紫球藻原生质体再生育种

Regeneration of Protoplasts from Porphyridium cruentum and Its Variation for Breeding

收集对数生长期紫球藻细胞,用50 mm o l/L EDTA+25 mm o l/L DTT溶液在30℃下恒温处理30m in,离心收集细胞.然后转入含1.5%蜗牛酶和2.0%的纤维素酶的混合液中,以0.2 m o l/L KC l为渗透压稳定剂,在pH 6.8,30℃下轻微振荡酶解去壁,6 h后收集紫球藻原生质体,原生质体制备率达60.23%.适当稀释后接入再生培养基再生,40%的陈旧培养上清液可提高再生率,再生率可达55.17%.25 d后从再生平板中分离到一株变异藻株,其叶绿素b含量比出发藻株提高53.13%,胞外多糖产量提高22.42%.

The logarithmic phase Porphyridium cruentum cells were pre-treated with mixture solution of the 50 mmol/L EDTA and 25 mmol/L DTT for 30 min, then hydrolyzed whh the mixture solution of 1.5% snailase and 2.0% cellulase （ contains 0.2 mol/L KCl ） at 30℃ and with gently shaking for 6 h, pH6.8. The protoplasts formation rate was 60. 23%. The regeneration rate of protoplasts on the hypertonic solid medium containing 40% supernatant of the culture mediun, and the protoplasts regeneration rate was 55.170%. A morphological variated train from regenerated algal clones was selected, which increased 53. 13% of chlorophyll b on average to the cell of parent strain and its extracellular polysaccharides was increased 22.42% too.