福建农业学报2024，Vol.39Issue(12)：1365-1373,9.DOI:10.19303/j.issn.1008-0384.2024.12.006
三红蜜柚肉桂酰辅酶A还原酶基因的克隆与表达分析
Cloning and Expressions of CmCCR1 in Citrus maxima Sanhongmiyou
摘要
Abstract
[Objective]Functions and expressions of cinnamoyl-CoA reductase genes(CmCCRs)associated with the lignin biosynthesis and juice vesicle granulation in Citrus maxima were investigated.[Method]Bioinformatics,gene cloning,subcellular localization,and fluorescence quantitative PCR were applied to select and clone CmCCRs from C.maxima Sanhongmiyou.Expressions of the genes in the juice sacs at different fruit development stages of Sanhongmiyou and Liuyuezaoyihao pomelos were examined.Lignin content in the sacs was determined using the acetyl bromide method.[Results]From the RNA-Seq data on Sanhongmiyou juice sacs,5 CmCCRs were identified.Since the lignin content in the mature juice sacs was shown to correlate with the gene transcription,CmCCR1 was supposed to be involved in the lignin formation,and thus further analyzed.The ORF of CmCCR1 was 1,017 bp long.The stable hydrophilic protein had a molecular weight of 37.5 kDa and a pI of 7.06 containing an NAD(P)H/NAD(P)(+)binding(NADB)domain and three shared common motifs of CcCCR1 in sweet orange with similar distribution patterns.Phylogenetically,it closely related to CcCCR1 and belonged to the dicotyledonous plants.The gene was located on the endoplasmic reticulum same as predicted previously by software.The expression of CmCCR1 in the Sanhongmiyou juice sacs increased significantly at maturation,but that of Liuyuezaoyihao remained relatively low throughout the entire developmental stages at significantly lower levels.However,when the Liuyuezaoyihao pomelo juice sacs were transiently transformed with CmCCR1,a significantly elevated expression of the gene resulted.Moreover,the phloroglucinol-stained sacs turned pink in color indicating lignin accumulation in them.[Conclusion]CmCCR1 was postulated to regulate the biosynthesis of secondary metabolites(e.g.,lignin)and the granulation of juice vesicles in pomelos.关键词
柚/肉桂酰辅酶A还原酶/基因克隆/表达分析Key words
Citrus maxima/cinnamoyl-CoA reductase/gene cloning/expression analysis分类
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蒋静怡,刘蕴暄,王欣佳,郭志雄,佘文琴,潘腾飞..三红蜜柚肉桂酰辅酶A还原酶基因的克隆与表达分析[J].福建农业学报,2024,39(12):1365-1373,9.基金项目
福建省自然科学基金项目(2020J01542) （2020J01542）
