In previous studies, a new IVF method of intracytoplasmic morphologically selected sperm injection (IMSI) was introduced, based on motile sperm organellar morphology examination (MSOME). It was concluded that microinjection of morphologically selected sperm cells with strictly normal nucleus, defined by MSOME, improves IVF-ICSI outcome. The aim of the present study was to confirm this conclusion in new, enlarged study groups. Comparison between 80 couples, who underwent an IVF-IMSI trial, with matched couples, who underwent a standard IVF-ICSI procedure, confirmed that pregnancy rate following IVF-IMSI was significantly higher, and abortion rate significantly lower than in the routine IVF-ICSI (60.0 versus 25.0%, and 14 versus 40% respectively, P
Protoplasts from two auxotrophic mutants of Trichoderma harzianum Rifai (ATCC 32173), obtained from young thalli following cell wall digestion by NovoZym 234, were fused in 33% PEG suspended in 10 mM Tris-HCl and 10 mM CaCl2, pH 7.5. The frequency of fusion between lysine- and arginine-requiring auxotrophs resulting in prototrophic strains was about 5%. These prototrophic strains were classified into parental and nonparental types. Colonies developed from single conidia of the nonparental phenotype exhibited prototrophic parental or recombinant phenotypes. The ability of both prototrophic and parental strains to overgrow the soil-borne pathogenic fungi Rhizoctonia solani, Sclerotium rolfsii, and Pythium aphanidermatum in dual cultures was used to evaluate their antagonistic capability. The antagonistic abilities of the prototrophic strains were found to vary with each pathogenic fungus. The prototrophic strain A2 overgrew all the pathogenic fungi more rapidly than the parental strains. Strain A2 effectively controlled Rhizoctonia damping-off of cotton seedlings, in the greenhouse, when compared with the parental strains. Protoplast fusion appears to be a useful tool for combining desirable traits from parental strains to produce improved biocontrol strains. Key words: Trichoderma harzianum, biocontrol, protoplast fusion.
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