【摘要】：The straw mushroom, Volvariella volvacea, is one of the most important cultivated edible mushrooms, representing a valuable source both in economy and in nutrition. But the straw mushroom has comparably lower individual yield and less substrate utilization efficiency. It is also unable to stand freeze preservation. Thus breeding novel strains (highly resistant and high- yield strain) is one important problem in straw mushroom production and research.
It is designed in this study to establish an efficient transformation system of straw mushroom with plasmid pCAMBIA1301 and recombinant plasmid pCTH823 by particle bombardment and Agrobacterium-mediated DNA entry. So that it can contribute to the genetic modification of straw mushroom varieties and can be helpful in the study of its functional genome.
The main content of the study as follows:
Optimization of parameters used in particle bombardment by visualization of the growth of putative transformants on selective medium.
Establishment of transformation conditions applied in Agrobacteriwn-mediated DNA entry.
Visualization of the expression of GUS gene in putative transformants using histochemical assay. The results show that GUS gene is integrated into the genome of straw mushroom.
The results of Cropping trial and GUS enzyme activity assay indicated that straw mushroom transformants can develop into normal fruiting body and GUS gene can be expressed in this kind of fruiting body.
The resistant strains of Straw mushroom obtained in the screening after bombardment were detected by Southern dot blotting. Positive signals were visualized after hybridization for 16 hours, which shows that GUS gene was integrated into the genome of straw mushroom from the level of gene.
Straw mushroom transformed by /Igroactermm-mediated DNA entry was assayed by PCR. 2 strains gave positive signals, which proves that transgenic straw mushroom was obtained.