The Effect Of Pseudolaric Acid B On Moisis Of Colletotrichum Gloeosporioides Based On RNA-seq

Mango anthracnose is a sever fungus disease prevalent in mango planting area,the massive use of selective synthesis chemicals presents a potential resistance risk and causes the low control efficiency.In our previous study on natural products with fungicidal activity,pseudolaric acid B(PAB)was found to exert good antifungal activities to many fungus,but its mechanism is unclear.Study on it may lead to discover a new type of antifungal mechanism.In order to gain a deeper understanding of the molecular mechanism of PAB,DAPI fluorecent staining and Illumina's Solexa sequencing technology was used to study the effect of PAB on the mitosis of Colletotrichum gloeosporioides(Penz.)Sacc.from the cellular and molecular level,carbendazim(CBM)as the fungicide control.The results are as follows:1.The results of nuclei behavior observation showed that most nuclei divided seven times,and the dynamic change of nuclear behavior was delayed about 3 h after the third nuclei division under treatment with 5 mg/L pseudolaric acid B within 12 h,which was similar to CBM treatment.2.By RNA-seq on C.gloeosporioides and its treatments,45351798 clean reads were obtained and assembled into 22865 unigenes.After annotation with Nr,Swissprot,KEGG,COG and GO,13999,79134,3931,5429 and 2468 unigenes were annotated respectively.The total annotation unigenes were 14100.3.Based on RPKM,there were 726 out of 1783 differentially expressed genes(DEGs)under PAB treatment,which were up-regulated,While 723 out of 1592 differentially expressed genesunder CBM treatment were up-regulated.Among them,25 genes were randomly selected to validate the results of transcriptome by using quantitative real time PCR(qRT-PCR).The results showed a good correlation between the transcriptome and quantitative real time PCR(R20.909).4.GO and KEGG functional enrichment analysis showed that there were significantly different in the fisrt two GO terms when annotated to celluar component.KEGG enrichment analysis showed that 5 pathways related to metabolite and nutrients were significantly enriched with most down-regulated genes and there was no target genes related to traditional fungicides under PAB treatment,while 4 pathways related to metabolite and ribosome synthesis were significantly enriched under CBM treatment.5.DEGs related to cell cycle,cell division and cell skeleton were found under PAB treatment: genes coded cell division control protein 6,type IIB DNA topoisomerase and?1-tubulin.In conclusion,both PAB and CBM can inhibit the mitosis progress by reducing the cell division times.However,there were significantly different in DEGs between PAB and CBM treatment.DEGs annotated into different biological functions and metabolic pathways showed that there were differences in molecular mechanism between PAB and CBM,the antifungal mechanism of PAB may be related to regulate the genes associated with microtube and cell cycle.