Plant Molecular Biology最新文献

Z. armatum is an economically valued crop known for its rich aroma and medicinal properties. This study identified 45 members of the SQUAMOSA-PROMOTER BINDING PROTEIN LIKE (SPL) gene family in the genome of Z. armatum. Phylogenetic and collinearity analyzes demonstrated a close relationship between ZaSPLs and ZbSPLs from B subgenomes of Zanthoxylum bungeanum. Our miRNA sequencing revealed a high degree of conservation of miR156a within Z. armatum, with the za-miR156a sequence identical to miR156-5p in Arabidopsis thaliana and Citrus sinensis. Of the 45 genes identified by ZaSPLs, 21 were targeted by za-miR156a, transient co-expression experiments in N. benthamiana demonstrated the targeting relationship between za-miR156 and ZaSPL21. Furthermore, RNA-seq and qRT-PCR analysis revealed that ZaSPL genes exhibited elevated expression levels in juvenile tissues of Z. armatum. The expression of nine representative ZaSPL genes were upregulated under polyethylene glycol (PEG) and abscisic acid (ABA). Overexpression of ZaSPL21 delayed the germination of transgenic tobacco and facilitated the flowering process in transgenic N. benthamiana. Significant up-regulation in the expression levels of flowering-related genes such as NbFT1, NbPIP2;1, NbTCP1, NbCOL1, NbGI2, NbGAI1, NbCKX2, and NbARR4 was observed in transgenic plants, suggesting that ZaSPL21 may stimulate plant flowering by regulation of these genes. Furthermore, ZaSPL21 also increased the germination speed of transgenic tobacco seeds during drought and salt stress conditions, and improved the salt tolerance of transgenic seedlings. In conclusion, our study contributes to understanding the functional analysis of the SPL gene family in Z. armatum and emphasizes the crucial role of ZaSPL21 in improving tolerance to salt and promoting flowering. The results offer potential strategies for the further utilization of these genes to improve the salt tolerance of Z. armatum.

Nitrogen (N) is a major plant nutrient and its deficiency can arrest plant growth. However, how low-N stress impair plant growth and its related tolerance mechanisms in peanut seedlings has not yet been explored. To counteract this issue, a hydroponic study was conducted to explore low N stress (0.1 mM NO₃^-) and normal (5.0 mM NO₃^-) effects on the morpho-physiological and molecular attributes of peanut seedlings. Low-N stress significantly decreased peanut plant height, leaf surface area, total root length, and primary root length after 10 days of treatment. Meanwhile, glutamate dehydrogenase, glutamine oxoglutarate aminotransferase activities, chlorophyll, and soluble protein contents were substantially decreased. Impairment in these parameters further suppressed photochemical efficiency (Fv/Fm), and chlorophyll fluorescence parameters (PI_ABS), under low-N stress. Transcriptome sequencing analysis showed a total of 2139 DEGs were identified between the two treatments. KEGG enrichment annotation analysis of DEGs revealed that 119 DEGs related to 10 pathways, including N assimilation, photosynthesis, starch, and sucrose degradation, which may respond to low-N stress in peanuts. Combined with transcriptome, small RNA, and degradome sequencing, we found that PC-3p-142756_56/A.T13EMM (CML3) and PC-5p-43940_274/A.81NSYN (YTH3) are the main modules contributing to low N stress tolerance in peanut crops. Peanut seedlings exposed to N starvation exhibited suppressed gene expression related to nitrate transport and assimilation, chlorophyll synthesis, and carbon assimilation, while also showing improved gene expression in N compensation/energy supply and carbohydrate consumption. Additionally, low N stress tolerance was strongly associated with the miRNA.

In a wake of shifting climatic scenarios, plants are frequently forced to undergo a spectrum of abiotic and biotic stresses at various stages of growth, many of which have a detrimental effect on production and survival. Naturally, microbial consortia partner up to boost plant growth and constitute a diversified ecosystem against abiotic stresses. Despite this, little is known pertaining to the interplay between endophytic microbes which release phytohormones and stimulate plant development in stressed environments. In a lab study, we demonstrated that an endophyte isolated from the Kargil region of India, a Fusarium equiseti strain K23-FE, colonizes the maize hybrid MAH 14 - 5, promoting its growth and conferring polyethylene glycol (PEG)-induced osmotic stress tolerance. To unravel the molecular mechanism, maize seedlings inoculated with endophyte were subjected to comparative transcriptomic analysis. In response to osmotic stress, genes associated with metabolic, photosynthesis, secondary metabolites, and terpene biosynthesis pathways were highly upregulated in endophyte enriched maize seedlings. Further, in a greenhouse experiment, maize plants inoculated with fungal endophyte showed higher relative leaf water content, chlorophyll content, and antioxidant enzyme activity such as polyphenol oxidase (PPO) and catalase (CAT) under 50% field capacity conditions. Osmoprotectant like proline were higher and malondialdehyde content was reduced in colonized plants. This study set as proof of concept to demonstrate that endophytes adapted to adverse environments can efficiently tweak non-host plant responses to abiotic stresses such as water deficit stress via physiological and molecular pathways, offering a huge opportunity for their deployment in sustainable agriculture.

Psa primarily utilises the type III secretion system (T3SS) to deliver effector proteins (T3Es) into host cells, thereby regulating host immune responses. However, the mechanism by which kiwifruit responds to T3SS remains unclear. To elucidate the molecular reaction of kiwifruit plants to Psa infection, M228 and mutant M228△hrcS strains were employed to inoculate Actinidia chinensis var. chinensis for performing comparative transcriptional and metabolomic analyses. Transcriptome analysis identified 973 differentially expressed genes (DEGs) related to flavonoid synthesis, pathogen interaction, and hormone signaling pathways during the critical period of Psa infection at 48 h post-inoculation. In the subsequent metabolomic analysis, flavonoid-related differential metabolites were significantly enriched after the loss of T3SS.Through multi-omics analysis, 22 differentially expressed genes related to flavonoid biosynthesis were identified. Finally, it was discovered that the transient overexpression of 3 genes significantly enhanced kiwifruit resistance to Psa. qRT-PCR analysis indicated that Ac4CL1, Ac4CL3 and AcHCT1 promote host resistance to disease, while Ac4CL3 negatively regulates host resistance to Psa. These findings enrich the plant immune regulation network involved in the interaction between kiwifruit and Psa, providing functional genes and directions with potential application for breeding kiwifruit resistance to canker disease.

Various biological processes are interconnected in plants. Transcription factors (TFs) often act as regulatory hubs to regulate plant growth and responses to stress by integrating various biological pathways. Despite extensive studies on TFs functions in various plant species, our understanding of the details of TFs regulation remains limited. In this study, clonal seedlings of Salvia miltiorrhiza were exposed to specific inhibitors for 12 h. Time-series transcriptome data, sampled hourly, were used to construct co-expression networks and gene regulatory networks (GRNs). Transcriptome dynamic analysis was utilized to capture the gene expression dynamics of various biological processes and decipher the potential molecular mechanisms that regulate these processes. The perturbation results showed the growth and development processes of S.miltiorrhiza were primarily affected at the early stage, whereas stress response-related biological processes were mainly influenced at the later stage. And there was a correlation between the series of key differentially expressed genes in terpenoid biosynthesis pathways and the topological distribution of these pathways. Furthermore, the GRNs based on TFs indicate that TFs play a crucial role in connecting various biological processes. In the cytoplasmic lysate gene regulatory module, SmWRKY48-SmTCP4-SmWRKY28 constituted a regulation hub regulating S.miltiorrhiza responses to perturbation of the MVA pathway. The regulation hub mediated various pathways, including pyruvate metabolism, glycolysis/gluconeogenesis, amino acid metabolism, and ubiquinone and other terpenoid-quinone biosynthesis.Our findings suggest that perturbation of a key biological pathway in S.miltiorrhiza has time-dependent effects on other biological processes. And SmWRKY48-SmTCP4-SmWRKY28 constitutes the regulatory hub in S.miltiorrhiza responses to perturbation of MVA pathway.

The accurate callose deposition plays important roles in pollen wall formation and pollen fertility. As a direct target of miRNA160, ARF17 participate in the formation of the callose wall. However, the impact of ARF17 misexpression in microsporocytes on callose wall formation and pollen fertility remains unknown. Here, the SDS promoter, which is capable of specifically driving gene expression in microsporocytes, was employed to drive the expression of 5mARF17. The pSDS:5mARF17#3 transgenic line were male sterile. TEM revealed that sporopollenin substance was embedded in a thicker callose layer, which resulted in the complete loss of exine structure and pollen abortion in the pSDS:5mARF17#3 line. Consistently, RT-qPCR revealed an increase in the expression of several Cals genes in pSDS:5mARF17#3. EMSA assay demonstrated that ARF17 could bind to the promoter of Cals4 gene, which further suggest that ARF17 could regulate several Cals genes expression. It is notable that the expression of several exine formation-related genes increased significantly in pSDS:5mARF17#3. In conclusion, our findings highlight that the regulation of miRNA160-ARF17 in microsporocytes modulates the thickness of the callose wall, which is crucial for pollen exine formation and intercellular communication.

The lipoxygenase (LOX) gene family is widely distributed in plants, and its activity is closely associated with seed viability and stress tolerance. In this study, we cloned the rice(Oryza sativa)lipoxygenase gene OsLOX1, a key participant in the 13-lipoxygenase metabolic pathway. Our primary focus was to investigate its role in mediating responses to drought stress and seed germination in rice. Histochemical staining and qPCR analysis indicated that the expression level of OsLOX1 was relatively high in leaves and early germinating seeds. Our findings revealed that mutant lines with CRISPR/Cas9-induced knockout of OsLOX1 exhibited reduced tolerance to drought stress compared with the wild-type. This was accompanied by elevated levels of H₂O₂ and malondialdehyde, and a decrease in the expression levels of genes associated with antioxidant enzymes. Furthermore, knockout of OsLOX1 reduced the longevity of rice seeds increased H₂O₂ and MDA levels, and decreased the activities of the antioxidant enzymes superoxide dismutase and catalase, compared with the wild-type. These findings demonstrated that OsLOX1 positively regulated rice seed vigor and drought stress.

Ensuring species integrity and successful reproduction is pivotal for the survival of angiosperms. Members of Brassicaceae family employ a "lock and key" mechanism involving stigmatic (sRALFs) and pollen RALFs (pRALFs) binding to FERONIA, a Catharanthus roseus receptor-like kinase 1-like (CrRLK1L) receptor, to establish a prezygotic hybridization barrier. In the absence of compatible pRALFs, sRALFs bind to FERONIA, inducing a lock state for pollen tube penetration. Conversely, compatible pRALFs act as a key, facilitating successful fertilization. Competing pRALFs reduce the sRALFs binding to FERONIA in a dose-dependent manner, enabling pollen tube penetration. Despite its crucial role in Brassicaceae hybridization, the structural basis of this binding remains elusive owing to the highly flexible nature of RALF peptides. Using advanced structural modeling techniques and flexible peptide molecular docking, this study reveals that pRALFs and sRALFs bind to negatively charged pockets in FERONIA with varying binding affinities. Our study unveils the structural basis of this binding, shedding light on the molecular mechanism underlying hybridization barriers in Brassicaceae.

Arabidopsis MYB transcription factor, AtDUO1 regulates generative cell body (GC) morphogenesis from round to semi and fully elongated forms before pollen mitosis-II (PM II). It was hypothesised that DUO1 might regulate morphogenesis through any of its direct target genes or components of the DUO1-DAZ1 network. The developmental analysis of plants harbouring T-DNA insertions in some DUO1 target genes using light and fluorescence microscopy revealed abnormal GC morphogenesis only in daz1 and daz2, but gcs1, trm16, mapkkk10, mapkkk20, tet11, and tip1 all undergo normal elongation indicating that these target genes have no important roles in morphogenesis or may be redundant. The important regulatory role of DUO1 was confirmed through the observed incomplete rescue of morphogenesis of mutant duo1-1 GCs by DAZ1 and independently by a C-terminally deleted version of DUO1 (DUO1∆C3) lacking activation sequences. The evidence supports the important role of DAZ1 in GC shape partial morphogenesis. The C-terminus of DUO1 may regulate some target genes that affect GC body elongation. Furthermore, an intact DUO1 is shown to be indispensable for GC shape and nucleus elongation and subsequently for timely division and sperm cell morphogenesis. The development of the GC cytoplasmic projection is regulated independently of DUO1, and all its target genes were able to form it.

In plants, cell fate determination is regulated temporally and spatially via a complex of signals consisting of a large number of genetic interactions. Trichome and root hair formation are excellent models for studying cell fate determination in plants. Nowadays, the mysteries underlying the reprograming of trichome and root hair and how nature programs the development of trichome and root hair is an interesting topic in the scientific field. In this review, we discuss the spatial and temporal regulatory networks and cross-talk between phytohormones and epigenetic modifications in the regulation of trichome and root hair initiation in Arabidopsis. The discussion in this review provides a good model for understanding the regulatory mechanism of cell differentiation processes in plants. Moreover, we summarize recent advances in the modulation of trichome and root hair initiation in plants and compare different regulatory mechanisms to help illuminate key goals for future research.