Transgenic plants of
Asparagus officinalis cv. Mary Washington 500W were generated by
Agrobacterium-mediated transformation. After induction of flowers by treatment with atrazine, calli were induced from male and female plants. Calli of male and female origin were co-cultivated with
Agrobacterium EHA101 (pIG121Hm) and EHA101 (pARK5), respectively. About thirty shoots developed from female calli and three shoots developed from male calli within six weeks on selective medium supplemented with 100mg/
l kanamycin and 500mg/
l claforan. Most of the shoots did not form roots. Seven shoots from female calli formed roots and were successfully transferred to soil in a greenhouse. But shoots of male calli did not form any roots. Stable integration and inheritance of transgenes were demonstrated by molecular and genetic analysis of transformants in the R
0 and R
1 generations. In an analysis using the polymerase chain reaction (PCR), fragments of both the
NPT II gene and the
bar gene were amplified from five R
0 plant fragments. Southern analysis showed that both
NPT II and
bar probes hybridized to products of PCR from five R
0 plants. One of the five R
0 plants was crossed with a non-transgenic male plant. The segregation ratio of transformed to non-transformed R
1 plants was about 1:1. These results indicated that drug-resistant genes were transferred to
Asparagus by
Agrobacterium-mediated transformation.
抄録全体を表示