Submitted to: In Vitro Biology
Publication Type: Abstract Only
Publication Acceptance Date: November 10, 2004
Publication Date: January 4, 2005
Citation: Moss, S.C., Dowd, M.K., Pelitre, S.M., Triplett, B.A. 2005. Culture parameters influencing the production of gossypol and related compounds in cotton hairy roots. In Vitro Biology-2005. poster-2023, p. 40-A. http://www.sivb.org/2005A_p1.pdf Technical Abstract: Previously, we established conditions for culturing hairy roots from Rhizobium rhizogenes-transformed Gossypium hirsutum and Gossypium barbadense. Significant levels of gossypol and related derivatives were produced in culture. Gossypol is a di-sesquiterpene that has antiviral activity against enveloped viruses such as HIV and inhibits the growth of numerous parasitic organisms, fungi, microbes, and insects. Two on-going clinical trials are testing the efficacy of gossypol as a human chemotherapeutic agent. In this study, several culture parameters were investigated to define the optimal biomass and gossypol production conditions for cotton hairy root cultures. Cultures were grown on Murashige-Skoog or Gamborg’s B5 media, pH 5.8 with 0, 10, 20, 30 g/L glucose or 0, 10, 20, 30 g/L sucrose. Gossypol levels were not greatly influenced by sugar type but were significantly affected by media composition. Cultures grown on Gamborg’s B5 media produced higher levels of gossypol and related compounds than cultures grown on Murashige-Skoog media. We are testing whether this difference is due to nitrogen content or form. Temperature effects (25º, 28º, 31º, 34ºC) on hairy root growth and gossypol production were also investigated. Gossypol productivity was greatest at 31ºC and production of methylated forms of gossypol was greatest at 34ºC. We will also report on the variability of gossypol production after sub-culturing transformants individually for several months and provide gossypol production data for 26 independently transformed lines. Establishing optimal growing conditions and gossypol levels produced by cotton hairy root cultures benefits our long-term goal of characterizing the gossypol biosynthetic pathway.