Evidence of parasexual exchange of DNA in the rice blast fungus challenges its exclusive clonality

ABSTRACT We applied DNA markers to determine whether parasexual recombination may contribute to the extreme genetic diversity and variability observed in Magnaporthe grisea, the causal agent of rice blast disease. Dispersed repetitive elements and mapped, low-copy restriction fragment length polymorphism (RFLP) probes were used to detect transfers of DNA between cultured isolates of M. grisea. Low-copy RFLP probes also were used to detect putative recombinants among isolates from well-characterized field populations of the pathogen. Microscopic examination of tufted mycelium between cocultured isolates revealed frequent hyphal fusions. Hyphal tips and conidia were recovered without selection from tufted zones in two separate vegetative pairings involving isolates with dissimilar haplotypes, based on the repetitive element MGR586. Haplotypic changes were observed at a higher frequency in tuft derivatives than in subcultures of each isolate alone. From 136 tuft derivatives analyzed, 5 putative recombinant haplotypes were identified. Introgression was demonstrated with two independent repetitive elements, fosbury and MGR586, as probes on DNA digested with several restriction enzymes. Introgressions were characterized by addition of 1 to 10 MGR586 bands, and 1 to 3 fosbury bands from one parent into the background of the other. Polymorphic single-copy probes were used to analyze putative recombinants. One probe detected an introgression event as predicted by analysis with MGR586. To assess the possible role of parasexual recombination in field populations of the pathogen, isolates in the Philippines previously grouped based on DNA fingerprinting were analyzed with low-copy RFLP markers. Polymorphism in single-copy loci typically was seen between, but not within, putative pathogen lineages. One lineage (designated lineage 4), however, was polymorphic for several probes. For some isolates, alleles at these loci comigrated with alleles characteristic of other lineages, suggesting the transfer of DNA fragments between lineages. One isolate was apparently a merodiploid, carrying an allele typical of lineage 4 plus another allele characteristic of a different lineage. In a survey of isolates from the Indian Himalayas, a merodiploid also was found with single- or low-copy probes. Examination of MGR586 profiles of the putative recombinant and its putative donor strains showed the expected introgression of MGR586 bands. The detection of parasexual DNA exchanges in wild-type strains under unselected conditions and the existence of merodiploids in nature suggest that parasexual recombination occurs in field populations of M. grisea. This raises questions concerning exclusive clonality in the blast fungus.     

Discovery of Eocene Sediments in Subsurface of Cape Cod

Spores, pollen, and charcoal taken from two wells drilled near the tip of Cape Cod have been identified as Eocene. These are the first Eocene rocks to be identified in Massachusetts. Interpretation of seismic records taken in the Gulf of Maine and Cape Cod Bay will be influenced by this discovery.     

稻瘟病菌致病突变株的分离

 利用4个分属宗亲群4,7,17 和44的菌株连续接种在C039的单基因近等基因系和基因累加系上, 以获得突变抗病基因新菌株。经13轮接种循环, 共分离到9个新菌株。其中8个针对C101LAC(Pi－1) 突变 , 另1个针对C101PKT (Pi－4a)和F128 (Pi－ta²) 的双位点突变。4个原始菌株, P06－6, 9248－6, 9239－4和101－7－2的突变频率分别为: 0, 2.82×10^－5, 2.05×10^－5 和9.09×10^－5。菌株针对不同抗病基因的突变频率不同,由Av－1变到Av－1^＋和由Avir－4a(t)突变到Av－4a^＋的频率分别为7.51×10^－5和6.14×10^－5。没有针对Pi－2突变体出现。     

Factors associated with shedding of Cryptosporidium parvum versus Cryptosporidium bovis among dairy cattle in New York State

OBJECTIVE: To isolate and speciate Cryptosporidium DNA from fecal samples obtained from dairy cattle in New York State and identify factors associated with whether cattle were shedding Cryptosporidium parvum versus Cryptosporidium bovis. DESIGN: Cross-sectional study. SAMPLE POPULATION: 115 fecal samples positive for DNA coding for the Cryptosporidium 18S rRNA gene from dairy cattle in New York State. PROCEDURES: A PCR assay was used to amplify DNA from fecal samples; amplification products were submitted for bidirectional DNA sequencing. Logistic regression was used to test for associations between various host factors and Cryptosporidium spp. RESULTS: 70 of the 115 (61%) fecal samples were found to have C parvum DNA, 42 (37%) were determined to have C bovis DNA, and 3 (3%) were found to have C parvum deer-type DNA. The presence of diarrhea at the time of fecal sample collection, oocyst count, and breed were associated with whether cattle were infected with C parvum or C bovis, with animals more likely to be infected with C parvum if they had diarrhea, had a high oocyst count, or were Holsteins. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that C parvum and C bovis can be isolated from dairy cattle in New York State and that various factors affect whether cattle infected with Cryptosporidium spp are infected with C parvum or C bovis. Findings also lend credence to the theory that C bovis may be more host adapted and thus less pathogenic to dairy cattle than C parvum.     

Demonstration that Feeds Containing <1% Fishmeal Can Support Grow‐out of Large Juvenile Red Drum,Sciaenops ocellatus,and Reduce Nutrient Waste

We conducted a 16‐wk feeding trial with large juvenile red drum, Sciaenops ocellatus. Four diets were randomly assigned to six replicate tanks per treatment. Three isonitrogenous (ca. 44.5% protein) and isolipidic (ca. 14.1% lipid) extruded diets were formulated to compare a fishmeal‐based diet with diets using alternative protein sources. Diet 1 contained 19.60% fishmeal and 21.42% poultry meals as primary protein sources. Two alternative diets were formulated reducing the fishmeal to 0.61% by substituting poultry meals (33.85%) and soybean protein concentrate (11.55% in Diet 2 and 11.70% in Diet 3). Diet 3 also included Allzyme Vegpro® and Allzyme® SSF at 0.04%. Diet 4, a natural diet consisting of chopped cigar minnows, squid, and shrimp, was used as a positive control to compare growth rates of formulated feeds to near maximum growth under these culture conditions. We found that reducing the amount of fishmeal to <1% by using alternative protein sources did not affect the growth rate, survival, or health of red drum but improved assimilation of phosphorus, reduced potential release of P to the environment, and significantly lowered the amount of feeder fish needed in feed. The control diet identified performance benchmarks for future feeds development work.