Marie Kočová

Terminal drought-tolerant pearl millet [Pennisetum glaucum (L.) R. Br.] have high leaf ABA and limit transpiration at high vapour pressure deficit

It was previously shown that pearl millet genotypes carrying a terminal drought tolerance quantitative trait locus (QTL) had a lower transpiration rate (Tr; g cm−2 d−1) under well-watered conditions than sensitive lines. Here experiments were carried out to test whether this relates to leaf abscisic acid (ABA) and Tr concentration at high vapour pressure deficit (VPD), and whether that leads to transpiration efficiency (TE) differences. These traits were measured in tolerant/sensitive pearl millet genotypes, including near-isogenic lines introgressed with a terminal drought tolerance QTL (NIL-QTLs). Most genotypic differences were found under well-watered conditions. ABA levels under well-watered conditions were higher in tolerant genotypes, including NIL-QTLs, than in sensitive genotypes, and ABA did not increase under water stress. Well-watered Tr was lower in tolerant than in sensitive genotypes at all VPD levels. Except for one line, Tr slowed down in tolerant lines above a breakpoint at 1.40–1.90 kPa, with the slope decreasing >50%, whereas sensitive lines showed no change in that Tr response across the whole VPD range. It is concluded that two water-saving (avoidance) mechanisms may operate under well-watered conditions in tolerant pearl millet: (i) a low Tr even at low VPD conditions, which may relate to leaf ABA; and (ii) a sensitivity to higher VPD that further restricts Tr, which suggests the involvement of hydraulic signals. Both traits, which did not lead to TE differences, could contribute to absolute water saving seen in part due to dry weight increase differences. This water saved would become critical for grain filling and deserves consideration in the breeding of terminal drought-tolerant lines.

Constitutive water-conserving mechanisms are correlated with the terminal drought tolerance of pearl millet [Pennisetum glaucum (L.) R. Br.]

Pearl millet, a key staple crop of the semi-arid tropics, is mostly grown in water-limited conditions, and improving its performance depends on how genotypes manage limited water resources. This study investigates whether the control of water loss under non-limiting water conditions is involved in the terminal drought tolerance of pearl millet. Two pairs of tolerant×sensitive pearl millet genotypes, PRLT 2/89-33–H77/833-2 and 863B-P2–ICMB 841-P3, and near-isogenic lines (NILs), introgressed with a terminal drought tolerance quantitative trait locus (QTL) from the donor parent PRLT 2/89-33 into H77/833-2 (NILs-QTL), were tested. Upon exposure to water deficit, transpiration began to decline at lower fractions of transpirable soil water (FTSW) in tolerant than in sensitive genotypes, and NILs-QTL followed the pattern of the tolerant parents. The transpiration rate (Tr, in g water loss cm−2 d−1) under well-watered conditions was lower in tolerant than in sensitive parental genotypes, and the Tr of NILs-QTL followed the pattern of the tolerant parents. In addition, Tr measured in detached leaves (g water loss cm−2 h−1) from field-grown plants of the parental lines showed lower Tr values in tolerant parents. Defoliation led to an increase in Tr that was higher in sensitive than in tolerant genotypes. The differences in Tr between genotypes was not related to the stomatal density. These results demonstrate that constitutive traits controlling leaf water loss under well-watered conditions correlate with the terminal drought tolerance of pearl millet. Such traits may lead to more water being available for grain filling under terminal drought.

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.

The Development of Chloroplast Ultrastructure and Hill Reaction Activity During Leaf Ontogeny in Different Maize (Zea Mays L.) Genotypes

Changes in Hill reaction activity (HRA) and ultrastructure of mesophyll cell (MC) chloroplasts were studied during the ontogeny of third leaf of maize plants using polarographic oxygen evolution measurement, transmission electron microscopy, and stereology. The chloroplast ultrastructure was compared in young (actively growing), mature, and senescing leaves of two different inbreds and their reciprocal F1 hybrids. Statistically significant differences in both HRA and MC chloroplast ultrastructure were observed between different stages of leaf ontogeny. Growth of plastoglobuli was the most striking characteristic of chloroplast maturation and senescence. The chloroplasts in mature and senescing leaves had a more developed system of thylakoids compared to the young leaves. Higher HRA was usually connected with higher thylakoid volume density of MC chloroplasts.

The influence of low-temperature on the photochemical activity of chloroplasts and activity of antioxidant enzymes in maize leaves

The effects of low growth temperature on the activities of photosystems (PS) 1 and 2 and antioxidant enzymes superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione reductase (GR) and catalase (CAT) in leaves of various maize inbred and hybrid genotypes (parental lines, F1 hybrids, F2 and backcross generations) were investigated. Considerable decrease of the PS 2 activity (contrary to the activity of PS 1) due to low-temperature stress was observed in the majority of genotypes/generations examined. The GR, APX and SOD activities markedly increased due to chilling, whereas the activity of CAT showed lesser changes which depended on the genotype/generation analysed. The higher susceptibility of the inbred line 2013 to low temperature was transmitted to further generations in case this line was used as the maternal parent. The intraspecific variability in photosynthetic and antioxidant parameters was caused particularly by the dominance (negative or positive), however, the level of the expression of this effect often changed after low-temperature stress and was probably the cause of the increase in the positive F1 heterosis observed in this case. Other genetic effects (e.g. the additivity, and particularly the additive or dominant maternal effects) were also found to contribute to the intraspecific variability in parameters analyzed. The dominant maternal effects possibly played an important role in maintaining positive heterosis in F2 generation.

The Exogenous Application of Brassinosteroids to Zea mays (L.) Stressed by Long-Term Chilling Does Not Affect the Activities of Photosystem 1 or 2

The effect of various concentrations of exogenously applied 24-epibrassinolide (E) and 2α,3α,17β-trihydroxy-5α-androstan-6-one (A) on the activities of Photosystem 1 and the Hill reaction, the contents of photosynthetic pigments, and the growth of plants was examined in young maize (Zea mays L.) plants subjected to long-term chilling stress or grown in normal-temperature conditions. Neither the activity of Photosystem 1 nor the Hill reaction activity of plants was in any way affected by the treatment with brassinosteroids (BRs), which suggests that the photosynthetic complexes of thylakoid membranes are not the primary site of the influence of BRs on photosynthesis. An extremely low (10−14 M) concentration of A applied to the nonstressed plants significantly increased the length of their 4th to the 7th leaves and their height, as well as the contents of chlorophylls a and b and total carotenoids. However, under chilling conditions, this positive effect was significant for the chlorophyll content only and higher concentrations of BRs (10−12, 10−10, 10−8 M) usually had no effect at all.

Photosynthetic parameters of maize (Zea mays L.) inbred lines and F1 hybrids: their different response to, and recovery from rapid or gradual onset of low-temperature stress

The activity of photosystems (PS) 1 and 2, together with the content and ratio of photosynthetic pigments, were measured in three inbred lines and two F1 hybrids of maize (Zea mays L.), grown in either optimum or low temperature (LT) conditions. The ability of chilling-stressed plants to deal with the negative effects of long-term exposure to LT and to recover the efficiency of photosynthetic apparatus after their return to optimum temperatures was examined during spring and autumn seasons. The aim was to analyse the possible differences between the rapid and gradual onset of LT on the response of young maize plants to chilling stress. The distinctive superiority of hybrids over their parental lines, found during the exposure of maize plants to LT, was not always retained after the return of chilling-stressed plants to optimum growth conditions. The response of individual genotypes to chilling stress, as well as their ability to recover the photosynthetic efficiency from the cold-induced damage, strongly depended also on the duration and the rapidity of the onset of LT.

The effect of brassinosteroids on the morphology, development and yield of field-grown maize.

The response of two field-grown inbred lines of maize (Zea mays L.) and their F1 hybrid to the application of 10−8–10−14 M solutions of 24-epibrassinolide or synthetic androstane analogue of castasterone in V3/4 and V6/7 developmental stages was followed during the vegetative and early reproductive phases of plant development. Brassinosteroids (BRs) significantly affected (either positively or negatively, depending on the genotype and the developmental stage they were applied) the height of plants during the early weeks after their application, but not the final plant height nor the number of leaves. Spraying of plants with BRs in V3/4 developmental stage usually also increased the length of the 7th to 10th leaf, whereas the application in V6/7 developmental stage had the opposite effect. The beginning of the reproductive phase of plant development and the course of flowering was strongly influenced by the application of BRs. Treatment of plants in V3/4 stage delayed and treatment of plants in V6/7 stage advanced the dates of anthesis and silking, regardless of the type of BR used, its concentration or plant genotype. The influence of BRs on the development of the secondary ear was the least pronounced in the F1 hybrid; in both inbred lines it strongly depended on the concentrations of BRs used. Various yield parameters were also affected by treatment of plants with BRs, but this effect depended on the developmental stage during which the application of BRs occured, the plant genotype, the type of BR and its concentration.

The ultrastructure of chloroplasts, content of photosynthetic pigments, and photochemical activity of maize (Zea mays L.) as influenced by different concentrations of the herbicide amitrole

The effect of three different concentrations of amitrole (AM), a bleaching herbicide affecting carotenogenesis, on chloroplast ultrastructure, photosynthetic pigment contents, and photochemical activity was studied in two maize genotypes differing in photosynthetic characteristics. The content of photosynthetic pigments in leaves of plants treated with low (20 μM) AM concentration was similar to control plants and no damaging effect of the herbicide on the ultrastructure of either mesophyll (MC) or bundle-sheath (BSC) cell chloroplasts was observed. Higher (60 and 120 μM) concentrations of AM caused a significant decrease in the content of carotenoids (especially xanthophylls), which was followed by photooxidative destruction of chlorophylls and some alterations of chloroplast ultrastructure. MC chloroplasts appeared more sensitive to the damaging effect of AM compared to BSC chloroplasts. A significant decrease in the amount of both granal and intergranal thylakoids in MC chloroplasts was observed with the increasing concentration of AM. As regards BSC chloroplasts, rapid decrease in the volume density of starch inclusions was found in plants treated with higher concentrations of AM. When 120 μM AM was used, both MC and BSC chloroplasts contained just a few thylakoid membranes that were strongly altered. The changes in the ultrastructure of MC chloroplasts were accompanied by the changes in their photochemical activity. The formation of chloroplast protrusions after treatment of plants with AM as well as in control plants was also observed.

The evaluation of photosynthetic parameters in maize inbred lines subjected to water deficiency: Can these parameters be used for the prediction of performance of hybrid progeny?

The response of selected photosynthetic and morphological parameters of plants to drought was examined in 5 inbred lines of maize (Zea mays L.) and their 10 F1 hybrids. The aim of the study was to establish whether the photosynthetic performance of parental genotypes under drought conditions correlates with the performance of their progeny and whether the net photosynthetic rate, the chlorophyll fluorescence parameters or the content of photosynthetic pigments could be used as reliable physiological markers for early breeding generations. The relative importance of the additive and the nonadditive (dominance, maternal) genetic effects in the inheritance of these parameters was also assessed by means of the quantitative genetics analysis. The results showed that the nonadditive genetic effects associated with a particular combination of genotypes or a particular direction of crossing are at least equally and often even more important as the additivity and that these genetic effects almost totally change with the exposure of plants to drought conditions. This was reflected in the inability to predict the response of F1 hybrids to drought on the basis of the photosynthetic performance of their parents, which indicates that the practical usability of such parameters in maize breeding programs is rather limited.