scholarly journals A new and fast method to obtain in vitro cultures of Huperzia selago (Huperziaceae) sporophytes, a club moss which is a source of huperzine A

2013 ◽  
Vol 82 (4) ◽  
pp. 313-320 ◽  
Author(s):  
Wojciech J. Szypuła ◽  
Paulina Mistrzak ◽  
Olga Olszowska
Keyword(s):  
Developmental Stages ◽  
Therapeutic Potential ◽  
Callus Formation ◽  
Axenic Culture ◽  
Growth Index ◽  
In Vitro Cultures ◽  
Huperzine A ◽  
Fast Method ◽  
Biomass Growth

This study presents a protocol for a fast and effective in vitro axenic culture of <em>Huperzia selago</em> (Huperziaceae Rothm.) sporophytes, a club moss which is a source of huperzine A, an alkaloid of a considerable therapeutic potential extensively investigated for its uses as treatment for some neurodegenerative diseases. The proposed procedure allowed approximately tenfold shortening of the species developmental stages with the omission of the gametophyte stage while the sporophyte mass could be increased tenfold within a 6-month period. The cultures were established using vegetative propagules (bulbils) procured from sporophytes growing in the wild without degrading the habitats of this endangered plant species. Explants underwent surface and internal disinfection to eliminate the epiphytic and endophytic bacteria and fungi. In in vitro cultures, the optimum results were achieved using Moore (Mr) medium without growth regulators or supplemented with 0.015 mg/l IBA and 0.3 mg/l kinetin. These media ensured both viability of the propagules and their further development. The biomass growth index for <em>H. selago</em> sporophytes grown from propagules, determined at 3 months of culture (1 passage) on Mr medium with IBA and kinetin was 650%. At 6 months, the biomass growth index increased to 1114%. Vigorous growth of adventitious roots, especially on Mr medium with the addition of 0.25 mg/l NAA, and callus formation on shoot apices were observed. At 6 months of culture, some sporophytes obtained from the bulbils were used as the initiating material for shoot subcultures, which developed best on Mr medium with IBA and kinetin.

scholarly journals Huperzine A and Huperzine B Production by Prothallus Cultures of Huperzia selago (L.) Bernh. ex Schrank et Mart

Molecules ◽  
2020 ◽  
Vol 25 (14) ◽  
pp. 3262
Author(s):  
Wojciech J. Szypuła ◽  
Beata Wileńska ◽  
Aleksandra Misicka ◽  
Agnieszka Pietrosiuk
Keyword(s):  
Pharmaceutical Industry ◽  
Current Source ◽  
Growth Index ◽  
Dry Weight ◽  
Huperzine A ◽  
Biomass Growth ◽  
Tissue Cultures ◽  
In The Wild ◽  
Very High

This is the first report of an efficient and effective procedure to optimize the biosynthesis of huperzine A (HupA) and huperzine B (HupB) in vitro from Huperzia selago gametophytes. Axenic tissue cultures were established using spores collected from the sporophytes growing in the wild. The prothalia were obtained after 7–18 months. Approximately 90 up to 100% of the gametophytes were viable and grew rapidly after each transfer on to a fresh medium every 3 months. The best biomass growth index for prothallus calculated on a fresh (FW) and dry weight (DW) basis, at 24 weeks of culture, was 2500% (FW) and 2200% (DW), respectively. The huperzine A content in the gametophytes was very high and ranged from 0.74 mg/g to 4.73 mg/g DW. The highest yield HupA biosynthesis at >4 mg/g DW was observed on W/S medium without growth regulators at 8 to 24 weeks of culture. The highest HupB content ranged from 0.10 mg/g to 0.52 mg/g DW and was obtained on the same medium. The results demonstrate the superiority of H. selago gametophyte cultures, with the level of HupA biosynthesis approximately 42% higher compared to sporophyte cultures and 35-fold higher than when the alkaloid was isolated from H. serrata, its current source for the pharmaceutical industry. Moreover, the biosynthesis of HupB was several-fold more efficient than in H. selago sporophytes growing in the wild. HPLC-HR-MS analyses of the extracts identified eight new alkaloids previously unreported in H. selago: deacetylfawcettine, fawcettimine, 16-hydroxyhuperzine B, deacetyllycoclavine, annopodine, lycopecurine, des-N-methylfastigiatine and flabelline.

scholarly journals Immature embryo is the potential source for in vitro plant regeneration in Jatropha curcas

2014 ◽  
Vol 20 ◽  
pp. 125-134
Author(s):  
MR Islam ◽  
MA Bari
Keyword(s):  
Developmental Stages ◽  
Callus Formation ◽  
Immature Embryo ◽  
Basal Medium ◽  
Coconut Water ◽  
Immature Embryos ◽  
Plantlet Formation ◽  
Potential Source ◽  
Wide Range

Context: Jatropha belongs the spurge family Euphorbiaceae. Special interest mounting for its biodiesel which has created enthusiasm in cultivation of the species for oil extraction. Objectives: The study was conducted to develop the protocol for tissue and callus culture in Bangladeshi Jatropha curcus plant particularly to identify the most suitable explants for its wide scale micropropagation. Materials and Methods: Immature embryos taken from four developmental stages of fruits were cultured on growth regulator free MS liquid medium. After fifteen days of germination, elongated hypocotyls and two cotyledonary leaves were used as explants. Results: Embryo derived seedlings acted as the potential source of explants both for callus and plantlets. The immature embryo of size 0.87cm produced highest callus formation (83.33%) on MS medium supplemented with lower concentration of 2, 4-D (0.5 mg/l) and coconut water 2% (v/v). Immature embryos grown on MS basal medium supplemented with 2,4-D (0.2 mg/l, 0.5 mg/l and 1.0 mg/l) alone or in combination with coconut water 2% (v/v) exhibited a wide range of callus induction percentage (26-100%) for hypocotyls and (20 - 40%) for cotyledonary leaves. Conclusion: The age of immature embryo and addition of growth adjuvants and growth additive to the culture medium played the role in promoting better callus and plantlet formation. DOI: http://dx.doi.org/10.3329/jbs.v20i0.17726 J. bio-sci.  20:  125-134, 2012

scholarly journals Linnaea borealis L. var. borealis—In Vitro Cultures and Phytochemical Screening as a Dual Strategy for Its Ex Situ Conservation and a Source of Bioactive Compounds of the Rare Species

Molecules ◽  
2021 ◽  
Vol 26 (22) ◽  
pp. 6823
Author(s):  
Barbara Thiem ◽  
Dariusz Kruszka ◽  
Natalia Turowska ◽  
Elwira Sliwinska ◽  
Viktor Berge ◽  
...  
Keyword(s):  
Plant Biomass ◽  
Shoot Culture ◽  
Growth Index ◽  
In Vitro Cultures ◽  
Raw Material ◽  
Ex Situ ◽  
Shoot Cultures ◽  
Triterpenoid Saponins ◽  
First Time

Linnaea borealis L. (Twinflower)—a dwarf shrub in the Linnaeeae tribe of Caprifoliaceae family—is distributed across the Northern Hemisphere. By means of this study, a reliable protocol for efficient micropropagation of uniform L. borealis L. var. borealis plantlets has been provided for the first time; callus culture was also established. Different initial explants, types of cultures, media systems, and plant growth regulators in Murashige and Skoog (MS) media were tested. Agitated shoot cultures in the liquid media turned out to be the best system for the production of sustainable plant biomass. After stabilization of the callus lines, the highest growth index (c.a. 526%) was gained for callus maintained on MS enriched with picloram. TLC and UHPLC-HESI-HRMS analysis confirmed the presence of phenolic acids and flavonoids, and for the first time, the presence of iridoids and triterpenoid saponins in this species. Multiplication of L. borealis shoot culture provides renewable raw material, allowing for the assessment of the phytochemical profile, and, in the future, for the quantitative analyses and the studies of the biological activity of extracts, fractions, or isolated compounds. This is the first report on in vitro cultures of traditionally used L. borealis rare taxon and its biosynthetic potential.

scholarly journals Roles and Mechanisms of Irisin in Attenuating Pathological Features of Osteoarthritis

2021 ◽  
Vol 9 ◽  
Author(s):  
Xiangfen Li ◽  
Xiaofang Zhu ◽  
Hongle Wu ◽  
Thomas E. Van Dyke ◽  
Xiaoyang Xu ◽  
...  
Keyword(s):  
Developmental Stages ◽  
Therapeutic Potential ◽  
Gene Knockout ◽  
Inflammatory Factors ◽  
Proliferative Potential ◽  
Opposite Pattern ◽  
Tissue Samples ◽  
Primary Chondrocytes ◽  
Cartilage Development

To investigate the effects and mechanisms of irisin, a newly discovered myokine, in cartilage development, osteoarthritis (OA) pathophysiology and its therapeutic potential for treating OA we applied the following five strategical analyses using (1) murine joint tissues at different developmental stages; (2) human normal and OA pathological tissue samples; (3) experimental OA mouse model; (4) irisin gene knockout (KO) and knock in (KI) mouse lines and their cartilage cells; (5) in vitro mechanistic experiments. We found that Irisin was involved in all stages of cartilage development. Both human and mouse OA tissues showed a decreased expression of irisin. Intra-articular injection of irisin attenuated ACLT-induced OA progression. Irisin knockout mice developed severe OA while irisin overexpression in both irisin KI mice and intraarticular injection of irisin protein attenuated OA progression. Irisin inhibited inflammation and promoted anabolism in chondrogenic ADTC5 cells. Proliferative potential of primary chondrocytes from KI mice was found to be enhanced, while KO mice showed an inhibition under normal or inflammatory conditions. The primary chondrocytes from irisin KI mice showed reduced expression of inflammatory factors and the chondrocytes isolated from KO mice showed an opposite pattern. In conclusion, it is the first time to show that irisin is involved in cartilage development and OA pathogenesis. Irisin has the potential to ameliorate OA progression by decreasing cartilage degradation and inhibiting inflammation, which could lead to the development of a novel therapeutic target for treating bone and cartilage disorders including osteoarthritis.

scholarly journals Preliminary study on in vitro shoot culture of Hibiscus coddii subsp. barnardii, an indigenous South African flowering plant

2021 ◽  
Vol 27 (3) ◽  
pp. 408-416
Author(s):  
Helena Jacoba du Plessis ◽  
Roumiana Vassileva Nikolova ◽  
Bronwyn Anne Egan ◽  
Riana Kleynhans
Keyword(s):  
Callus Formation ◽  
Shoot Culture ◽  
Shoot Multiplication ◽  
In Vitro Cultures ◽  
Flowering Plant ◽  
Ms Medium ◽  
Axillary Shoots ◽  
Short Shoots

Abstract In vivo and in vitro grown plants of Hibiscus coddii subsp. barnardii were used as explant source for establishment of in vitro cultures. Nodal shoot explants derived from in vivo grown plants, both naturally and under controlled environmental conditions, showed high sensitivity to the surface disinfection treatment and poor survival in in vitro culture. In vitro grown seedlings proved successful as aseptic source of apical and basal shoot explants to establish contamination-free in vitro cultures. Sprouting of axillary buds was observed on 90% of apical shoot explants after four weeks of culture on full strength, plant growth regulator (PGR)-free Murashige and Skoog (MS) medium. However, further proliferation of short shoots, limited to the bud sprout at the explant base, occurred on only 50% of these explants. In contrast, all basal shoot explants attained 3-5 single primary axillary shoots (30-40 mm in length) while a clump of short (5-10 mm) shoots also formed at the base in 60% of these explants. In both explant types, addition of 0.25-1 mg L-1 6-Benzylaminopurine (BAP) to the MS medium resulted in a low frequency (10%-60%) of explants with short shoots (5-10 mm) that showed no further elongation. Moreover, explants cultured in the presence of BAP showed a high frequency of callus formation (up to 90%) and low survival (20%-60%). A lower frequency of callus formation (30%-40%) and higher survival (90%-100%) of both explant types occurred on BAP-free medium. Further subculturing of primary and secondary axillary shoots onto fresh MS medium (with and without BAP) did not improve shoot multiplication. Regenerated plantlets from PGR-free MS medium were successfully acclimatized and hardened-off.

Correlated Studies of Cellular Interactions Using TEM, Freeze Etching, and SEM

1974 ◽  
Vol 32 ◽  
pp. 320-321
Author(s):  
J. P. Revel
Keyword(s):  
Developmental Stages ◽  
Three Dimensional ◽  
Cell Movement ◽  
Cellular Interactions ◽  
Serial Sections ◽  
Cell Sheets ◽  
Freeze Etching ◽  
Early Developmental

Movement of individual cells or of cell sheets and complex patterns of folding play a prominent role in the early developmental stages of the embryo. Our understanding of these processes is based on three- dimensional reconstructions laboriously prepared from serial sections, and from autoradiographic and other studies. Many concepts have also evolved from extrapolation of investigations of cell movement carried out in vitro. The scanning electron microscope now allows us to examine some of these events in situ. It is possible to prepare dissections of embryos and even of tissues of adult animals which reveal existing relationships between various structures more readily than used to be possible vithout an SEM.

Lignans from in vitro cultures of transgenic roots of Taxus x media var. Hicksii

Planta Medica ◽  
2012 ◽  
Vol 78 (11) ◽  
Author(s):  
K Sykłowska-Baranek ◽  
A Pietrosiuk ◽  
M Grech-Baran ◽  
M Bonfill ◽  
P Mistrzak
Keyword(s):  
In Vitro Cultures ◽  
Transgenic Roots ◽  
Taxus X Media

Phenolic compounds from in vitro cultures of Rindera graeca Boiss. & Feldr.

Planta Medica ◽  
2013 ◽  
Vol 79 (13) ◽  
Author(s):  
K Sykłowska-Baranek ◽  
A Pietrosiuk ◽  
K Graikou ◽  
H Damianakos ◽  
M Jeziorek ◽  
...  
Keyword(s):  
Phenolic Compounds ◽  
In Vitro Cultures
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