Anti-SYN1 monoclonal antibody, clone NFHT4-20B

Outbred forage species syn1 polycross seed production has a major impact on expected inbreeding depression in later seed increase generations. In this study, we examined the relationship between seed-parent and pollenparent fecundity and how this relationship relates to other polycross mating metrics. Twelve bumble bee (Bombus impatience Cr.)-pollinated red clover (Trifolium pratense L.) polycrosses ranging in size from 13 to 96 parents were evaluated for median seed yield per plant, seed-parent fecundity Weibull distribution shape parameter lambda-values, pollen-parent fecundity distribution lambda-values, and seed-pollen-parent fecundity correlations. Seed-pollen-parent fecundity correlations were significant and varied from r(2) = 0.187 to 0.879 with a median value of 0.464. Seed-pollen-parent fecundity was negatively correlated with seed-parent fecundity distribution lambda-values (r = -0.633), indicating that less panmictic polycross mating was associated with more correlated seed-and pollen-parent fecundity. Multiple regression of a polycross's median seed yield per plant and seed-parent fecundity distribution l-values onto seed-pollen-parent fecundity correlations revealed a positive slope with median seed yield per plant (partial r(2) = 0.255) and a negative slope with seed-parent fecundity distribution l-values (partial r(2) = 0.400). This study reveals that seedand pollen-parent fecundity are correlated and that this correlation is higher when median seed yield per plant is lower and when polycross mating is less panmictic. To mitigate the impact of increased inbreeding due to this correlation, we recommend creating a stress-free polycross environment that maximizes seed production per plant in the polycross.

Current DNA assembly methods for preparing highly purified linear subassemblies require complex and time-consuming in vitro manipulations that hinder their ability to construct megabase-sized DNAs (e.g. synthetic genomes). We have developed a new method designated 'CasHRA (Cas9-facilitated Homologous Recombination Assembly)' that directly uses large circular DNAs in a one-step in vivo assembly process. The large circular DNAs are co-introduced into Saccharomyces cerevisiae by protoplast fusion, and they are cleaved by RNA-guided Cas9 nuclease to release the linear DNA segments for subsequent assembly by the endogenous homologous recombination system. The CasHRA method allows efficient assembly of multiple large DNA segments in vivo; thus, this approach should be useful in the last stage of genome construction. As a proof of concept, we combined CasHRA with an upstream assembly method (Gibson procedure of genome assembly) and successfully constructed a 1.03 Mb MGE-syn1.0 (Minimal Genome of Escherichia coli) that contained 449 essential genes and 267 important growth genes. We expect that CasHRA will be widely used in megabase-sized genome constructions.