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Mutants as Tools for the Analytical Dissection of Cell Differentiation in Physcomitrella Patens Gametophytes
Published in R. N. Chopra, Satish C. Bhatla, Bryophyte Development: Physiology and Biochemistry, 2019
The majority of tropically abnormal P. patens strains11,12,19-21 have been obtained, following mutagenesis, by means of nonselective (i.e., total) isolation procedures.22,30 Mutants affected in cell differentiation have been isolated both nonselectively3,4,22 and selectively.4,22 In the nonselective isolation procedure mutagenized spores have been allowed to germinate and grow into gametophytes, which have been examined visually to detect mutants with abnormal morphologies. Many morphologically abnormal strains obtained in this way have been shown subsequently to have altered sensitivities to exogenous auxin and/or cytokinin.4,22 Many mutants isolated selectively by their resistance to concentrations of the synthetic auxin 1-naphthaleneacetic acid (NAA) or the synthetic cytokinin 6-benzylaminopurine (BAP), which cause profound changes in the growth and development of the wild-type, have been found to be altered developmentally even when grown in the absence of exogenous hormones.4,22
In Vitro Plant Regeneration, Comparative Biochemical and Antioxidant Potential of Calli and Seeds of Sesbania grandiflora (L.) Poiret
Published in Parimelazhagan Thangaraj, Medicinal Plants, 2018
Krishnamoorthy Vinothini, Masilamani Sri Devi, Sudharshan Sekar, Blassan P. George, Heidi Abrahamse, Bettine van Vuuren, Arjun Pandian
Surface sterilized leaf explants were cut into small pieces (0.5–1.0 cm) barring the cut ends and transferred to MS basal medium. Three different auxins [(indole-3-acetic acid (IAA), α-naphthalene acetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D)] cytokinins and 6-benzylaminopurine (BAP) at the same concentration (0.5 mg/L) were used in the callus induction. Leaf explants cultured on the MS medium without any growth regulators were used as controls for callus induction. All treatments were conducted thrice with replicates in each treatment.
Biotechnology
Published in Massimo Maffei, Vetiveria, 2002
Marco Mucciarelli, Ruth E. Leupin
A very simple procedure for shoot culture and propagation of vetiver is given as follows: Young growing sprouts are dissected from plants growing in the field and thoroughly washed in running tap water, external two-three pairs of leaf sheaths being carefully removed and discarded.Remaining shoots are incubated for no more than 1–2 days, at room temperature, on wet paper, in order to favour mould growth and spore germination.After the shoots have been washed and sterilized with ethanol 70% (3–5 min) the next pairs of leaves are removed.Complete sterilization is attained by soaking shoots in NaOCl 20% plus Tween 20 or other detergent (0.1%) for 15 min. For older and denser culms, the outermost sheathing leaf pairs can be still discarded and sterilization repeated before washing the shoots with sterile distilled water.Shoots are cut at the base and incubated on full strength MS medium (Murashige and Skoog, 1962) supplemented with 3.0 mg l−1 6-benzylaminopurine (BAP) for 2–6 weeks in order to obtain clumping and sprouting (Figure 7.1a). In vitro grown clumps are then transferred to an MS medium supplemented with 1.5 mg l−1 BAP and 0.1 mg l−1 indolebutyrric acid (IBA) in order to increase clumping and sprouting (Figure 7.1b).Rooting can be enhanced by incubation on MS with 0.1 mg l−1 indoleacetic acid (IAA).Rooted shoots are separated from clumps, washed with water to remove agar and then treated with a fungicidal solution before acclimatization in a greenhouse. To prevent infections and dehydration, plantlets must be placed in half bottles filled with a 1:1 compost:sand sterile mixture and top wrapped with a transparent plastic seal.
Interactive effects of zinc oxide nano particles and different light regimes on growth and silymarin biosynthesis in callus cultures of Silybum marianum L.
Published in Artificial Cells, Nanomedicine, and Biotechnology, 2021
Muhammad aamir Shehzad, Mubarak Ali Khan, Amir Ali, Sher Mohammad, Ahmed Noureldeen, Hadeer Darwish, Asif Ali, Ayaz Ahmad, Tariq Khan, Raham Sher Khan
For the establishment of callus cultures, three different explants (root, hypocotyl and cotyledon) sections of size approximately 1 to 1.5 cm were cut out from four weeks old in vitro germinated seedlings. The excised explants were inoculated on MS medium supplemented with a combination of plant growth hormones 2,4-Dichlorophenoxyacetic acid (2,4-D) and 6-Benzylaminopurine(BA) added at various concentrations (1.1 mg/L, 2.2 mg/L and 3.3 mg/L), respectively. For each treatment, an explant was cultured on MS medium in a single test tube. Therefore, to obtain three replicate, the experiment was repeated thrice (1 explant × 3 culture tubes). Total 27 test tubes were subjected for each explant inoculation with various concentrations of the selected hormones. The data collection for callus culture was initiated after the 4th day of inoculation till the 35th day. Fifth weeks old calli were then sub-cultured on respective hormonal media composition for callus proliferation.
Green synthesis of silver nanoparticles using transgenic Nicotiana tabacum callus culture expressing silicatein gene from marine sponge Latrunculia oparinae
Published in Artificial Cells, Nanomedicine, and Biotechnology, 2018
Yuri N. Shkryl, Galina N. Veremeichik, Dmitriy G. Kamenev, Tatiana Y. Gorpenchenko, Yulia A. Yugay, Dmitriy V. Mashtalyar, Aleksander V. Nepomnyaschiy, Tatiana V. Avramenko, Aleksandr A. Karabtsov, Vladimir V. Ivanov, Victor P. Bulgakov, Sergey V. Gnedenkov, Yury N. Kulchin, Yury N. Zhuravlev
Agrobacterium tumefaciens carrying pPZP-RCS2-nptII, pPZP-RCS2-nptII/LoSilA1, pPZP-RCS2-nptII/LoSilA1-EGFP and pPZP-RCS2-nptII/EGFP plasmids were used to inoculate leaf discs of sterile, clonally cultivated plantlets of N. tabacum L. (cv Xanthi) according to the previously described conditions [42]. Leaf discs were co-cultivated with A. tumefaciens for 48 h and transferred to the WB/A agarized medium supplemented with 0.5 mg/L 6-benzylaminopurine and 2.0 mg/L α-naphthaleneacetic acid [43] containing 500 mg/L cefotaxime and 100 mg/L kanamycin. Transgenic callus cultures expressing the selectable marker (nptII) gene alone or together with the LoSilA1 gene, LoSilA1-EGFP fusion gene or EGFP gene, namely Nt-cV, Nt-cS, Nt-cSG and Nt-cG, were obtained after four months of selection with kanamycin. Plant regeneration was achieved by placing the 4–5 week-old primary calli on hormone-free W medium containing 100 mg/L kanamycin under a 16/8 h light/dark cycle. Thus, transgenic plantlets expressing the selectable marker (nptII) gene alone or together with the LoSilA1 gene, LoSilA1-EGFP fusion gene or EGFP gene, namely Nt-pV, Nt-pS, Nt-pSG and Nt-pG, were obtained. The control non-transformed callus culture and plants were established from the same plantlets and cultivated under the same conditions as the transformed ones.
Marginalized zero-inflated generalized Poisson regression
Published in Journal of Applied Statistics, 2018
Felix Famoye, John S. Preisser
Ridout et al. [18] fitted Poisson, ZIP, NB and ZINB models to the number of roots from 270 shoots of apple cultivar Trajan. For more detailed description of the data, see also [19]. The frequency distributions of the number of roots produced were classified by two experimental conditions: four concentrations of the cytokinin 6-benzylaminopurine (BAP) and two levels of photoperiod. In our evaluation, we fitted NB, MZINB, GP and MZIGP for comparisons with regard to overall exposure effects of BAP concentration and photoperiod on the mean number of roots produced. The first level of each experimental condition, 8 h photoperiod and 2.2 BAP concentration, respectively, is the reference level. Thus, x1 is an indicator variable for photoperiod 16 h while x2, x3 and x4 are indicators for BAP = 4.4, 8.8 and 17.6, respectively. For the marginalized models, we used Table 3.