香花槐愈伤组织的诱导与细胞悬浮培养技术

Callus induction and cell suspension culture of Robinia pseudoacacia 'Idaho'

香花槐是集绿化、固土、饲料等于一体的多用途林木,其细胞培养则是充分利用该树种优良特性的生物工程技术基础.该文以香花槐叶片为外植体,研究愈伤组织诱导和细胞悬浮培养的技术体系.结果表明,愈伤组织诱导最适培养基为MS+6-BA 0.5 mg/L+NAA 0.5 mg/L+蔗糖30 g/L.愈伤组织生长符合S型曲线,在固体培养基上,培养7 d进入指数生长期,到28 d,愈伤组织鲜重从1.329 3 g增长到10.763 7 g.悬浮培养的细胞,最大生长量为鲜重8.496 g或干重0.474 g.S型生长曲线分析表明,细胞生长迟滞期为3.6 d,最大生长量为14.6 g.

Robinia pseudoacacia ‘Idaho＇ is one of the multiple purpose trees used in virescence,soil and water conservation,fodder of animals,etc.Its cell somatic culture is the fundamental biotechnology maintaining the tree＇s super characteristics.Leaflets of R.pseudoacacia Idaho＇ were taken as the explants in this paper for studying the techniques of callus induction and cell suspension culture.As the result,the optimal medium for callus induction was MS ＋ 6-BA 0.5 mg/L ＋ NAA 0.5 mg/L ＋ sucrose 30 g/L.The callus growth was in trend of logistic curve（S type）,and on solid medium,the calli were in the lag phase of seven days,were in exponential phase and reached the stationary phase on 28 days when the fresh weight growing from 1.329 3 to the maximum of 10.763 7 g.The suspension cultured cells were at stationary phase in 12 days and the weight growing from 1.037 7 g in fresh to the maximum of 8.496 or 0.474 g in dry weight.Theoretically （eg in the model of S growth curve）,the lag phase was 3.6 days and the cell maximum growth was 14.6 g,respectively.