Clark Joanna I M, Hall J L
School of Biological Sciences, University of Southampton, Southampton SO16 7PX, UK.
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New Phytol. 1998 Oct;140(2):261-269. doi: 10.1046/j.1469-8137.1998.00263.x.
The transport of sugars and amino acids into the mycelium of Erysiphe pisi DC. was investigated using two different systems, intact leaf discs and mycelial suspensions. Of the sugars tested, glucose was preferentially taken up by both uninfected and mildew-infected leaf discs, whereas glutamine was taken up by both tissues at a higher rate than lysine or aspartic acid. Leaf discs from infected tissue had a greater uptake capacity than those from healthy tissue for both sugars and amino acids. The uptake of glucose was inhibited more markedly than that of sucrose and fructose by 10 μm carbonyl cyanide m-chlorophenylhydrazone (CCCP), 1 mmN-ethylmaleimide (NEM), 1 mm diethyl pyrocarbonate (DEPC) and 1 mm phenylglyoxal, whereas 1 mm PCMBS (p-chloro-mercuribenzenesulphonic acid) inhibited sucrose uptake to the greatest extent. Uptake of glutamine, lysine and aspartic acid was inhibited similarly by CCCP (80%), NEM (20%), DEPC (70%) and PCMBS (60%). Additionally, leaf discs were used to determine which solutes could be taken up from leaf tissue by the fungus. The uptake of sugars into the mycelium was greater than that of amino acids. Suspensions of powdery mildew mycelium accumulated glucose at about three times the rate of sucrose or fructose, and the amino acid glutamine was taken up at three times the rate of lysine or aspartic acid. Spores separated from the suspension had a low uptake capacity. When the reducing sugar concentration of leaf apoplastic fluid was estimated, leaves infected by powdery mildew had much higher amounts in the apoplast, whereas the activity of acid invertase also appeared to be higher in apoplastic fluids from infected leaves. When apoplastic fluid samples were run on SDS gels, an invertase antibody detected two bands in samples from infected tissues that were not found in the uninfected samples.
利用完整叶盘和菌丝体悬浮液这两种不同体系,对豌豆白粉菌(Erysiphe pisi DC.)菌丝体中糖和氨基酸的转运进行了研究。在所测试的糖类中,葡萄糖无论是未感染还是感染白粉病的叶盘都优先吸收,而谷氨酰胺在两种组织中的吸收速率都高于赖氨酸或天冬氨酸。感染组织的叶盘对糖和氨基酸的吸收能力均高于健康组织的叶盘。10μm羰基氰化物间氯苯腙(CCCP)、1mM N-乙基马来酰亚胺(NEM)、1mM焦碳酸二乙酯(DEPC)和1mM苯乙二醛对葡萄糖吸收的抑制作用比对蔗糖和果糖更为显著,而1mM对氯汞苯磺酸(PCMBS)对蔗糖吸收的抑制作用最大。CCCP(80%)、NEM(20%)、DEPC(70%)和PCMBS(60%)对谷氨酰胺、赖氨酸和天冬氨酸吸收的抑制作用相似。此外,还利用叶盘来确定真菌能够从叶片组织中吸收哪些溶质。糖类进入菌丝体的吸收量大于氨基酸。白粉病菌丝体悬浮液积累葡萄糖的速率约为蔗糖或果糖的三倍,氨基酸谷氨酰胺的吸收速率是赖氨酸或天冬氨酸的三倍。从悬浮液中分离出的孢子吸收能力较低。当估算叶片质外体汁液中还原糖浓度时,感染白粉病的叶片质外体中的还原糖含量要高得多,而感染叶片质外体汁液中的酸性转化酶活性似乎也更高。当质外体汁液样品在SDS凝胶上进行电泳时,转化酶抗体在感染组织的样品中检测到两条未感染样品中未出现的条带。