交易激励
转录因子
生物
基因
电泳迁移率测定
互补DNA
下调和上调
蛋白质生物合成
贮藏蛋白
核糖核酸
细胞生物学
基因表达
分子生物学
生物化学
作者
Anirban Chakraborty,B. M. Singh,Vimal Pandey,Swarup K. Parida,Sabhyata Bhatia
摘要
Summary Development of protein‐enriched chickpea varieties necessitates an understanding of specific genes and key regulatory circuits that govern the synthesis of seed storage proteins (SSPs). Here, we demonstrated the novel involvement of Ca‐miR164e ‐CaNAC100 in regulating SSP synthesis in chickpea. Ca‐miRNA164e was significantly decreased during seed maturation, especially in high‐protein accessions. The miRNA was found to directly target the transactivation conferring C‐terminal region of a nuclear‐localized transcription factor, CaNAC100 as revealed using RNA ligase‐mediated‐rapid amplification of cDNA ends and target mimic assays. The functional role of CaNAC100 was demonstrated through seed‐specific overexpression ( NAC OE) resulting in significantly augmented seed protein content (SPC) consequential to increased SSP transcription. Further, NAC OE lines displayed conspicuously enhanced seed weight but reduced numbers and yield. Conversely, a downregulation of CaNAC100 and SSP transcripts was evident in seed‐specific overexpression lines of Ca‐miR164e that culminated in significantly lowered SPC. CaNAC100 was additionally demonstrated to transactivate the SSP‐encoding genes by directly binding to their promoters as demonstrated using electrophoretic mobility shift and dual‐luciferase reporter assays. Taken together, our study for the first time established a distinct role of CaNAC100 in positively influencing SSP synthesis and its critical regulation by CamiR164e, thereby serving as an understanding that can be utilized for developing SPC‐rich chickpea varieties.
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