Biochemical events during somatic embryogenesis in Coffea arabica L.

Several biochemical components associated with different stages of somatic embryogenesis in coffee (Coffea arabica L.) are investigated using foliar explants. Soluble sugar, starch, free amino acids and total proteins were extracted and quantified at different stages of somatic embryogenesis, such as foliar segments (initial explants), primary calluses, embryogenic calluses, globular embryos, torpedoes, cotyledonary embryos and mature fruit zygotic embryos. Total soluble sugar levels increased sixfold at the initial stages of somatic embryogenesis induction. During this period, total soluble sugar in the cultures contained approximately 99.3% glucose and fructose. At 67.4 µg/mg MS, no significant changes were observed in total sugar content during the embryo's somatic maturation and regeneration. During this stage, total soluble sugar was composed of 60% sucrose. After primary callus formation, starch contents increased gradually until the culture's conclusion. Total free amino acids, particularly arginine, lysine, methionine, asparagine, glutamine and histidine, revealed a higher synthesis until the formation of the primary callus, after which they remain statistically constant up to the end of the process. During the induction of calluses, a gradual increase of total proteins occurred, which, in the differentiating and maturing of somatic embryos, did not differ statistically till the formation of a cotyledonary embryo, when rates decreased 21.8%.

Organic acid metabolism and root excretion of malate in wheat cultivars under aluminium stress.

The effects of aluminium (Al) on the metabolism of organic acids synthesised via nonphotosynthetic carbon fixation in the roots and on malate exudation were investigated in Al-tolerant Shirosanjyaku (SH) and Al-sensitive Chikushikomugi (CK) wheat cultivars labelled with bicarbonate-(14)C. Aluminum triggered the excretion of (14)C into the solution, especially in the SH that excreted 2.5 times more (14)C than the CK. The loss of radioactivity (about 10%) into the solution represented a small drain in the (14)C reserve found in the roots. In the organic acid fraction within the roots, malate contained the greatest amount of (14)C, and this amount decreased rapidly with time in both cultivars. The disappearance of radioactivity in the malate resulted from metabolism and translocation rather than to root efflux. Aluminium decreased the malate concentrations in roots of both cultivars. The Al-sensitive cultivar had higher concentrations of malate regardless of the presence of Al. It was therefore assumed that the decrease of malate concentration in roots under Al stress did not result from the decline in malate synthesis but due to an increase in malate decomposition. This response was interpreted as the result of the Al-induced stress and not as the cause of a differential Al-tolerance between the wheat cultivars. An important component of the differential Al tolerance between SH and CK is the greater ability of the Al-tolerant cultivar to excrete malate from the roots, which is independent of its internal concentration in the roots.