A Quick Enzyme Squash Technique for Detailed Studies on Female Meiosis in Solanum: Stain Technology

A simple enzyme squash technique that enables detailed studies of meiosis in potato ovules has been developed. Fixation of ovules in iron-propionic-ethanol followed by enzymatic maceration and squashing in acetocarmine yielded numerous well preserved meg-asporocytes with nicely spread chromosomes. Resolution was sufficient, allowing detailed analysis of chromosome pairing and chiasms formation and readily permitting distinction between normal and desynaptic mutant plants. Whereas the use of previously developed ovule squash techniques has been restricted to cytogenetic analyses of plant species with relatively large megasporocytes and large chromosomes, the present technique is potentially more useful for analyses of species with small megasporocytes and small chromosomes.     

Engineering virus resistance in agricultural crops

Plant viral genomes are relatively small and in the past decade many have been characterized at the molecular level. This has prompted research into the development of virus resistance based on interference with the viral multiplication cycle by the introduction of viral sequences into the plant genome. Several strategies have been tested. The most successful one so far involves the constitutive expression of the coat protein gene of the virus against which resistance is desired. In this review we describe progress made in engineering virus resistance into potato, an important agricultural crop. To this end the molecular structure of the potato viruses X and Y and leafroll is discussed as well as the introduction of resistance against potato virus X into potato. In addition, we address the question of preservation of cultivar-specific characteristics, an important prerequisite for commercial application. Finally, recent investigations for alternative forms of virus resistance are described against the background of the results of coat protein-mediated protection.     

Formation of first division restitution (FDR) 2n-megaspores through pseudohomotypic division in ds-1 (desynapsis) mutants of diploid potato: routine production of tetraploid progeny from 2xFDR × 2xFDR crosses

Summary The level and mode of 2n megaspore formation was studied in full-sib diploid potato clones with either normal or desynaptic (ds-1ds-1) meiosis. Cytological analysis revealed that functional 2n megaspores produced by normal and desynaptic clones originate exclusively from second division restitution (SDR) and first division restitution (FDR), respectively. SDR 2n megaspores resulted from the omission of the second meiotic division following chromosome doubling after anaphase I, whereas FDR 2n megaspores resulted from a direct equational division of univalent chromosomes at anaphase I (pseudohomotypic division). Comparative data strongly indicated that the observed mechanisms of SDR and FDR 2n megaspore formation are extremes of a continuum that is being brought about by common genes for precocious chromosome division. Depending on the relative timing of cell cycle and chromosome division, this precocious chromosome division may impose postreductional (SDR) or prereductional (FDR) restitution of the sporophytic chromosome number under normal synaptic and desynaptic conditions, respectively. The observed frequencies of 2n megaspores closely correlated with seed set, following pollination by tetraploid varieties and by desynaptic diploid clones with exclusive FDR 2n pollen formation. Up to 54.0 and 21.5 seeds/ fruit were obtained from normal synaptic (SDR) and desynaptic (FDR) progeny, respectively. The high frequency of segregants with either SDR or FDR 2n megaspore formation (78.0 and 45.2%, respectively) supports the hypothesis that sexual polyploidization is the driving force behind the origin and evolution of polyploid Solanum species. The present identification of diploid potato clones with consistent FDR 2n megaspore formation extends the opportunities for direct transfer of enhanced diploid germ plasm to tetraploids, and particularly advocates the feasibility of 2x(ds-1; FDR)×2x(ds-1; FDR) breeding schemes in cultivar development and the production of relatively vigorous and uniform true potato seed (TPS) varieties. Its potential value and limitations for breeding and the experimental induction of diplosporic apomixis are discussed.     

A quick enzyme squash technique for detailed studies on female meiosis in Solanum

A simple enzyme squash technique that enables detailed studies of meiosis in potato ovules has been developed. Fixation of ovules in iron-propionic-ethanol followed by enzymatic maceration and squashing in acetocarmine yielded numerous well preserved megasporocytes with nicely spread chromosomes. Resolution was sufficient, allowing detailed analysis of chromosome pairing and chiasma formation and readily permitting distinction between normal and desynaptic mutant plants. Whereas the use of previously developed ovule squash techniques has been restricted to cytogenetic analyses of plant species with relatively large megasporocytes and large chromosomes, the present technique is potentially more useful for analyses of species with small megasporocytes and small chromosomes.