光照与温度对黄独带芽茎段和茎尖离体保存的影响

Effects of Light and Temperature on in vitro Conservation of Dioscorea bulbifera Stem Segments with a Bud and Shoot Tips

为弄清光照与不同温度条件下黄独种质的保存时间以及保存期间马铃薯Y病毒(PVY)含量的变化,以黄独带芽茎段和茎尖为外植体,采用单因子试验方法研究了光照条件(光照和黑暗)和培养温度(15℃、25℃和35℃)对黄独种质离体保存的影响,并对离体保存后的试管苗进行了ISSR分析。结果表明:黑暗培养有利于黄独带芽茎段的离体保存,而光照培养有利于黄独茎尖的离体保存;光照下15℃的培养温度有利于提高黄独6个月后的成活率。经ISSR分析,离体保存后的试管苗没有发生遗传变异,经PVY的RT-PCR检测,离体保存后的茎尖较带芽茎段病毒含量显著减少;光照条件对带芽茎段和茎尖离体保存过程中病毒含量没有显著影响;培养温度对带芽茎段离体保存过程中病毒含量也没有显著影响,但可导致茎尖离体保存过程中病毒含量的显著减少。

The D.bulbifera stem segments with a bud and shoot tips were cultured under different light and temperature condition to study the effect on in vitro conservation of D.bulbifera germplasm and change of PVY content during the conservation process.The results showed that dark culture was beneficial to in vitro conservation of D.bulbifera stem segments with a bud,light culture was beneficial to in vitro conservation of D.bulbifera shoot tips,and light culture at 15℃ helped to improve survival rate of D.bulbifera stem segments with a bud and shoot tips after six months.The ISSR analysis showed that there was no genetic variation in D.bulbifera plantlets after in vitro conservation.The RT-PCR analysis indicated that the PVY content in D.bulbifera tips after in vitro conservation was significantly lower than that in D.bulbifera buds after in vitro conservation,light had no obvious effect on PVY content in D.bulbifera stem segments with a bud and shoot tips during their conservation process,and temperature had no significant effect on PVY content in D.bulbifera stem segments with a bud during the conservation process but temperature could result in significant reduction of PVY content in D.bulbifera shoot tips during their conservation process.