In Vitro Establishment of Powdery Mildew (Microsphaera pulchra) on Microshoots of Flowering Dogwood

Microshoot and callus cultures of Cornus florida (flowering dogwood), which were grown on woody plant medium amended with BA, were inoculated with Microsphaera pulchra (an obligate plant parasite) by gently shaking infected leaves bearing numerous conidia over the tissue. Culture dishes were sealed with parafilm and incubated at 24 °C with 25 mol·m–2·s–1 provided by cool fluorescent bulbs for 15 h. Cultures were examined with a dissecting scope every 24 h and cultures transferred when contaminating fungi were present. Specimens were prepared light microscopy and SEM. The fungus infected individual callus cells, but did not sporulate. In contrast, powdery mildew was well-established (both primary and secondary hyphae) in 70% of the microshoot cultures after 6 days and sporulated on 20% by 7 to 8 days. The cellular relationship between host and pathogen in vitro was similar to that found in greenhouse-grown plants. This technique has possible applications in maintaining fungal culture collections and studying host–pathogen relationships under more stringently controlled conditions.

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In vitro establishment and proliferation of red currant cultivars

Horticultural Science ◽

10.17221/40/2011-hortsci ◽

2012 ◽

Vol 39 (No. 1) ◽

pp. 21-25 ◽

Cited By ~ 2

Author(s):

J. Sedlák ◽

F. Paprštein

Keyword(s):

Woody Plant ◽

Adventitious Bud ◽

Mineral Salts ◽

The Czech Republic ◽

Shoot Number ◽

Adventitious Bud Formation ◽

Woody Plant Medium ◽

In Vitro Establishment ◽

Red Currant

The goal of this study was to investigate in vitro multiplication protocols for use with red currant cultivars grown in the Czech Republic. Cultivars Detvan, Vitan and Rotte Höllandische were successfully established in vitro using mercuric chloride in a concentration of 0.15% as a sterilization solution. The overall rate of contamination was 25.7%. Two proliferation media Murashige and Skoog medium (MS) and McCown woody plant medium (WPM) containing 1 or 2 mg/l of 6-benzylaminopurine (BAP) were tested. Initial explants produced new plants in the form of rosettes. Rosettes arose from the base of the initial explants in the form of adventitious bud formation. The shoot number was relatively low and varied between 1.0 and 2.1. Generally, the highest number was obtained for cultivar Rotte Holländische that produced 2.1 ± 0.1 new rosettes on MS medium containing lower concentration 1 mg/l BAP. In contrary, Vitan cv. had significantly lower shoot number ranging from 1.0 to 1.3. WPM medium with a lower concentration of mineral salts proved to be unsuitable for the multiplication of tested cultivars.

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In Vitro Establishment of ‘Delite’ Rabbiteye Blueberry Microshoots

Horticulturae ◽

10.3390/horticulturae5010024 ◽

2019 ◽

Vol 5 (1) ◽

pp. 24 ◽

Cited By ~ 1

Author(s):

Carolina Schuchovski ◽

Luiz Biasi

Keyword(s):

Woody Plant ◽

Shoot Formation ◽

Initial Study ◽

Shoot Length ◽

In Vitro Studies ◽

Basic Medium ◽

Mass Propagation ◽

Woody Plant Medium ◽

In Vitro Establishment

Micropropagation is an important technique for clonal mass propagation and a tool for in vitro studies. One of the first steps to overcome in this process is the establishment of new explants in vitro. ‘Delite’ rabbiteye blueberry was cultured in vitro with four cytokinins (zeatin (ZEA), 6-(γ-γ-dimethylallylamino)-purine (2iP), 6-benzylaminopurine (BAP), and kinetin (KIN)) at eight concentrations (0, 2.5, 5, 10, 20, 30, 40, and 50 µM). Additionally, nine combinations of nitrogen salts were tested, using Woody Plant Medium (WPM) and a modified WPM as the basic medium. ZEA and 2iP showed better responses, but ZEA was superior at lower (2.5 µM) concentrations (89.7% survival, 81.3% shoot formation, 1.3 shoots, 13.8 mm shoot length, 10.0 leaves). BAP and KIN showed very low responses. In the combinations of salts with modified WPM, no differences were observed. However, the original WPM with treatments of 0.5 × NH4NO3 and 1 × Ca(NO3)2, 0.5 × NH4NO3 and 0.5 × Ca(NO3)2, and the modified WPM alone showed the lowest rates of survival and shoot formation and the shortest shoot lengths. The highest shoot lengths were observed in treatments with the original WPM, 1.5 × NH4NO3 and 0.5 × Ca(NO3)2, and 1.5 × NH4NO3 and 1.5 × Ca(NO3)2. This initial study with ‘Delite’ can be the basis for further experiments with different combinations of salts, 2iP, and ZEA.

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In Vitro Colonization of Micropropagated Pieris floribunda by Ericoid Mycorrhizae. I. Establishment of Mycorrhizae on Microshoots

HortScience ◽

10.21273/hortsci.36.2.353 ◽

2001 ◽

Vol 36 (2) ◽

pp. 353-356 ◽

Cited By ~ 3

Author(s):

Mark C. Starrett ◽

Frank A. Blazich ◽

Steven R. Shafer ◽

Larry F. Grand

Keyword(s):

Root Development ◽

Adventitious Roots ◽

Woody Plant ◽

Hymenoscyphus Ericae ◽

Ericoid Mycorrhizae ◽

Woody Plant Medium ◽

In Vitro Establishment ◽

Shoot Necrosis

Selected isolates of Hymenoscyphus ericae (Read) Korf and Kernan, Oidiodendron griseum Robak, O. maius Barron, and a second O. Robak species were evaluated as inocula for in vitro establishment of micropropagated plantlets of Pieris floribunda (Pursh ex Sims) Benth. and Hook. Severity of shoot necrosis on microshoots differed for each isolate of Oidiodendron. Inoculation of micropropagated plantlets with isolates of H. ericae benefited initial shoot and root development on agar-solidified Woody Plant Medium (WPM) supplemented with sucrose and covered by a layer of autoclaved 1 peat: 1 vermiculite (v/v). Inoculation of microshoots with H. ericae or isolates of Oidiodendron did not stimulate production of adventitious roots.

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Comparative Investigation on Formation and Accumulation of Rare Phenylpropanoids in Plants and in vitro Cultures of Pimpinella major

Zeitschrift für Naturforschung C ◽

10.1515/znc-1990-0607 ◽

1990 ◽

Vol 45 (6) ◽

pp. 602-606 ◽

Cited By ~ 5

Author(s):

B. Merkel ◽

J. Reichling

Keyword(s):

Callus Culture ◽

Nutrient Medium ◽

Callus Cultures ◽

Comparative Investigation ◽

In Vitro Cultures ◽

Liquid Nutrient Medium ◽

Whole Plants ◽

Plant Seedlings

Abstract Unorganized callus and leaf/root-differentiating callus cultures of Pimpinella major have been established in liquid nutrient medium. Their capacity to accumulate rare phenylpropanoids such as epoxy-pseudoisoeugenol tiglate, epoxy-anol tiglate and anol tiglate was compared with that of seedlings and whole plants. The unorganized callus cultures were not able to accumulate any phenylpropanoids. In comparison, the leaf/root-differentiating callus culture promoted the accumulation of epoxy-pseudoisoeugenol tiglate (up to 90 mg/100 g fr.wt.) but not that of anol-derivatives. The accumulated amount of EPT in PMD-SH was comparable with that in plant seedlings.

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In vitro metabolism of Tebuconazole, Flurtamone, Fenhexamid, Metalaxyl‐M and Spirodiclofen in Cannabis sativa L. (hemp) callus cultures

Pest Management Science ◽

10.1002/ps.6575 ◽

2021 ◽

Author(s):

Leonie Hillebrands ◽

Marc Lamshoeft ◽

Andreas Lagojda ◽

Andreas Stork ◽

Oliver Kayser

Keyword(s):

Cannabis Sativa ◽

Callus Cultures ◽

In Vitro Metabolism

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Analysis of a Preliminary microRNA Expression Signature in a Human Telangiectatic Osteogenic Sarcoma Cancer Cell Line

International Journal of Molecular Sciences ◽

10.3390/ijms22031163 ◽

2021 ◽

Vol 22 (3) ◽

pp. 1163

Author(s):

Gaia Palmini ◽

Cecilia Romagnoli ◽

Simone Donati ◽

Roberto Zonefrati ◽

Gianna Galli ◽

...

Keyword(s):

Cell Line ◽

Molecular Mechanisms ◽

Osteogenic Sarcoma ◽

Cancer Cell Line ◽

Microscopic Features ◽

In Vitro Establishment ◽

Profile Characteristic ◽

First Time

Telangiectatic osteosarcoma (TOS) is an aggressive variant of osteosarcoma (OS) with distinctive radiographic, gross, microscopic features, and prognostic implications. Despite several studies on OS, we are still far from understanding the molecular mechanisms of TOS. In recent years, many studies have demonstrated not only that microRNAs (miRNAs) are involved in OS tumorigenesis, development, and metastasis, but also that the presence in high-grade types of OS of cancer stem cells (CSCs) plays an important role in tumor progression. Despite these findings, nothing has been described previously about the expression of miRNAs and the presence of CSCs in human TOS. Therefore, we have isolated/characterized a putative CSC cell line from human TOS (TOS-CSCs) and evaluated the expression levels of several miRNAs in TOS-CSCs using real-time quantitative assays. We show, for the first time, the existence of CSCs in human TOS, highlighting the in vitro establishment of this unique stabilized cell line and an identification of a preliminary expression of the miRNA profile, characteristic of TOS-CSCs. These findings represent an important step in the study of the biology of one of the most aggressive variants of OS and the role of miRNAs in TOS-CSC behavior.

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In vitro somatic embryogenesis in callus cultures ofAzadirachta indicaA. Juss.—a multipurpose tree

Journal of Forest Research ◽

10.1007/s10310-004-0130-y ◽

2005 ◽

Vol 10 (4) ◽

pp. 263-267 ◽

Cited By ~ 13

Author(s):

Gyana Ranjan Rout

Keyword(s):

Somatic Embryogenesis ◽

Callus Cultures ◽

Multipurpose Tree

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Period of harvest, sprouting ability of cuttings, and in vitro plant regeneration in Fraxinus excelsior

Canadian Journal of Botany ◽

10.1139/b94-035 ◽

1994 ◽

Vol 72 (2) ◽

pp. 261-267 ◽

Cited By ~ 9

Author(s):

Conceição Eneida Silveira ◽

Alain Cottignies

Keyword(s):

Chromosome Number ◽

In Vitro Culture ◽

Adventitious Roots ◽

Woody Plant ◽

Fraxinus Excelsior ◽

Stem Cuttings ◽

Natural Conditions ◽

Apical Bud ◽

Woody Plant Medium

Propagation by stem cuttings and in vitro culture of apical bud explants were studied on Fraxinus excelsior L. Stem cuttings from 4- to 7-year-old trees growing under natural conditions sprouted only when cuttings were taken from dormant material. Only 6% of those that had sprouted developed roots by the 7th month of culture. Similarly, only apical bud explants harvested during the dormant period sprouted in vitro. Up to 87% of these sprouts developed two to four branching adventitious roots after 5 months of culture. During the initial phase of in vitro culture, the Quoirin and Lepoivre medium and the woody plant medium favoured sprout lengthening. During the phase of multiplication, up to three sprouts per explant developed with the woody plant medium in the presence of a combination of high 6-benzylaminopurine (3.0–4.0 mg∙L−1) and low indole-3-butyric acid (0.01–0.03 mg∙L−1) concentrations. Rooting was obtained in a medium without any growth regulators. Microscopic analysis showed a direct connection between the vascular elements of adventitious roots and stem of plantlet. Chromosome number in root apices of ash plantlets and ash trees grown under natural conditions was 2n = 46. Key words: chromosome number, Fraxinus excelsior L., in vitro plants, micropropagation, stem cuttings.

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