Seeds of Coreopsis leavenworthii Torr. & Gray (Asteraceae) are being commercially produced but the lack of genetic diversity information has hindered growers and end users from addressing several critical issues affecting wild collection, commercial production, distribution, and the use of seeds. In this study, the genetic diversity and differentiation among natural, production, and introduced populations were analyzed at the molecular level using 320 amplified fragment length polymorphism (AFLP) markers. A high level of diversity [68.6% average polymorphism; total genetic diversity (H t) = 0.309] and a moderate level of genetic differentiation [total genetic diversity residing among populations (G st) = 0.226; Φst = 0.244; Bayesian analog of Nei's G st (G st-B) = 0.197] was detected among six natural populations—two each from northern, central, and southern Florida. Two distance-based clustering analyses, based on an individual's AFLP phenotypes or a population's allele frequencies, grouped natural populations into three clusters, concordant with our previous results from a common garden study of phenotypic variation. Clustering of populations was mostly according to their respective geographical origin within Florida. The correlation between geographical distances and pairwise F st values between populations was very significant (r = 0.855, P < 0.0001). Two central Florida natural populations were divergent and grouped into separate clusters, indicating that the existence of factors other than physical distance alone were contributing to genetic isolation. Three production populations maintained a level of genetic diversity comparable to that in the natural populations and were grouped with the natural populations from which the production populations were derived, suggesting that the genetic identity of the seed origin was maintained under production practices. The genetic diversity of the introduced population was comparable to that of the source populations (central Florida natural populations), but genetic shift seems to have occurred, causing the introduced population to cluster with local (northern Florida) populations where planted. The observed genetic differentiation among natural populations may indicate a need to develop appropriate zones within Florida for preservation of genetic diversity during seed collection, increase, and distribution. This high level of population differentiation also suggests a need to collect and analyze more natural populations across Florida and from Alabama for a better understanding of the species' genetic diversity and population structure across its distribution range.
David M. Czarnecki II, Madhugiri Nageswara Rao, Jeffrey G. Norcini, Frederick G. Gmitter Jr and Zhanao Deng
Chunxian Chen, Qifa Zheng, Xu Xiang, Jaya R. Soneji, Shu Huang, Young A Choi, Madhugiri Nageswara Rao and Fred G. Gmitter Jr.
Eight new green fluorescent protein (GFP) binary vectors were developed by inserting gfp reporter gene cassettes into pGreen vectors. We chose one of them, pG52KF, with the nptII selection and gfp reporter gene and one recombinant construct, pG52KFp, for a preliminary evaluation in citrus using Agrobacterium-mediated transformation. High-transformation efficiency was observed, whereas green fluorescence greatly facilitated the early in vivo screening and categorizing of the transformants. These pGreen-derived GFP binary vectors, freely available on request, provide more and flexible options for genetic transformation in citrus and other woody plants.