元麦叶肉原生质体细胞壁再生过程中的扫描电镜观察及过氧化物酶活性

元麦叶肉原生质体在MS培养基(附加2,4-D 1mg/L,6-BA 0.25 mg/L)中,进行液体浅层培养。用荧光增白剂(VBL)染色,培养1天出现再生壁。通过扫描电镜观察,发现随着培养时间的延长,原生质体表面逐渐出现短棒状突出物和纤维状结构;培养第5天,原生质体表面覆盖较厚的纤维层,与未脱壁的元麦叶肉细胞表面形态结构相似。用愈创木酚作氢供体测定原生质体胞壁再生过程中过氧化物酶活性,发现随着壁再生率提高,过氧化物酶活性明显下降。用聚丙烯酰胺凝胶电泳分离阳极向过氧化物酶同工酶酶谱,酶带也随着培养时间的延长而减少。由刚分离的原生质体中的8条减少到培养4天的2条,反映胞壁再生和过氧化物酶活性呈负相关。

SCANNING ELECTRON MICROSCOPY OBSERVATION AND PEROXIDASE ACTIVITY STUDY ON CELL WALL REGENERATION PROCESS IN ISOLATED NAKED BARLEY MESOPHYLL PROTOPLASTS

Protoplasts released by EA_3-867 cel-lulase from naked barley(Hordeum vul-gare Var.medum(Arp)Hook,f.)(Zhe-114)leaves were cultured in MS mediumfor observation of cell wall regeneration.The cell wall was formed after one dayculturation. The morphological character of cellwall regeneration has been studied withscanning electron microscopy.With suc-cessive culture,short stick projectionsand fibres were seen on membrane sur-face.The fibres were often associatedwith projections.After 5 days,large num-ber of fibrous components were presenton protoplast surface. The peroxidase activity in proto-plasts was determined with guaiacol asHdonors during cell wall regeneration. With the increasing of cell wall re-generation,the peroxidase activity inprotoplasts decreased.The zymogram ofisoperoxidases from protoplasts was ob-tained by polyacrylamide gel electrophore-sis.Isoperoxidase bands of the newlyisolated protoplasts were eight,and therewere only two bands after 4 days. The results obtained this experimentindicate that negative interrelation existsbetween cell wall regeneration and isoper-oxidases activity.