Traditional and in vitro development of new clover (Trifolium spp.) plants^©

The objectives of the study were to cultivate and breed Trifolium repens, grow and micropropagate various species of Trifolium, and develop protocols for the genetic manipulation of T. repens in vitro. Because white clover is a self-sterile hermaphrodite, cross-pollination is necessary to create viable seed from genetically different parents. As a result of exposing T. repens to 6-benzylaminopurine (BAP) in vitro, adventitious shoot formation was initiated and it was observed that a concentration of 1 mg L^-1 BAP is optimal for adventitious shoot initiation. Other species of Trifolium responded similarly to that of T. repens while cultivated in vitro. Colchicine and Surflan^® (chemical mutagens) were used successfully to produce mutations in T. repens. The plants exposed to these mutagens demonstrated physical mutations such as an increase in leaflets per clover and thicker petiole tissue. This research provides evidence that plant tissue culture can be used to micropropagate endangered Trifolium species and chemically induced mutations which resulted from this study.