Studies on somatic embryogenesis and plant regeneration from anthers of Guangxi-2,a line of Chinese wild grape (Vitis pseudoreticulata)
ZHI Yu-xi,ZHANG Jian-xia,WANG Yue-jin (Northwest China's Key Laboratory of Horticultural Plant Germplasm Resources Utilization,Ministry of Agriculture,Shaanxi Key Laboratory of Agricultural Molecular Biology,College of Horticulture,Northwest A & F University,Yangling,Shaanxi 712100 China)
In order to offer a regeneration system for functional characterization of resistant genes in grapevine genetic trans-formation,anthers from powdery mildew-susceptible line Guangxi-2 of Chinese wild Vitis pseudoreticulata were used to induce somatic embryos and establish regeneration system. The induction of secondary embryos and the normal seedling from abnormal germinated somatic embryo were also explored in this study. The results indicated Guangxi-2 anthers could be induced and formed embryo callus on B5 medium containing 6-BA 4.0 mg·L-1+ 2,4-D 0.7 mg·L-1+ CH 1.0 g·L-1+ PVP 1.0 g· L-1+sucrose 30.0 g·L-1 after 3-month subculture. The B5 medium containing 2,4-D 0.7 mg·L-1+ CH 1.0 g·L-1+ PVP 1.0 g·L-1+ sucrose 15.0 g·L-1 was beneficial to form embryos. The germination of embryos on the1/2 B5 liquid medium containing CH1.0 g·L-1+PVP 1.0 g·L-1+sucrose 15.0 g·L-1 was 14 days earlier than on the B5 solid medium. The medium for root formation on the germinated embryo was WPM medium containing IBA 0.15 mg·L-1+ AC 0.5 g·L-1+ sucrose 30.0 g·L-1. The MS medium containing 6-BA 1.5 mg·L-1+ IBA 0.2 mg·L-1+ sucrose 30.0 g·L-1 was favorable to the normal seedling formation from mal-formed embryos. The embryos in early developmental stage could be generated secondary somatic embryos on the B5 medium containing 6-BA 1.0 mg·L-1+2,4-D 0.5 mg·L-1+ CH 1.0 g·L-1+ PVP 1.0 g·L-1+ sucrose 30.0 g·L-1.