scholarly journals The effect of cytokinins on shoot proliferation, biochemical changes and genetic stability of Rhododendron ‘Kazimierz Odnowiciel’ in the in vitro cultures

Author(s):  
Karolina Nowakowska ◽  
Anna Pińkowska ◽  
Ewa Siedlecka ◽  
Andrzej Pacholczak

AbstractShoot proliferation is a very important micropropagation phase, decisive for economic efficiency of this method for a given taxon. To obtain a high multiplication ratio and a good quality of microshoots a detailed propagation protocol must be developed for particular species or even cultivars. Rhododendron ‘Kazimierz Odnowiciel’ is a relatively new cultivar distinguished by large, beautiful flowers and high frost resistance so there is a need to develop an efficient method of its propagation to satisfy a growing demand for this plant. The aim of the experiment was to evaluate effects of cytokinins: meta-Topolin (mT), zeatin (ZEA), 6-benzyladenine (BA), thidiazuron (TDZ), 2-isopentenyladenine (2iP), or the combination of 2iP+ZEA on proliferation of shoots in R. ‘Kazimierz Odnowiciel’ cultured on Anderson’s medium (AN). Biochemical changes in plant material affected by cytokinins during this phase of micropropagation were determined and occurrence of genetical changes was followed using ISSR markers. TDZ, ZEA or the combination of ZEA+2iP resulted in 100% explant regeneration. On the medium with TDZ or ZEA over two new shoots per explant were produced but the highest proliferation was attained on the medium containing ZEA+2iP – over three shoots per explant. Microshoots developed in this treatment had also the highest contents of chlorophyll, carotenoids and soluble sugars as well as the highest catalase activity. Microshoots formed on the medium with zeatin showed the lowest polymorphism (below 4%) relative to a stock plant.

Plants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 206 ◽  
Author(s):  
Michał P. Maliński ◽  
Małgorzata Anna Kikowska ◽  
Agata Soluch ◽  
Mariusz Kowalczyk ◽  
Anna Stochmal ◽  
...  

Lychnis flos-cuculi L., a species with potential medicinal value, contains flavonoids, phenolic acids, triterpenoid saponins and ecdysteroids. In this study, the antioxidant activity of plant material of L. flos-cuculi obtained from in vitro cultures compared to that of intact plants from the natural site has been evaluated for the first time. Phytochemical screening of the in-vitro-derived material by ultra-high-performance liquid chromatography mass spectrometry (UHPLC-MS) confirmed the presence of the aforementioned metabolite classes. The aqueous methanolic extracts from in-vitro-derived plant material and the organs of intact plants were analyzed using spectrophotometric methods to quantify total phenolics, phenolic acids and flavonoids, and determine the preliminary antioxidant activity by ferric reducing antioxidant potential (FRAP) and DPPH radical scavenging activity assays. The results showed that the inflorescence (Ns-F), and flowering herb of both plants gathered from natural habitat (Ns-H) and in-vitro-derived plants from the experimental plot (ExV-H) are the materials richest in polyphenols (195.4, 113.47, 112.1 mg GAE g−1 d.w., respectively), and demonstrate the highest antioxidant activity (20.14, 11.24, and 11.46 mg AAE g−1 d.w.). The extract from callus exhibited the lowest polyphenol content and antioxidant potential. The contents of total phenolics, flavonoids and phenolic acids correlate with the results of the antioxidant capacity of L. flos-cuculi extracts.


Author(s):  
Danuta Kucharska ◽  
Robert Maciorowski ◽  
Małgorzata Kunka ◽  
Angelika Niewiadomska-Wnuk

Short storage of in vitro cultures under slow-growth conditions is included in the commercial large-scale micropropagation process. It is dictated by the organizational scheme that provides temporary stop multiplication of shoots for some months. To avoid subculturing to fresh media every 4 weeks, which is obligatory for gooseberry, they can be kept in conditions that protect them from ageing, by slowing down their metabolism. To develop a rational schedule of gooseberry micropropagation, two experiments were used to adopt a temperature and length of time for storage. The best results were obtained with storage conditions at 2 °C for two or four months for proliferating cultures. Under these conditions, the percentage of necrotic shoots was low (< 10%), and shoot proliferation in the subsequent passages was at a level similar to proliferation cultures incubated in the growth room and sub-cultured monthly. The rate of shoots > 1 cm was higher than in the control in the growth room. Storage at 4 °C increased the probability of necrotic shoots up to 80% and decreased the number of all shoots and shoots > 1 cm in subsequent passages.


Author(s):  
Břetislav Křižan ◽  
Eva Ondrušiková ◽  
Jana Moudrá

The current demand for in vitro cultures of grape rootstocks, not only for mass production of plants, but also for genetic engineering is evident. The study on micropropagation of grape rootstock genotypes namely Kober 5BB, Kober 125AA and Teleki 5C was performed. The aim of the study was to develop an optimized protocol to obtain large quantity of plant material. Protocol is based on regeneration via organogenesis, considering that grape embryogenic calluses are laborious to establish and the genotype of the regenerated plants can be altered. Using of Driver and Kuniyuki Walnut media for the establishing of proliferating cultures gave better results than Murashige Skoog media in case of all used rootstocks. Subsequent cultivation on modified Murashige Skoog media with 1-naphtalene acetic acid and increased concentration of cytokynin was characterized by multiplication of cultures and formation of clusters with high multiplication capability. The clusters obtained from rootstock genotypes were suitable for mass propagation as well as for genetic transformation due to their high ability of regeneration.


1994 ◽  
Vol 4 (3) ◽  
pp. 315-317 ◽  
Author(s):  
Kenneth W. Mudge ◽  
Chin-Chang Chu

In vitro asymbiotic seed germination, subculture, and outplanting of orchids is presented as a laboratory exercise suitable for students of plant propagation or tissue culture. Dendrobium antennatum (Lindley), Phalaenopsis (Blume) white hybrid, or both, are used in this exercise because they flower predictably in the greenhouse, are reliable for seed production, and germinate and grow rapidly in vitro. The exercises can be used to instruct students in the skills involved in orchid seed sterilization, sowing, and culture, as well as instruct students in the unique features of orchid reproductive biology and symbiosis. A schedule is suggested for stock plant flower pollination, capsule harvest, seed sowing, and seedling subculture so that the necessary plant material is available for students to sow, subculture, and outplant seedlings during a single laboratory session.


2012 ◽  
Vol 40 (1) ◽  
pp. 113 ◽  
Author(s):  
Snjezana KERESA ◽  
Anita MIHOVILOVIC BOSNJAK ◽  
Marijana BARIC ◽  
Ivanka HABUS JERCIC ◽  
Hrvoje SARCEVIC ◽  
...  

'Topaz' is a modern Czech apple cultivar well accepted by consumers and scab-resistant, providing reasons for the significant spreadof cv. 'Topaz' in European orchards, especially in the organic fruit production industry. Growing the apple trees on their own rootsprovides some advantages in comparison with grafted trees. Micropropagation is the method of choice for plantlet production for thispurpose as well as for the establishment of healthy mother stock trees as a source of scions. The efficiency of axillary shoot proliferationwas examined on four media differing in plant growth regulators and their concentrations, and from three explant types: intact ordecapitated and defoliated microshoots placed vertically and one-nodal segments placed horizontally. All media consisted of Quoirin and Lepoivre (QL) macroelements and Murashige and Skoog (MS) microelements. Furthermore, rooting efficiency on six different media/treatments was analyzed. Media with 1 mg/L 6-benzylaminopurine (BA) or BA (0.5 mg/L) + 1.5 mg/L kinetin (Kin) produced similarnumber of microshoots per inoculated one (2.5 and 2.4, respectively). Medium with 1 mg/L thidiazuron (TDZ) produced significantlyhigher number of shoots (3.6) but they were fasciated. Three different explant types also produced similar numbers of microshoots.High rooting efficiency (68.7%), a high number of roots per shoot (6.6) and the best quality of shoots were obtained in rooting mediumcontaining 2 mg/L of indole-3-butyric acid (IBA). An efficient method of shoot proliferation was established, and, since rooting was themost critical step, an efficient procedure for rooting apple cv. 'Topaz' was established.


Viruses ◽  
2020 ◽  
Vol 12 (1) ◽  
pp. 70 ◽  
Author(s):  
Kiyoaki Maeda ◽  
Teppei Kikuchi ◽  
Ichiro Kasajima ◽  
Chungjiang Li ◽  
Noriko Yamagishi ◽  
...  

Apple latent spherical virus (ALSV) was successfully used in promoting flowering (virus-induced flowering, VIF) in apple and pear seedlings. In this paper, we report the use of ALSV vectors for VIF in seedlings and in vitro cultures of grapevine. After adjusting experimental conditions for biolistic inoculation of virus RNA, ALSV efficiently infected not only progeny seedlings of Vitis spp. ‘Koshu,’ but also in vitro cultures of V. vinifera ‘Neo Muscat’ without inducing viral symptoms. The grapevine seedlings and in vitro cultures inoculated with an ALSV vector expressing the ‘florigen’ gene (Arabidopsis Flowering locus T, AtFT) started to set floral buds 20–30 days after inoculation. This VIF technology was successfully used to promote flowering and produce grapes with viable seeds in in vitro cultures of F1 hybrids from crosses between V. ficifolia and V. vinifera and made it possible to analyze the quality of fruits within a year after germination. High-temperature (37 °C) treatment of ALSV-infected grapevine disabled virus movement to newly growing tissue to obtain ALSV-free shoots. Thus, the VIF using ALSV vectors can be used to shorten the generation time of grapevine seedlings and accelerate breeding of grapevines with desired traits.


HortScience ◽  
2000 ◽  
Vol 35 (3) ◽  
pp. 397C-397
Author(s):  
Guochen Yang ◽  
Marihelen Kamp-Glass

Exochorda racemosa is an ornamental shrub with white flowers that is spiraea-like, deciduous, and hardy. The buds resemble pearls. Normally it is propagated by seeds, layers, and cuttings of softwood. However, it is a slow process that takes a few years to produce a reasonable size plant for the demanding market. Our objective was to establish a successful in vitro culture and to rapidly multiply this ornamental species. Softwood explant materials were collected from a local nursery and were disinfested with 15% bleach solution and rinsed three times with sterile distilled and deionized water. In vitro cultures were established and maintained in woody plant medium (WPM) supplemented with BA at 0.1 mg·L-1, 3% sucrose, and 0.7% agar with the pH adjusted to 5.8. Then shoots were transferred to the multiplication medium containing BA, CPPU, or thidiazuron (TDZ) at various concentrations. Preliminary results show that explants cultured on medium containing TDZ produced the best shoot proliferation.


Sign in / Sign up

Export Citation Format

Share Document