2 mg or less. Culture of pituitary tissue completely submerged in medium was not compatible with optimum hormone production or AP survival. Pituitary pieces supported by stainless steel platforms with filter paper wicks resulted in better tissue survival and significantly greater prolactin production that when cultured under similar conditions but without a wick.


HortScience ◽  
1994 ◽  
Vol 29 (5) ◽  
pp. 514e-514
Author(s):  
Barbara M. Reed

Cultures of 49 Pyrus species and cultivars and one Pyronia (Pyrus × Cydonia hybrid) selection were screened in vitro to determine a rooting method suitable for a wide range of germplasm. Auxin treatment was required for rooting in most cases. Eighteen of the 50 accessions rooted with a 15 sec. 10 mM indole-3-butyric acid (IBA) dip followed by growth on medium with no growth regulators (NCR). Medium with 10 μM IBA for one week followed by NCR medium produced 12 rooted accessions, but NCR medium alone produced little or no rooting. A 15 sec. dip in 10 mM naphthaleneacetic acid (NAA) followed by NCR medium was tested on 29 accessions which rooted poorly on the other three treatments. Twice as many (28%) rooted on NAA as on either IBA treatment (14% each). Additional treatments combining IBA with darkness or higher temperature were also tested and were successful for some cultivars. P. calleryana, P. koehnei, P. pashia, P. hondoensis, P. ussuriensis, P. betulifolia, P. regelii, P. pyrifolia hybrid cv. Shinseiki and the Pyronia selection failed to root. Twenty two of the 32 P. communis cultivars rooted on at least one treatment.


2020 ◽  
Vol 73 (1) ◽  
pp. 9039-9046
Author(s):  
Rodrigo Alberto Hoyos Sanchez ◽  
Diego Chicaíza Finley ◽  
Juan Carlos Zambrano Arteaga

Carludovica palmata Ruíz & Pavón is a plant that belongs to the Cyclanthaceae family. Its commercial interest is related to the production of fibers for the manufacture of handicrafts, mainly the Panama hat, so it is important to study its propagation. This investigation aimed to determine the effect of 6-benzylaminopurine (BAP) in the formation of new shoots and 1-naphthaleneacetic acid (NAA) in the formation of roots, as well as the adaptation in greenhouse conditions of Carludovica palmata Ruíz & Pavón. In order to find the optimal multiplication rate, 0.5 cm length explants were planted in glass jars with 15 mL of semisolid MS with different concentrations of BAP and cultured under in vitro conditions for 90 days. The multiplication parameters in this stage were number of shoots per explant (NSE), length of shoots (LS), and length of roots (LR) as multiplication parameters. In a similar procedure, the number of roots per explant (NRE), length of roots (LR), and length of plantlets (LP) was determined using different concentrations of NAA. Finally, different substrates were evaluated for the adaptation of plantlets of C. palmata produced in vitro, under greenhouse conditions for 80 days. The highest multiplication rate (17±3 shoots per explant) was obtained with 2.0 mg L-1 of BAP. Root formation occurred efficiently in all treatments, without significant statistical differences between them. On the other hand, the use of substrate soil-t15 was the best treatment for the growth of C. palmata under greenhouse conditions. From the results obtained, it is concluded that C. palmata can be efficiently multiplied under in vitro conditions and did not present problems during the in vivo rooting process.


1974 ◽  
Vol 52 (3) ◽  
pp. 473-476 ◽  
Author(s):  
H. Chlyah

The histological study of the de novo formation of shoot and root meristems from inter-nodal segments of Torenia fournieri showed that buds are formed exclusively from epidermal cells and roots from endogenous perivascular tissue. It seems that there are no particular epidermal cells which are preferentially involved in the initiation of buds. This study showed the different and apparently specific potentialities of superficial and endogenous tissues.


1983 ◽  
Vol 10 (1) ◽  
pp. 21-25 ◽  
Author(s):  
R. N. Pittman ◽  
D. J. Banks ◽  
J. S. Kirby ◽  
E. D. Mitchell ◽  
P. E. Richardson

Abstract Green immature leaflets (2-5 mm in length from shoots of germinated seeds or greenhouse grown plants) from species representing seven taxonomic sections of the genus Arachis (Ambinervosae, Arachis, Caulorhizae, Erectoides, Extranervosae, Rhizomatosae, and Triseminalae), were cultured aseptically, in vitro, on a medium composed of Murashige and Skoog salts, Gamborg's B5 vitamins, 0.8% Difco agar, and supplemented with 1 mg/L each of naphthaleneacetic acid and N-6 benzyladenine. Histological examination of the cultures revealed that the meristematic areas originated from epidermal cells. Embryoids and meristematic shoots developed after lysis of the surrounding cells. All species of Arachis tested produced callus. Genotypic differences for the production of callus, shoots, and roots were observed with cultivated peanuts. Organogenesis occurred in the leaflet cultures, and plants were recovered from sections Arachis and Extranervosae of the genus Arachis.


2018 ◽  
Vol 47 (1) ◽  
pp. 152-161 ◽  
Author(s):  
Athanasios TSAFOUROS ◽  
Peter A. ROUSSOS

Krymsk® 5 (VSL-2) is a semi-dwarf cherry rootstock adaptable to a range of climates. The present study aimed to establish the first efficient in vitro propagation protocol for this rootstock. Therefore, six cytokinines, four adenine type (6-benzyladenine, 2-isopentenyladenine, kinetin and meta-topolin) and two phenylureas (thidiazuron and forchlorfenuron) at three (2.4 μΜ, 4.8 μΜ and 9.6 μΜ) concentrations plus three (0.24 μΜ, 0.48 μΜ, 0.96 μΜ) for thidiazuron only were tested during the multiplication stage. 6-Benzyladenine was the most efficient cytokinin, based on the number of shoots produced (3.5 shoots at 9.6 μΜ) and the number of nodes per explant (10 nodes at 9.6 μΜ) whereas the other aromatic adenine tested, i.e. meta-topolin, presented the highest number of nodes per cm and node per shoot. During the rooting stage the synthetic auxins 1-naphthaleneacetic acid (1-NAA) and indolebutyric acid (IBA) were tested at concentrations of 0, 2.5, 5, 10 and 20 μΜ both separately and in all possible combinations. The percentage of successfully rooted explants reached 95% under the combination of 20 μΜ IBA plus 5 μΜ 1-NAA, whereas the highest number of roots recorded was 8.5 roots for the treatment 20 μΜ ΙΒΑ plus 2.5 μΜ 1-NAA. Furthermore, two different carbon sources were compared, the widely used sucrose and the endogenous sugar ratio of mother plant softwood shoot, sampled during late of May. Endogenous sugar ratio proved to be the preferable carbon source, since it increased the number of shoots produced and almost doubled the number of produced nodes per explant.


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