Identification and Analysis of NBS-LRR Family Genes in Dioscorea opposita Based on Transcriptome Sequencing
To explore the role of NBS-LRR resistance genes in response to different fungal infections in'Tiegun'yam,NBS-LRR genes DoRGA2 and DoRGA4 were amplified from the genome of'Tiegun'yam by homologous cloning.RT-qPCR analysis revealed that the infection of the black spot disease pathogenic fungus Alternaria alternata and the endophytic fungus Penicillium halotolerans could induce the up-regulated expression of DoRGA2 and DoRGA4 in different degrees.Meanwhile,RNA-seq was performed to analyze the NBS-LRR family genes of'Tiegun'callus infected by A.alternata and P.halotolerans,the results showed that a total of 47 NBS-LRR family genes were identified,including 5 types of N,NL,CN,CNL and TN,with 25,15,3,3 and 1 gene,respectively.Furthermore,different subfamilies of NBS-LRR proteins had different evolutionary paths,and these proteins were mainly acidic proteins with hydrophilic properties,and were mainly located in the cell membrane or cytoplasm.In addition,among these NBS-LRR family genes,the expressions of TRINITY_DN9146_c0_g1,TRINITY_DN8009_c1_g1,TRINITY_DN49830_c0_g1 and TRINITY_DN39475_c0_g1 were significantly up-regulated after inoculation with A.alternata,while TRINITY_DN18700_c0_g3 was significantly up-regulated after infection with P.halotolerans.
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