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MiRNAs and their targets regulate plant architecture in Brassica napus

ABSTRACT: Background: MicroRNAs (miRNAs), a class of non-coding small RNAs, are crucial to the regulation of various developmental processes. Plant architecture is a collection of genetically controlled agronomic traits that determine crop production and mechanized harvesting. Although several genes had been found to regulate plant architecture, the mechanisms whereby miRNAs regulate plant architecture in the rapeseed Brassica napus remain unknown. Results: In this study, we characterized a rod-like rapeseed mutant with an ideal plant architecture that substantially enhanced its breeding potential. To explore miRNAs that contribute to the rapeseed plant architecture, backcross progenies that developed into small plants (rod-like) and tall plants (normal) were used for study. Four small RNA (sRNA) libraries and two degradome libraries from the shoot apex of normal and rod-like plants were sequenced. A total of 925 non-redundant B. napus miRNA precursors were identified, representing 315 precursors for 74 known miRNAs and 610 precursors for 327 novel miRNAs. Expression analysis revealed that 10 known miRNAs and 7 novel miRNAs were differentially expressed between the normal and rod-like plants. In addition, 408 targets were identified through degradome sequencing and 14 targets were further validated via RNA ligase-mediated 5′ rapid amplification of cDNA ends. Furthermore, the functions of miR319 and its target gene TCP4 were studied and provided a novel insight into how miR319 regulates plant architecture. Conclusions: Correlation analysis between differentially expressed miRNAs and their targets demonstrated that nutrition and metal deprivation, energy supply deficiency, senescence and TEOSINTE BRANCHED1/CYCLOIDEA/PCF (TCPs) contributed to the premature termination of shoot development in rod-like mutant. The work further elucidates the mechanism of miRNAs participate in the regulation of plant architecture.

ORGANISM(S): Brassica napus

PROVIDER: GSE102050 | GEO | 2024/10/31

REPOSITORIES: GEO

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Project description:Chemical hybridization agent (CHA)-induced male sterility is becoming one of the most useful tools for the crop heterosis in seed production. We had previously discovered monosulfuron ester sodium (Mes), an acetolactate synthase (ALS) inhibitor of the herbicide sulfonylurea family, could induce rapeseed (Brassica napus L.) male sterility. To investigate the mechanism of Mes inducing male sterility, a cytological analysis and comparative transcriptome analysis were performed between Mes-treated plants leaves and different developmental stage anthers and those of the Mock-treated plants. Cytological analysis exhibited that Mes destroyed plastids ultrastructure and depressed the materials accumulation during anther development and maturation process. Comparative transcriptome analysis identified a total of 1501 transcripts differentially expressed in the leaves and different developmental stage anthers. Subcellular localization analysis, functional analysis and pathway analysis were carried out to obtain the clues that plastid was the seriously damaged organelle, and many genes involved in carbon and lipid metabolism and cellular transport differentially expressed. Detailed expression pattern analysis of genes related with these functions were conducted and verified by RT-PCR. In addition, several transcription factors, protein kinases and hormone related genes were identified. Carbohydrate content analysis confirmed carbon metabolism were influenced by Mes. Taken together, we proposed a putative action mode that Mes inhibited activity of acetolactate synthase, which localized in plastid in plants, and then disturbed the normal supply of carbon and lipid metabolite for anther development, finally displayed male sterility. This results have important significance for uncovering the metabolic gene regulation during anther development and may provide more potential targets for developing new male sterility inducing CHAs in rapeseed breeding. Rapeseed (Brassica napus L.) plants were foliar sprayed with 0.1Î¼g mL-1 Monosulfuron ester sodium (Mes) solution containing 50ppm DMF and 5ppm Tween 80 for about 15 ml per plant. Young leaves (C2Y1, C2Y2, C2Y3; T2Y1, T2Y2, T2Y3) of the main inflorescences and developing anthers (including small buds (C2S1, C2S2, C2S3; T2S1, T2S2, T2S3), middle anthers (C2M1, C2M2, C2M3; T2M1, T2M2, T2M3), and large anthers (C2L1, C2L2, C2L3; T2L1, T2L2, T2L3)) from Mes-treated and Mock-treated plants were collected for each experiment. Each tissue has three independent biological replicates.

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MiRNAs and their targets regulate plant architecture in Brassica napus

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