激素对黑松愈伤组织褐变和增殖的作用

HORMONES APPLICATION AFFECTS THERATES OF BROWNING AND PROPAGATION OF CALLI IN BIACK PINE AND MASSON TISSUE CULTURE

用成熟胚接种于 1 2MS +2 ,4 -D10 .0mg L +KT4 .0mg L +6 -BA4 .0mg L的培养基中 ,在 2 7± 1℃黑暗条件下诱导 ,成功地诱导出黑松和马尾松的愈伤组织 ,且诱导率分别高达 89%和 90 %。附加激素对黑松愈伤组织褐变和增殖作用研究的结果表明 ,在原有培养基[8] 的基础上 ,附加GA30 .2mg L能促进愈伤组织的生长 ,而附加0 .6mg L或 1.0mg L的GA3能明显改善褐变程度 ;加入LH能提高愈伤组织的增殖倍数 ,且当加入LH 10 0mg L或5 0 0mg L时 ,可降低褐变指数 ;加入 6 -BA既不能降低褐变度 ,也不能提高增殖率 ;2 ,4 -D用量为 1.0mg L时 ,褐变指数为 0 ,这是所试培养基配方中最低的 ,且其增殖倍数在该组试验中最高。结合褐变指数和增殖倍数两项指标 ,培养基组成以 1 2MS +NAA 0 .5mg L +IBA 0 .1mg L +GA31.0mg L +LH 10 0mg L +2 ,4 -D 1.0mg L最佳。使用该配方继代 6代诱导出的黑松愈伤组织没有明显褐变 ,10d增殖倍数保持在 1.

Calli of black pine and masson pine were successfully induced in the tissue culture medium,1/2MS+2,4-D10.0mg/1+KT4.0mg/1+6-BA4.0mg/1,when the mature embryos were inoculated and cultured on the dark condition under 27±1℃.The calli inducing rate were reached 89% and 90% respectively.The results showed that adding GA 3 in 2.0mg/l could promote callus growth,whereas adding GA 3 in 0.6mg/l or 1.0mg/l could enhance the callus browing obviously.Adding LH could increase callus propagating rate.When adding LH in 100mg/l or 500mg/l,the callus browning rate could be reduced.Adding 6-BA had no effect on the rate of callus browning and propagation.When adding 2,4-D in 1.0mg/l,the rate of callus browning was reduced to zero,which was the lowest in the tests,and at the same time the propagating rate was the highest in the tests.The tissue culture medium,1/2MS+NAA0.5mg/l+IBA0.1mg/l+GA 31.0mg/l+LH100mg/l+2,4-D1.0mg/l,was selected as the best medium.No visible browning has been found in the six generation and the propagating rate has been kept at over 1.5times per 10 days.