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Premature abscission of flowers and fruits triggered by low light stress can severely reduce crop yields. However, the underlying molecular mechanism of this organ abscission is not fully understood. Here, we show that a gene (SlCLV3) encoding CLAVATA3 (CLV3), a peptide hormone that regulates stem cell fate in meristems, is highly expressed in the pedicel abscission zone (AZ) in response to low light in tomato (Solanum lycopersicum). SlCLV3 knockdown and knockout lines exhibit delayed low light-induced flower drop. The receptor kinases SlCLV1 and BARELY ANY MERISTEM1 function in the SlCLV3 peptide-induced low light response in the AZ to decrease expression of the transcription factor gene WUSCHEL (SlWUS). DNA affinity purification sequencing identified the transcription factor genes KNOX-LIKE HOMEDOMAIN PROTEIN1 (SlKD1) and FRUITFULL2 (SlFUL2) as SlWUS target genes. Our data reveal that low light reduces SlWUS expression, resulting in higher SlKD1 and SlFUL2 expression in the AZ, thereby perturbing the auxin response gradient and causing increased ethylene production, eventually leading to the initiation of abscission. These results demonstrate that the SlCLV3-SlWUS signaling pathway plays a central role in low light-induced abscission by affecting auxin and ethylene homeostasis.

Plant organ abscission, a process that is important for development and reproductive success, is inhibited by the phytohormone auxin and promoted by another phytohormone, jasmonic acid (JA). However, the molecular mechanisms underlying the antagonistic effects of auxin and JA in organ abscission are unknown. We identified a tomato (Solanum lycopersicum) class III homeodomain-leucine zipper transcription factor, HOMEOBOX15A (SlHB15A), which was highly expressed in the flower pedicel abscission zone and induced by auxin. Knocking out SlHB15A using clustered regularly interspaced short palindromic repeats-associated protein 9 technology significantly accelerated abscission. In contrast, overexpression of microRNA166-resistant SlHB15A (mSlHB15A) delayed abscission. RNA sequencing and reverse transcription-quantitative PCR analyses showed that knocking out SlHB15A altered the expression of genes related to JA biosynthesis and signaling. Furthermore, functional analysis indicated that SlHB15A regulates abscission by depressing JA-isoleucine (JA-Ile) levels through inhabiting the expression of JASMONATE-RESISTANT1 (SlJAR1), a gene involved in JA-Ile biosynthesis, which could induce abscission-dependent and abscission-independent ethylene signaling. SlHB15A bound directly to the SlJAR1 promoter to silence SlJAR1, thus delaying abscission. We also found that flower removal enhanced JA-Ile content and that application of JA-Ile severely impaired the inhibitory effects of auxin on abscission. These results indicated that SlHB15A mediates the antagonistic effect of auxin and JA-Ile during tomato pedicel abscission, while auxin inhibits abscission through the SlHB15A-SlJAR1 module.

Auxin plays a central role in control of organ abscission, and it is thought that changes in the auxin gradient across the abscission zone are the primary determinant of the onset of abscission. The nature of this gradient, whether in concentration, flow, or perhaps in the response system has not conclusively been determined. We employed a DR5::GUS auxin response reporter system to examine the temporal and spatial distribution of the auxin response activity in response to developmental and environmental cues during pedicel abscission in tomato. In pedicels of young and fully open flowers, auxin response, as indicated by GUS activity, was predominantly detected in the vascular tissues and was almost entirely confined to the abscission zone (AZ) and to the distal portion of the pedicel, with a striking reduction in the proximal tissues below the AZ-a 'step', rather than a gradient. Following pollination and during early fruit development, auxin response increased substantially throughout the pedicel. Changes in GUS activity following treatments that caused pedicel abscission (flower removal, high temperature, darkness, ethylene, or N-1-naphthylphthalamic acid (NPA) treatment) were relatively minor, with reduced auxin response in the AZ and some reduction above and below it. Expression of genes encoding some auxin efflux carriers (PIN) and influx carriers (AUX⁄LAX) was substantially reduced in the abscission zone of NPA-treated pedicels, and in pedicels stimulated to abscise by flower removal. Our results suggest that changes in auxin flow distribution through the abscission zone are likely more important than the auxin response system in the regulation of abscission.

In many fruiting plant species, flower abscission is induced by low light stress. Here, we elucidated how signaling mediated by the peptide INFLORESCENCE DEFICIENT IN ABSCISSION (IDA) controls low light-induced flower drop in tomato (Solanum lycopersicum). We analyzed the expression patterns of an IDA-Like gene (SlIDL6) during low light-induced flower abscission, and used tandem mass spectrometry to identify and characterize the mature SlIDL6 peptide. Tomato knockout lines were created to investigate the in vivo function of SlIDL6. In addition, yeast one-hybrid assays were used to investigate the binding of the SlWRKY17 transcription factor to the SlIDL6 promoter, and silencing of SlWRKY17 expression delayed low light-induced flower abscission. SlIDL6 was specifically expressed in the abscission zone and at high levels during low light-induced abscission and ethylene treatment. SlIDL6 knockout lines showed delayed low light-induced flower drop, and the application of SlIDL6 peptide accelerated abscission. Overexpression of SlIDL6 rescued the ida mutant phenotype in Arabidopsis (Arabidopsis thaliana), suggesting functional conservation between species. SlIDL6-mediated abscission was via an ethylene-independent pathway. We report a SlWRKY17-SlIDL6 regulatory module that functions in low light promoted abscission by increasing the expression of enzymes involved in cell wall remodeling and disassembly.

High temperatures are a major threat to plant growth and development, leading to yield losses in crops. Calcium-dependent protein kinases (CPKs) act as critical components of Ca2+ sensing in plants that transduce rapid stress-induced responses to multiple environmental stimuli. However, the role of CPKs in plant thermotolerance and their mechanisms of action remain poorly understood. To address this issue, tomato (Solanum lycopersicum) cpk28 mutants were generated using a CRISPR-Cas9 gene-editing approach. The responses of mutant and wild-type plants to normal (25°C) and high temperatures (45°C) were documented. Thermotolerance was significantly decreased in the cpk28 mutants, which showed increased heat stress-induced accumulation of reactive oxygen species (ROS) and levels of protein oxidation, together with decreased activities of ascorbate peroxidase (APX) and other antioxidant enzymes. The redox status of ascorbate and glutathione were also modified. Using a yeast two-hybrid library screen and protein interaction assays, we provide evidence that CPK28 directly interacts with cytosolic APX2. Mutations in APX2 rendered plants more sensitive to high temperatures, whereas the addition of exogenous reduced ascorbate (AsA) rescued the thermotolerance phenotype of the cpk28 mutants. Moreover, protein phosphorylation analysis demonstrated that CPK28 phosphorylates the APX2 protein at Thr-59 and Thr-164. This process is suggested to be responsive to Ca2+ stimuli and may be required for CPK28-mediated thermotolerance. Taken together, these results demonstrate that CPK28 targets APX2, thus improving thermotolerance. This study suggests that CPK28 is an attractive target for the development of improved crop cultivars that are better adapted to heat stress in a changing climate.

Abscission of plant organs is induced by developmental signals and diverse environmental stimuli and involves multiple regulatory networks, including biotic or abiotic stress-impaired auxin flux in the abscission zone (AZ). Depletion of auxin activates AZ ethylene (ETH) production and triggers acceleration of abscission, a process that requires hydrogen peroxide (H2O2). However, the interaction between these networks and the underlying mechanisms that control abscission are poorly understood. Here, we found that expression of tonoplast intrinsic proteins, which belong to the aquaporin (AQP) family in the AZ was important for tomato (Solanum lycopersicum) pedicel abscission. Liquid chromatography-tandem mass spectrometry and in situ hybridization revealed that SlTIP1;1 was most abundant and specifically present in the tomato pedicel AZ. SlTIP1;1 localized in the plasma membrane and tonoplast. Knockout of SlTIP1;1 resulted in delayed abscission, whereas overexpression of SlTIP1;1 accelerated abscission. Further analysis indicated that SlTIP1;1 mediated abscission via gating of cytoplasmic H2O2 concentrations and osmotic water permeability (Pf). Elevated cytoplasmic levels of H2O2 caused a suppressed auxin signal in the early abscission stage and enhanced ETH production during abscission. Furthermore, we found that increasing Pf was required to enhance the turgor pressure to supply the break force for AZ cell separation. Moreover, we observed that SlERF52 bound directly to the SlTIP1;1 promoter to regulate its expression, demonstrating a positive loop in which cytoplasmic H2O2 activates ETH production, which activates SlERF52. This, in turn, induces SlTIP1;1, which leads to elevated cytoplasmic H2O2 and water influx.

Interactions between endophytes (endophytic bacteria and fungi) and plants are crucial in maintaining crop fitness in agricultural systems, particularly in relation to abundant and rare subcommunities involved in community construction. However, the influence of long-term fertilization on heterogeneous rhizosphere nitrogen and phosphorus environments and how these conditions affect the key subcommunities of root endophytes and their community assembly mechanisms remain unclear. We studied the 26th year of a field experiment conducted in a greenhouse with varying levels of nitrogen and phosphorus (CKP0, CKP1, CNP0, CNP1, ONP0, and ONP1) to assess the composition of tomato root endophytes and their impact on yield. We employed 16S rRNA and fungal ITS region amplicon sequencing to investigate the assembly mechanisms of abundant and rare endophytic subcommunities, network correlations, core subcommunity structures, and key species that enhance crop yield. The results indicated that organic manure and phosphorus fertilizers significantly increased the rhizosphere soil nitrogen content, phosphorus content, and phosphorus availability (labile P, moderately labile P, and non-labile P). These fertilizers also significantly affected the composition (based on Bray-Curtis distance) and community assembly processes (βNTI) of endophytic microbial subcommunities. The assembly of both bacterial and fungal subcommunities was primarily governed by dispersal limitation, with community structures being significantly regulated by the content of rhizosphere soil available nitrogen (AN) and moderately labile P (MLP). Rare bacterial and fungal subcommunities complemented the ecological niches of abundant subcommunities in the co-occurrence network, supporting community functions and enhancing network stability. Nitrogen-adapting abundant and rare bacterial subcommunities provided a stronger predictive correlation for tomato yield than phosphorus-adapting fungal subcommunities. Additionally, three core genera of rare endophytic bacteria such as Arthrobacter , Microbacterium , and Sphingobium were identified as potentially involved in improving crop yield improvement. These findings revealed the distinct assembly mechanisms of endophytic microbial subcommunities affected by fertilization, enhancing our understanding of better management practices and controlling endophytes to improve crop yield in intensive agricultural ecosystems. [ABSTRACT FROM AUTHOR]

Siberian apricot (Prunus sibirica) is an important ecological and commercial woody plant that is negatively affected by spring frosts. However, the mechanisms that control gene expression in adaptation to freezing remain largely unknown. In this work, we investigated the physiological, molecular, and phenotypic characteristics of pistils of two P. sibirica clones that differ in their ability to withstand freezing stress. A total of 14,717 unigenes categorized into 38 functional groups were identified. Additionally, the two P. sibirica clones included 3931 up-regulated and 2070 down-regulated differentially expressed genes (DEGs). Many DEGs are related to Ca2+ and MAPK signaling, carbohydrate biosynthesis and metabolism, plant hormone signal transduction, biosynthesis of amino acids, and photosynthesis. The metabolism of carbohydrates, amino acids, lipids, secondary metabolites, plant hormone signal transduction, and terpenoid metabolism were the transcriptome modifications most significantly altered by freezing stress. Real-time quantitative PCR (RT-qPCR) was used to verify the precision of the RNA-seq data. PsbHLH18, PsMYB4, PsMYB44, PsPOD1, and PsCDPK5 may play important roles in the freezing tolerance of the P. sibirica floral organ. This study provides a foundation for further studies on the complex mechanisms of freezing stress response in P. sibirica. [ABSTRACT FROM AUTHOR]