Eukaryotic-like Ser/Thr Protein Kinases SpkC/F/K Are Involved in Phosphorylation of GroES in the Cyanobacterium Synechocystis

Serine/threonine protein kinases (STPKs) are the major participants in intracellular signal transduction in eukaryotes, such as yeasts, fungi, plants, and animals. Genome sequences indicate that these kinases are also present in prokaryotes, such as cyanobacteria. However, their roles in signal transduction in prokaryotes remain poorly understood. We have attempted to identify the roles of STPKs in response to heat stress in the prokaryotic cyanobacterium Synechocystis sp. PCC 6803, which has 12 genes for STPKs. Each gene was individually inactivated to generate a gene-knockout library of STPKs. We applied in vitro Ser/Thr protein phosphorylation and phosphoproteomics and identified the methionyl-tRNA synthetase, large subunit of RuBisCO, 6-phosphogluconate dehydrogenase, translation elongation factor Tu, heat-shock protein GrpE, and small chaperonin GroES as the putative targets for Ser/Thr phosphorylation. The expressed and purified GroES was used as an external substrate to screen the protein extracts of the individual mutants for their Ser/Thr kinase activities. The mutants that lack one of the three protein kinases, SpkC, SpkF, and SpkK, were unable to phosphorylate GroES in vitro, suggesting possible interactions between them towards their substrate. Complementation of the mutated SpkC, SpkF, and SpkK leads to the restoration of the ability of cells to phosphorylate the GroES. This suggests that these three STPKs are organized in a sequential order or a cascade and they work one after another to finally phosphorylate the GroES

Improvement of regeneration in pepper: a recalcitrant species

[EN] Organogenesis is influenced by factors like genotype, type of explant, culture medium components, and incubation conditions. The influence of ethylene, which can be produced in the culture process, can also be a limiting factor in recalcitrant species like pepper. In this work, bud induction was achieved from cotyledons and hypocotyls-from eight pepper cultivars-on Murashige and Skoog (MS) medium supplemented with 22.2 mu M 6-benzyladenine (6BA) and 5.71 mu M indole-3-acetic acid (IAA), in media with or without silver nitrate (SN) (58.86 mu M), a suppressor of ethylene action. In the SN-supplemented medium, the frequencies of explants with buds and with callus formation were lower in both kinds of explant, but higher numbers of developed shoots were isolated from explants cultured on SN. Bud elongation was better in medium with gibberellic acid (GA(3)) (2.88 mu M) than in medium free of growth regulators or supplemented with 1-aminocyclopropane-1-carboxylic acid (ACC) at 34.5 mu M. However, isolation of shoots was difficult and few plants were recovered. The effect of adding SN following bud induction (at 7 d) and that of dark incubation (the first 7 d of culture) was also assessed in order to improve the previous results. When SN was added after bud induction, similar percentages of bud induction were found for cotyledons (average frequency 89.37% without SN and 94.37% with SN) whereas they doubled in hypocotyls (50% without SN and 87.7% with SN). In addition, in both kinds of explant, the number of developed plants able to be transferred to soil (developed and rooted) was greatly increased by SN. Dark incubation does not seem to improve organogenesis in pepper, and hypocotyl explants clearly represent a better explant choice-with respect to cotyledonary explants-for the pepper cultivars assayed.We thank the COMAV germplasm bank at Universitat Politecnica de Valencia and the Arid Lands Institute for pepper seeds and the Tunisian Ministry of Higher Education and Scientific Research who fund N. Gammoudi's stay.Gammoudi, N.; San Pedro-Galan, T.; Ferchichi, A.; Gisbert Domenech, MC. (2018). Improvement of regeneration in pepper: a recalcitrant species. In Vitro Cellular & Developmental Biology - Plant. 54(2):145-153. https://doi.org/10.1007/s11627-017-9838-1S145153542Ashrafuzzaman M, Hossain MM, Razi Ismail M, Shahidul Haque M, Shahidullah SM, Uz Zaman S (2009) Regeneration potential of seedling explants of chilli (Capsicum annuum). Afr J Biotechnol 8:591–596Bortesi L, Fischer R (2015) The CRISPR/Cas9 system for plant genome editing and beyond. 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Three approaches to data analysis

In this book, the following three approaches to data analysis are presented:  - Test Theory, founded by Sergei V. Yablonskii (1924-1998); the first publications appeared in 1955 and 1958, -           Rough Sets, founded by Zdzisław I. Pawlak (1926-2006); the first publications appeared in 1981 and 1982, -           Logical Analysis of Data, founded by Peter L. Hammer (1936-2006); the first publications appeared in 1986 and 1988. These three approaches have much in common, but researchers active in one of these areas often have a limited knowledge about the results and methods developed in the other two. On the other hand, each of the approaches shows some originality and we believe that the exchange of knowledge can stimulate further development of each of them. This can lead to new theoretical results and real-life applications and, in particular, new results based on combination of these three data analysis approaches can be expected