Rumpho M E, Kennedy R A
Department of Horticulture and Landscape Architecture, Washington State University, Pullman, Washington 99164.
Plant Physiol. 1983 May;72(1):44-9. doi: 10.1104/pp.72.1.44.
Tolerance to ethanol and the ability to metabolize key intermediary substrates under anaerobiosis were studied in Echinochloa crus-galli (L.) Beauv. var oryzicola seeds to further characterize the mechanisms which enable it to germinate and grow without O(2).Our results indicate that E. crus-galli var oryzicola possesses an inherently high tolerance to ethanol and is able to metabolize low levels of ethanol in the absence of O(2). Concentrations of ethanol 45-fold greater than endogenous levels did not prove toxic to germinating seeds.Five-day anaerobically grown seedlings of E. crus-galli var oryzicola metabolized added [(14)C]sucrose primarily to CO(2) and ethanol. Of the soluble compounds labeled, the phosphorylated intermediates of glycolysis and the oxidative pentose phosphate pathway predominated more under anaerobiosis than in air. In addition, organic acids and lipids were labeled from [(14)C]sucrose, the latter indicating that metabolism of carbohydrate via acetyl-CoA occurred in the absence of O(2). Lipids were also labeled when seeds were supplied with [(14)C]ethanol or [(14)C]acetate. Labeling experiments using the above compounds plus [(14)C]NaHCO(3), showed further labeling of organic acids; succinate and citrate being labeled under nitrogen, while fumarate was formed in air.The above metabolic characteristics would allow for the maintenance of an active alcoholic fermentation system which, along with high alcohol dehydrogenase activity, would continue to recycle NAD and result in continued energy production without O(2). In addition, Echinochloa's ability to metabolize carbohydrate intermediates and to synthesize lipids indicates that mechanisms exist for providing the carbon intermediates for biosynthesis, particularly membrane synthesis for growth, even in the absence of O(2).
为了进一步阐明稗草变种稻稗种子在无氧条件下对乙醇的耐受性以及代谢关键中间底物的能力,从而明确其在无氧环境下发芽和生长的机制,开展了相关研究。我们的研究结果表明,稻稗对乙醇具有内在的高耐受性,并且能够在无氧条件下代谢低水平的乙醇。浓度比内源性水平高45倍的乙醇对发芽种子并无毒性。无氧培养5天的稻稗幼苗将添加的[¹⁴C]蔗糖主要代谢为二氧化碳和乙醇。在标记的可溶性化合物中,糖酵解和氧化戊糖磷酸途径的磷酸化中间产物在无氧条件下比在空气中更为占主导地位。此外,有机酸和脂质也被[¹⁴C]蔗糖标记,后者表明在无氧条件下通过乙酰辅酶A进行了碳水化合物的代谢。当种子供应[¹⁴C]乙醇或[¹⁴C]乙酸盐时,脂质也被标记。使用上述化合物加上[¹⁴C]碳酸氢钠的标记实验表明,有机酸进一步被标记;琥珀酸和柠檬酸在氮气环境下被标记,而富马酸则在空气中形成。上述代谢特征有助于维持活跃的酒精发酵系统,该系统与高乙醇脱氢酶活性一起,将继续循环利用NAD,并在无氧条件下持续产生能量。此外,稗草代谢碳水化合物中间产物和合成脂质的能力表明,即使在无氧条件下,也存在为生物合成提供碳中间产物的机制,特别是为生长所需的膜合成提供碳中间产物。