| Anthura {Anthnrium andraeanum)is a perennial evergreen monocotyledon plant and a famous tropical flower.It has gorgeous colors and many kinds of flower colors.In recent years,researches on the flower color of Anthura at home and abroad mainly focus on pigment types and contents,pigment anabolism and gene expression regulation.There are few reports on the molecular mechanism of flower color variation in Anthura at the protein level.Over the years,our laboratory has created and preserved many types of Anthura color mutants,which are ideal materials for studying the mechanism of flower color variation.In this study,proteomic methods were used to isolate and identify differential proteins related to flower color variation in Anthura mutants.Molecular biology techniques were used to clone the cDNA sequence of the gene encoding the protein related to flower color variation.And qRT-PCR were used to analyze expression characteristics of 6 genes in Anthura flower color mutants,to explore the molecular mechanism of flower color variation in Anthura.The main results are as follows:1.The many types of Anthura color mutants are used as materials,and the 1-DE expression profiles of leaves,inflorescences and spathes in Anthura were preliminarily analyzed by SDS-PAGE(1-DE)technologies.It was found that there were no significant differences in protein bands of leaves and inflorescences in different color mutants,but there were significant differences in protein bands of spathes.Cuting differential protein bands to mass spectrometry analysis,and 21 proteins were detected in the differentially expressed bands of wild type and rose red mutant in 'Madural'.9 of them were specifically expressed in wild type and 4 in rose red mutant.Their functions involved metabolic regulation,cytoskeleton formation,resistance,gene regulation,transportation and signal transduction.138,112,71 and 431 proteins were detected in 4 different bands of wild and white mutants in 'pink champion'.Their molecular functions mainly involved catalytic activity and binding activity.The identified proteins were related to flavonoid synthesis and metabolism,resistance,sugar metabolism and anthocyanin transfer.2.The wild type,pink mutant and pale pink mutant in 'Turenza' are used as materials,1109 proteins were separated and identified by TMT/MS technologies.96,94 and 96 different proteins were detected by comparing the three groups of materials,including flavonoid metabolism-related proteins,stress-related proteins,resistance-related proteins and photosynthesis-related proteins.The molecular functions of differential proteins mainly involved catalytic activity and binding activity.They participate in biological processes such as cell processes and metabolic processes,and are distributed in cells,organelles and membranes.3.The spathes in Anthura were used as materials.Based on the results of differential protein identification,the 4 cDNA sequences of the genes encoding the related differential protein were cloned by molecular biology technologies,which were respectively named as AnCAT(555bp,185 amino acid residues,GenBank accession number:MK574928),AnAPX(441 bp,147 amino acid residues,GenBank accession number:MK574930),AnCCP(714bp,238 amino acid residues,GenBank accession number:MK574931),AnLAC(663bp,221 amino acid residues,GenBank accession number:MK574929).Homology analysis showed that the sequence of genes encoding differential proteins was conservative.4.The expression characteristics of AnCAT,AnAPX,AnCCP,AnHSP70,AnLAC and AnCHI in wild type,pink mutant,pale pink mutant and white mutant spathe were analyzed by qRT-PCR.The results showed that with the fading of the color of spathe,the expression levels of AnCCP,AnLAC and AnCHI was increased,the expression levels of AnCCP,AnLAC and AnCHI was decreased.Among them,the expression of AnAPX increased in the spathe of white mutant,there was no significant difference between the wild type and the mutant.And the expression of AnHSP70 changed slightly. |
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