Genetic variability in growth, reproductive and photosynthetic parameters of plants and its changes by exogenously applied steroids

Živočišné steroidní hormony jsou velmi dobře známy a jejich výzkum probíhá již dlouhou dobu, v rostlinách však dlouho žádné steroidní látky s biologickou funkcí nebyly identifikovány. Teprve v druhé polovině minulého století byly objeveny brassinosteroidy, u nichž byla později potvrzena hormonální funkce v rostlinách. Stále je však při výzkumu jejich funkce mnoho neznámých. Tato práce předkládá ve své první části důkazy, že brassinosteroidy regulují u kukuřice (Zea mays L.) pěstované v polních podmínkách nejen morfologii a výnos, ale i některé vývojové a reprodukční charakteristiky, jako například počet samičích květenství či rychlost vývoje samčích květenství. Konkrétní reakce rostliny však závisí na typu použitého brassinosteroidu, na jeho koncentraci, a v neposlední řadě i na konkrétním genotypu kukuřice a na fázi vývoje rostliny v době postřiku. Vliv brassinosteroidů na primární fotosyntetické procesy v rostlině za těchto podmínek pěstování nebyl prokázán, a to ani na aktivitu fotosystému (PS) 1, ani na aktivitu Hillovy reakce. Nebyly nalezeny ani statisticky průkazné rozdíly v obsahu fotosyntetických pigmentů. Dalším tématem, řešeným v této práci, byl možný ochranný vliv brassinosteroidů na rostliny vystavené chladu. U kontrolních rostlin došlo při exogenní aplikaci velmi nízkých koncentrací...While animal steroid hormones are very well known and have been studied for a long time, in plants no steroid substances were known until relatively recently. Only in the second half of the past century brassinosteroids were discovered; later on, their hormonal function in plants was confirmed. Still a lot of unknown remains as regards their function in plant cells. This paper presents in its first part the evidence that brassinosteroids control in maize (Zea mays L.) grown under field conditions not only its morphology and yield but also some developmental/reproduction characteristics like e.g. number of female inflorescences or speed of the development of male inflorescences. Particular response of a plant depends, however, on the type of applied brassinosteroid, its concentration, and last but not least also on a particular maize genotype and developmental stage of the plant during applicatin. Impact of brassinosteroids on primary photosynthetic processes in plants has not been proven under these conditions, neither on the activity of photosystem (PS) I nor on the Hill reaction. No statistically significant differences in the content of photosynthetic pigments have been found either. Another topic dealt with in this thesis is the possible protective influence of brassinosteroids on plants...Katedra genetiky a mikrobiologieDepartment of Genetics and MicrobiologyPřírodovědecká fakultaFaculty of Scienc

Contents of endogenous brassinosteroids and the response to drought and/or exogenously applied 24-epibrassinolide in two different maize leaves

Exogenously applied brassinosteroids (BRs) improve plant response to drought. However, many important aspects of this process, such as the potential differences caused by different developmental stages of analyzed organs at the beginning of drought, or by BR application before or during drought, remain still unexplored. The same applies for the response of different endogenous BRs belonging to the C27, C28-and C29- structural groups to drought and/or exogenous BRs. This study examines the physiological response of two different leaves (younger and older) of maize plants exposed to drought and treated with 24-epibrassinolide (epiBL), together with the contents of several C27, C28-and C29-BRs. Two timepoints of epiBL application (prior to and during drought) were utilized to ascertain how this could affect plant drought response and the contents of endogenous BRs. Marked differences in the contents of individual BRs between younger and older maize leaves were found: the younger leaves diverted their BR biosynthesis from C28-BRs to C29-BRs, probably at the very early biosynthetic steps, as the levels of C28-BR precursors were very low in these leaves, whereas C29-BR levels vere extremely high. Drought also apparently negatively affected contents of C28-BRs (particularly in the older leaves) and C29-BRs (particularly in the younger leaves) but not C27-BRs. The response of these two types of leaves to the combination of drought exposure and the application of exogenous epiBL differed in some aspects. The older leaves showed accelerated senescence under such conditions reflected in their reduced chlorophyll content and diminished efficiency of the primary photosynthetic processes. In contrast, the younger leaves of well-watered plants showed at first a reduction of proline levels in response to epiBL treatment, whereas in drought-stressed, epiBL pre-treated plants they were subsequently characterized by elevated amounts of proline. The contents of C29- and C27-BRs in plants treated with exogenous epiBL depended on the length of time between this treatment and the BR analysis regardless of plant water supply; they were more pronounced in plants subjected to the later epiBL treatment. The application of epiBL before or during drought did not result in any differences of plant response to this stressor

The Physiology and Proteomics of Drought Tolerance in Maize: Early Stomatal Closure as a Cause of Lower Tolerance to Short-Term Dehydration?

Understanding the response of a crop to drought is the first step in the breeding of tolerant genotypes. In our study, two maize (Zea mays L.) genotypes with contrasting sensitivity to dehydration were subjected to moderate drought conditions. The subsequent analysis of their physiological parameters revealed a decreased stomatal conductance accompanied by a slighter decrease in the relative water content in the sensitive genotype. In contrast, the tolerant genotype maintained open stomata and active photosynthesis, even under dehydration conditions. Drought-induced changes in the leaf proteome were analyzed by two independent approaches, 2D gel electrophoresis and iTRAQ analysis, which provided compatible but only partially overlapping results. Drought caused the up-regulation of protective and stress-related proteins (mainly chaperones and dehydrins) in both genotypes. The differences in the levels of various detoxification proteins corresponded well with the observed changes in the activities of antioxidant enzymes. The number and levels of up-regulated protective proteins were generally lower in the sensitive genotype, implying a reduced level of proteosynthesis, which was also indicated by specific changes in the components of the translation machinery. Based on these results, we propose that the hypersensitive early stomatal closure in the sensitive genotype leads to the inhibition of photosynthesis and, subsequently, to a less efficient synthesis of the protective/detoxification proteins that are associated with drought tolerance

Genetic variability in growth, reproductive and photosynthetic parameters of plants and its changes by exogenously applied steroids

While animal steroid hormones are very well known and have been studied for a long time, in plants no steroid substances were known until relatively recently. Only in the second half of the past century brassinosteroids were discovered; later on, their hormonal function in plants was confirmed. Still a lot of unknown remains as regards their function in plant cells. This paper presents in its first part the evidence that brassinosteroids control in maize (Zea mays L.) grown under field conditions not only its morphology and yield but also some developmental/reproduction characteristics like e.g. number of female inflorescences or speed of the development of male inflorescences. Particular response of a plant depends, however, on the type of applied brassinosteroid, its concentration, and last but not least also on a particular maize genotype and developmental stage of the plant during applicatin. Impact of brassinosteroids on primary photosynthetic processes in plants has not been proven under these conditions, neither on the activity of photosystem (PS) I nor on the Hill reaction. No statistically significant differences in the content of photosynthetic pigments have been found either. Another topic dealt with in this thesis is the possible protective influence of brassinosteroids on plants..

Genetic variability in growth, reproductive and photosynthetic parameters of plants and its changes by exogenously applied steroids

While animal steroid hormones are very well known and have been studied for a long time, in plants no steroid substances were known until relatively recently. Only in the second half of the past century brassinosteroids were discovered; later on, their hormonal function in plants was confirmed. Still a lot of unknown remains as regards their function in plant cells. This paper presents in its first part the evidence that brassinosteroids control in maize (Zea mays L.) grown under field conditions not only its morphology and yield but also some developmental/reproduction characteristics like e.g. number of female inflorescences or speed of the development of male inflorescences. Particular response of a plant depends, however, on the type of applied brassinosteroid, its concentration, and last but not least also on a particular maize genotype and developmental stage of the plant during applicatin. Impact of brassinosteroids on primary photosynthetic processes in plants has not been proven under these conditions, neither on the activity of photosystem (PS) I nor on the Hill reaction. No statistically significant differences in the content of photosynthetic pigments have been found either. Another topic dealt with in this thesis is the possible protective influence of brassinosteroids on plants..

Contents of total and individual brassinosteroids (BRs) in two maize genotypes (2023 and CE704).

<p>Plants were either subjected to normal watering (control; grey columns) or to 14 days of withholding water (stress; black columns). Mean values ± SEM are shown (n = 3). Asterisks indicate significant (<i>p</i>≤0.05; *) or highly significant (<i>p</i>≤0.01; **) differences between mean values according to Tukey's tests made separately for each genotype (in case of the differences between control and stress treatment) or for each treatment (in case of the differences between both genotypes). BL … brassinolide, CS … castasterone, DS … dolichosterone, FM … leaf fresh mass, TY … typhasterol.</p

The difference kinetics and the relative variable fluorescences calculated from OJIP analysis of two maize genotypes (2023 and CE704).

<p>The difference kinetics ΔW_OJ (<b>A</b>) reveals the K-band; ΔW_OK (<b>B</b>) reveals the L-band. Only the part between the I and P points of the OJIP curve is shown for the relative variable fluorescence W_OI (<b>C</b>). The normalization of OJIP curve between the I and P points with the maximum amplitude fixed as 1 is shown as the relative variable fluorescence W_IP (<b>D</b>). Plants were subjected either to normal watering (control) or to 14 days of withholding water (stress). ΔW_OJ and ΔW_OK were calculated from the comparisons of the stressed and control plants; the latter are represented by the zero point of the respective y axes in graphs <b>A</b> and <b>B</b>. Mean values (n = 8) are shown. r.u. … relative units.</p

Drought-tolerant and drought-sensitive genotypes of maize (<i>Zea mays</i> L.) differ in contents of endogenous brassinosteroids and their drought-induced changes

<div><p>The contents of endogenous brassinosteroids (BRs) together with various aspects of plant morphology, water management, photosynthesis and protection against cell damage were assessed in two maize genotypes that differed in their drought sensitivity. The presence of 28-norbrassinolide in rather high quantities (1–2 pg mg^-1 fresh mass) in the leaves of monocot plants is reported for the first time. The intraspecific variability in the presence/content of the individual BRs in drought-stressed plants is also described for the first time. The drought-resistant genotype was characterised by a significantly higher content of total endogenous BRs (particularly typhasterol and 28-norbrassinolide) compared with the drought-sensitive genotype. On the other hand, the drought-sensitive genotype showed higher levels of 28-norcastasterone. Both genotypes also differed in the drought-induced reduction/elevation of the levels of 28-norbrassinolide, 28-norcastasterone, 28-homocastasterone and 28-homodolichosterone. The differences observed between both genotypes in the endogenous BR content are probably correlated with their different degrees of drought sensitivity, which was demonstrated at various levels of plant morphology, physiology and biochemistry.</p></div

Selected parameters of plant morphology in two maize genotypes (2023 and CE704).

<p>Plants were either subjected to normal watering (control; grey columns) or to 14 days of withholding water (stress; black columns). Mean values ± SEM are shown (n = 8). Asterisks indicate significant (<i>p</i>≤0.05; *) or highly significant (<i>p</i>≤0.01; **) differences between mean values according to Tukey's tests made separately for each genotype (in case of the differences between control and stress treatment) or for each treatment (in case of the differences between both genotypes).</p