The polymerase chain reaction (PCR) and RAPD fragments are potentially useful methods for identifying turfgrass cultivar breeding lines. RAPD markers were studied in 25 vegetatively propagated buffalograss lines using oligonucleotide random primers and agarose-gel electrophoresis to determine their potential for identifying cultivar breeding lines. The variation of RAPD markers was extensive. The RAPD markers produced by one random primer were sufficient to separate the 25 buffalograss lines. Cluster analysis baaed on' the RAPD markers produced by two random primers revealed that the 25 buffalograss lines generally fell into two groups: diploid and hexaploid. Three DNA extraction methods—sarcosyl lysis-chloroform extraction-isopropanol precipitation, sodium dodecyl sulfate (SDS) lysine-isopropanol precipitation, and boiling in the presence of Chelex-100 resin—and fresh or oven-dried tissues were tested for reproducibility of RAPD markers. The three DNA extraction methods, using dry or fresh plant tissues, produced highly comparable RAPD marker profiles. More than 80%1 of the RAPD markers was consistently detected in six replicate analyses. The above studies demonstrate that small quantities (5 mg) of oven-dried leaf tissue and several DNA extraction methods can be used for buffalograss fingerprint studies.
The most obvious effects of a low leaf:fruit (LF) ratio [two leaves for one cluster per shoot (LF2)] on grape (Vitis vinifera) berries are suppressed anthocyanin biosynthesis in the berry skin, decreased berry weight and soluble solids concentration, and increased titratable acidity. In this study, proteins isolated from berry skins grown under low and high LF ratio conditions, LF2 and LF12, respectively, were characterized by two-dimensional gel electrophoresis coupled to mass spectrometry. A survey of ≈600 to 700 spots from berry skin yielded 77 proteins with differential expression between LF12 and LF2 treatments. Of these, the 59 proteins that were identified consisted of 47 proteins that were down-regulated and 12 that were up-regulated under LF2 conditions compared with LF12 conditions. Most proteins involved in metabolism, energy, transcription, protein synthesis, binding function, signal transduction, and cell defense were down-regulated in LF2 berries, whereas two important enzymes of anthocyanin biosynthesis, chalcone synthase and dihydroflavonol reductase, were not detected. Only a few proteins (e.g., two heat shock proteins related to protein fate and nutrient reservoir storage protein) were found to be up-regulated in LF2 berries. This suggested that, with the exception of secondary metabolism, many proteomic events may have an effect on anthocyanin synthesis in the skins responding to LF.