用添加不同蔗糖与甘露醇配比的培养基对切花菊品种'神马'(Jinba)试管苗进行离体保存,并对保存材料再生后代的遗传稳定性进行了研究.结果表明,在温度(23±2)℃、光照强度2 000~3 000 lx、光照时间12 h/d的培养条件下,MS+0.3 mg·L^-1 6-BA+0.1 mg·L^-1NAA培养基中分别添加10 g·L^-1蔗糖和10 g·L^-1甘露醇、10 g·L^-1蔗糖和20 g·L^-1甘露醇、20 g·L^-1蔗糖和20 g·L^-1甘露醇或30 g·L^-1蔗糖和10 g·L^-1甘露醇均能使菊花试管苗连续保存12个月,其中10 g·L^-1蔗糖和20 g·L^-1甘露醇、20 g·L^-1蔗糖和20 g·L^-1甘露醇组合处理保存12个月后成活率较高,分别达到50.00%和60.00%,且均保持最高的增殖倍数2.67.保存12个月的试管苗在增殖、生根培养基上均能正常恢复生长,且其再生后代的田间生物学性状、过氧化物酶(POD)及酯酶(EST)同工酶酶谱、ISSR扩增图谱与对照株无差异,保持了良好的遗传稳定性,说明利用上述方法保存'神马'种质是可行的.
In vitro conservation of ‘Jinba’plantlets in medium by adding different concentrations of sucrose and mannitol was carried out. Genetic stability of regenerated plantlets was also studied. Results showed that MS+0.3 mg· L^1 6-BA+0.1 mg· L^1 NAA medium supplemented with 10 or 30 mg· L^1 sucrose+ 10 mg· L^1 mannitol,10 or 20 g · L^-1 sucrose+20 g · L^-1 mannitol could prolong plantlets‘ conservation time to 12 month at (23±2)℃ ,2 000-3 000 lx and 12 h photoperiod culture conditions,especially,the last two treatments had higher survival rate of 50.00% and 60.00 % and the two treatments also had the highest proliferation rate of 2.67. After cultured in recovering medium,all plantlets which had being conserved for 12 months recovered normal growth ability and the regenerated plantlets showed no obvious difference in biological characteristics, isoenzyme zymogram of peroxidase and esterase, amplified profiles of ISSR with control, remained well genetic stability. We suggested that it is a feasible method to conserve ‘Jinba’ germplasm.