Su-Sheng Gan

Whether somatically acquired traits can contribute to heritable variation has remained an open question in plant biology for over a century. Here, using graft-induced periclinal chimeras between Brassica juncea and Brassica oleracea , we identified extrachromosomal circular DNA (eccDNA) as mobile genetic elements capable of crossing histological boundaries and entering the germline. We demonstrated that grafting could promote the horizontal transfer of eccDNA between somatic cell layers, enabling its stable maintenance in recipient tissues and permitting transmission through sexual reproduction across multiple generations. We found that transmitted eccDNA is non-randomly distributed across the genome, preferentially originating from gene-dense regions, and enriched for features associated with molecular persistence, such as inverted repeats, hairpin-forming sequences, and autonomously replicating sequence (ARS) consensus motifs. These genetic regions conferred replication competence, as evidenced by autonomous propagation in an ARS-less heterologous yeast system. Sexual progeny carrying graft-acquired eccDNA exhibited reproducible and lineage-dependent alterations in leaf morphology and drought tolerance, accompanied by coordinated transcriptional reprogramming. Notably, a subset of inherited eccDNA remained transcriptionally active in progeny, producing transcripts absent from self-grafted controls. Our findings establish eccDNA as heritable extrachromosomal elements that link graft-mediated somatic genetic transfer with stable germline transmission, thereby expanding the molecular scope of heritable variation in plants and providing a conceptual framework for graft-based trait transmission.

Gaseous phytohormone ethylene regulates various aspects of plant development. Ethylene is perceived by ER membrane-localized receptors, which are inactivated upon binding with ethylene molecules, thereby initiating ethylene signal transduction. Here, we report that a novel E3 ligase RING finger for Ethylene receptor Degradation (RED) and its E2 partner UBC32 ubiquitinate ethylene-bound receptors for degradation through an ER associated degradation (ERAD) pathway in both Rosa hybrida and Solanum lycopersicum. The depletion of RED or UBC32 leads to hypersensitivity to ethylene, which is manifested as premature leaf abscission and petal shedding in roses, as well as the dwarf plants and accelerated fruit ripening in tomatoes. Disruption of the conserved ethylene binding site of receptors prevents RED-mediated degradation of the receptors. Our study discovers an ERAD branch that facilitates the ethylene-induced degradation of receptors, and provides insights into how the plant’s response to ethylene can be controlled by modulating the turnover of ethylene receptors.

We previously reported that ABA inhibits stomatal closure through AtNAP-SAG113 PP2C regulatory module during leaf senescence. The mechanism by which this module exerts its function is unknown. Here we report the identification and functional analysis of SAG114, a direct target of the regulatory module. SAG114 encodes SnRK3.25. Both bimolecular fluorescence complementation (BiFC) and yeast two-hybrid assays show that SAG113 PP2C physically interacts with SAG114 SnRK3.25. Biochemically the SAG113 PP2C dephosphorylates SAG114 in vitro and in planta. RT-PCR and GUS reporter analyses show that SAG114 is specifically expressed in senescing leaves in Arabidopsis. Functionally, the SAG114 knockout mutant plants have a significantly bigger stomatal aperture and a much faster water loss rate in senescing leaves than those of wild type, and display a precocious senescence phenotype. The premature senescence phenotype of sag114 is epistatic to sag113 (that exhibits a remarkable delay in leaf senescence) because the sag113 sag114 double mutant plants show an early leaf senescence phenotype, similar to that of sag114. These results not only demonstrate that the ABA-AtNAP-SAG113 PP2C regulatory module controls leaf longevity by dephosphorylating SAG114 kinase, but also reveal the involvement of the SnRK3 family gene in stomatal movement and water loss during leaf senescence. Graphical Abstract

Summary: The vascular cambium is the main secondary meristem in plants that produces secondary phloem (outside) and xylem (inside) on opposing sides of the cambium. The phytohormone ethylene has been implicated in vascular cambium activity, but the regulatory network underlying ethylene‐mediated cambial activity remains to be elucidated. Here, we found that PETAL MOVEMENT‐RELATED PROTEIN1 (RhPMP1), an ethylene‐inducible HOMEODOMAIN‐LEUCINE ZIPPER I transcription factor in woody plant rose (Rosa hybrida), regulates local auxin biosynthesis and auxin transport to maintain cambial activity. Knockdown of RhPMP1 resulted in smaller midveins and reduced auxin content, while RhPMP1 overexpression resulted in larger midveins and increased auxin levels compared with the wild‐type plants. Furthermore, we revealed that Indole‐3‐pyruvate monooxygenase YUCCA 10 (RhYUC10) and Auxin transporter‐like protein 2 (RhAUX2), encoding an auxin biosynthetic enzyme and an auxin influx carrier, respectively, are direct downstream targets of RhPMP1. In summary, our results suggest that ethylene promotes an auxin maximum in the cambium adjacent to the xylem to maintain cambial activity.

Increasingly warming temperature impacts on all aspects of growth and development in plants. Flower development is a complex process that is very sensitive to ambient temperature, and warming temperatures often lead to abnormal flower development and remarkably reduce the quality and yield of inflorescent vegetables and many other crops, which can be exemplified by Brassica oleracea cv. Green Harmony F1, a broccoli cultivar, whose floral development is ceased at inflorescence meristem (at 28 °C) or floral primordium stage (at 22 °C), forming a cauliflower-like curd (28 °C) or intermediate curd (22 °C) instead of normal broccoli head at 16 °C. However, the underlying molecular regulatory mechanisms are not well understood. Here we report that warming temperature (28 °C or 22 °C) induced hypermethylation of the genome, especially the promoter regions of such sets of genes as ribosome biogenesis-related and others, leading to the suppression of the apex-highly-expressed distinctive genes, subsequently resulting in the abnormal floral development, as revealed by methylome and transcriptome co-profiling. The regulation of warming-induced abnormal floral development in broccoli was further verified by the fact that the DNA methylation inhibitor 5-azacytidine (5-azaC) released the expression of genes from the warming temperature-induced suppression, and restored the broccoli development to normalcy at warming temperature. The research provided new approaches to breeding broccoli and other crops for growing in wider or warmer temperature zones.

Salicylic acid (SA) is an important plant hormone that regulates defense responses and leaf senescence. It is imperative to understand upstream factors that regulate genes of SA biosynthesis. SAG202/SARD1 is a key regulator for isochorismate synthase 1 (ICS1) induction and SA biosynthesis in defense responses. The regulatory mechanism of SA biosynthesis during leaf senescence is not well understood. Here we show that AtNAP, a senescence-specific NAC family transcription factor, directly regulates a senescence-associated gene named SAG202 as revealed in yeast one-hybrid and in planta assays. Inducible overexpreesion of AtNAP and SAG202 lead to high levels of SA and precocious senescence in leaves. Individual knockout mutants of sag202 and ics1 have markedly reduced SA levels and display a significantly delayed leaf senescence phenotype. Furthermore, SA positively feedback regulates AtNAP and SAG202. Our research has uncovered a unique positive feedback regulatory loop, SA-AtNAP-SAG202-ICS1-SA, that operates to control SA biosynthesis associated with leaf senescence but not defense response.

Cytokinins (CKs) are a class of adenine-derived plant hormones that plays pervasive roles in plant growth and development including cell division, morphogenesis, lateral bud outgrowth, leaf expansion and senescence. CKs as a “fountain of youth” prolongs leaf longevity by inhibiting leaf senescence, and therefore must be catabolized for senescence to occur. AtNAP, a senescence-specific transcription factor has a key role in promoting leaf senescence. The role of AtNAP in regulating CK catabolism is unknown. Here we report the identification and characterization of AtNAP-AtCKX3 (cytokinin oxidase 3) module by which CKs are catabolized during leaf senescence in Arabidopsis. Like AtNAP, AtCKX3 is highly upregulated during leaf senescence. When AtNAP is chemically induced AtCKX3 is co-induced; and when AtNAP is knocked out, the expression of AtCKX3 is abolished. AtNAP physically binds to the cis element of the AtCKX3 promoter to direct its expression as revealed by yeast one-hybrid assays and in planta experiments. Leaves of the atckx3 knockout lines have higher CK concentrations and a delayed senescence phenotype compared with those of WT. In contrast, leaves with inducible expression of AtCKX3 have lower CK concentrations and exhibit a precocious senescence phenotype compared with WT. This research reveals that AtNAP transcription factor˗AtCKX3 module regulates leaf senescence by connecting two antagonist plant hormones abscisic acid and CKs.