菌草绿洲3号快繁及遗传转化体系的初步研究

Establishment of plant tissue culture system and preliminary study on genetic transformation of Lvzhou No.3

以芦竹属(Arundo)菌草绿洲3号诱导的愈伤组织为材料,进行不定芽、不定根诱导以及移栽试验。以愈伤组织为受体材料,对绿洲3号的遗传转化体系进行初步研究。结果表明,不定芽最佳分化培养基为MS+1.0 mg·L^(-1)6-BA+0.1 mg·L^(-1)NAA,不定芽诱导率为100%。不定根诱导的最适培养基为MS,不定根的诱导率为100%。在9种移栽基质中,泥炭基质中组培苗的移栽成活率最高,为100%。以绿洲3号愈伤组织预培养3 d,于农杆菌OD600值为0.1侵染10 min为宜,共培养基中添加300μmol·L^(-1)AS的条件下,共培养1 d的GUS表达率最高,为1.5%。

The calli derived from Lvzhou No. 3( Arundo) were used as experimental material to induce adventitious buds and roots and conduct apot experiment. The calli were then used as receptors to examine the genetic transformation of Lvzhou No. 3. The results demonstrated that the most effective formula for adventitious bud induction was MS + 1. 0 mg·L^(-1)6-BA + 0. 1 mg·L^(-1) NAA,which exhibited an adventitious bud induction rate of 100%. The best formula for root induction was MS; the induction rate was 100%. Among nine different transplanting media,the rate of living seedling was 100% after being transplanted in peaty soil. The results showed that calli of Lvzhou No. 3( Arundo) that were precultured for 3 d,inoculated for 10 minutes,OD600= 0. 1,and cocultured for 1 d with 300 μmol·L^(-1) AS are adapted for genetic transformation. The GUS expression rate might reach 1. 5%.