Farmer E E
Lehrstuhl für Biochemie der Pflanzen, Institut für Biologie II, Albert-Ludwigs-Universität, Schänzlestrasse 1, D-7800 Freiburg, Federal Republic of Germany.
Plant Physiol. 1985 Jun;78(2):338-42. doi: 10.1104/pp.78.2.338.
Soybean (Glycine max L.) cells cultured in B5 medium produce extremely low amounts of lignin. However, modification in the growth medium, by lowering the concentration of NO(-) (3) and PO(2-) (4), results in the lignification of these cells without affecting levels of cell wall-esterified 4-coumaric and ferulic acid. The production of an extracellular, macromolecular complex by the cultured soybean cells (Moore TS Jr 1973 Plant Physiol 51: 529-536) allows a rapid, nondestructive solubilization of the lignin which can be estimated by reaction with phloroglucinol in free solution. This system has been used to study the effects of fungal elicitor on the synthesis of lignin in soybean cells. The inclusion of very low levels of an elicitor fraction from the cell walls of Phytophthora megasperma in the medium in which lignification of the soybean cells occurs suppressed both the accumulation of extracellular lignin and phloroglucinol staining of the cell walls without affecting the levels of bound hydroxycinnamic acids. The activity profiles of phenylalanine ammonia-lyase (EC 4.3.1.5) and isoenzymes of 4-coumarate:CoA ligase (EC 6.2.1.12) were compared in lignifying and elicitor-treated cell cultures as was the activity of chalcone synthase, an enzyme of flavonoid biosynthesis. The measured activities of these enzymes in cell cultures treated with elicitor were considerably lower than in untreated cells.
在B5培养基中培养的大豆(Glycine max L.)细胞产生的木质素极少。然而,通过降低培养基中NO₃⁻和PO₄²⁻的浓度来改变生长培养基,会导致这些细胞木质化,且不影响细胞壁酯化的4-香豆酸和阿魏酸水平。培养的大豆细胞会产生一种细胞外大分子复合物(Moore TS Jr 1973《植物生理学》51: 529 - 536),它能使木质素快速、无损地溶解,可通过与游离溶液中的间苯三酚反应来估算。该系统已用于研究真菌激发子对大豆细胞中木质素合成的影响。在大豆细胞发生木质化的培养基中加入极低水平的来自大豆疫霉细胞壁的激发子组分,会抑制细胞外木质素的积累以及细胞壁的间苯三酚染色,同时不影响结合态羟基肉桂酸的水平。在木质化和激发子处理的细胞培养物中比较了苯丙氨酸解氨酶(EC 4.3.1.5)的活性谱以及4-香豆酸:辅酶A连接酶(EC 6.2.1.12)的同工酶,还比较了黄酮类生物合成酶查尔酮合酶的活性。在用激发子处理的细胞培养物中测得的这些酶的活性明显低于未处理的细胞。