Identification of protoplast fusion strain Fhhh by randomly amplified polymorphic DNA

Abstract

A functional strain Fhhh was constructed through protoplast fusion of three parental strains (Phanerochaete chrysosporium, Saccharomyces cerevisiae and native bacterium YZ) to improve the degradation efficiency of purified terephthalic acid wastewater. Randomly amplified polymorphic DNA (RAPD) and scanning electron microscope (SEM) analysis were applied to identify and confirm the fusant Fhhh through phenotypic and genetic relationship. The result of SEM analysis demonstrated that the cell shape of fusant Fhhh differed from all three parental strains. RAPD analysis of 40 arbitrary primers generated a total of 1,135 bands. The genetic similarity indices between Fhhh and parental strains Phanerochaete chrysosporium (PC), Saccharomyces cerevisiae (SC) and native bacterium (YZ) were 34.01%, 33.16%, and 35.97%, respectively. The targeted-gene PCR results showed that Fhhh inherited the DNA fragments of mnp and lip genes from parental strain PC and FLO1 gene fragment from parental strain SC. Our results suggested protoplast fusion technique may be considered as a promising technique in environmental pollution control.

Construction and Characterization of an Intergeneric Fusant That Degrades the Fungicides Chlorothalonil and Carbendazim

“…Scanning electron microscopy (SEM) has been extensively used in biological experiments and has become an effective way to confirm protoplast fusion ( Zhao et al, 2009 ; Sun et al, 2013 ). As shown in Figure 2 , the cell morphology of BD2 is somewhat different from that of the two parents.…”

“…One hundred thirty-three clear and consistent bands were obtained through analysis. The results showed that there were both common bands and characteristic bands between the fusion bacteria and the parents, indicating that the genetic basis of each strain was roughly the same and the existence of genetic differentiation, respectively (Zhao et al, 2009). The genetic similarity index between BD2 and its two parents The genetic similarity indexes between CTN-16 and fusion strain BD2 and MBC-3 and fusion strain BD2 were 0.571 and 0.428, respectively, indicating fusion strain BD2 was more inclined to CTN-16 in terms of genetic inheritance (Figure 4).…”

“…A stable putative fusion strain that grew after eight rounds of transfer was identified and designated as fusant-BD2, which produced a distinct transparent circle on a selective agar plate containing 200 mg•L −1 CTN. Scanning electron microscopy (SEM) has been extensively used in biological experiments and has become an effective way to confirm protoplast fusion (Zhao et al, 2009;Sun et al, 2013). As shown in Figure 2, the cell morphology of BD2 is somewhat different from that of the two parents.…”

Pursuing the Perfect Performer of Fermented Beverages: GMMs vs. Microbial Consortium

“…In this methodology, the cell wall is disrupted generating spheroplasts that will spontaneously fuse to other cells, integrating their DNA into a single cell and, then, recombination occurs. The insertion of genetic material could be done even from microorganisms of other kingdoms [89].…”

Development and application of RAPD-SCAR markers to identify intra-species hybrids of industrial Saccharomyces cerevisiae

“…To this end, Random Amplified Polymorphic DNA (RAPD) has been widely used to detect extensive polymorphisms because of its simplicity, rapidity, and lack of any requirement for radioisotope usage (Echeverrigaray et al 2000;Zhao et al 2009). However, the sensitivity of RAPD profiles to a number of reaction conditions, makes them less useful as markers for the analysis of industrial strains (Li et al 2008).…”