四倍体葡萄诱导技术的研究

Study on Induction of Tetraploid Grapes

以秋水仙素为诱变剂,利用茎段浸泡和秋水仙素培养基两种方法对二倍体葡萄(Vitis.vinifera)京秀和红地球2个品种的组培苗进行诱导,并采用流式分析仪对诱变植株的细胞DNA含量进行鉴定。结果表明,茎段浸泡秋水仙素法诱导四倍体葡萄的效果优于秋水仙素培养基法。在低浓度秋水仙素(2000mg·L-1或低于2000mg·L-1)溶液中浸泡时间短于48h的所有处理,仅在继代2～5次后获得了纯合四倍体植株;在秋水仙素浓度2000mg·L-1对茎段浸泡3～4d或3000mg·L-1和4000mg·L-1对茎段浸泡2～4d,在茎段扦插的当代就获得了纯合四倍体植株,诱变四倍体植株的适宜条件为3000mg·L-1秋水仙素茎段浸泡3～4d,纯合四倍体植株获得的比率为16.7%～23.3%。

Tetraploids were induced successfully from in vitro Vitis vinifera cv. Jingxiu and cv Red Globe by soaking shoot segments in colchicine solution and adding colchicine in the culture medium. Ploidy levels were determined by flow cytometry. The results showed that the soaking-treatment increased the efficiency of induced tetraploids than one of adding in medium. Tetraploid plants were obtained from the shoot segments soaked in colchicine solution at concentrations of 2 000 mg·L^-1 or lower during 48 h or shorter time after two to four regenerations of micropropagation. However, when soaking was extended to 3 to 4 d at colchicines concentrations of 2 000 mg·L^-1 or when colchicine concentration increased to 3 000 mg·L^-1 for 3 to 4 d, or 4 000 mg·L^-1 for 2 to 4 d, pure tetraploids were easily obtained during the first micropropagation regeneration. The optimal conditions for inducing tetraploids were under soaking shoot segments for 3 to 4 d with colchicine at a concentration of 3 000 mg·L^-1. The rate of induction was 16.7%-23.3%.