期刊
JOURNAL OF EXPERIMENTAL BOTANY
卷 73, 期 18, 页码 6170-6185出版社
OXFORD UNIV PRESS
DOI: 10.1093/jxb/erac248
关键词
Callus; citrus; miR156; miR172; somatic embryogenesis; starch
资金
- Ministry of Science and Technology of China [2018YFD1000106]
- National Natural Science Foundation of China [32072528, 31872051]
- Foundation of Hubei Hongshan Laboratory [2021hszd009]
This study reveals the regulatory pathway of miR156-SPLs and miR172-TOEs in somatic embryogenesis (SE) in citrus callus. CsAGL15 and CsFUS3 activate csi-miR156a, which represses CsSPL genes and further regulates csi-miR172d and CsTOEs. This leads to alterations in starch accumulation in callus cells and affects SE efficiency in citrus.
Somatic embryogenesis (SE) is a major regeneration approach for in vitro cultured tissues of plants, including citrus. However, SE capability is difficult to maintain, and recalcitrance to SE has become a major obstacle to plant biotechnology. We previously reported that miR156-SPL modules regulate SE in citrus callus. However, the downstream regulatory pathway of the miR156-SPL module in SE remains unclear. In this study, we found that transcription factors CsAGL15 and CsFUS3 bind to the CsMIR156A promoter and activate its expression. Suppression of csi-miR156a function leads to up-regulation of four target genes, SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (CsSPL) genes, and reduction of SE efficiency. In the short tandem target mimic (STTM)-miR156a overexpression callus (MIM156), the number of amyloplasts and starch content were significantly reduced, and genes involved in starch synthesis and transport were down-regulated. csi-miR172d was down-regulated, whereas the target genes, CsTOE1.1 and CsTOE1.2, which inhibit the expression of starch biosynthesis genes, were up-regulated. In our working model, CsAGL15 and CsFUS3 activate csi-miR156a, which represses CsSPLs and further regulates csi-miR172d and CsTOEs, thus altering starch accumulation in callus cells and regulating SE in citrus. This study elucidates the pathway of miR156-SPLs and miR172-TOEs-mediated regulation of SE, and provides new insights into enhancing SE capability in citrus. miR156-SPLsand miR172-TOEsaffect starch content and thus alter somatic embryo formation capacity in citrus callus.
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