Fertile Transgenic Asparagus Plants Produced by Agrobacterium-mediated Transformation.

Abstract

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 100 mg/l kanamycin and 500 mg/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 Ro and R1 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 R0 plant fragments. Southern analysis showed that both NPT II and bar probes hybridized to products of PCR from five R0 plants. One of the five R0 plants was crossed with a non-transgenic male plant. The segregation ratio of transformed to non-transformed R1 plants was about 1:1. These results indicated that drug-resistant genes were transferred to Asparagus by Agrobacterium-mediated transformation.

Agrobacterium-mediated production of transgenic plants of Muscari armeniacum Leichtl. ex Bak.

“…Until recently, however, monocotyledonous plant species were considered to be recalcitrant to Agrobacterium-mediated transformation and, therefore, major efforts in these species have previously been directed to the production of transgenic plants via direct gene transfer. However, in the past few years successful application of the Agrobacterium-mediated method has been reported for some Liliaceous plants including Asparagus officinalis (Kisaka and Kameya 1998), Allium sativum (Kondo et al 2000), Allium cepa (Eady et al 2000) and Agapanthus praecox , indicating that transformation with this method should be applicable to other Liliaceous species. The objective of the investigation reported here was to establish an efficient system for producing transgenic plants of the Liliaceous ornamental plant M. armeniacum using A. tumefaciens.…”

Production of transgenic lily plants by Agrobacterium -mediated transformation

“…In the past few years, successful application of Agrobacterium-mediated transformation has been reported for some Liliaceous species, including Asparagus officinalis (Kisaka and Kameya 1998), Allium sativum (Kondo et al 2000), Allium cepa (Eady et al 2000), Agapanthus praecox (Suzuki et al 2001), and Muscari armeniacum (Suzuki and Nakano 2002), suggesting that transformation with this method could be applicable to other Liliaceous species. However, in the genus Lilium, Agrobacterium-mediated production of transgenic plants has not yet been reported, although transient expression of the gus reporter gene following cocultivation with Agrobacterium has been demonstrated for an interspecific hybrid cv.…”

Regeneration and production of transgenic Lilium longiflorum via Agrobacterium tumefaciens

“…'Harmony') is a host of Agrobacterium based on the formation of tumors after the inoculation of stem internodes with the wild-type C58 strain. Since then, Agrobacteriummediated transformation has been successfully performed in some Liliaceous species, including Asparagus officinalis (Kisaka and Kameya 1998), Allium sativum (Kondo et al 2000), Allium cepa (Eady et al 2000), Agapanthus praecox (Suzuki et al 2001), Muscari armeniacum (Suzuki and Nakano 2002), and the Oriental hybrid lily, Lilium cv. Acapulco (Hoshi et al 2004).…”