Genome-wide characterization of soybean RALF genes and their expression responses to Fusarium oxysporum.

RALFs (RAPID ALKALINIZATION FACTORs) are small peptides required for plant growth, development and immunity. RALF has recently been discovered to regulate plant resistance to fungal infection. However, little is known in crops, particularly in soybean. Here, 27 RALFs were identified in the genome of Glycine max. All Glycine max RALFs (GmRALFs) and 34 Arabidopsis RALFs were classified into 12 clades via the phylogenetic analyses. Gene structures, conserved motifs, chromosome distribution and cis-elements were analyzed in this study. Furthermore, 18 GmRALFs were found in response to Fusarium oxysporum (F. oxysporum) infection in soybean and to have distinct expression patterns. Among them, secretory function of two GmRALFs were identified, and three GmRALFs were detected to interact with FERONIA in Glycine max (GmFERONIA, GmFER). Our current study systematically identified and characterized GmRALFs in the soybean genome, laying a groundwork for further functional analyses and soybean breeding.

Multifaceted roles of the ERECTA family in plant organ morphogenesis.

Receptor-like kinases (RLKs) can participate in multiple signalling pathways and are considered one of the most critical components of the early events of intercellular signalling. As an RLK, the ERECTA family (ERf), which comprises ERECTA (ER), ERECTA-Like1 (ERL1), and ERECTA-Like2 (ERL2) in Arabidopsis, regulates multiple signalling pathways in plant growth and development. Despite its indispensability, detailed information on ERf-manipulated signalling pathways remains elusive. In this review, we attempt to summarize the essential roles of the ERf in plant organ morphogenesis, including shoot apical meristem, stem, and reproductive organ development.

Gibberellins modulate shade-induced soybean hypocotyl elongation downstream of the mutual promotion of auxin and brassinosteroids.

Plants and crops are widely suffered by shade stress in the natural communities or in the agricultural fields. The three main phytohormones auxin, gibberellins (GAs) and brassinosteroids (BRs) were found essential in shade avoidance in Arabidopsis. However, their relationship have been seldom reported in plant shade avoidance control. Here, we report our investigation of the crosstalk of auxin, GAs and BRs in shade-induced hypocotyl elongation of soybean. Exogenous feeding of indol-3-acetic acid (IAA), GA3 or 24-epibrassinolide (EBL) distinctly promoted hypocotyl elongation in the white light, while the potent biosynthesis inhibitors of GA3, IAA, BRs severely diminished shade-induced hypocotyl elongation. Synergistic treatment of their biosynthesis inhibitors showed that GA3 fully, while EBL slightly, restored the hypocotyl elongation that was efficiently repressed by IAA biosynthesis inhibitor, GA3 and IAA dramatically suppressed the hypocotyl growth inhibition by BR biosynthesis inhibitor in the shade, whereas both IAA and EBL feeding cannot suppress the elongation inhibition by GA biosynthesis inhibitor. Further analyses revealed that shade remarkably upregulated expression of key genes of IAA, GA and BR biosynthesis in the soybean hypocotyls, and GA biosynthesis genes were effectively blocked by IAA, GA and BR biosynthesis inhibitors in the shade. Taken together, these results suggest that GAs modulate shade-induced hypocotyl elongation downstream of mutual promotion of auxin and BRs in soybean.

Quantitative proteomic analyses identified multiple sugar metabolic proteins in soybean under shade stress.

Soybean-based intercropping is important for sustainable agricultural practice on a regional and global scale. However, most soybean varieties use shade avoidance strategy to acquire more light absorption when suffered in canopy shade in intercropping systems, thus reduced the yield of the whole population on a farmland. The mechanisms underlying early response of soybean in shade avoidance is still largely unknown. Here we report our identification of differentially accumulated proteins in shade-sensitive soybean seedlings by global quantitative proteome analysis under white light (WL) and shade conditions. By using Tandem Mass Tag (TMT) labelling and HPLC fractionation followed by high-resolution LC-MS/MS analysis, 29 proteins were found up-regulated and 412 proteins were found down-regulated in soybean seedlings by 2-h shade stress than that by 2-h WL treatment. Multiple differentially expressed proteins are enriched in carbohydrate metabolic process especially in the biosynthetic pathways of cell wall polysaccharides in soybean seedlings by shade stress comparing to those in WL growth conditions. Physiological assays showed that saccharides were rapidly declined in shoot apex of soybean seedlings under a short-term shading. Our results would provide new insights into the mechanisms of shade avoidance responses in soybean seedlings.

Characterization of a splice variant of soybean ERECTA devoid of an intracellular kinase domain in response to shade stress.

The receptor-like kinase ERECTA (ER) plays vital roles in plant adaptation under environmental stress including shade avoidance in Arabidopsis. In our previous study, we identified four ER paralogues in soybean (GmERs) that showed high similarities to Arabidopsis ER. Each GmER was predicted to generate diverse alternative splicing variants. However, whether soybean GmERs contribute to shade avoidance is unknown. Here we report our characterization of GmERs in response to shading. Promoter::GUS staining analysis shows that expression of GmER paralogous genes was differentially induced under shade stress. Further analyses show that GmERa.1 and GmERa.2 exhibit a larger distinction in length than the other GmER variants. GmERa.2 has the shortest length of amino acid with only 15 leucine-rich repeats which is the part of the extracellular domain of GmERa.1. Overexpression of GmERa.2 fully rescued the hypocotyl length, leaf area and petiole length, and the sensitivity of the hypocotyl of Arabidopsis mutant er-3 to shading, suggesting that the truncated extracellular domain of GmERa might contribute importantly to shade avoidance.

Auxin and Gibberellins Are Required for the Receptor-Like Kinase ERECTA Regulated Hypocotyl Elongation in Shade Avoidance in Arabidopsis.

Plants use shade avoidance strategy to escape the canopy shade when grown under natural conditions. Previous studies showed that the Arabidopsis receptor-like kinase ERECTA (ER) is involved in shade avoidance syndrome. However, the mechanisms of ER in modulating SAR by promoting hypocotyl elongation are unknown yet. Here, we report that ER regulated hypocotyl elongation in shade avoidance requires auxin and gibberellins (GAs). The T-DNA insertional ER mutant er-3 shows a less hypocotyl length than that in Col-0 wild type. Promoter::GUS staining analysis shows that ER and its paralogous genes ERECTA-LIKE1 (ERL1) and ERECTA-LIKE2 (ERL2) are differentially expressed in the seedlings, of which only ER is most obviously upregulated in the hypocotyl by shade treatment. Exogenous feeding assay by using media-application with vertical-grown of Arabidopsis seedlings showed that the hypocotyl length of er-3 is partially promoted by indol-3-acetic acid (IAA), while it is relatively insensitive of er-3 to various concentrations of IAA than that of Col-0. Hypocotyl elongation of er-3 is promoted similar to that of Col-0 by high temperature in the white light condition, but the elongation was not significantly affected by the treatment of the auxin transport inhibitor 1-N-naphthylphthalamic acid (NPA). Exogenous GA3 increased the hypocotyl elongation of both er-3 and the wild type in the shade condition, and the GA3 biosynthesis inhibitor paclobutrazol (PAC) severely inhibits the hypocotyl elongation of Col-0 and er-3. Further analysis showed that auxin biosynthesis inhibitors yucasin and L-kynurenine remarkably inhibited the hypocotyl elongation of er-3 while yucasin shows a more severe inhibition to er-3 than Col-0. Relative expression of genes regulating auxin homeostasis and signaling, and GA homeostasis is less in er-3 than that in Col-0. Furthermore, genetic evidences show that ER regulated hypocotyl elongation is dependent of PHYTOCHROME B (PHYB). Overall, we propose that ER regulated shade avoidance by promoting hypocotyl elongation is PHYB-dependent and requires auxin and GAs.