Preliminary Identification Of Transcription Factors AanWRKY1, AaMYB3 And AabHLH1 In Anthurium Andraeanum

Anthurium andraeanum is a perennial herb, native to Columbia and Central America tropical rainforest area.It has become an important cut flower and potted plant, which is the second largest tropical flower merchandise after tropical floss because of its characteristics such as floral inflorescence, flower-like peculiar,bright and gorgeous color, rich in color, ornamental and annual flowering.Anthurium production is generally threatened by bacterial leaf blight (pathogen is Xanthomonas axonopodispv. Dieffenbachiae), which is mainly invaded by leaf margin, from mesophyll cells to transport tissue , Affecting the water and nutrient transport, resulting in the death of the red palm, the red palm caused a devastating harm. The current production is still the lack of high resistance varieties and effective prevention and control measures.The study of the molecular pathology of pathogens-plant interactions will help to develop targeted disease resistance molecular breeding strategies.Based on the analysis of RNA-seq analysis of Anthurium andraeanum Infection in the early stage of the research group, the transcription factor gene of three disease responses was validated and identified in order to lay the foundation for the molecular pathology mechanism of the disease response. Below are key research findings:1.In this study, a full-length cDNA of 1 658 bp was amplified and contained a 1035 bp open reading frame encoding 344 amino acids. Bioinformatics analysis predicted that this gene was a typical WRKY transcription factor named AanWRKY1(NCBI No. KY597634 ). QRT-PCR confirmed that the gene expression was up-regulated in the leaves of the gene, and could be induced by hormone signal molecules such as SA, JA, ABA, ETH and GA, and the highest expression in pedicels and stems under untreated conditions. The expression of AanWRKY1-eGFP was transiently expressed by Agrobacterium tumefaciens, which indicated that the gene was located in the nucleus and further yeast transactivation activity assay showed that the gene had inactivated transcriptional activity. The results showed that the AanWRKY1 gene was able to respond to bacterial epidemics and other stress responses, and it had the effect of stress transcription regulation. The identification of this gene laid the foundation for further analysis of its molecular pathology mechanism.2.The expression of AaMYB3 gene in the leaves of Anthurium andraeanum was analyzed by qRT-PCR. The expression of SA, JA, ABA, ETH and GA was regulated by The highest expression was observed in pedicels, stems and petioles under untreated conditions. AaMYB3-eGFP was constructed by Agrobacterium tumefaciens-mediated onion epidermal infection. The results showed that the gene was in vitro and proved to be in vitro. The results showed that the AaMYB3 gene was able to respond to bacterial epidemics and other stress responses, and it had the effect of stress transcription regulation. The results showed that the gene was identified by the method of stress transcription. Further analysis of its molecular pathology mechanism laid the foundation.3.The expression of AabHLH1 transcription factor was confirmed by qRT-PCR.The expression of SA, JA, ABA, ETH and GA was regulated by qRT-PCR. Under the condition of non-treatment, the pedicels, stems and petioles The highest expression level. AabHLH1-eGFP was constructed by Agrobacterium tumefaciens-mediated onion epidermal infection, and the AabHLH1 gene was found to be located in the nucleus. Further, the AabHLH1 gene was not isolated. The results showed that the AabHLH1 gene of Anthurium andraeanum could respond to bacterial epidemiology and other stress response, and it was the role of stress transcription regulation. The gene was identified as Further analysis of its molecular pathology mechanism laid the foundation.4.1n order to help the research of target gene in Anthurium organ development and in response to stress, it is necessary to establish the molecular genetic basis of Anthurium trait controlled, so the establishment of Anthurium genome database is of great significance. Continuous growth stage based on Illumina HiSeqTM2000/MiSeqTM sequencing method of Anthurium ornamental organ leaves, bracts and inflorescence of transcriptome sequencing, the transcriptome database information,and the transcriptome database of GO, KOG and KEGG enriched classification and ornamental genes in different developmental stages of organ expression, provides the reference background the significance for the study of gene function follow-up.The above results confirmed that the three genes were regulated by the infection of the red palm, and their expression characteristics and the identification of the transcription factor activity were helpful to reveal the mechanism of molecular pathology.