Effects of photoperiod and temperature on the developmental duration and diapause in Dolycoris baccarum (Heteroptera: Pentatomidae) from Hohhot, Inner Mongolia, China.

The shield bug, Dolycoris baccarum (L.) (Heteroptera: Pentatomidae), is widely distributed across Asia and Europe. At high latitudes, it overwinters, as adult in diapause, which then becomes the insect source for the following year. To fully understand the developmental duration and diapause characteristics of D. baccarum, the effects of photoperiod and temperature were studied in a population from Hohhot, Inner Mongolia, China. The results indicated that the developmental duration was significantly prolonged at temperatures of 20 or 25 °C, with a prolonged light period; however, when the light period was prolonged to 16L:8D and 18L:6D, the developmental duration was shortened significantly. Furthermore, the developmental duration was also shortened significantly with increasing temperature, when the photoperiod was 12L:12D for short days and 16L:8D for long days. All individuals entered diapause under short-day conditions of 10L:14D and 12L:12D at a temperature of 20 °C; however, the diapause rate decreased significantly under 14L:10D and 16L:8D photoperiods, and the diapause rate decreased significantly at a temperature of 25 °C with prolonged photoperiod. Interestingly, when the photoperiod was fixed at 12L:12D, the diapause rates at different temperatures (20, 25, 28, and 30 °C) exceeded 95%; while the effect of temperature on diapauses was nonsignificant under this photoperiod, it was still sensitive to the photoperiod; at a photoperiod of 16L:8D, the effect of temperature on the diapause rate was noticeable, and the diapause rate decreased significantly with increasing temperature.

A novel transcriptional factor important for pathogenesis and ascosporogenesis in Fusarium graminearum.

Fusarium head blight or scab caused by Fusarium graminearum is an important disease of wheat and barley. The pathogen not only causes severe yield losses but also contaminates infested grains with mycotoxins. In a previous study, we identified several pathogenicity mutants by random insertional mutagenesis. One of these mutants was disrupted in the ZIF1 gene, which encodes a b-ZIP transcription factor unique to filamentous ascomycetes. The Δzif1 mutant generated by gene replacement was significantly reduced in deoxynivalenol (DON) production and virulence on flowering wheat heads. It was defective in spreading from inoculated florets to the rachis and other spikelets. Deletion of the ZIF1 ortholog MoZIF1 in the rice blast fungus also caused reductions in virulence and in invasive growth. In addition, the Δzif1 mutant is defective in sexual reproduction. Although it had normal male fertility, when selfed or mated as the female in outcrosess, the Δzif1 mutant produced small, pigmented perithecia that were sterile (lack of asci and ascospores), suggesting a female-specific role for ZIF1 during fertilization or ascus development. Similar female-specific defects in sexual reproduction were observed in the ΔMozif1 mutant. When mated as the female, the ΔMozif1 perithecia failed to develop long necks and asci or ascospores. The ZIF1 gene is well conserved in filamentous ascomycetes, particularly in the b-ZIP domain, which is essential for its function. Expression of ZIF1 in Magnaporthe oryzae complemented the defects of the ΔMozif1 mutant. These results indicate that this b-ZIP transcription factor is functionally conserved in these two fungal pathogens for plant infection and sexual reproduction.

Cryptic promoter activity in the coding region of the HMG-CoA reductase gene in Fusarium graminearum.

Head blight or scab disease caused by Fusarium graminearum poses a major threat to wheat and barley production in North America and other countries. To better understand the molecular mechanisms of F. graminearum pathogenesis, we have generated a collection of random insertional mutants. In mutant 222, one of the transformants significantly reduced in virulence, the transforming vector was inserted at amino acid 269 of the hydroxymethyl-glutaryl CoA reductase gene (HMR1) that encodes a key enzyme in sterol and isoprenoid biosynthesis. The N-terminal transmembrane domains of HMR1 were disrupted, but the C-terminal catalytic domain was intact in mutant 222. We failed to isolate mutants deleted of the HMR1 gene, suggesting that HMR1 is an essential gene. Mutants deleted of the N-terminal 254 amino acids of HMR1 were viable and phenotypically similar to mutant 222. In both mutant 222 and the hmr1Delta254 mutants, a 3-kb truncated HMR1 transcript was detectable by northern blot analyses. In the wild-type strain, only the 5-kb messenger was observed. The initiation site of truncated HMR1 transcripts was determined by 5'-RACE to be 507bp upstream from the catalytic subunit. When a HMR1 fragment corresponding to the DNA sequence of HMR1269-641 was translationally fused to a promoter-less GFP construct, green fluorescent signals were detectable in vegetative hyphae of the resulting transformants. These data indicate that this region of HMR1 ORF has cryptic promoter activity and can express the catalytic domain in hmr1 mutants deleted of its N-terminal portion. Our results also illustrate the importance of the HMR1 gene and the function of its transmembrane domains in F. graminearum.

Random Insertional Mutagenesis Identifies Genes Associated with Virulence in the Wheat Scab Fungus Fusarium graminearum.

ABSTRACT Fusarium graminearum is an important pathogen of small grains and maize in many areas of the world. To better understand the molecular mechanisms of F. graminearum pathogenesis, we used the restriction enzyme-mediated integration (REMI) approach to generate random insertional mutants. Eleven pathogenicity mutants were identified by screening 6,500 hygromycin-resistant transformants. Genetic analyses indicated that the defects in plant infection were tagged by the transforming vector in six of these mutants. In mutant M8, the transforming plasmid was integrated 110-bp upstream from the start codon of the cystathionine betalyase gene (CBL1). Gene replacement mutants deleted for CBL1 and the methionine synthase gene MSY1 were also obtained. Both the cbl1 and msy1 deletion mutants were methionine auxotrophic and significantly reduced in virulence on corn silks and wheat heads. We also identified genes disrupted by the transforming DNA in three other REMI mutants exhibiting reduced virulence. In mutants M68, the transforming vectors were inserted in the NADH: ubiquinone oxidoreductase. The putative b-ZIP transcription factor gene and the transducin beta-subunit-like gene disrupted in mutants M7 and M75, respectively, had no known homologs in filamentous fungi and were likely to be novel fungal virulence factors.

A mitogen-activated protein kinase gene (MGV1) in Fusarium graminearum is required for female fertility, heterokaryon formation, and plant infection.

Fusarium graminearum is an important pathogen of small grains and maize in many areas of the world. Infected grains are often contaminated with mycotoxins harmful to humans and animals. During the past decade, F. graminearum has caused several severe epidemics of head scab in wheat and barley. In order to understand molecular mechanisms regulating fungal development and pathogenicity in this pathogen, we isolated and characterized a MAP kinase gene, MGV1, which is highly homologous to the MPS1 gene in Magnaporthe grisea. The MGV1 gene was dispensable for conidiation in F. graminearum but essential for female fertility during sexual reproduction. Vegetative growth of mgv1 deletion mutants was normal in liquid media but reduced on solid media. Mycelia of the mgv1 mutants had weak cell walls and were hypersensitive to cell wall degrading enzymes. Interestingly, the mgv1 mutants were self-incompatible when tested for heterokaryon formation, and their virulence was substantially reduced. The ability of the mutants to accumulate trichothecene mycotoxins on inoculated wheat was also greatly reduced. Our data suggest that MGV1 in F. graminearum is involved in multiple developmental processes related to sexual reproduction, plant infection, and cell wall integrity.