scholarly journals Preliminary results of in vitro culture of pea and lupin embryos for the reduction of generation cycles in single seed descent technique

2013 ◽  
Vol 82 (3) ◽  
pp. 231-236 ◽  
Author(s):  
Maria Surma ◽  
Tadeusz Adamski ◽  
Wojciech Święcicki ◽  
Paweł Barzyk ◽  
Zygmunt Kaczmarek ◽  
...  

The aim of the studies was to establish in vitro conditions for the culture of pea and lupin embryos as the first step in the development of an in vitro assisted single seed descent technique for the attainment of homozygous populations. Materials for the study included of pea, and narrow-leafed and yellow lupin cultivars. Embryos dissected from mature but still-green seeds were cultured in vitro on two modified MS media and under three temperature regimes. Shoot and root lengths of regenerated plants were measured after 7, 14 and 21 days of culture. For pea plants full-strength MS medium with 4 g l<sup>−1</sup> agar and temperature 22/ 20°C (day/night) appeared to be the most conducive to shoot and root development, whereas for lupin plants lower temperatures were more propitious: 12°C in the dark for narrow-leafed lupin and 16/ 12°C (day/night) for yellow lupin. Almost all the cultured embryos developed into plants, but not all the regenerated plants survived acclimation to ex vitro conditions.

2011 ◽  
Vol 72 (4) ◽  
pp. 295-302 ◽  
Author(s):  
Tomasz Pniewski ◽  
Joanna Wachowiak ◽  
Józef Kapusta ◽  
Andrzej B. Legocki

The complete protocol for regeneration and long-term micropropagation of several Polish cultivars of pea (<em>Pisum sativum </em>L.) has been elaborated. The shoots were the most likely regenerated via de novo organogenesis. The adventitious buds formed in callus derived from cotyledons tissue adjacent to the axillary meristems of immature embryos. All cultivars' calli regenerated several shoots per explant on the MS medium supplemented with B5 vitamins and 4.5 mgl<sup>-1</sup> of BAP, however some differences in regeneration capacity among cultivars were observed. The plantlets were subsequently micropropagated with slightly higher efficiency and preserving a good viability over the long-term culture on a medium containing 2.0 mgl<sup>-1</sup> than one with 4.5 mgl<sup>-1</sup> of BAP. The additional step of the pre-conditioning culture of multiplicated shoots on a medium with very low BAP concentration i.e. 0.02 mgl<sup>-1</sup> was applied and appeared to be beneficial before rooting in vitro or grafting. The modified MS-derived medium with the half-strength of MS macroelements but with the full original dose of calcium and supplemented with B5 vitamins and 1.0 mgl<sup>-1</sup> of NAA was developed for effective rooting. The shoots were also sufficiently transferred into ex vitro conditions using grafting. The majority of the regenerated plants had adapted to in vivo conditions in a greenhouse and subsequently has set seeds. The presented protocol provides relatively efficient rate of de novo pea regeneration and would be useful for <em>Agrobacterium</em>-mediated transformation purposes.


2019 ◽  
pp. 57-67
Author(s):  
T.M. Tabatskaya ◽  
N.I. Vnukova

A technique for the long-term (up to 27 years) in vitro storage of valuable birch genotypes under normal (25 °C, 2.0 klx, 16-h day and 8-h night) and low temperature (4 °C, 0.5 klx, 6-h day and 18-h night) growing conditions on hormone-free media has been described. The study explored for the first time the influence of different strategies to store the clones of Betula pubescens and B. pendula var. сarelica (6 genotypes) on the regenerative capacity of collection samples, adaptive potential of regenerated plants and plant production by the in vitro and ex vitro techniques. It was established that both storage strategies provided a persistently high survival rate (82-100%) and regenerative capacity of in vitro shoots (the multiplication coefficient of 4.2-6.3 and rhizogenic activity of 90-100%). The clones retained their characteristics of height growth under the in vitro and ex vitro conditions, and demonstrated intraclonal homogeneity and lack of signs of somaclonal variability. The plants showed substantial interspecific differences at the stage of multiplication and transfer to the greenhouse. The highest percentage of acclimated plants (75-98% depending on the clone genotype) was obtained after planting of micro plants straight in the greenhouse, which simplified the technology and made plant production less costly. long-term in vitro storage, birch, species, genotype, micropropagation, ex vitro adaptation, plant material


2018 ◽  
Vol 77 (1) ◽  
pp. 80-87 ◽  
Author(s):  
Mahipal S. Shekhawat ◽  
M. Manokari

AbstractHybanthus enneaspermusis a rare medicinal plant. We defined a protocol for micropropagation,ex vitrorooting of cloned shoots and their acclimatization. Surface-sterilized nodal segments were cultured on Murashige and Skoog (MS) medium with different concentrations of 6-benzylaminopurine (BAP) and kinetin (Kin). Medium supplemented with 1.5 mg L−1BAP was found optimum for shoot induction from the explants and 6.4±0.69 shoots were regenerated from each node with 97% response. Shoots were further proliferated maximally (228±10.3 shoots per culture bottle with 7.5±0.43 cm length) on MS medium augmented with 1.0 mg L−1each of BAP and Kin within 4–5 weeks. The shoots were rootedin vitroon half strength MS medium containing 2.0 mg L−1indole-3 butyric acid (IBA). The cloned shoots were pulse-treated with 300 mg L–1 of IBA and cultured on soilrite® in a greenhouse. About 96% of the IBA-pulsed shoots rootedex vitroin soilrite®, each shoot producing 12.5±0.54 roots with 5.1±0.62 cm length. Theex vitrorooted plantlets showed a better rate of survival (92%) in a field study thanin vitrorooted plantlets (86%). A comparative foliar micromorphological study ofH. enneaspermuswas conducted to understand the micromorphological changes during plant developmental processes fromin vitrotoin vivoconditions in terms of variations in stomata, vein structures and spacing, and trichomes. This is the first report onex vitrorooting inH. enneaspermusand the protocol can be exploited for conservation and large-scale propagation of this rare and medicinally important plant.


2008 ◽  
Vol 43 (10) ◽  
pp. 1325-1330 ◽  
Author(s):  
Lucymeire Souza Morais-Lino ◽  
Janay Almeida dos Santos-Serejo ◽  
Sebastião de Oliveira e Silva ◽  
José Raniere Ferreira de Santana ◽  
Adilson Kenji Kobayashi

The objective of this study was to establish cell suspension culture and plant regeneration via somatic embryogenesis of a Brazilian plantain, cultivar Terra Maranhão, AAB. Immature male flowers were used as explant source for generating highly embryogenic cultures 45 days after inoculation, which were used for establishment of cell suspension culture and multiplication of secondary somatic embryos. Five semisolid culture media were tested for differentiation, maturation, somatic embryos germination and for plant regeneration. An average of 558 plants per one milliliter of 5% SCV (settled cell volume) were regenerated in the MS medium, with 11.4 µM indolacetic acid and 2.2 µM 6-benzylaminopurine. Regenerated plants showed a normal development, and no visible somaclonal variation was observed in vitro. It is possible to regenerate plants from cell suspensions of plantain banana cultivar Terra using MS medium supplemented with 11.4 µM of IAA and 2.2 µM of BAP.


2019 ◽  
Vol 11 (2) ◽  
Author(s):  
POPY HARTATIE HARDJO ◽  
DANNY PUTRA SENTOSA SUSANTO ◽  
WINA DIAN SAVITRI ◽  
MARIA GORETTI MARIANTI PURWANTO

Abstract. Hardjo PH, Susanto DPS, Savitri WD, Purwanto MGM. 2019. Shoot multiplication of Pogostemon cablin var. Sidikalang and patchouli oil profile. Nusantara Bioscience 11: 123-127. Pogostemon cablin Benth. is a plant producing patchouli oil, which mostly consists of patchouli alcohol compound. Patchouli oil has great potentials in the world market because of its stability and high price. In this study, in vitro multiplication of Sidikalang variety of Acehnese patchouli shoots was done on solid and liquid Murashige & Skoog (MS) medium. This study aimed to determine the effect of cytokinins in various combinations of shoot multiplication and to compare the patchouli oil yield of in vitro and ex vitro culture. In vitro multiplication of Acehnese patchouli shoots by using solid MS medium with addition of 0.2 ppm benzyl aminopurine (BAP) and 0.2 ppm Kinetin resulted in shoot explants with an average growth index of 82.198 ± 0.690. Patchouli oil extraction was done on 7 weeks old in vitro shoot explants cultured on solid MS medium + 0.2 ppm BAP + 0.2 ppm Kinetin using water distillation method. In vitro shoots yielded 2.5% patchouli oil and contained ± 35% patchouli alcohol compound, whereas ex vitro shoots produced 4% patchouli oil and contained ± 25% patchouli alcohol compound. The qualitative analysis by using thin layer chromatography (TLC) showed that there were similarities in the number of spot and Rf value for each spot of ex vitro and in vitro patchouli oil.


2015 ◽  
pp. 99-112
Author(s):  
Marija Markovic ◽  
Dragana Skocajic ◽  
Mihailo Grbic ◽  
Matilda Djukic ◽  
Dragica Obratov-Petkovic ◽  
...  

The aim of this study was to determine the possibility of micropropagation of the medicinal plant A. millefolium on half-strength MS medium and ex vitro rooting and acclimatization of the obtained microshoots in hydroculture in order to establish an efficient production method. Two explant types were used: basal and terminal cuttings, and better results were achieved when terminal cuttings were used. The development of shoots in the multiplication phase was successful with a regeneration percentage of 100%. Ex vitro rooting in a modified Hoagland nutrient solution was successful (83%), but the percentage of in vitro rooting on half-strength MS medium without hormones was higher (95%). However, bearing in mind that mass production of A. millefolium is more efficient when the phase of in vitro rooting is excluded, this method could be recommended for commercial propagation of this medicinal plant. It is necessary to conduct additional research in order to optimize the composition, EC and pH value of the hydroponic nutrient solution.


2019 ◽  
Vol 13 (2) ◽  
pp. 269-278
Author(s):  
Adriano Bortolotti Silva ◽  
Ligiane Aparecida Florentino ◽  
Dalvana De Sousa Pereira ◽  
Paulo Roberto Correa Landgraf ◽  
Ana Carolina Rodrigues Alves ◽  
...  

Ornamental pineapple is a hardy plant with significant landscaping value. Tissue culture of plants is viable for producing plants with a high phytosanitary quality. However, one of the difficulties with this cultivar is the acclimatization process, which is slow and can cause losses. The objective of the present study was to verify the potential of inoculation with diazotrophic bacteria for in vitro and ex vivo growth of ornamental pineapple. A group of diazotrophic bacterial strains selected at the Universidade José do Rosário Vellano (UNIFENAS) was prioritized in this study, and the treatments included bacterial strains UNIFENAS (100-13, 100-60, 100-68, 100-153, 100-167 and 100-198). These strains were evaluated in terms of their capacity to produce indole 3-acetic acid. Subsequently, plants were cultivated in a medium composed of MS medium salts (1/4), adding 1 mL of the bacterial strain. In the control treatment, the plants were maintained in 2 mL of MS medium. 7 days after inoculation, the plants were transplanted into the MS, where they were maintained for 30 days. After in vitro cultivation, the plants were transferred to pots containing commercial Plantmax® substrate and maintained under these conditions for 60 days. The diazotrophic bacteria were able to synthesize auxins, and their inoculation promoted greater growth in vitro and ex vitro in the plants. In the acclimatization phase, the plants inoculated with UNIFENAS strains (100-60, 100-68 and 100-153) promoted a higher shoot growth, chlorophyll content and nitrate reductase enzyme activity.


1970 ◽  
Vol 45 (1) ◽  
pp. 75-78 ◽  
Author(s):  
Shahina Islam ◽  
Mosfequa Zahan ◽  
Shahina Akter ◽  
Tanjina Akhtar Banu ◽  
Ahashan Habib ◽  
...  

An efficient mass propagation method for Feronia limonia was developed from excised shoot tips and nodal explants of in vitro grown seedlings. Explants were cultured on MS medium with different conc. of NAA, Kn, IAA and BAP singly or in combinations. Highest number of micro shoots and better plant growth were obtained from these two explants on MS medium supplemented with 0.2 mg/l BAP alone. The regenerated shoots were successfully rooted on MS medium supplemented with 0.5 mg/l NAA. The in vitro raised plantlets were successfully established in soil following the formation of roots with 100% survivability under ex vitro condition. Key words: Feronia limonia; Mass propagation; Node; Shoot tips; Multiple shoot DOI: 10.3329/bjsir.v45i1.5186 Bangladesh J. Sci. Ind. Res. 45(1), 75-78, 2010


Author(s):  
Alexandru Fira ◽  
Nirmal Joshee ◽  
Victoria Cristea ◽  
Manuela Simu ◽  
Monica Harta ◽  
...  

Micropropagation of Lycium barbarum cv. 'Ningxia N1' was achieved. The cultures were by initiated by axenical seed germination. The highest shoot proliferation was obtained on the MS media with 1.33 or 2.22 µM benzyl adenine, gelled with wheat starch as an agar alternative. The treatments with 2.22 µM benzyl adenine ensured proliferation rates superior to the ones with 1.33 μM benzyl adenine, but the latter provided longer and more robust shoots. Use of large microcuttings as an explant onto the multiplication media ensured higher in vitro explant survival, higher number of shoots regeneration and more vigorous plantlets. The microcuttings inserted vertically into the media yielded superior growth and multiplication as compared to the microcuttings placed horizontally. The non-rooted, elongated shoots from the treatment 1.33 μM benzyl adenine were either rooted in vitro on a hormone-free MS medium with starch or used for direct ex vitro rooting and acclimatization. The optimal number of microcuttings/vessel for in vitro rooting was 40 and the rooted plantlets were efficiently acclimatized ex vitro by three methods: float hydroculture in floating cell trays, floating perlite, and in Jiffy7 pellets.


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