Publications

Rice blast is one of the main diseases of rice, causing severe economic losses to agricultural production; thus, the search for blast resistance is a top priority for rice breeding. When challenged by the blast causal fungus Magnaporthe oryzae the expression level of OsMAPKKK69 gene in rice cultivar Nipponbar was found to increase significantly. Such an induction was also found in a different genetic material, cultivar Shufanggaonuo, indicating that OsMAPKKK69 plays an important role in blast disease response. However, the function of OsMAPKKK69 remains unclear. In this study, wild type ZH11 was selected as the background material to investigate the expression and functions of OsMAPKKK69 in rice disease resistance by constructing knockout mutants. The results showed that OsMAPKKK69 is mainly expressed in four-week-old shoots and localized in cell membrane, cytoplasm, and nucleus. The two allelic knockout mutants, osmapkkk69-1 and osmapkkk69-2, were more resistant to M. oryzae and bacterial blight Xanthomonas oryzae pv. Oryzae (Xoo). Further agronomic trait analysis revealed that the osmapkkk69-1 and osmapkkk69-2 mutants had reduced plant height, smaller grain size, a significant increase in tillering number, but also a significant increase in yield per plant. Our results show that OsMAPKKK69 is involved in the immune response of rice by negatively regulating the resistance to rice blast and blight diseases, and in regulating important agronomic traits. This study lays a foundation for revealing the molecular mechanism of OsMAPKKK69 in the immune response to rice diseases and provides novel genetic resources for rice breeding.

Alternative polyadenylation (APA) generates transcript diversity by producing mRNA isoforms with distinct 3' ends. Despite the critical roles that APA plays in various biological processes, the mechanisms regulating APA in response to stresses have remained poorly understood in plants. Here, we perform comprehensive analysis of APA in tomato, and focus on a phosphate (Pi)- regulated APA gene SlSPX5, encoding a putative Pi sensor protein. SlSPX5 interacts with and sequesters the transcription factor SlPHL1 in the cytosol, thereby inhibiting the expression of Pi starvation inducible genes. We discover that a cis-natural antisense RNA (cis-NAT) is activated from SlSPX5 to promote its proximal polyadenylation under Pi-depleted conditions. The transcription of this cis-NAT induces RNA Polymerase II pausing, generating Ser2 phosphorylation signals that recruit polyadenylation machinery to the 5' end of SlSPX5. Our findings demonstrate that a cis-NAT regulates APA of its cognate gene in response to Pi starvation.

Alternative polyadenylation of pre-mRNA generates mRNAs with alternative 3' ends, thereby increasing the complexity of the transcriptome and proteome. This process is carried out by pre-mRNA 3' end processing complexes, with the specificity and site selection of polyadenylation primarily defined by the interaction between these complexes and conserved pre-mRNA cis-elements. However, the cis-element recognized by the polyadenylation CFII complex and the downstream cis-element of the poly(A) site remain unknown. Here, FPA, a conserved Spen family RNA-binding protein, is identified to physically interacts with the CFII complex in Arabidopsis. FPA specifically binds to a GA-rich cis-element downstream of the proximal poly(A) site, which is required for the CFII complex to promote proximal poly(A) site usage and prevent 3' extension. Thus, the CFII complex recognizes the downstream cis-element of the poly(A) site through its interaction with FPA, highlighting the role of RNA-binding proteins as accessory factors in alternative polyadenylation.

Co-transcriptional cleavage and transcription termination are closely related processes during mRNA maturation, yet their coordination remains poorly understood due to difficulties in detecting these transient events. Here, we applied single-molecule nascent RNA sequencing to simultaneously capture the cleavage status and readthrough distance on the same nascent RNA molecules and characterize 14 mutants of various pre-mRNA processing factors in Arabidopsis. Our results reveal diverse roles for these processing factors in coordinating cleavage and termination: core components of CPSF and CstF complex stimulate both cleavage and termination, facilitating access to exoribonuclease AtXRN3; mutations in nuclear poly(A) polymerase PAPS1 and AtXRN3 caused delayed termination with minimal effects on cleavage, suggesting their roles are further downstream; BORDER proteins facilitate termination while simultaneously inhibiting cleavage, suggesting a complex interplay between these two actions; the phosphatase SSU72 specifically promotes efficient termination without affecting cleavage. Our method also enables us to distinguish cleaved readthrough transcripts from full-length readthrough, and we found termination factor FPA specifically promotes termination of cleaved readthrough, suggesting FPA facilitates access of AtXRN3 to the 3' cleavage product. Our comprehensive datasets reveal cleavage and termination are highly coordinated during pre-mRNA processing.

Photosynthesis in mangroves contributes to one of the most carbon-rich ecosystems on Earth and plays a significant role in mitigating global climate change. However, the mechanisms underlying the high productivity of mangroves remain largely unexplored. Through anatomical analyses, we found that mangrove species with higher biomass production, such as Sonneratia apetala, exhibit isobilateral leaves, which enhance light harvesting and reduce light inhibition, resulting in higher photosynthetic yields. Transcriptomic and genomic analyses revealed the molecular processes underlying the formation of isobilateral leaves. We found that auxin is rapidly synthesized and works in coordination with gibberellin and brassinosteroid in the isobilateral leaves of S. apetala. Interestingly, we identified a group of genes related to adaxial-abaxial leaf polarity in S. apetala, with upregulated genes associated with chlorophyll synthesis, adaxial cell identity, and erect leaf growth, while genes related to the recognition of adaxial cell boundaries - possibly related to the lower palisade tissues - were downregulated. Additionally, we identified amino acid substitutions and changes in promoter cis-acting elements in Indole-3-acetic acid carboxylmethyltransferase 1 (IAMT1) in Sonneratia species. These findings provide new insights into the formation of isobilateral leaves in mangroves and their adaptation to intertidal high-light coastal conditions.

Seed germination is a critical developmental stage in the lifecycle of plants, and its regulation is essential for ensuring crop productivity, particularly under adverse environmental conditions. Here, we find that the Arabidopsis thaliana Pre-mRNA PROCESSING FACTOR 21 (PRP21) is crucial for regulating the abscisic acid (ABA) response and seed germination. Our RNA deep sequencing and poly(A) tag sequencing analyses reveal that PRP21 is involved in pre-mRNA splicing, genome-wide gene expression, and mRNA 3' end processing, highlighting its multifunctional role in gene regulation. Furthermore, PRP21 interacts with various splicing factors and small nuclear ribonucleoproteins, confirming its involvement in spliceosome assembly. Additionally, we demonstrate that PRP21 negatively regulates the expression of ABA-responsive genes, such as ABI3, ABI5, EM1, and EM6, thereby modulating ABA response and seed germination. Our findings underscore the importance of PRP21 in coordinating transcriptional and post-transcriptional processes and provide insights into the molecular mechanisms underlying seed germination, potentially guiding crop improvement for stress tolerance.

Parental or ancestral environments can induce heritable phenotypic changes, but whether such environment-induced heritable changes are a common phenomenon remains unexplored. Here, we subject 14 genotypes of Arabidopsis thaliana to 10 different environmental treatments and observe phenotypic and genome-wide gene expression changes over four successive generations. We find that all treatments caused heritable phenotypic and gene expression changes, with a substantial proportion stably transmitted over all observed generations. Intriguingly, the susceptibility of a genotype to environmental inductions could be predicted based on the transposon abundance in the genome. Our study thus challenges the classic view that the environment only participates in the selection of heritable variation and suggests that the environment can play a significant role in generating of heritable variations.

Precise regulation of gene expression is crucial for plant survival. As a cotranscriptional regulatory mechanism, pre-mRNA polyadenylation is essential for fine-tuning gene expression. Polyadenylation can be alternatively projected at various sites of a transcript, which contributes to transcriptome diversity. Epigenetic modification is another mechanism of transcriptional control. Recent studies have uncovered crosstalk between cotranscriptional polyadenylation processes and both epigenomic and epitranscriptomic markers. Genetic analyses have demonstrated that DNA methylation, histone modifications, and epitranscriptomic modification are involved in regulating polyadenylation in plants. Here we summarize current understanding of the links between epigenetics and polyadenylation and their novel biological efficacy for plant development and environmental responses. Unresolved issues and future directions are discussed to shed light on the field.