Cloning and functional analysis of MYB protein target gene NnTOM1 in Nelumbo nucifera
[Objective]The study aimed to clone and analyze the bioinformatics,expression patterns and tobacco genetic transformation of Nn-TOM1 gene in lotus(Nelumbo nucifera),which would provide a foundation for further study on the function of NnTOM1 gene.[Method]Nn-TOM1 gene was obtained by cloning,and its protein physicochemical properties,and conserved domain etc.were analyzed using bioinformatics tools.The tissue-specific expression of NnTOM1 gene was analyzed by quantitative real-time polymerase chain reaction(qRT-PCR).The bio-logical function of NnTOM1 gene was preliminarily verified by tobacco genetic transformation.[Result]The study revealed that the coding se-quence(CDS)of NnTOM1 gene was 1191 bp,which encoded 396 amino acids,and the protein molecular formula was C1918H3130N560O589S16 The secondary structure of the protein was mainly composed of α-helix and random coil.It contained the TOM family conserved domains(VHS-ENTH-ANTH and GAT),which was localized in the cytoplasm;Multiple sequences comparison and phylogenetic tree analysis showed that NnTOM1 protein was closely related to Macadamia integrifolia,Telopea speciosissima and Macleaya cordata,and the sequence identity was high.The results of qRT-PCR showed that the expression of NnTOM1 gene was the highest in petalized stamens and the lowest in roots,moreover,it was higher in multiple-petal varieties than single-petal varieties.The phenotype of NnTOM1 positive transgenic plants showed that compared with the wild type,the color of the petals was deepened,the tip of the petals was pointed,the spacing of the petals was widen,and the filaments were longer and obviously longer than the style.The OE-1 strain with a phenotype of 6 petals had one more petal than the wild type.[Conclusion]The NnTOM1 gene in lotus might be involved in flower organs development.The study provides a preliminary basis for fur-ther study on the function of NnTOM1 gene,and provides a new gene resource for breeding of new lotus varieties.
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