Selection of reference genes and expression analysis of photosynthesis-related genes responding to Aureobasidium pullulans in Chenopodium album
[Objective]The paper aimed to screen the suitable reference genes and to analyze and verify the expression of photosynthesis-relat-ed genes in Chenopodium album responding to Aureobasidiumpullulans PA-2.[Method]The leaves of C.album were treated for 0,1,3,5 and 7 days under PA-2 infection and the root,stem and leave of normal growth C.album were used as materials,and qRT-RCR technology and Ct value were used to assess the expression stability of three candidate reference genes(GAPDH,β-ACTIN and β-TUBULIN).The expression levels of photosynthesis-related genes(LFRN,MDH,PetC,psaA,CAB40,psaN,BHY and MO2)were quantitatively analyzed by the optimal selected reference genes.[Result]GAPDH was the optimal reference gene of C.album responding to A.pullulans PA-2,and could be used for analyzing the related genes expression level to photosynthesis pathway.Using GAPDH as internal reference,the expression levels of 8 photo-synthesis-related genes showed varying degrees of down-regulation,except for MDH,which reached the lowest level at 5 days.Among them,PetC,psaN,CAB40,BHY and MO2 was down-regulated C.album infected 0-1 day,reaching a high level at 3 days,and increased in the later stage.The expression levels of pasA was firstly up-regulated and then down-regulated during the infection.The expression level of LFRN was down-regulated continuously after PA-2 infection.The expression level of MDH was significantly down-regulated at first,reached the lowest level at 3 days,and increased in the later stage.In different tissues infected with PA-2,8 photosynthesis-related genes were highly expressed in leaves,among which MDH gene was highly expressed in stems,while PctC and CAB40 were the lowest expression levels in roots.Except for MDH gene,the expression levels of 7 photosynthesis-related genes in different tissues were ranked as:leaves＞stems＞roots.[Conclusion]The study provides foundation for the analysis of C.album genes by using qRT-RCR and the infection mechanism of A.pullulans PA-2.
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