Molecular characterization and expression analysis of Triticum aestivum squamosa-promoter binding protein-box genes involved in ear development

Wheat （Triticum aestivum L.） is one of the most important crops in the world. Squamosa-promoter binding protein （SBP）-box genes play a critical role in regulating flower and fruit development. In this study, 10 novel SBP-box genes （TaSPL genes） were isolated from wheat （（Triticum aestivum L.） cultivar Yanzhan 4110）. Phylogenetic analysis classified the TaSPL genes into five groups （G1-G5）. The motif combinations and expression patterns of the TaSPL genes varied among the five groups with each having own distinctive characteristics： TaSPL20/21 in G1 and TaSPL17 in G2 mainly expressed in the shoot apical meristem and the young ear, and their expression levels responded to development of the ear; TaSPL6/15 belonging to G3 were upregulated and TaSPL1/23 in G4 were downregulated during grain development; the gene in G5 （TaSPL3） expressed constitutively. Thus, the consistency of the phylogenetic analysis, motif compositions, and expression patterns of the TaSPL genes revealed specific gene structures and functions. On the other hand, the diverse gene structures and different expression patterns suggested that wheat SBP-box genes have a wide range of functions. The results also suggest a potential role for wheat SBP-box genes in ear development. This study provides a significant beginning of functional analysis of SBP-box genes in wheat.

Low-affinity SPL binding sites contribute to subgenome expression divergence in allohexaploid wheat

Expression divergence caused by genetic variation and crosstalks among subgenomes of the allohexaploid bread wheat(Triticum aestivum.L.,BBAADD)is hypothesized to increase its adaptability and/or plasticity.However,the molecular basis of expression divergence remains unclear.Squamosa promoter-binding protein-like(SPL)transcription factors are critical for a wide array of biological processes.In this study,we constructed expression regulatory networks by combining DAP-seq for 40 SPLs,ATACseq,and RNA-seq.Our findings indicate that a group of low-affinity SPL binding regions(SBRs)were targeted by diverse SPLs and caused different sequence preferences around the core GTAC motif.The SBRs including the low-affinity ones are evolutionarily conserved,enriched GWAS signals related to important agricultural traits.However,those SBRs are highly diversified among the cis-regulatory regions(CREs)of syntenic genes,with less than 8%SBRs coexisting in triad genes,suggesting that CRE variations are critical for subgenome differentiations.Knocking out of Ta SPL7A/B/D and Ta SPL15A/B/D subfamily further proved that both high-and low-affinity SBRs played critical roles in the differential expression of genes regulating tiller number and spike sizes.Our results have provided baseline data for downstream networks of SPLs and wheat improvements and revealed that CRE variations are critical sources for subgenome divergence in the allohexaploid wheat.

Complex regulatory network of Betula BplSPL8 in planta

SQUAMOSA-promoter binding protein-like (SPL) proteins are plant-specific transcription factors and participate in different pathways, including the vegetative to reproductive transition, male sterility, biosynthesis of gibberellic acid (GA), plant morphogenesis and response to environmental stress. In this study, we generated transgenic Arabidopsis that overexpressed Betula BplSPL8 and confirmed that BplSPL8 is a transcription factor with transcriptional activation activity and is located in the nucleus. Functional analysis of BplSPL8 showed that it is involved in regulating different development processes: (1) BplSPL8 can delay flowering by reducing sensitivity to GA under short days; (2) BplSPL8 controls the number and morphogenesis of leaves, including up-rolling leaves under long days and folded leaves mediated by GA under short days; (3) BplSPL8 can promote root elongation during late development of roots and inhibit lateral root formation; (4) BplSPL8 may be involved in regulating carotenoid biosynthesis and secretion metabolism. These results show that there is a complex regulatory network for the SPL family genes that is mediated by other components and may provide a new insights for the functional research of SPL genes.

The miR156/SPL Module, a Regulatory Hub and Versatile Toolbox, Gears up Crops for Enhanced Agronomic Traits

In the past two decades, members of the SQUAMOSA-PROMOTER BINDING PROTEIN-LIKE （SPL） family of transcription factors, first identified in Antirrhinum majus, have emerged as pivotal regulators of diverse biological processes in plants, including the timing of vegetative and reproductive phase change, leaf development, tillering/branching, plastochron, panicle/tassel architecture, fruit ripening, fertility, and response to stresses. Transcripts of a subset of SPLs are targeted for cleavage and/or translational repres- sion by microRNA156s （miR156s）. The levels of miR156s are regulated by both endogenous developmental cues and various external stimuli. Accumulating evidence shows that the regulatory circuit around the miR156/SPL module is highly conserved among phylogenetically distinct plant species, and plays impor- tant roles in regulating plant fitness, biomass, and yield. With the expanding knowledge and a mechanistic understanding of their roles and regulatory relationship, we can now harness the miR156/SPL module as a plethora of tools to genetically manipulate crops for optimal parameters in growth and development, and ultimately to maximize yield by intelligent design of crops.

Genome-wide analysis and molecular dissection of the SPL gene family in Salvia miltiorrhiza

SQUAMOSA promoter binding protein-likes （SPLs） are plant-specific transcription factors playing vital regulatory roles in plant growth and development. There is no information about SPLs in Salvia miltiorrhiza （Danshen）, a significant medicinal plant widely used in Traditional Chinese medicine （TCM） for＆gt;1,700 years and an emerging model plant for TCM studies. Through genome-wide identification and subsequent molecular cloning, we identified a total 15 SmSPLs with divergent sequence features, gene structures, and motifs. Comparative analysis showed sequence conservation between SmSPLs and their Arabidopsis counterparts. A phylogenetic tree clusters SmSPLs into six groups. Many of the motifs identified commonly exist in a group/subgroup, implying their functional redundancy. Eight SmSPLs were predicted and experimental y validated to be targets of miR156/157. SmSPLs were differen-tial y expressed in various tissues of S. milltiorrhiza. The expression of miR156/157-targeted SmSPLs was increased with＆amp;nbsp;the maturation of S. miltiorrhiza, whereas the expression of miR156/157 was decreased, confirming the regulatory roles of miR156/157 in SmSPLs and suggesting the functions of SmSPLs in S. miltiorrhiza development. The expression of miR156/157 was negatively correlated with miR172 during the maturation of S. miltiorrhiza. The results indicate the significance and complexity of SmSPL-, miR156-, and miR172-mediated regula-tion of developmental timing in S. miltiorrhiza.