Objective To identify the MADS-box gene family of Lonicera macranthoides,analyze its bioinformatics,validate the expression patterns,and clone the full-length sequences of CMB1,a member of the MADS-box family of Lonicera macranthoides of Xianglei and wild types.Methods Based on the transcriptome data,the bioinformatics of Lonicera macranthoides MADS-box was analyzed with online tools,the expression patterns of MADS-box genes in different cultivars were verified using qRT-PCR,and the full-length sequences of Lonicera macranthoides CMB1 gene of the Xianglei and wild types were cloned by RT-PCR and RACE techniques.Results The sequence lengths of 28 MADS-box proteins of Lonicera macranthoides ranged from 89 aa to 359 aa,with alkali proteins accounting for 85.7%,unstable proteins accounting for 96.4%,and hydrophilic proteins accounting for 96.4%,which were localized in the nucleus,and were non-secretory proteins without transmembrane structure.The transcriptome showed that compared with the wild type,17.86%of the 28 MADS-box genes in the Xianglei type and 18.75%of the genes of MIKCC type were downregulated.The full-length sequences of CMB1 genes,which were highly specifically expressed in two varieties of flowers of Lonicera macranthoides,were cloned.Both of them contained a 738 bp ORF,encoding 245 amino acids.There was a significant difference in expression level of CMB1 gene between the two varieties of flowers(P<0.01),and it was highly and specifically expressed in flowers compared to that in stems and leaves(P<0.01).Conclusion Based on the transcriptome data of Lonicera macranthoides,28 MADS-box family genes were identified,and the full-length sequences of CMB1 gene of Xianglei and wild types were cloned,which provides the research foundation and theoretical basis for the further study of molecular mechanism of development and superior phenotype formation of Lonicera macranthoides.
维普
万方数据