Basic helix-loop-helix(bHLH)transcription factors regulate diverse plant processes,particularly anthocyanin biosynthesis through the MYB-bHLH-WD40 complex.Despite snapdragon(Antirrhinum majus)serving as a classical mo...Basic helix-loop-helix(bHLH)transcription factors regulate diverse plant processes,particularly anthocyanin biosynthesis through the MYB-bHLH-WD40 complex.Despite snapdragon(Antirrhinum majus)serving as a classical model for studying flower pigmentation genetics,its bHLH gene family has rarely been comprehensively characterized.Here,we performed a genome-wide identification and systematic characterization of the bHLH gene family in A.majus,with a focus on candidates involved in anthocyanin biosynthesis.A total of 150 AmbHLH genes were identified and subjected to in-silico analyses,including phylogenetic classification,structural analysis,and promoter cis-element characterization.Comparative transcriptomic profiling between anthocyanin-poor(“SIPPE50”,Green)and anthocyanin-rich(“JI2R”,Red)snapdragon lines highlighted eight differentially expressed AmbHLHs.AmbHLH001,AmbHLH002,and AmbHLH042 showed significant upregulation in the anthocyanin-rich line and showed positive correlations with the expression of key anthocyanin biosynthetic genes.Among these,AmbHLH002 was prioritized as a candidate and was assessed via heterologous overexpression in tomatoes.Notably,AmbHLH002 is a newly identified regulator whose overexpression in tomato resulted in visible purple pigmentation and increased anthocyanin accumulation.These findings support the view that AmbHLH002 acts as a positive regulator,with phylogenetic evidence for conservation of anthocyanin biosynthesis,presenting valuable potential for engineering pigmentation traits in ornamental plants and serving as a candidate visible marker for plant genetic transformation.展开更多
Flower color is an essential trait in ornamental plant breeding. Lycoris longituba is a popular ornamental plant native to central eastern China. The decrease in anthocyanin accumulation causes L. longituba petal colo...Flower color is an essential trait in ornamental plant breeding. Lycoris longituba is a popular ornamental plant native to central eastern China. The decrease in anthocyanin accumulation causes L. longituba petal color fading during flower development, which considerably affects the ornamental value of L. longituba. However, mechanisms underlying anthocyanin biosynthesis inhibition during L. longituba petal development remain unclear. In this study, three LlDFR genes were confirmed to be involved in anthocyanin biosynthesis and LlDFRc exerted the strongest promoting effect on anthocyanin accumulation. According to the correlation analysis results, LlbHLH12 exhibited the strongest negative correlation with LlDFRc. Quantitative real-time PCR analysis showed that LlbHLH12 was highly expressed during the medium bud and full bloom stages of flower development. LlbHLH12 was identified as a member of subgroup XII of bHLH transcription factor family. Subcellular localization and transcriptional activation ability assay revealed that LlbHLH12 was located in the nucleus without transcriptional activation activity. Overexpression of LlbHLH12 in Nicotiana tabacum and L. longituba inhibited anthocyanin accumulation by suppressing the expression of anthocyanin biosynthetic pathway genes. Furthermore, yeast one-hybrid, dual-luciferase, and β-glucuronidase activity assays showed that LlbHLH12 directly bound to the promoters of LlPAL and LlDFRc and suppressed their expression to inhibit anthocyanin biosynthesis. Overall, our study identified a novel bHLH repressor negatively regulating anthocyanin biosynthesis and provided new insights into the molecular mechanisms underlying color fading in L. longituba petals.展开更多
基金funded by the USDANIFA grant 2019-67013-29236the USDA HATCH program FLA-MFC-006387,awarded to Heqiang Huo.
文摘Basic helix-loop-helix(bHLH)transcription factors regulate diverse plant processes,particularly anthocyanin biosynthesis through the MYB-bHLH-WD40 complex.Despite snapdragon(Antirrhinum majus)serving as a classical model for studying flower pigmentation genetics,its bHLH gene family has rarely been comprehensively characterized.Here,we performed a genome-wide identification and systematic characterization of the bHLH gene family in A.majus,with a focus on candidates involved in anthocyanin biosynthesis.A total of 150 AmbHLH genes were identified and subjected to in-silico analyses,including phylogenetic classification,structural analysis,and promoter cis-element characterization.Comparative transcriptomic profiling between anthocyanin-poor(“SIPPE50”,Green)and anthocyanin-rich(“JI2R”,Red)snapdragon lines highlighted eight differentially expressed AmbHLHs.AmbHLH001,AmbHLH002,and AmbHLH042 showed significant upregulation in the anthocyanin-rich line and showed positive correlations with the expression of key anthocyanin biosynthetic genes.Among these,AmbHLH002 was prioritized as a candidate and was assessed via heterologous overexpression in tomatoes.Notably,AmbHLH002 is a newly identified regulator whose overexpression in tomato resulted in visible purple pigmentation and increased anthocyanin accumulation.These findings support the view that AmbHLH002 acts as a positive regulator,with phylogenetic evidence for conservation of anthocyanin biosynthesis,presenting valuable potential for engineering pigmentation traits in ornamental plants and serving as a candidate visible marker for plant genetic transformation.
基金supported by the National Natural Science Foundation of China(Grant Nos.31870695,32071828)the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD).
文摘Flower color is an essential trait in ornamental plant breeding. Lycoris longituba is a popular ornamental plant native to central eastern China. The decrease in anthocyanin accumulation causes L. longituba petal color fading during flower development, which considerably affects the ornamental value of L. longituba. However, mechanisms underlying anthocyanin biosynthesis inhibition during L. longituba petal development remain unclear. In this study, three LlDFR genes were confirmed to be involved in anthocyanin biosynthesis and LlDFRc exerted the strongest promoting effect on anthocyanin accumulation. According to the correlation analysis results, LlbHLH12 exhibited the strongest negative correlation with LlDFRc. Quantitative real-time PCR analysis showed that LlbHLH12 was highly expressed during the medium bud and full bloom stages of flower development. LlbHLH12 was identified as a member of subgroup XII of bHLH transcription factor family. Subcellular localization and transcriptional activation ability assay revealed that LlbHLH12 was located in the nucleus without transcriptional activation activity. Overexpression of LlbHLH12 in Nicotiana tabacum and L. longituba inhibited anthocyanin accumulation by suppressing the expression of anthocyanin biosynthetic pathway genes. Furthermore, yeast one-hybrid, dual-luciferase, and β-glucuronidase activity assays showed that LlbHLH12 directly bound to the promoters of LlPAL and LlDFRc and suppressed their expression to inhibit anthocyanin biosynthesis. Overall, our study identified a novel bHLH repressor negatively regulating anthocyanin biosynthesis and provided new insights into the molecular mechanisms underlying color fading in L. longituba petals.
