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The membrane damage and changes of some related enzyme activities in protoplasts of sugarcane and tobacco during enzymatic isolation of leaf protoplsts

He Ruotian, Qin Wei, Li Renqiang

Research Centre, Guangxi Agricultural University. Nanning,Guangxi

Abstract

During the isolation of mesophyll protoplasts of sugarcane(saccharum officinarum)and tobacco(Nicotiana tabacum)by cell wall degrading enzyme mixture (1.5% cellulase, 0.5% pectinase and in 0.5mol/L of mannitol), the contents of O_(2) in protoplasts (containing some undigested cells) of both species increased gradually, and the level of malondialdehybe (MDA), as an index of peroxidation of membrane lipids, also increased in protoplasts of both species. The electrolyte leakage from freshly isolated protoplasts of both species were markedly higher than intact leaf tissue. The activities of superoxide dismutase (SOD) and peroxidase (POD) in protoplasts of both species increased gradually in varying degrees (Fig.3 and 4), and the activity of catalase (CAT) remained unchanged during enzymatic isolation of leaf protoplasts. In comparison with that in tobacco protoplasts, the content of O_(2) in sugarcane protoplasts was far higher but the acticities of SOD, POD and CAT were lower. The level of MDA in and the electrolytes leakafe from sugarcane protoplasts was markedly higher than tobacco proto-plasts. The above results showed that membrane damage of leaf protoplasts of sugarcane was...

Key words: Protoplast;Superoxide anion free radical[O_(2)~(-)];Membrane damage;Sugarcane;Tobacco

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Citing This Article：

He Ruotian, Qin Wei, Li Renqiang. The membrane damage and changes of some related enzyme activities in protoplasts of sugarcane and tobacco during enzymatic isolation of leaf protoplsts. Acta Physiol Sin 1994; 46 (1): (in Chinese with English abstract).