Di-monad Breeding And Genetic Analysis Of Flammulina Velutipes

Flammulina velutipes is a kind of mushroom cultivar in China.New species selection and its application is very singificant to the development of the mushroom industry. The purpose of this study was to build a system of Di-monad breeding to provide scientific basis for new species selection by di-monad breeding and genetic analysis of Flammulina velutipes. In this paper, we studied the mating type of mononuclear protoplast from Flammulina velutipes parent strains, the agronomic character of the first and second filial from fully compatible and hemi-compatible di-monad breeding, and the mating type of the first filial generation from mononuclear protoplast di-monad breeding. The results were obtained as below.1 Mononuclear protoplast, mating type and mycelial growth rate analysisIn this study, we obtained mononuclear protoplast regeneration strains from F32 (jinsi), F21 (Sanming1) and F2121 and two mating types were also obtained. Respectively, the two mating types ratio of F32 (jinsi) and F21 (Sanming 1) are fitting to 1:1 according to test except F2121, which existed deviation. The mycelial growth rates exhibited significantly differences between two different mating type strains obtained from F32 and F2121 by mononuclear protoplast regeneration, but there was no significant differences between.the pairing of a dikaryon strain of F21 (Sanming 1) and theirs parent strains. This is indicated that the mating type of the tested strains as follow: F32 (jinsi) mating type was A1B1, A2B2; F21 (Sanming 1) mating type was A3B3, A1B4; F2121 mating type was A1B1 , A2B2.2 ISSR(inter-simple sequence repeat) analysis of different mating type ofmononuclear protoplast strains, original strains and paired of dual-core strains. The genetic relationship is close between dual-core strains which paired from two different mating type of mononuclear protoplast strains and the original strains , the genetic similarity coefficient ranged from 0.75 to 0.97. and there were genetic differece among two different mating type of mononuclear protoplast strains, the genetic similarity coefficient between F32's dual-core strain and the original strains reached coincidence but not F2121 and F21. 3 Di-monad breeding and antagonistic identify of hybrid strainsTwo dual-core strains F2315(A1B1+A3B3)and F2715(A2B2+A3B3)which were obtained from the two different mating type parents strain F32 and F21 as the donator, and the mononuclear parents F2121P1 (A2B2) as a receptor. We got 16 combinations through full compatible di-monad breeding of (F2315×F2121P1),and got another 16 combinations through semi-compatible di-monad breeding of (F2715×F2121P1). Full compatible di-monad breeding and semi-compatible di-monad breeding have antagonistic with theirs dual-core parents, and there wasn't antagonistic among completely affinity hybrids and semi affinity hybrids.4 Analysis of mating type of mononuclear protoplast strains from the first fifial (F1)of Di-monad breedingThere were two mating types of the F1 obtained from full compatible di-monad breeding and semi-compatible di-monad breeding and theirs mating types fitting to 1:1 according to test. The mating type of the completely affinity hybrids and semi- affinity hybrids,tested through the standard of parental mating type of mononuclear protoplast, were A2B2 and A3B3. One mating type was the same as the one of dual-core donatar, another was the same as monoclear receptor.5 ISSR(inter-simple sequence repeat) analysis of Di-monad hybrids and mononuclear protoplast strains of the two mating types and theirs parentsThere was close genetic relationship among full compatible di-monad hybrids and receptor; dual-core and two different mating type of mononuclear protoplast, also the semi affinity di-monad hybrids and mononuclear receptor. The genetic relationship of two different mating types of mononuclear protoplast strains from di-monad hybrids accorded protoplast receptor but far away from dual-core mononuclear protoplast strains same as the semi affinity di-monad hybrids and dual-core. Di-monad hybrids of mononulear from two mating types have much genetic variation, one is close to dual-core donator and mononuclear receptor atrains. But another is far away from them.6 Agronomic traits the first and second fifial from di-monad breedingThe mycelial growth rate of the completely affinity hybrids and semi-affinity hybrids are quicker than binuclear parents, they differs much in the yield, but there are no obvious difference in fruit body's morphology, They are both yellow. In the second filial generation of completely affinity and semi-affinity di-monad hybrids are different in yield. Among the second filial generation of completely affinity di-monad hybrids, 27.78% of filial generation exhibited higher yield than parents, the hightest increase rate is up to 25.22%; while these two data of the semi-affinity di-monad hybrids are 25.53% and 17.58 respectively. The yellow and white types both appeared in the second filial generation of completely affinity and semi-affinity di-monad hybrids, and the ratio of yellow and white is fitting to 3:1 according to detection.