MORPHOLOGY,INHERITANCE, AND EVOLUTIONARY SIGNIFICANCE OF SEX REVERSAL IN TRIPSACUM DACTYLOIDES (POACEAE)

Inflorescences of Tripsacum dactyloides (L.) L. (Andropogoneae) are characterized by single female spikelets at one to several basal nodes and paired male spikelets at several nodes above them on each raceme. Female spikelets are one-flowered and male spikelets are two-flowered. A sex form variant was found in a wild population in north central Kansas and classified as T. dactyloides (L.) L. forma prolificum Dayton et Dewald. The variant of this native distant relative of maize (Zea mays L. spp. mays) differs from the normal form by having both pistillate and perfect rather than staminate spikelets in the terminal (tassel) portion of the inflorescence and by having two functional pistillate florets in the basal spikelets instead of one. A recessive major gene at a single locus regulates the change of the inflorescence from monoecious to gynomonoecious.     

COUNTERFEIT HYBRIDS BETWEEN TRIPSACUM AND ZEA (GRAMINEAE)

Diploid (2n = 36) Tripsacum australe Cutler and Anderson var. hirsutum de Wet and Timothy, T. cundinamarce de Wet and Timothy, T. dactyloides (L.) L. var. dactyloides and var. meridonale de Wet and Timothy, and T. laxum Nash were crossed with Zea mays L. (2n = 20) as the pollen parent. True hybrids combine the cytologically nonreduced genome of Tripsacum (36 chromosomes) with the haploid (10 chromosomes) or more rarely diploid (20 chromosome) genome of Zea. Maternal offspring with 2n = 36 Tripsacum chromosomes commonly result from parthenogenetic development of cytologically nonreduced eggs. Some individuals with 2n = 36 Tripsacum chromosomes, however, resemble true hybrids in phenotype. These counterfeit hybrids incorporated Zea genetic material into their Tripsacum genomes without true fertilization having taken place. Offspring of counterfeit hybrids that were grown to maturity resembled their mothers in phenotype, and must have originated parthenogenetically. It is proposed that counterfeit hybrids are also produced in nature, and that this process contributes to origins of variation in gametophytic apomicts, and perhaps also in sexually reproducing species.     

Asymmetric Switching in a Homodimeric ABC Transporter: A Simulation Study

ABC transporters are a large family of membrane proteins involved in a variety of cellular processes, including multidrug and tumor resistance and ion channel regulation. Advances in the structural and functional understanding of ABC transporters have revealed that hydrolysis at the two canonical nucleotide-binding sites (NBSs) is co-operative and non-simultaneous. A conserved core architecture of bacterial and eukaryotic ABC exporters has been established, as exemplified by the crystal structure of the homodimeric multidrug exporter Sav1866. Currently, it is unclear how sequential ATP hydrolysis arises in a symmetric homodimeric transporter, since it implies at least transient asymmetry at the NBSs. We show by molecular dynamics simulation that the initially symmetric structure of Sav1866 readily undergoes asymmetric transitions at its NBSs in a pre-hydrolytic nucleotide configuration. MgATP-binding residues and a network of charged residues at the dimer interface are shown to form a sequence of putative molecular switches that allow ATP hydrolysis only at one NBS. We extend our findings to eukaryotic ABC exporters which often consist of two non-identical half-transporters, frequently with degeneracy substitutions at one of their two NBSs. Interestingly, many residues involved in asymmetric conformational switching in Sav1866 are substituted in degenerate eukaryotic NBS. This finding strengthens recent suggestions that the interplay of a consensus and a degenerate NBS in eukaroytic ABC proteins pre-determines the sequence of hydrolysis at the two NBSs.     

Firefly luciferase gene: structure and expression in mammalian cells.

The nucleotide sequence of the luciferase gene from the firefly Photinus pyralis was determined from the analysis of cDNA and genomic clones. The gene contains six introns, all less than 60 bases in length. The 5' end of the luciferase mRNA was determined by both S1 nuclease analysis and primer extension. Although the luciferase cDNA clone lacked the six N-terminal codons of the open reading frame, we were able to reconstruct the equivalent of a full-length cDNA using the genomic clone as a source of the missing 5' sequence. The full-length, intronless luciferase gene was inserted into mammalian expression vectors and introduced into monkey (CV-1) cells in which enzymatically active firefly luciferase was transiently expressed. In addition, cell lines stably expressing firefly luciferase were isolated. Deleting a portion of the 5'-untranslated region of the luciferase gene removed an upstream initiation (AUG) codon and resulted in a twofold increase in the level of luciferase expression. The ability of the full-length luciferase gene to activate cryptic or enhancerless promoters was also greatly reduced or eliminated by this 5' deletion. Assaying the expression of luciferase provides a rapid and inexpensive method for monitoring promoter activity. Depending on the instrumentation employed to detect luciferase activity, we estimate this assay to be from 30- to 1,000-fold more sensitive than assaying chloramphenicol acetyltransferase expression.     

The long and the short of gene flow and reproductive isolation: Inter-Simple Sequence Repeat (ISSR) markers support the recognition of two floral forms in Pelargonium reniforme (Geraniaceae)

The current taxonomy of Pelargonium reniforme recognises two subspecies on the basis of habit and vegetative characters, but excludes floral characters. However, populations of P. reniforme in the wild tend to belong to easily discernable floral groups based on floral colour and hypanthium length. The aim of this study was to determine the genetic diversity within and between populations of both subspecies using the Inter-Simple Sequence Repeat (ISSR) method of DNA fingerprinting. Ninety individuals from eight populations were sampled. Both phenetic and Neighbor-Net analyses reveal that populations are genetically discernable, but that there is no genetic evidence to support the recognition of the two currently defined subspecies of P. reniforme. Instead, the analyses resolved all individuals that possess long hypanthia into a single group, and the role of different pollinators in driving reproductive isolation of this long-tubed form is suggested.