| Stemphylium eturmiunum,a member of Stemphylium,can carry out sexual reproduction,which is very important in filamentous fungi.Asf1,which is a molecular chaperone of histones H3 and H4,can affect the expression of related genes by affecting the structure of nucleosomes.We found that ?asf1 strain is completely sterile.Transcriptome analysis showed that other related genes have undergone large up-and-down changes after knocking out the asf1 gene.To further study the effects of asf1 and its related genes on the reproductive development of S.eturmiunum,we mainly did the following research:Firstly,the proline dehydrogenase(04320)and allantoic acid permease(10206)gene,which may be associated with asf1,was cloned from the whole genome of S.eturmiunum.Then,we performed gene silencing and screened for successful silent strains by qRT-PCR.The biological characteristics of the silenced strains were observed.The mycelial growth rate of the silenced strain was slower,the colony area was smaller,the color was lighter,and the colonies were more fluffy compared with wild type.Moreover,the distribution of nuclei in hyphae is more sparse,and the spacing of nuclei is more irregular.After silencing the 04320 gene,the development was delayed,and the strain was completely sterile after silencing the10206 gene.These results indicated that these two genes have important effects on the sexual and asexual reproduction of S.eturmiunum.We infered that 04320 and 10206 interact with asf1 or H3 and H4,thereby regulating the expression of related genes and affecting the Biological characteristics of S.eturmiunum.Yeast two-hybrid and pull-down experiments indeed demonstrated that 04320 interacted with H4.In summary,it is speculated that 04320 is indirectly involved in the evolution and development of S.eturmiunum regulated by asf1.The insights gained in our research can betransferred to higher eukaryotes and are important for understanding the development of eukaryotic cells. |
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