Ploidy dimorphism and reproductive biology in Stenodrepanum bergii (Leguminosae), a rare South American endemism

This is the first report on chromosome numbers and the reproductive behaviour in Stenodrepanum Harms, a rare endemic and monotypic legume genus from the arid and salty areas of central–western Argentina. Sixty individuals belonging to two populations from two salty areas (“salinas”) were surveyed and included mostly triploid (2n = 3x = 36) and only two diploid (2n = 2x = 24) plants. Meiosis in diploids is regular, with bivalent pairing and uniform and viable pollen. In contrast, meiosis in triploids is characterized by high trivalent pairing, with irregularly shaped pollen and variation in cytoplasm content and stainability, which is in agreement with an unbalanced segregation occurring in anaphases I and II. However, different triploid plants/individuals showed various degrees of pollen fertility, which may be attributed to particular genotypes. Research on reproductive biology events indicates sexual cross-pollinated reproduction enhanced by protogyny in both cytotypes. All plants produced seeds, but seedlings were only recovered from diploid plants pollinated with triploids, and even those eventually perished. Chromosome counts in these seedlings revealed aneuploid chromosome numbers owing to the combination of unbalanced gametes.

Mitogen-Activated Protein Kinase Cascade Required for Regulation of Development and Secondary Metabolism in Neurospora crassa

ABSTRACT Mitogen-activated protein kinase (MAPK) signaling cascades are composed of MAPK kinase kinases (MAPKKKs), MAPK kinases (MAPKKs), and MAPKs. In this study, we characterize components of a MAPK cascade in Neurospora crassa (mik-1, MAPKKK; mek-1, MAPKK; and mak-1, MAPK) homologous to that controlling cell wall integrity in Saccharomyces cerevisiae. Growth of basal hyphae is significantly reduced in mik-1, mek-1, and mak-1 deletion mutants on solid medium. All three mutants formed short aerial hyphae and the formation of asexual macroconidia was reduced in Δmik-1 mutants and almost abolished in Δmek-1 and Δmak-1 strains. In contrast, the normally rare asexual spores, arthroconidia, were abundant in cultures of the three mutants. Δmik-1, Δmek-1, and Δmak-1 mutants were unable to form protoperithecia or perithecia when used as females in a sexual cross. The MAK-1 MAPK was not phosphorylated in Δmik-1 and Δmek-1 mutants, consistent with the involvement of MIK-1, MEK-1, and MAK-1 in the same signaling cascade. Interestingly, we observed increased levels of mRNA and protein for tyrosinase in the mutants under nitrogen starvation, a condition favoring sexual differentiation. Tyrosinase is an enzyme that catalyzes production of the secondary metabolite l-DOPA melanin. These results implicate the MAK-1 pathway in regulation of development and secondary metabolism in filamentous fungi.

A New and Improved Automated Technology for Early Sex Determination of Ginkgo biloba

Random amplified polymorphic DNA (RAPD) technique with male associated decamer primer S1478 was used to amplify DNA from 72 leaf samples collected from Ginkgo biloba trees with known sexual determinism in the canton of Geneva, Switzerland. This marker was found to be male-specific and was lacking in all female plants. Automated random polymorphic DNA analysis (ARPA), a new automated technology developed in the frame of this work, proved highly effective in distinguishing males and females with 100% efficiency and successful in male and female discrimination from a collection of young seedlings derived from a sexual cross. Our findings provide unambiguous evidence that ARPA combined with the male-associated decamer primer S1478 could be considered an efficient, rapid, and easy method to make an early sex determination in the dioecious tree Ginkgo biloba.

Evidence of Genetic Exchange by Parasexual Recombination and Genetic Analysis of Pathogenicity and Mating Type of Parasexual Recombinants in Rice Blast Fungus, Magnaporthe oryzae

A selectable marker gene conferring resistance to bialaphos (BI) was introduced into rice blast isolate Y90-71BI and another conferring resistance to blasticidin S (BS) into isolate 3514-R-2BS of Magnaporthe oryzae to demonstrate exchange of DNA. Colonies obtained from co-cultures of these two isolates were resistant to both BI and BS and had both resistance genes as shown by Southern blot analysis of their genomic DNA. Conidia from these BI-BS-resistant isolates had only one nucleus per cell after staining with 4′,6-diamidino-2-phenylindole (DAPI). Using flow cytometry, however, these BI-BS-resistant isolates were found to be haploid. Segregation of BI-BS-resistant isolates for pathogenicity (avirulence to virulence) on rice line K59-1 was consistent with a 1:1 ratio, as was segregation for mating type. These BI-BS-resistant isolates were thus apparently derived from parasexual exchange of DNA and the segregation of pathogenicity and of mating type of the parasexual recombinants might correspond to that of the progeny of the offspring of the sexual cross.

PCR-Based Identification of MAT-1 andMAT-2 in the Gibberella fujikuroi Species Complex

ABSTRACT All sexually fertile strains in the Gibberella fujikuroi species complex are heterothallic, with individual mating types conferred by the broadly conserved ascomycete idiomorphsMAT-1 and MAT-2. We sequenced both alleles from all eight mating populations, developed a multiplex PCR technique to distinguish these idiomorphs, and tested it with representative strains from all eight biological species and 22 additional species or phylogenetic lineages from this species complex. In most cases, either an ∼800-bp fragment from MAT-2 or an ∼200-bp fragment from MAT-1 is amplified. The amplified fragments cosegregate with mating type, as defined by sexual cross-fertility, in a cross of Fusarium moniliforme (Fusarium verticillioides). Neither of the primer pairs amplify fragments from Fusarium species such as Fusarium graminearum, Fusarium pseudograminearum, andFusarium culmorum, which have, or are expected to have,Gibberella sexual stages but are thought to be relatively distant from the species in the G. fujikuroi species complex. Our results suggest that MAT allele sequences are useful indicators of phylogenetic relatedness in these and otherFusarium species.

Synergistic epistasis between loci affecting fitness: evidence in plants and fungi

Information on the nature of epistasis between alleles affecting fitness is hardly available, but relevant for, among other issues, our understanding of the evolution of sex and recombination. Evidence of synergistic epistasis between deleterious mutations is support for the Mutational Deterministic hypothesis of the evolution of sex, while finding antagonistic epistasis between beneficial alleles would support the Environmental Deterministic hypotheses. Both types of epistasis are expected to cause negatively skewed fitness distributions of full-sib offspring from a sexual cross. Here, we have studied the form of the distribution of a variety of quantitative characters related to fitness by searching the literature. The fitness traits encountered include the mycelial growth rate in fungi, and earliness, resistance against pathogens, seed number, and pollen fitness in plants. Fitness-related traits in plants show almost exclusively negative skewness, while the results for fungal species are more ambiguous. Possible sources of negative skewness other than epistasis, such as recessiveness of deleterious alleles or a negatively skewed error variance, were tested and found to be unimportant. We argue that these results suggest the existence of synergistic epistasis between deleterious alleles or antagonistic epistasis between beneficial alleles in plants, which is general support for the currently popular hypotheses of sex and recombination, but does not distinguish between them.

Metschnikowia agaveae sp.nov., a heterothallic haploid yeast from blue agave

Several strains of a new haploid, heterothallic species of Metschnikowia have been isolated from Agave tequilana var. azul in two agave-growing localities of Jalisco, Mexico. The new yeast species forms two aciculate ascospores per ascus after conjugation of enlarged and elongated compatible cells. Named after its host, Metschnikowia agaveae resembles superficially Metschnikowia hawaiiensis, but differs from the latter by some physiological and morphological characteristics. These two species exhibited no signs of sexual cross-reactivity. Strain UWO(PS)92-207.1 (h+, CBS 7744, ATCC 90148) is the type culture, and strain UWO(PS)92-210.1 (h−, CBS 7745, ATCC 90147) has been designated as isotype.Key words: agave, yeast, Metschnikowia, new species.

The genetic basis of isozyme variation in the wheat stem rust fungus (Puccinia graminis tritici)

The pattern of inheritance of isozyme alleles was examined in the F2 progeny of a sexual cross between two different physiologic races of the wheat stem rust pathogen, Puccinia graminis Pers. f. sp. tritici Eriks. &Henn. Segregation occurred at five isozyme loci (Got, Lap, Nadhd, Pgi-2 and Pgm-1). In all of these, the observed number of F2 progeny of each of the three possible isozyme phenotypes did not differ significantly from a simple 1:2:1 ratio. Joint dihybrid segregation indicated that three of the loci formed a single linkage group: Lap–Nadhd–Pgi-2.Key words: linkage, segregation.