The production and role of reactive oxygen species (ROS) in the expanding zone of maize (Zea mays) leaf blades were investigated. ROS release along the leaf blade was evaluated by embedding intact seedlings in 2Ј,7Ј-dichlorofluoresceincontaining agar and examining the distribution of 2Ј,7Ј-dichlorofluorescein fluorescence along leaf 4, which was exposed by removing the outer leaves before embedding the seedling. Fluorescence was high in the expanding region, becoming practically non-detectable beyond 65 mm from the ligule, indicating high ROS production in the expansion zone. Segments obtained from the elongation zone of leaf 4 were used to assess the role of ROS in leaf elongation. The distribution of cerium perhydroxide deposits in electron micrographs indicated hydrogen peroxide (H 2 O 2 ) presence in the apoplast. 2Ј,7Ј-Dichlorofluorescein fluorescence and apoplastic H 2 O 2 accumulation were inhibited with diphenyleneiodonium (DPI), which also inhibited O ⅐ 2 Ϫ generation, suggesting a flavin-containing enzyme activity such as NADPH oxidase was involved in ROS production. Segments from the elongation zone incubated in water grew 8% in 2 h. KI treatments, which scavenged H 2 O 2 but did not inhibit O ⅐ 2 Ϫ production, did not modify growth. DPI significantly inhibited segment elongation, and the addition of H 2 O 2 (50 or 500 m) to the incubation medium partially reverted the inhibition caused by DPI. These results indicate that a certain concentration of H 2 O 2 is necessary for leaf elongation, but it could not be distinguished whether H 2 O 2 , or other ROS, are the actual active agents.

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Buffel grass (Cenchrus ciliaris L.) is an important apomictic grass used as forage for ruminant livestock. Biotechnological methods provide opportunities for producing new germplasm. Mature embryos of fourteen buffel grass apomictic cultivars (2n = 4x = 36) were used to induce embryogenic callus formation using a basal medium supplemented with 3% sucrose and with the testing of five concentrations of 2,4‐dichlorophenoxyacetic acid (2,4‐D) and four concentrations of 6‐benzylaminopurine (BAP). The effects of cultivar and culture medium on callus induction and plant regeneration were evaluated. Significant differences were observed among the fourteen cultivars and the five concentrations of 2,4‐D (P < 0·01). Values for embryogenic callus production varied from 0 to 86·7. Most cultivars showed the highest level of embryogenic callus production on the medium with the concentration of 3 mg L−1 2,4‐D. The addition of different BAP concentrations in combination with 2,4‐D in the medium inhibited embryogenic callus growth and did not permit plant regeneration. The data clearly demonstrated that the genotype and concentrations of 2,4‐D had significant effects both on the frequency of embryonic callus formation from mature embryos and on the subsequent efficiency of plant regeneration of apomictic cultivars of buffel grass. Cultivars Biloela and Nunbank showed the greatest efficiency in in vitro culture response.

Buffel grass (Cenchrus ciliaris L.) is an important forage grass in arid and semiarid regions. As part of a genetic improvement programme, four genotypes [Biloela (Bl), Americana (Am), Texas (Tx) and Sexual (Sx)] were categorized by tolerance to heat stress in a greenhouse experiment. At 30 d after sowing, half of the plants (control plants) were transferred to growth chambers (28°C day per night), and the other half (pre-treated plants, Prt) were exposed to heat stress treatment (0, 24, 48 and 72 h; 45°C day per night). Malondialdehyde (MDA) content, an indicator of oxidative damage, was determined from foliar samples. During heat stress, Sx showed the earliest increase in MDA (at 24 h) followed by Tx (48 h) and Am and Bl (72 h). Results were compared with heat-stress tolerance evaluated as morphological traits at the end of recuperation (60 d after sowing). Fresh weight and aerial plant height were lowest in the Prt-Sx genotype and highest in Am and Bl genotypes; Tx showed intermediate tolerance. Results suggest that tolerance to heat stress in C. ciliaris genotypes could be related to the capacity for regulating the oxidative damage increase. Foliar MDA content might therefore be used in a genetic improvement programme of C. ciliaris as a potential biochemical indicator for a rapid, simple and low-cost identification of heat-stress tolerant genotypes.

Rhodes grass (Chloris gayana Kunth) is widely cultivated in the semi-arid tropics and favoured for salt tolerance; nevertheless, productivity decreases significantly under saline conditions, especially in tetraploid cultivars. The purpose of this work was to explore, in the tetraploid cultivar Boma, the physiological causes for such decrease. Experiments were carried out in vegetative plants in the greenhouse. At high salinity (200 mM NaCl), an early reduction in leaf area expansion was observed, and, later, decreased dry matter accumulation, suggesting that the central effect of salinity was a limitation of leaf growth. This was evaluated in 2 closely related Boma clones, exhibiting different degrees of salt tolerance. Growth, ion accumulation and excretion, sugars, and proline concentrations were measured under a range of salt concentrations (0–200 mM NaCl). Differences between the clones in leaf expansion were expressed only at high salinity, but were not associated with differences in water potential or hydraulic conductance in the expanding region. At all salinity levels, the proportion of dry leaves was higher in the clone which also had lower salt gland density and Na excretion rates. Less efficient Na extrusion, associated with high oxidative stress, may be the main cause for leaf senescence and differences in productivity between these clones.