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miR171-targeted SCARECROW-LIKE genes CsSCL2 and CsSCL3 regulate somatic embryogenesis in citrus
Meng-Qi Feng1, Nan Jiang1, Peng-Bo Wang1
1National Key Laboratory for Germplasm Innovation & Utilization of Horticultural Crops, College of Horticulture and Forestry Sciences, Huazhong Agricultural University, Wuhan 430070, China.
Plant Physiology
|May 19, 2023
Summary
Scientists uncovered a key regulatory pathway involving CsSCL2/3 genes and csi-miR171c in citrus somatic embryogenesis (SE). Suppressing CsSCL2 enhanced SE, revealing a mechanism for improving crop regeneration.
Area of Science:
- Plant Biotechnology
- Molecular Biology
- Developmental Biology
Background:
- Somatic embryogenesis (SE) is crucial for crop improvement, particularly in woody perennials like citrus.
- Maintaining SE capability is a significant bottleneck in plant biotechnology.
- Understanding the molecular mechanisms regulating SE is essential for efficient crop regeneration.
Purpose of the Study:
- To identify key regulators of SE in citrus.
- To elucidate the molecular mechanism by which CsSCL2/3 genes influence SE.
- To explore potential strategies for enhancing SE in citrus.
Main Methods:
- RNA interference (RNAi) to suppress CsSCL2 expression.
- Identification of CsSCL2/3 interacting proteins using co-immunoprecipitation.
- Overexpression of CsClot to study its effect on SE and ROS homeostasis.
- Chromatin immunoprecipitation sequencing (ChIP-Seq) and RNA-Seq to identify target genes.
- Analysis of gene expression and promoter binding.
Main Results:
- Suppression of CsSCL2 using RNAi significantly enhanced SE in citrus callus.
- CsSCL2/3 genes were found to positively regulate csi-miR171c expression.
- An interacting protein, CsClot, was identified, which modulates reactive oxygen species (ROS) homeostasis and enhances SE upon overexpression.
- CsSCL2 directly suppresses regeneration-related genes, including CsWOX2, CsWOX13, and LBD40.
- A total of 660 genes were identified as directly suppressed by CsSCL2, enriched in development and auxin signaling pathways.
Conclusions:
- A novel regulatory pathway involving csi-miR171c-targeted CsSCL2/3 genes in citrus SE was uncovered.
- CsSCL2/3 modulate SE by interacting with CsClot to regulate ROS homeostasis and by directly repressing key regeneration-associated genes.
- This study provides critical insights into the mechanism of SE and regeneration capability maintenance in citrus, offering potential targets for crop improvement.
