The Study On Spheroplast Vacuoles In Cyanobacteria

Anabaena sp. 595 was treated in the presence of 0.15ml/l KNO^ KCK K2SO4> KH2PO4, K2HPO4, NaNO3, NaCK Na2SO4, NaH2PO4, Na2HPO4, (NH4)2SO4, MgSO4 and CaCl2 separately mixed with 0.1%(W/V) lysozyme for 4 hours.The results show that protoplasts rather than vacuolized protoplasts could be easily induced by KH2PO4, K2SO4, Na2SO4, NaH2PO4 , (NH4)2SO4, MgSO4 and CaCl2 and the vacuolized protoplasts could be induced by KNO3, NaNO3 and NaCl.The inducing effects were not evident under the treatment of two salts of similar concentration like KNO3 and NaCK KNO3 and (NH4)2SO4, NaCl and (NH4)2SO4 mixed with 0.1%(w/v) lysozyme.However, the induction rate of both the protoplasts and vacuolized protoplasts was markedly raised by three salts of similar concentration like NaCl, KNO3 and (NH4)2S04 with 0.1%(w/v) mixed with lysozyme; and by five salts of NaCl , KNO3 , (NH4)2SO4, KH2PO4 and Na2HPO4 mixed with lysozyme.Respectively the percentage of vacuolized protoplasts could reached up to 66.90% and 79.97%.Anabaena sp.595 and Nostoc sp.96 were treated in the presence of 0.15ml/L NaCl , KNO3 , (NH4)2SO4 and buffer solution (KH2P04 and Na2HPO4, 1/15 mol/L,pH=6.8) with 0.1%(w/v) mixed with lysozyme.After 24 hours vacuolized spheroplasts broke and released a great deal of vacuoles.The vacuole yield of Anabaena sp. 595 could reached up to 6.5 ×105 per mg and the release rate was 81.49%.The vacuole yield of Nostoc sp.96 could reached up to 6.1 × 105 per mg and the release rate was 71.79%.The trichomes of Anabaena sp.595 and Nostoc sp.96 were treated in the presence of 0.15ml/L NaCl > KNO3 ^ (NH4)2SO4 and buffer solution (KH2PO4 and Na2HPO4,l/15 mol/L,pH=6.8) with 0.1%(w/v) mixed with lysozyme.After 24 hours of treatment vacuolized spheroplasts were noticed to break and vacuoles werereleased.The property of the spheroplast vacuoles were subsequently preliminarily investigated.Vacuoles behaved themselves amalgamation and phagotrophy.The vocuoles in distilled water were evidently larger in volume than those in salt solution mixed with lysozyme.The results indicate that with regard to the preservation of vocuoles 50% of vacuoles of Anabaena sp. 595 remains intact after 96 hours in the distilled water.The weakly acid conditions kept the stability but the acid and alkali conditions were unfavorable to the low stability.The vacuole presented itself intact under the mild osmotic pressure and was more stable in NaCl than that in CaCl2 and MgCl2. The vacuoles of Anabaena sp. 595 showed more stable than those of Nostoc sp.96.