| Typha orientalis Presl is a perennial herbaceous plants,has developed rhizomes,can form various network and is a kind of new type Pb2+ enrichment plant,but the current of cattail,on Pb2+ tolerance mechanism is not yet clear.Therefore,this article selects Ty pha orientalis Presl as experimental material,using hydroponics Hoagland nutrient solutio n on the basis of preliminary experiment,for the treatment of 0.1mmol—L-1,0.25mmol· L-1 Pb2+ and through the transcriptome sequencing Pb2+ different processing group of Typha orientalis Presl roots of differently expressed genes,the purpose of the genes asso ciated with Pb2+ patience;The expression of 13 differently expressed genes was verified by qRT-PCR.The total length of glutathione S transferase gene(ToGST)associated with Pb2+ tolerance was obtained by RACE technology,and bioinformatics analysis was carrie d out on the gene.The abiotic stresses(drought and salt stress)other ToGST gene expre ssion under the amount of change,in order to clarify Typha orientalis Presl the tolerance mechanism of Pb2+ and for use of Typha orientalis Presl provides the theoretical basis of Pb2+ pollution on environment restoration.The main results as follows:1.The transcriptome sequencing:based on the control tendency for 0.1mmol·L-1,0.25 mm ol·L-1 Pb2+,the root of the samples the transcriptome sequencing,joining together after a ssembly to get 84530 Unigenes,average length is 806.04 nt,N50 length is 1422 nt.Aft er Unigene function annotation,Unigene with annotation information is 44,307,accountin g for 52.41%of Unigenes totally.By GO,COG,KEGG annotation results,under Pb2+stress of differently expressed genes is involved in the styrene acrylic biosynthesis,styren e acrylic pigment metabolism,GSH metabolism as well as process of plant pathogen inte ractions.2.Expression of differently validation genes:Ubiquitin gene(ToUBQ)with stable expressi on in each experimental group was determined as the internal reference gene after PCR verification,.Using the qRT-PCR technology for 13 candidates differently expressed genes for expression quantity verification,validation results show that the 3 gene expression up-regulated,down-regulated gene expression by 10,then consistent with the results of the transcriptome sequencing,prove the transcriptome sequencing and reliable results.3.The glutathione S transferase gene cloning and analysis:using the RACE technology in Pb2+ treatment group significantly increase glutathione S transferase gene(ToGST)clo ning,finally get the total length of 1211 bp sequence,coding district of 840 bp.ToGST for bioinformatics analysis found that the formula of the gene encoding protein C1388H2192 N382O388S8,relative molecular weight of 30.695 kDa,theory of isoelectric point pI = 10.35,according to the protein GRAVY value infer the protein for hydrophobic protein,no s ignal peptide,across a membrane structure,which is a transmembrane protein,there are several O-glycosylation sites and phosphorylation sites,probably for phosphorylated modif ication after translation.4.Under drought and salt stress ToGST expression quantity changes:research under drou ght and salt stress treatment typha biomass,root system activity in order to found that t he quantity of ToGST expression of drought and salt stress are led to the decrease of th e biomass and root activity decrease;In PEG treatment group,with the increase of PEG treatment concentration,ToGST expression level gradually increased,and the expression 1 evel reached the peak at 15%PEG treatment.In the PEG complex water treatment grou p,the expression level of ToGST was significantly increased after direct exposure to 15%PEG.There was no significant change in ToGST in the pretreatment group and the c ontrol group,but the expression level of ToGST in the pretreatment group decreased sign ificantly.In the salt stress treatment group,the expression of ToGST gene gradually incre ased with the increase of treatment concentration,and reached the maximum at 150mMc oncentration. |
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