红三叶新品系组织培养和植株再生

摘要

Red clover new line(Trifolium pratense L. )is a cold-resistant material domesticated from natural distributed red clover in Daxinganling. It was cultivated and selected systematically by Inner Mongolia Agricultural University over 10 years. It had good adaptability and cold resistance in the central and western of Inner Mongolia. Regeneration plants were propagated by means of callus induction and differentiation using different explants. Results showed that cotyledons, hypocotyl, petiole, leaf and root callus were easily induced in a growth medium of MS containing 0. 5 mg · L-1 6-BA and 1~2 mg · L-1 2,4-D. The best callus subculture medium was MS containing 0. 5 mg · L-1 6-BA and 1. 0 mg · L-1 2,4-D. Adding 2,4-D was more conducive to callus growth and preservation than NAA. The best medium for differentiation was MS plus 0. 75 mg · L-1 6-BA and 0. 5 mg · L-1 NAA with up to 16. 67% differentiation rate. The differentiation effect was better with 6-BA than KT, while the combination of 6-BA and NAA was better than that of 6-BA and IBA. Corm-bud could be multiplicated in MS medium plus 0. 5 mg · L-1 6-BA and 1. 0 mg · L-1 IBA, and 2,4-D was not conducive to the growth of stem and buds. IBA was better than NAA for regenerated seedling proliferation. Regenerated plants were immersed in 50 mg · L-1 IBA for 1 h then subcultured in MS medium plus IBA 0. 5 mg · L-1, the rooting rate was 80%. The maximum concentration of IBA for rooting was 0. 75 mg · L-1, otherwise the high concentration is harmful to root growth.%利用红三叶(Trifolium pratense L.)新品系无菌苗不同部位,通过愈伤组织诱导及分化途径获得再生植株.结果表明:子叶、下胚轴、叶柄、叶片及根段在MS+0.5 mg·L-1 6-BA+1～2 mg·L-1 2,4-D培养基上都极易诱导出愈伤组织.最佳继代培养基为MS+0.5 mg·L-1 6-BA+1.0 mg·L-12,4-D,添加2,4-D比NAA更利于愈伤的增长和保存；最佳分化培养基为MS+0.75 mg·L-1 6-BA+0.5 mg·L-1 NAA,最高分化率为16.67％,且分裂素6-BA优于KT,6-BA与NAA组合比与IBA组合更有利于红三叶愈伤组织的分化；茎芽增殖最佳培养基为MS+0.5 mg·L-1 6-BA+1.0 mg· L-1 IBA,再生苗的增殖效果IBA＞ NAA,2,4-D不利于茎芽生长；最佳生根条件为50 mg·L-1浓度的IBA中浸泡1h后植入含0.5 mg·L-1 IBA的MS中,生根率达80％,红三叶新品系IBA生根最大极限浓度为0.75 mg·L-1,浓度过高表现出对根系生长的毒害.