Xu Z Q, Ma H J, Hao J G, Jia J F
Shi Yan Sheng Wu Xue Bao. 2000 Mar;33(1):63-8.
Hypocotyl segments of Onobrychis viciaefolia were transformed by Agrobacterium rhizogenes LBA9402 which harboured pBin19 and pRi1855. Seedling age and preculture time of hypocotyl segments influenced the transformation frequency. Paper electrophoresis revealed that 70% of single hairy root cultures could synthesize agropine. Calli were induced from hairy root segments on MS medium containing 0-9.05 mumol/L 2,4-D and 0-2.22 mumol/L 6-BA at first, then they were transferred onto MS0 medium without kanamycin for regeneration. Constitution and concentration of phytohormones in callus induction media affected subsequent regeneration of calluses on MS0 medium remarkably. Regeneration frequency and shoot number per callus declined when 2,4-D concentration in callus induction media increased from 4.52 to 9.05 mumol/L, while they ascended when 6-BA in callus induction media increased from 0 to 2.22 mumol/L. On MS medium supplemented with 4.52 mumol/L 2,4-D and 2.22 mumol/L 6-BA, only 14.2% hairy root segments could produce calluses, but the regeneration frequency reached 58.1% and the shoot number per callus was 37.2. In 32 analysed plants regenerated from 8 kanamycin resistant hairy root lines, 25 were nptII positive and showed different copy numbers.
用携带pBin19和pRi1855的发根农杆菌LBA9402转化红豆草下胚轴切段。下胚轴切段的苗龄和预培养时间影响转化频率。纸电泳显示，70%的单毛状根培养物能够合成农杆碱。首先在含有0 - 9.05 μmol/L 2,4 - D和0 - 2.22 μmol/L 6 - BA的MS培养基上从毛状根切段诱导愈伤组织，然后将它们转移到不含卡那霉素的MS0培养基上进行再生。愈伤组织诱导培养基中植物激素的组成和浓度对随后在MS0培养基上愈伤组织的再生有显著影响。当愈伤组织诱导培养基中2,4 - D浓度从4.52 μmol/L增加到9.05 μmol/L时，再生频率和每个愈伤组织的芽数下降，而当愈伤组织诱导培养基中6 - BA从0增加到2.22 μmol/L时，它们上升。在添加4.52 μmol/L 2,4 - D和2.22 μmol/L 6 - BA的MS培养基上，只有14.2%的毛状根切段能够产生愈伤组织，但再生频率达到58.1%，每个愈伤组织的芽数为37.2。在从8个抗卡那霉素毛状根系再生的32株分析植物中，25株nptII呈阳性，且显示出不同的拷贝数。
