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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.

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Buffel grass is a fodder grass that reproduces mainly via apomixis. Using in vitro tissue culture in apomictic species provides alternatives that can be applied in a breeding program. The aims of this work were to establish a protocol for in vitro generation of apomictic buffel grass genotypes using anthers as explants and to evaluate the genetic stability of regenerated plants via flow cytometry (FCM). Three genotypes were able to induce embryogenic calli in Murashige and Skoog (MS) medium supplemented with 6 mg/l of 2,4-dichlorophenoxyacetic acid. Seedling regeneration occurred in a MS medium supplemented with 0.5 mg/l napthaleneacetic acid ? 1 mg/l 6-benzylaminopurin. Seedlings were derived from somatic embryos and the morphogenic process was induced using the somatic tissue of the stamens. Induction and regeneration efficiency depended on the genotype and was affected by date of tiller collection, different pretreatments or the interaction of these variables. FCM analyses in in vitro regenerated plants showed genetic instability in their nuclear DNA content. Plants with lower nuclear DNA content may indicate DNA aneuploids (8.6 %), whereas plants that had twice the value of nuclear DNA content (4.7 %) suggest in vitro polyploidization. This variation observed in apomictic genotypes provides an opportunity to obtain new variants, which may then be included as sources of genetic variability in buffel grass breeding programs.

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