Aluminum effects on citric and malic acid excretion in roots and calli of rice cultivars

Abstract

Citric and malic acid excretion in the medium and malic acid accumulation in seedling roots and embryo-derived calli as possible mechanisms of aluminum (Al) resistance and the effects of a 17-h Al stress period on root growth in Oryza sativa have been studied. Four-day-old seedlings and embryo-derived calli of Al-resistant (IRAT 112 and IR6023) and Al-sensitive (Aiwu and IKP) cultivars were treated with 250 and 500 µM {Al 2 (S0 4 ) 3 .18H 2 0 }of total aluminum or without Al for 36 hours. After 3 to 36 hours of stress, seedlings and calli were removed from the flasks and concentration of citric and malic acids was estimated in the Al and control solutions. Malic acid was also assayed in roots tips and in callus tissues. After 17-h of Al stress, inhibition of root growth was a typical effect of Al in rice and the extent of the inhibition depended on both cultivar and Al concentration. At 500 µM of Al, strong reduction of root elongation occurred in all cultivars while at 250 µM of Al, only IRAT was unaffected, when compared to their control. In the absence of Al, all varieties excreted comparable amounts of citric and malic acid. Al treatments, were without effect upon citrate excretion in both Al-resistant and Alsensitive cultivars. Al treatment, for periods from 3 to 24h, slightly stimulated the excretion of malic acid from seedlings, in all cultivars. Malic acid concentrations in root apices, in the presence or absence of aluminum, were not correlated with aluminum resistance. No differences in malic excretion and internal concentrations were detected between Al-treated and untreated rice calli of the same four cultivars. It is therefore concluded that, in our experimental conditions, differences in Al resistance in our rice cultivars cannot be attributed to citric and malic acids. Further research needs to be carried out to examine other possible mechanisms of Al-resistance in rice and to determine whether organic acids such as succinic and oxalic acid are implicated. RESUMO -A excreção dos ácidos málico e cítrico no meio de cultura, assim com a acumulação do ácido málico nas raízes e em calos derivados de embriões, foram estudadas como um possível mecanismo de resistência ao alumínio em arroz. Plântulas de 4 dias e calos derivados de embriões das cultivares resistentes ao alumínio (IRAT 112 e IR6023) e das cultivares sensíveis (Aiwu e IKP) foram tratadas com 0, 250 e 500µM de alumínio {Al 2 (S0 4 ) 3 .18H 2 0 }. Em seguida, de 3 a 36 horas de estresse, as plântulas e os calos foram removidos dos frascos e as concentrações dos ácidos cítrico e málico, determinadas. A concentração do ácido málico foi também determinada nos ápices das raízes e nos calos. Após 17 horas de estresse, o crescimento radicular foi inibido, mostrando um efeito tipico do Al em arroz. Entretanto, a extensão da inibição depende da cultivar e da concentração em Al. Na presença de 500 µM de Al, ocorreu uma forte redução no alongamento radicular em todos as cultivares, ao passo que a 250 µM de Al, a cultivar IRAT não foi afetada. Na ausência...