Cloning and spatiotemporal expression analysis of BoLMI gene in kale
[Objective]The study aimed to investigate the molecular mechanism of phenotype formation in kale cleavage leaves,and to investi-gate the role of BoLMI gene in split leaf formation.[Method]Using the DH line of split leaf kale as the test material,the complete cDNA se-quence of the gene was successfully obtained using homologous cloning technology,and it was named BoLMI.[Result]The sequence analysis showed that the full-length of BoLMI was 531 base pairs and encoded 176 amino acids.The protein had a molecular weight of 20 789.35 Da and a theoretical isoelectric point of 7.24.According to Pfam conserved domain analysis,BoLMI protein contained a homeobox conserved do-main.Phylogenetic tree analysis showed that the BoLMI of kale belonged to the same branch as that of Brassica napus and B.oleracea,and the genetic relationship was relatively close.BoLMI protein was a soluble protein located in the nucleus,containing one transmembrane domain and no signal peptide sequence.The qRT-PCR analysis showed that the expression level of BoLMI was higher that in the seedling stage of split leaf kale and lower that in the rosette stage.During the lotus stage of kale,the expression levels in different parts of the leaves of cracked leaf varieties were higher than those of round leaf varieties,with the highest expression level in the first leaf.There was a significant difference in expression levels between cracked leaf varieties and round leaf varieties.[Conclusion]It is speculated that the BoLMI gene plays an impor-tant role in the formation of split leaves in kale,providing a theoretical basis for accelerating kale leaf shape breeding.
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