scholarly journals PROLINE IN WINTER WHEAT SHOOTS, OBTAINED AFTER GENETIC TRANSFORMATION

2020 ◽  
pp. 106-114
Author(s):  
Л. Є. Сергєєва ◽  
Л. І. Броннікова
Keyword(s):  
Osmotic Stress ◽  
Winter Wheat ◽  
Open Data ◽  
Numerical Data ◽  
Salt Resistance ◽  
Free Proline ◽  
Stress Reactions ◽  
In Planta ◽  
National Academy Of Sciences ◽  
Initial Forms

Gene engineering is one of the most appropriate methods for obtaining plants with higher tolerance to osmotic stresses. Osmotic stress stimulates the synthesis of compatible solutions that protect plants. The free proline was suggested as one of the possible means for overcoming osmotic stress. Its degradation after stress can provide nitrogen, carbon energy. The enzyme connected with proline degradation is proline dehydrogenase, (ProDH). ProDH serves important functions of stress reactions and the development of plants. Agrobacterium-mediated winter wheat transformation in planta using the strain LBA4404 was performed. The primary forms, genotypes UK 95/17 and UK 322/17, were selected in the Institute of Plant Physiology and Genetics of the National Academy of Sciences of Ukraine. The seeds were gathered and considered to be T0 generation, but till the experiment insertions of the transgene were not verified by PCR. The seeds were germinated on filter paper soaked with a 0.5M solution of mannitol. Germination frequencies were scored after a week of incubation. Mannitol affected seed germination in all tested types. At the same time genotype differences were observed. Under stress condition, the germination level of 95/17 initial form exceeded this parameter of T0 variants. At the same time, the 322/17 genotype demonstrated the opposite tendency. To study the salt resistance of the seeds, they were germinated in 0.5 diluted Murashige-Skuga solution with the addition of 20.0 g / l of seawater salts for 10 days. Free proline levels were estimated in the leaves of 10-day shoots. The winter wheat genotypes demonstrated peculiar characteristics. Salinity provoked the growth of free proline levels. For initial forms of UK95/17 and UK322/17, the proline levels were 1.77 and 4.53 times higher than normal parameters. At the same time under salinity the proline levels in T0 shoots of genotype 95/17 were 0.28–1.43 times and in T0 shoots of genotype 322/17 were 2.67–3.70 times of control marks. However, the proline numerical data of T0 forms of both genotypes were lower than the stress figures of their initial forms. Under osmotic stresses, the increase of proline is usually due to the growth of its synthesis. The events of transgene insertions were not verified by PCR. So we have no open data about transgene activity. But the peculiar features that we observed can be indicators of the indirect influence of transgene. The plant proline level even under normal conditions is not a constant feature but it changes during the vegetation. Proline is not only a compatible osmolyte but regulates the gene expression. In our opinion, the effectiveness of such a construction for obtaining plant forms with higher stress tolerance can be estimated during changes in stress/restoration conditions.

scholarly journals Reactions of corn cell cultures during hard osmotic stresses action

1970 ◽  
Vol 21 ◽  
pp. 178-182
Author(s):  
L. E. Sergeeva ◽  
M. O. Dykun ◽  
L. I. Bronnikova
Keyword(s):  
Osmotic Stress ◽  
Cell Cultures ◽  
Sea Water ◽  
Free Proline ◽  
In Planta ◽  
Agrobacterium Mediated Transformation ◽  
Protein Levels ◽  
Osmotic Stress Tolerance ◽  
Cultural Medium ◽  
High Level

Aim. There are corn cell cultures, obtained from plants of maize inbred line L-390 (control) and from T2 progeny (L-390-T) of plants transformed via in planta Agrobacterium-mediated transformation with LBA4404 strain harboring pBi2E with double-stranded RNA-suppressor of the proline dehydrogenase gene. The reactions of cell variants, cultivated under hard osmotic stress pressure were investigated. Methods. 25.0 g/l of sea water salts or 0.8 M of mannitol were added to F1 cultural medium. Corn cell variants were tested under osmotic stress pressure. The free proline and protein levels were estimated on 14-th and 34-th days of the experiment. Results. L-390-T cell cultures maintained viability and wild type cultures died at the end of experiment. The levels of free proline rose in calli tissues, cultivated on nutrition medium with the addition of mannitol or salinity. At the same time the proline levels of L-390-T cells were products of biosynthesis. While the proline content in control cultures elevated after the degradation of proline rich proteins (PRPs). Conclusions. The L-390-T high level of osmotic stress tolerance is a possible result of transgene activity. Keywords: Zea mays, cell cultures, Agrobacterium-mediated transformation salinity, water stress, proline.

scholarly journals Regulation of Nicotiana tabacum osmotic stress-activated protein kinase and its cellular partner GAPDH by nitric oxide in response to salinity

2010 ◽  
Vol 429 (1) ◽  
pp. 73-83 ◽  
Author(s):  
Izabela Wawer ◽  
Maria Bucholc ◽  
Jéremy Astier ◽  
Anna Anielska-Mazur ◽  
Jennifer Dahan ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Nicotiana Tabacum ◽  
Protein Kinase ◽  
Osmotic Stress ◽  
Time Course ◽  
Biological Effects ◽  
Glycolytic Enzyme ◽  
In Planta ◽  
No Donor ◽  
Cellular Partner

Several studies focusing on elucidating the mechanism of NO (nitric oxide) signalling in plant cells have highlighted that its biological effects are partly mediated by protein kinases. The identity of these kinases and details of how NO modulates their activities, however, remain poorly investigated. In the present study, we have attempted to clarify the mechanisms underlying NO action in the regulation of NtOSAK (Nicotiana tabacum osmotic stress-activated protein kinase), a member of the SNF1 (sucrose non-fermenting 1)-related protein kinase 2 family. We found that in tobacco BY-2 (bright-yellow 2) cells exposed to salt stress, NtOSAK is rapidly activated, partly through a NO-dependent process. This activation, as well as the one observed following treatment of BY-2 cells with the NO donor DEA/NO (diethylamine-NONOate), involved the phosphorylation of two residues located in the kinase activation loop, one being identified as Ser158. Our results indicate that NtOSAK does not undergo the direct chemical modifications of its cysteine residues by S-nitrosylation. Using a co-immunoprecipitation-based strategy, we identified several proteins present in immunocomplex with NtOSAK in salt-treated cells including the glycolytic enzyme GAPDH (glyceraldehyde-3-phosphate dehydrogenase). Our results indicate that NtOSAK directly interacts with GAPDH in planta. Furthermore, in response to salt, GAPDH showed a transient increase in its S-nitrosylation level which was correlated with the time course of NtOSAK activation. However, GADPH S-nitrosylation did not influence its interaction with NtOSAK and did not have an impact on the activity of the protein kinase. Taken together, the results support the hypothesis that NtOSAK and GAPDH form a cellular complex and that both proteins are regulated directly or indirectly by NO.

scholarly journals Agrobacterium-mediated in planta genetic transformation of winter wheat (Triticum aestivum L.)

2018 ◽  
Vol 22 ◽  
pp. 293-298
Author(s):  
S. I. Mykhalska ◽  
A. G. Komisarenko ◽  
V. M. Kurchii ◽  
O. M. Tishchenko
Keyword(s):  
Triticum Aestivum ◽  
Transgenic Plants ◽  
Genetic Transformation ◽  
Winter Wheat ◽  
Natural Frequency ◽  
Triticum Aestivum L ◽  
Transformation Method ◽  
In Planta ◽  
Agrobacterium Mediated Transformation ◽  
Pollination Methods

Aim. To optimize the agrobacterium-mediated method of winter wheat transformation (Triticum aestivum L.); to select the conditions and period of inoculation to effectively transfer the genes during pollination. Methods. Agrobacterium-mediated in planta genetic transformation of winter wheat (Triticum aestivum L.) during pollination. Results. The conditions for agrobacterium-mediated transformation method of winter wheat during natural (frequency pollination was 1 %) and non-natural (frequency pollination was 4 %) pollination were defined. Conclusions. The possibility of integrating transgenes into the genome of winter wheat plants by the method of Agrobacterium-mediated transformation in planta in the process of forced and natural pollination is demonstrated. It is found that the transformation efficiency to a large extent depends on the plant genotype and the method of carrying out the transformation procedure. The selection of transgenic plants under water deficit conditions allowed to identify the plants with functional transgene. The signs of functioning transgene have been remaining in the next generation of genetically modified winter wheat. Keywords: Triticum aestivum L., Agrobacterium-mediated transformation in planta, transgenic plants, seeds.

scholarly journals Early-season mapping of winter wheat in China based on Landsat and Sentinel images

2020 ◽  
Vol 12 (4) ◽  
pp. 3081-3095
Author(s):  
Jie Dong ◽  
Yangyang Fu ◽  
Jingjing Wang ◽  
Haifeng Tian ◽  
Shan Fu ◽  
...  
Keyword(s):  
Winter Wheat ◽  
Spatial Resolution ◽  
Vegetation Index ◽  
Open Data ◽  
Geographical Location ◽  
Crop Growth ◽  
Data Repository ◽  
Large Area ◽  
Early Season ◽  
Distribution Maps

Abstract. Early-season crop identification is of great importance for monitoring crop growth and predicting yield for decision makers and private sectors. As one of the largest producers of winter wheat worldwide, China outputs more than 18 % of the global production of winter wheat. However, there are no distribution maps of winter wheat over a large spatial extent with high spatial resolution. In this study, we applied a phenology-based approach to distinguish winter wheat from other crops by comparing the similarity of the seasonal changes of satellite-based vegetation index over all croplands with a standard seasonal change derived from known winter wheat fields. Especially, this study examined the potential of early-season large-area mapping of winter wheat and developed accurate winter wheat maps with 30 m spatial resolution for 3 years (2016–2018) over 11 provinces, which produce more than 98 % of the winter wheat in China. A comprehensive assessment based on survey samples revealed producer's and user's accuracies higher than 89.30 % and 90.59 %, respectively. The estimated winter wheat area exhibited good correlations with the agricultural statistical area data at the municipal and county levels. In addition, the earliest identifiable time of the geographical location of winter wheat was achieved by the end of March, giving a lead time of approximately 3 months before harvest, and the optimal identifiable time of winter wheat was at the end of April with an overall accuracy of 89.88 %. These results are expected to aid in the timely monitoring of crop growth. The 30 m winter wheat maps in China are available via an open-data repository (DOI: https://doi.org/10.6084/m9.figshare.12003990, Dong et al., 2020a).

scholarly journals A Comparison of Geovisualizations and Data Tables for Transparency Enablement in the Open Government Data Landscape

2018 ◽  
Vol 14 (4) ◽  
pp. 39-64 ◽  
Author(s):  
Auriol Degbelo ◽  
Jonas Wissing ◽  
Tomi Kauppinen
Keyword(s):  
User Study ◽  
Open Data ◽  
Numerical Data ◽  
Spatial Knowledge ◽  
Open Government ◽  
Open Government Data ◽  
Hidden Knowledge ◽  
Actual Effectiveness ◽  
Government Data ◽  
Data Tables

Recent years have witnessed progress of public institutions in making their datasets available online, free of charge, for re-use. There have been however limited studies which assess the actual effectiveness of different communication media in making key facts visible to citizens. This article analysed and systematically compared two representations which are relevant in the context of open government data: geovisualizations and data tables. An empirical user study (N=16) revealed that both types of representations have their strengths: geovisualizations make spatial knowledge and the attractiveness of open government data more visible, while data tables are more adequate for the communication of numerical data. The ideas presented are relevant to open data publishers interested in strategies to effectively put the hidden knowledge in current open government datasets into the hands of citizens.

scholarly journals The Mitogen-Activated Protein Kinase BcSak1 of Botrytis cinerea Is Required for Pathogenic Development and Has Broad Regulatory Functions Beyond Stress Response

2012 ◽  
Vol 25 (6) ◽  
pp. 802-816 ◽  
Author(s):  
Jens Heller ◽  
Nadja Ruhnke ◽  
José Juan Espino ◽  
Michelli Massaroli ◽  
Isidro Gonzalez Collado ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Stress Response ◽  
Protein Kinase ◽  
Osmotic Stress ◽  
Botrytis Cinerea ◽  
Fluorescent Protein ◽  
Mitogen Activated Protein Kinase ◽  
In Planta ◽  
Mitogen Activated Protein

The mitogen-activated protein kinase (MAPK) BcSak1 of Botrytis cinerea is activated upon exposure to H2O2 and, hence, might be involved in coping with oxidative stress during infection. However, beside osmotic and oxidative stress sensitivity, Δbcsak1 mutants have a pleiotropic phenotype, as they do not produce conidia and are unable to penetrate unwounded host tissue. In this study, the role of BcSak1 was investigated in the stress response and during infection of French beans by Botrytis cinerea. Using a macroarray approach, it was shown that BcSak1 is only marginally involved in the specific oxidative stress response. In fact, the induction of several genes after oxidative stress treatment is BcSak1-dependent, but most of these genes are also induced under conditions of osmotic stress. The majority of genes regulated by BcSak1 are not involved in the stress response at all. Using a translational fusion of BcSak1 to green fluorescent protein, it was shown clearly that the localization of this MAPK depends on the type of stress being applied; it associates rapidly to the nucleus only under osmotic stress. Therefore, a model is proposed in which BcSak1 acts in the cytosol by activation of one or more transcription factors under oxidative stress and, at the same time, it reacts to osmotic stress by migrating to the nucleus. Interestingly, the MAPK is also involved in the regulation of secondary metabolism, as the major phytotoxins secreted by this fungus are reduced in the Δbcsak1 deletion mutant. Experiments done in planta underlined the essential role of BcSak1 in the early stages of infection, when it translocates to the nucleus and then changes to cytosolic distribution during hyphal growth within the tissue.

Investigating the role of polyols in Cladosporium fulvum during growth under hyper-osmotic stress and in planta

Planta ◽  
2003 ◽  
Vol 216 (4) ◽  
pp. 614-619 ◽  
Author(s):  
Anthony J. Clark ◽  
Kerry J. Blissett ◽  
Richard P. Oliver
Keyword(s):  
Osmotic Stress ◽  
Cladosporium Fulvum ◽  
In Planta

scholarly journals Modeling of asymmetric division of somatic cell in protoplasts culture of higher plants

2020 ◽  
Vol 11 (2) ◽  
pp. 255-265
Author(s):  
S. I. Kondratenko ◽  
T. P. Pasternak ◽  
O. P. Samovol ◽  
O. M. Mogilna ◽  
O. V. Sergienko
Keyword(s):  
Cell Division ◽  
Osmotic Stress ◽  
Growth Regulators ◽  
Nutrient Medium ◽  
Higher Plants ◽  
Asymmetric Division ◽  
Cycle Phase ◽  
In Planta ◽  
Selection Of

The key result of the work is the selection of factors for the cultivation of protoplasts of higher plants in vitro, which allowed induction of asymmetrical cell division during the first cell cycle phase. Gibberellin has been proved to be one of the main cofactors of asymmetric division of plant cells. The objects of research were plants of the following cultivars aseptically grown in hormone-free MS medium: tobacco (Nicotiana tabacum L.), SR-1 line; Arabidopsis thaliana var. columbia (L.) Heynh; potato (Solanum tuberosum L.), Zarevo cultivar; cultivated white head cabbage (Brassica oleraceae var. capitata L.) of the following varieties: Kharkivska zymnia, Ukrainska osin, Yaroslavna, Lika, Lesya, Bilosnizhka, Dithmarscher Früher, Iyunskarannya; rape (Brassica napus L.) of Shpat cultivar; winter radish (Raphanus sativus L.) of Odessa-5 cultivar. In experiments with mesophilic and hypocotyl protoplasts of the above-mentioned plant species it has been proved that short-term osmotic stress within 16–18 hours being combined with subsequent introduction of high doses of gibberellin GK3 (1 mg/L) into the modified liquid nutrient media TM and SW led to the occurrence of pronounced morphological traits of cytodifferentiation already at the initial stages of the development of mitotically active cells in a number of higher plants. Meanwhile, in all analyzed species, there was observed the division of the initial genetically homogeneous population of mitotically active cells into two types of asymmetric division: by the type of division of the mother cell into smaller daughter cells and by the type of the first asymmetric division of the zygotic embryo in planta. In this case, the first type of asymmetric division occurred during unusual cytomorphism of the mother cells: a pronounced heart-shaped form even before the first division, which is inherent in the morphology of somatic plant embryo in vitro at the heart-shaped stage. A particular study of the effect of osmotic stress influencing protoplasts of various cultivars of white cabbage, isolated from hypocotyls of 7–9 day etiolated seedlings, revealed quite a typical consistent pattern: the acquisition and maintenance of the axis of symmetry in growing microcolonies occurred without extra exogenous gibberellin (GK3), which was added to the nutrient medium earlier. While analyzing the effect of growth regulators on the formation of microcolonies with traits of structural organization, the conclusion was made regarding the commonality of the revealed morphogenetic reactions of cells within the culture of protoplasts of higher plants in vitro with similar reactions studied earlier on other plants, both in vitro and in planta. Modeling of asymmetric cell division in protoplast culture in vitro has become possible by carrying out a balanced selection of growth regulators as well as their coordinated application through time along with changes in osmotic pressure of a nutrient medium.

scholarly journals Corn plant comparative reactions to artificial dehydration

1970 ◽  
pp. 252-255
Author(s):  
L. Ye. Serhieieva ◽  
S. I. Mykhalska ◽  
V. M. Kurchii ◽  
O. M. Tyshchenko
Keyword(s):  
Water Stress ◽  
Free Proline ◽  
In Planta ◽  
Short Term ◽  
Agrobacterium Mediated Transformation ◽  
Water Stress Tolerance ◽  
Dehydrogenase Gene ◽  
Maize Plants ◽  
The Stability ◽  
High Level

Aim. There are maize plants of inbred line L-370 (control) and from T4 progeny of plants transformed via in planta Agrobacterium-mediated transformation with LBA4404 strain harboring pBi2E with double-stranded RNA-suppressor of the proline dehydrogenase gene. The free proline and sucrose levels, sucrose/fructose ratio during period of desicca-tion/rehydration were investigated. Methods. Maize T4 progeny and initial plants of L-370 line were cultivated in con-tainers. Variants were tested under short term (4 days) dehydration and after 3.5 hours of rehydration. The free proline and carbohydrates levels were estimated at those times of the experiment. Results. The short-term water deficit did not create pathological changes in plants. But the free proline levels rose in leaves of all variants. At the same time the ami-no acid levels under stress condition in T4 plans were higher than in L-370-plants. After 3.5 hours of rehydration the proline content in control plants decreased and stayed without changes in T4 plans. In T4 plants the carbohydrate meta-bolism systems energy maintained the stability of the sucrose/fructose ratio during the whole time of experiment. Conclusions. The T4 plants high level of water stress tolerance is a possible result of transgene activity.Keywords: Zea mays, Agrobacterium-mediated transformation, T4-progeny, short-term water stress, dehydration, proline sucrose.

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