二甲基亚砜对海带配子体克隆低温冻存的影响

Effects of Different Concentrations of Dimethyl Sulfoxide on Gametophyte Clones from Kelpinthe Cryopreservation

以海带配子体克隆为试验对象,采用4种体积分数二甲基亚砜(DMSO)冷冻保护液,对10个品种(系)20个克隆系进行了低温冷冻保存试验。结果表明,海带配子体克隆在-80℃冻存10 d,复苏后培养,DMSO各体积分数组均有海带配子体细胞存活,其中10%和15%体积分数组冻存的20个克隆系均有存活细胞,5%和20%体积分数组仅部分克隆系有存活细胞。对于相同克隆系,10%和15%体积分数组配子体存活率高于5%和20%体积分数组。指出,在海带配子体低温冻存中,10%和15%DMSO可作为冷冻保护液,为降低成本,优选10%DMSO作为冷冻保护液。提出,经-80℃冻存的海带配子体复苏后培养,进行繁育能力验证,能正常发育,形成孢子体,低温冻存不影响其繁育能力。

Twenty gametophyte clones of Kelp from 10 varieties or strains were cryopreserved with four concentrations of DMSO cryoprotectantat cryogenic temperatures of-80℃degrees for 10 days,and the gametophyte clones of Kelp werecultured after resuscitation.The results showed that Living cells were found in all DMSO concentration groups.In 10%and 15%concentration groups,living cells were found in 20 clone lines cryopreserved,while only some clone lines had living cellsin 5%and 20%concentration groups.For the same clonal lines,the survival rate of gametophytes in 10%and 15%concentration groups were higher than that in 5%and 20%concentration groups.According to the results,10%and 15%DMSO can be used as cryoprotectants in cryopreservationof kelp gametophytes.To reduce the cost,10%DMSO is preferred as cryoprotectants.The gametophytes of Kelp stored at-80℃were resuscitated and cultured to verify their breeding ability.It showed that the gametophytes of Kelp could develop into sporophytes normally,and the cryopreservation did not affect their breeding ability.