Small secreted peptides (SSPs) are important signals for cell-to-cell communication in plant, involved in a variety of growth and developmental processes, as well as responses to stresses. While a large number of SSPs have been identified and characterized in various plant species, little is known about SSPs in wheat, one of the most important cereal crops. In this study, 4,981 putative SSPs were identified on the wheat genome, among which 1,790 TaSSPs were grouped into 38 known SSP families. The result also suggested that a large number of the putaitive wheat SSPs, Cys-rich peptides in particular, remained to be characterized. Several TaSSP genes were found to encode multiple SSP domains, including CLE, HEVEIN and HAIRPININ domains, and two potentially novel TaSSP family DYY and CRP8CI were identified manually among unpredicted TaSSPs. Analysis on the transcriptomic data showed that a great proportion of TaSSPs were expressed in response to abiotic stresses. Exogenous application of the TaCEPID peptide encoded by TraesCS1D02G130700 enhanced the tolerance of wheat plants to drought and salinity, suggesting porential roles of SSPs in regulating stress responses in wheat.

As a major component of the plant primary cell wall, structure changes in pectin may affect the formation of the secondary cell wall and lead to serious consequences on plant growth and development. Pectin-modifying enzymes including pectate lyase-like proteins (PLLs) participate in the remodeling of pectin during organogenesis, especially during fruit ripening. In this study, we used Arabidopsis as a model system to identify critical PLL genes that are of particular importance for vascular development. Four PLL genes, named AtPLL15, AtPLL16, AtPLL19, and AtPLL26, were identified for xylem-specific expression. A knock-out T-DNA mutant of AtPLL16 displayed an increased amount of pectin, soluble sugar, and acid-soluble lignin (ASL). Interestingly, the atpll16 mutant exhibited an irregular xylem phenotype, accompanied by disordered xylem ray cells and an absence of interfascicular phloem fibers. The xylem fiber cell walls in the atpll16 mutant were thicker than those of the wild type. On the contrary, AtPLL16 overexpression resulted in expansion of the phloem and a dramatic change in the xylem-to-phloem ratios. Altogether, our data suggest that AtPLL16 as a pectate lyase plays an important role during vascular development in Arabidopsis.

As one of the major groups of small post-translationally modified peptides, the CLV3/EMBRYO SURROUNDING REGION-RELATED (CLE)-like (CLEL) peptide family has been reported to regulate root growth, lateral root development and plant gravitropic responses in Arabidopsis thaliana. In this study, we identified 12 CLEL genes in Populus trichocarpa and performed a comprehensive bioinformatics analysis on these genes. Among them, five P. trichocarpa CLELs (PtrCLELs) were revised with new gene models. All of these PtrCLEL proteins were structurally similar to the A. thaliana CLELs (AtCLELs), including an N-terminal signal peptide, a conserved C-terminal 13-amino-acid CLEL motif and a variable intermediate region. In silico and quantitative real-time PCR analyses showed that PtrCLELs were widely expressed in various tissues, including roots, leaves, buds and stems. Exogenous application of chemically synthesized PtrCLEL peptides resulted in wavy or curly roots and reduced lateral root formation in A. thaliana. Moreover, germinating Populus deltoides seedlings on a growth medium containing these peptides caused the roots to thicken and to form abnormal lateral roots, in many cases in clusters. Anatomical and histological changes in thickened roots were further investigated by treating Populus 717 cuttings with the PtrCLEL10 peptide. We observed that root thickening was mainly due to an increased number of cells in the epidermis, hypodermis and cortex. The results of our study suggested that PtrCLEL and AtCLEL genes encode proteins with similar protein structures, sequences of peptide motif and peptide activities on developing roots. The activities of PtrCLEL peptides in root development were species-dependent.

Holotrichia parallela is a globally distributed soil-dwelling pest that poses a major threat to peanut cultivation in China. Neuropeptides, as critical signaling molecules, regulate multiple physiological and behavioral processes in insects and represent highly promising targets for pest management. To date, the functional characteristics of neuropeptides in H. parallela remain unreported. In this study, we isolated and cloned one NPF and one NPFR gene, respectively. Bioinformatics analysis revealed that alternative splicing of the NPF gene produces two transcript variants, NPFa (255 bp) and NPFb (369 bp). The NPFR gene spans a length of 1188 bp, encoding 395 amino acids that contain seven α-helical transmembrane domains, indicating that it belongs to the family A G protein-coupled receptor (GPCR) family. Spatiotemporal expression profiles demonstrated that NPF was most abundant in the adult brain, whereas NPFR was highly enriched in the brain and antennae. NPF expression peaked in second-to-third-instar larvae, while NPFR was highly expressed in eggs. Starvation stress significantly upregulated the expression of both genes. RNA interference (RNAi)-mediated silencing of NPF and NPFR significantly reduced food intake, female fecundity, and glycogen content in adults. These findings enhance our understanding of insect neuropeptides signaling networks and support the development of behavior-based pest control strategies.

Phytosulfokines (PSKs) are sulfated peptide hormones involved in plant growth, development, and stress responses. However, direct genetic evidence for the involvement of PSK peptide in cambium development and secondary vascular development in woody plants remains largely lacking. Here, we identified nine PSK family members in Populus trichocarpa. Phylogenetic, gene structure, conserved motif, and local synteny analyses indicated that the poplar PSK family is evolutionarily conserved, and some members may have arisen from the whole-genome duplication (WGD) event. Subsequently, we generated overexpression and CRISPR/Cas9-mediated knockout lines of PtrPSK2a in hybrid poplar 717. PtrPSK2a is preferentially expressed in stems, particularly in the cambium and developing xylem. Both gain-of-function and loss-of-function of PtrPSK2a exhibited growth defects, characterized by reduced plant height and stem diameter, with more severe overall growth defects in OE lines. Consistently, histological analysis demonstrated that cambium activity and secondary xylem development were significantly inhibited in both transgenic lines, as reflected by fewer cambium cell layers, narrower xylem, and fewer vessels in young stems. In addition, genes associated with cambial development, vessel differentiation, and secondary cell wall formation were broadly downregulated, accompanied by altered expression of several hormone-related marker genes in transgenic lines. Together, these findings demonstrate that PtrPSK2a is essential for cambial activity and xylem development during secondary growth in poplar.