Takashi Yamanoshita

Aluminum distribution and reactive oxygen species accumulation in root tips of two Melaleuca trees differing in aluminum resistance

To elucidate the mechanism of the high aluminum (Al) resistance of a Myrtaceae tree, Melaleuca cajuputi Powell, we investigated the responses of root tips to Al and compared them with those of an Al-sensitive species, M. bracteata F. Muell. Roots of seedlings of both species were treated with a calcium solution (pH 4.0) containing 0 or 1 mM AlCl3. After 3 h of Al treatment, inhibition of root elongation and deposition of callose and lignin in root tips, typical signs of Al injury, were induced in M. bracteata but not in M. cajuputi, yet Al accumulation in root tips was similar in both species. These results indicate that internal Al tolerance mechanisms, not Al exclusion mechanisms, are responsible for the Al resistance of M. cajuputi. After 3 h of Al treatment, amount of Al tightly bound to root tips, Al remaining after washing with a desorbing solution, was less in M. cajuputi than in M. bracteata. In M. bracteata, 6 h of Al treatment triggered the accumulation of hydrogen peroxide (H2O2) in root tips despite the upregulation of antioxidant mechanisms, activity of peroxidase and concentration of reduced glutathione. In M. cajuputi, 6 h of Al treatment did not affect the concentration of H2O2, but decreased activity of peroxidase, and increased concentration of reduced glutathione in root tips. These results suggest that the less Al tightly bound to root tips is involved in the suppressing the H2O2 accumulation and the internal Al tolerance in M. cajuputi, and that the H2O2 accumulation or changes in cellular environment that bring about H2O2 accumulation despite the upregulation of antioxidant mechanisms results in Al-induced inhibition of root elongation in M. bracteata.

Effects of flooding on downstream processes of glycolysis and fermentation in roots of Melaleuca cajuputi seedlings

We investigated the energy metabolism in roots of flooded Melaleuca cajuputi Powell, a tropical flood-tolerant tree species, by measuring adenylate concentrations and activities of glycolytic and fermentative enzymes under flooded conditions. Adenylate energy charge (AEC) decreased slightly to 0.72 on the second day of flooding and recovered to around 0.8 by the fourth day of flooding. Activities of pyruvate decarboxylase (EC 4.1.1.1) and alcohol dehydrogenase (EC 1.1.1.1) increased initially and then decreased to the control level after 14 days of flooding. On the other hand, activities of pyruvate kinase (EC 2.7.1.40), phosphoenolpyruvate phosphatase (EC 3.1.3.2), and a series of phosphoenolpyruvate carboxylase (EC 4.1.1.31), malate dehydrogenase (EC 1.1.1.37), and NADP dependent malic enzyme (EC 1.1.1.40), which can convert PEP into pyruvate, were not induced in flooded roots throughout the experiment. These results suggest that neither the downstream reactions of glycolysis nor ATP production via glycolysis was enhanced by flooding, whereas alcohol fermentation was enhanced. With the low ATP yield of the glycolysis–alcohol fermentation pathway and no induction of glycolytic enzymes, the glycolysis–alcohol fermentation pathway itself contributes little to ATP production in flooded roots of M. cajuputi. These physiological responses of M. cajuputi to flooding may have the advantages of surviving flooded conditions because they can avoid exhaustion of sugar and accumulation of ethanol, a toxic end product of alcohol fermentation.

Growth Response of Melaleuca cajuputi to Flooding in a Tropical Peat Swamp

We investigated the growth response ofMelaleuca cajuputi Powell to flooding at 3 sites in a tropical swamp in southern Thailand. The relative growth rate (RGR) in height tended to be higher where the water level was higher. The maximumRGR in height was achieved during the flood period at the 2 sites where water levels were higher than at the other site. The height growth ofM. cajuputi was not reduced by flooding, but enhanced. No decline in growth due to post-anoxic injury was observed after flooding.Melaleuca cajuputi may have mechanisms to tolerate rhizospheric oxygen deficiency and to avoid post-anoxic injury.

Steady sucrose degradation is a prerequisite for tolerance to root hypoxia

We investigated the role of glycolysis and sucrolysis in the difference in tolerance to root hypoxia between two Myrtaceae tree species, Melaleuca cajuputi (which shows superior tolerance to root hypoxia) and Eucalyptus camaldulensis (which does not). Analysis of the adenylate energy charge (AEC) in roots subjected to a 4-day hypoxic treatment (HT) in hydroponic culture revealed that the interspecies difference in tolerance corresponds to the ability to maintain energy status under root hypoxia: AEC was reduced by HT in E. camaldulensis, but not in M. cajuputi. The energy status in HT roots of E. camaldulensis was restored by feeding of glucose (Glc) but not sucrose (Suc). These data provide evidence that low substrate availability for glycolysis resulting from an impairment of sucrolysis suppresses ATP production under hypoxic conditions in this species. Measurements of the rates of O2 consumption and CO2 production in roots indicated that E. camaldulensis, but not M. cajuputi, failed to activate fermentation in HT roots. These results cannot be attributed to enzymatic dysfunction, because no inhibition of main glycolytic and fermentative enzymes was observed in both species, and Glc feeding had a beneficial effect on AEC of HT roots of E. camaldulensis. The impairment of sucrolysis was demonstrated by inhibited soluble acid invertase activity in HT roots of E. camaldulensis. In contrast, there was no inhibition in all sucrolytic enzymes tested in HT roots of M. cajuputi, suggesting that steady Suc degradation is essential for maintaining high energy status under root hypoxia. We conclude that root sucrolysis is one of the essential factors that determines the extent of tolerance to root hypoxia.