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Title: Development of an efficient in vitro regeneration system for leafy spurge (Euphorbia esula L.)

Author
item XU, BIN - NORTH DAKOTA STATE UNIV
item DAI, WENHAO - NORTH DAKOTA STATE UNIV
item Chao, Wun

Submitted to: In Vitro Cellular and Developmental Biology - Plants
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/23/2008
Publication Date: 6/14/2008
Citation: Xu, B., Dai, W., Chao, W.S. 2008. An Efficient In Vitro Regeneration of Leafy Spurge (Euphorbia esula L.). In Vitro Cellular and Developmental Biology - Plants. 44:548-556.

Interpretive Summary: Leafy spurge is an invasive, perennial weed. Molecular tools such as an expressed sequence tag (EST) database and DNA microarrays have been developed to study its invasive nature. However, the lack of genetic transformation systems has hampered molecular approaches to study leafy spurge. Here, we described an efficient tissue regeneration system, which is a requisite for the development of genetic transformation systems for leafy spurge. Three high regenerative lines were selected by screening the regeneration capabilities of stem pieces (explants) of 162 seedlings. The effects of various culture conditions on regeneration were then evaluated based on explants’ competence to form calli and shoots. The results indicated that high rates of shoot regeneration can be obtained using a growth medium containing 1x WPM and 1x MS basal salts, 1x MS vitamins, 0.25 mg l-1 BA, 0.4 mg l-1 IBA, and 3% sucrose, pH 5.6-5.8. After 30 d of culture, multiple shoots were formed either directly from the outer cell layers of the stem or indirectly from the callus. This paper reports the first successful endeavor at developing an efficient regeneration system for leafy spurge.

Technical Abstract: Leafy spurge (Euphorbia esula L.) is an invasive, perennial weed. Molecular tools such as a deep EST-database and DNA microarrays have been developed to study its invasive nature. However, the lack of transformation systems has hampered molecular approaches to study leafy spurge. Here, we described an efficient in vitro regeneration system, which is a requisite for the development of genetic transformation systems for leafy spurge. Three high regenerative lines were selected by screening the in vitro regeneration capabilities of stem pieces (explants) of 162 seedlings. The effects of various culture conditions on in vitro regeneration were then evaluated based on explants’ competence to form calli and shoots. The results indicated that high rates of shoot regeneration can be obtained using a growth medium containing 1x WPM and 1x MS basal salts, 1x MS vitamins, 0.25 mg l-1 BA, 0.4 mg l-1 IBA, and 3% sucrose, pH 5.6-5.8. After 30 d of culture, multiple shoots were formed either directly from the outer cell layers of the stem or indirectly from the callus. This paper reports the first successful endeavor at developing an efficient in vitro regeneration system for leafy spurge.