Peptide REF1 is a local wound signal promoting plant regeneration.

Plants frequently encounter wounding and have evolved an extraordinary regenerative capacity to heal the wounds. However, the wound signal that triggers regenerative responses has not been identified. Here, through characterization of a tomato mutant defective in both wound-induced defense and regeneration, we demonstrate that in tomato, a plant elicitor peptide (Pep), REGENERATION FACTOR1 (REF1), acts as a systemin-independent local wound signal that primarily regulates local defense responses and regenerative responses in response to wounding. We further identified PEPR1/2 ORTHOLOG RECEPTOR-LIKE KINASE1 (PORK1) as the receptor perceiving REF1 signal for plant regeneration. REF1-PORK1-mediated signaling promotes regeneration via activating WOUND-INDUCED DEDIFFERENTIATION 1 (WIND1), a master regulator of wound-induced cellular reprogramming in plants. Thus, REF1-PORK1 signaling represents a conserved phytocytokine pathway to initiate, amplify, and stabilize a signaling cascade that orchestrates wound-triggered organ regeneration. Application of REF1 provides a simple method to boost the regeneration and transformation efficiency of recalcitrant crops.

Tomato MED25 regulates fruit ripening by interacting with EIN3-like transcription factors.

Fruit ripening relies on the precise spatiotemporal control of RNA polymerase II (Pol II)-dependent gene transcription, and the evolutionarily conserved Mediator (MED) coactivator complex plays an essential role in this process. In tomato (Solanum lycopersicum), a model climacteric fruit, ripening is tightly coordinated by ethylene and several key transcription factors. However, the mechanism underlying the transmission of context-specific regulatory signals from these ripening-related transcription factors to the Pol II transcription machinery remains unknown. Here, we report the mechanistic function of MED25, a subunit of the plant Mediator transcriptional coactivator complex, in controlling the ethylene-mediated transcriptional program during fruit ripening. Multiple lines of evidence indicate that MED25 physically interacts with the master transcription factors of the ETHYLENE-INSENSITIVE 3 (EIN3)/EIN3-LIKE (EIL) family, thereby playing an essential role in pre-initiation complex formation during ethylene-induced gene transcription. We also show that MED25 forms a transcriptional module with EIL1 to regulate the expression of ripening-related regulatory as well as structural genes through promoter binding. Furthermore, the EIL1-MED25 module orchestrates both positive and negative feedback transcriptional circuits, along with its downstream regulators, to fine-tune ethylene homeostasis during fruit ripening.

The genomic route to tomato breeding: Past, present, and future.

Over the past 10,000 years, tomato species have undergone both unintentional and intentional selection to enhance their favorable traits for human consumption and manufacturing. These selection processes have significantly influenced the genomes of tomato species and have played a critical role in improving tomato varieties. In this review, we summarize recent advances in tomato genome sequencing, explore the impact of human-driven selection, and recapitulate key genes associated with important agronomic traits in tomato breeding. We provide several examples of genomics-guided tomato breeding to highlight the potential of genome resources in facilitating tomato improvement. Furthermore, we elaborate the progress and strategies of tomato breeding through genomic designing, and present how such efforts can help future enhancements of tomato to align with the demands of sustainability and evolving human societies.

Self-powered deep ultraviolet photodetector based on p-CuI/n-ZnGa2O4 heterojunction with high sensitivity and fast speed.

Self-powered deep ultraviolet photodetectors (DUV PDs) are essential in environmental monitoring, flame detection, missile guidance, aerospace, and other fields. A heterojunction photodetector based on p-CuI/n-ZnGa2O4 has been fabricated by pulsed laser deposition combined with vacuum thermal evaporation. Under 260 nm DUV light irradiation, the photodetector exhibits apparent self-powered performance with a maximum responsivity and specific detectivity of 2.75 mA/W and 1.10 × 1011 Jones at 0 V. The photodetector exhibits high repeatability and stability under 260 nm periodic illumination. The response and recovery time are 205 ms and 133 ms, respectively. This work provides an effective strategy for fabricating high-performance self-powered DUV photodetectors.

Rapid generation of a tomato male sterility system and its feasible application in hybrid seed production.

KEY MESSAGE: A practical approach for the rapid generation and feasible application of green hypocotyl male-sterile (GHMS) tm6 dfr lines in tomato hybrid breeding was established. Male sterility enables reduced cost and high seed purity during hybrid seed production. However, progress toward its commercial application has been slow in tomato due to the disadvantages of most natural male-sterile mutants. Here, we developed a practical method for efficient tomato hybrid seed production using a male-sterile system with visible marker, which was rapidly generated by CRISPR/Cas9-mediated gene editing. Two closely linked genes, TM6 and DFR, which were reported to be candidates of ms15 (male sterile-15) and aw (anthocyanin without) locus, respectively, were knocked out simultaneously in two elite tomato inbred lines. Mutagenesis of both genes generated green hypocotyl male-sterile (GHMS) lines. The GHMS lines exhibited male sterility across different genetic backgrounds and environmental conditions. They also showed green hypocotyl due to defective anthocyanin accumulation, which serves as a reliable visible marker for selecting male-sterile plants at the seedling stage. We further proposed a strategy for multiplying the GHMS system and verified its high efficiency in stable male sterility propagation. Moreover, elite hybrid seeds were produced using GHMS system for potential side effects evaluation, and no adverse influences were found on seed yield, seed quality as well as important agronomic traits. This study provides a practical approach for the rapid generation and feasible application of male sterility in tomato hybrid breeding.

Mediator complex subunit MED25 physically interacts with DST to regulate spikelet number in rice.

Grain number is a flexible trait and contributes significantly to grain yield. In rice, the zinc finger transcription factor DROUGHT AND SALT TOLERANCE (DST) controls grain number by directly regulating cytokinin oxidase/dehydrogenase 2 (OsCKX2) expression. Although specific upstream regulators of the DST-OsCKX2 module have been identified, the mechanism employed by DST to regulate the expression of OsCKX2 remains unclear. Here, we demonstrate that DST-interacting protein 1 (DIP1), known as Mediator subunit OsMED25, acts as an interacting coactivator of DST. Phenotypic analyses revealed that OsMED25-RNAi and the osmed25 mutant plants exhibited enlarged panicles, with enhanced branching and spikelet number, similar to the dst mutant. Genetic analysis indicated that OsMED25 acts in the same pathway as the DST-OsCKX2 module to regulate spikelet number per panicle. Further biochemical analysis showed that OsMED25 physically interacts with DST at the promoter region of OsCKX2, and then recruits RNA polymerase II (Pol II) to activate OsCKX2 transcription. Thus, OsMED25 was involved in the communication between DST and Pol II general transcriptional machinery to regulate spikelet number. In general, our findings reveal a novel function of OsMED25 in DST-OsCKX2 modulated transcriptional regulation, thus enhancing our understanding of the regulatory mechanism underlying DST-OsCKX2-mediated spikelet number.

Mediator Subunit MED25 Physically Interacts with PHYTOCHROME INTERACTING FACTOR4 to Regulate Shade-Induced Hypocotyl Elongation in Tomato.

Shade triggers important adaptive responses such as the shade-avoidance syndrome, which enable plants to respond to the depletion of photosynthetically active light. The basic helix-loop-helix transcription factors PHYTOCHROME INTERACTING FACTORS (PIFs) play a key role in the shade-avoidance syndrome network by regulating the biosynthesis of multiple phytohormones and the expression of cell expansion-related genes. Although much has been learned about the regulation of PIFs in response to shade at the protein level, relatively little is known about the PIF-dependent transcriptional regulation of shade-responsive genes. Mediator is an evolutionarily conserved transcriptional coactivator complex that bridges gene-specific transcription factors with the RNA polymerase II (Pol II) machinery to regulate gene transcription. Here, we report that tomato (Solanum lycopersicum) PIF4 plays an important role in shade-induced hypocotyl elongation by regulating the expression of genes that encode auxin biosynthesis and auxin signaling proteins. During this process, Mediator subunit25 (MED25) physically interacts with PIF4 at the promoter regions of PIF4 target genes and also recruits Pol II to induce gene transcription. Thus, MED25 directly bridges the communication between PIF4 and Pol II general transcriptional machinery to regulate shade-induced hypocotyl elongation. Overall, our results reveal a novel role of MED25 in PIF4-mediated transcriptional regulation under shade.

Gene duplication confers enhanced expression of 27-kDa γ-zein for endosperm modification in quality protein maize.

The maize opaque2 (o2) mutant has a high nutritional value but it develops a chalky endosperm that limits its practical use. Genetic selection for o2 modifiers can convert the normally chalky endosperm of the mutant into a hard, vitreous phenotype, yielding what is known as quality protein maize (QPM). Previous studies have shown that enhanced expression of 27-kDa γ-zein in QPM is essential for endosperm modification. Taking advantage of genome-wide association study analysis of a natural population, linkage mapping analysis of a recombinant inbred line population, and map-based cloning, we identified a quantitative trait locus (qγ27) affecting expression of 27-kDa γ-zein. qγ27 was mapped to the same region as the major o2 modifier (o2 modifier1) on chromosome 7 near the 27-kDa γ-zein locus. qγ27 resulted from a 15.26-kb duplication at the 27-kDa γ-zein locus, which increases the level of gene expression. This duplication occurred before maize domestication; however, the gene structure of qγ27 appears to be unstable and the DNA rearrangement frequently occurs at this locus. Because enhanced expression of 27-kDa γ-zein is critical for endosperm modification in QPM, qγ27 is expected to be under artificial selection. This discovery provides a useful molecular marker that can be used to accelerate QPM breeding.

Genetic analysis and major QTL detection for maize kernel size and weight in multi-environments.

KEY MESSAGE: Twelve major QTL in five optimal clusters and several epistatic QTL are identified for maize kernel size and weight, some with pleiotropic will be promising for fine-mapping and yield improvement. Kernel size and weight are important target traits in maize (Zea mays L.) breeding programs. Here, we report a set of quantitative trait loci (QTL) scattered through the genome and significantly controlled the performance of four kernel traits including length, width, thickness and weight. From the cross V671 (large kernel) × Mc (small kernel), 270 derived F2:3 families were used to identify QTL of maize kernel-size traits and kernel weight in five environments, using composite interval mapping (CIM) for single-environment analysis along with mixed linear model-based CIM for joint analysis. These two mapping strategies identified 55 and 28 QTL, respectively. Among them, 6 of 23 coincident were detected as interacting with environment. Single-environment analysis showed that 8 genetic regions on chromosomes 1, 2, 4, 5 and 9 clustered more than 60 % of the identified QTL. Twelve stable major QTLs accounting for over 10 % of phenotypic variation were included in five optimal clusters on the genetic region of bins 1.02-1.03, 1.04-1.06, 2.05-2.07, 4.07-4.08 and 9.03-9.04; the addition and partial dominance effects of significant QTL play an important role in controlling the development of maize kernel. These putative QTL may have great promising for further fine-mapping with more markers, and genetic improvement of maize kernel size and weight through marker-assisted breeding.

Tomato CYP94C1 inactivates bioactive JA-Ile to attenuate jasmonate-mediated defense during fruit ripening.

As the master regulators of the ET signaling pathway, EIL transcription factors directly activate the expression of CYP94C1 to inactivate bioactive JA-Ile, thereby attenuating JA-mediated defense during fruit ripening. Knockout of CYP94C1 improves tomato fruit resistance to necrotrophs without compromising fruit quality.

Recoloring tomato fruit by CRISPR/Cas9-mediated multiplex gene editing.

Fruit color is an important horticultural trait, which greatly affects consumer preferences. In tomato, fruit color is determined by the accumulation of different pigments, such as carotenoids in the pericarp and flavonoids in the peel, along with the degradation of chlorophyll during fruit ripening. Since fruit color is a multigenic trait, it takes years to introgress all color-related genes in a single genetic background via traditional crossbreeding, and the avoidance of linkage drag during this process is difficult. Here, we proposed a rapid breeding strategy to generate tomato lines with different colored fruits from red-fruited materials by CRISPR/Cas9-mediated multiplex gene editing of three fruit color-related genes (PSY1, MYB12, and SGR1). Using this strategy, the red-fruited cultivar 'Ailsa Craig' has been engineered to a series of tomato genotypes with different fruit colors, including yellow, brown, pink, light-yellow, pink-brown, yellow-green, and light green. Compared with traditional crossbreeding, this strategy requires less time and can obtain transgene-free plants with different colored fruits in less than 1 year. Most importantly, it does not alter other important agronomic traits, like yield and fruit quality. Our strategy has great practical potential for tomato breeding and serves as a reference for improving multigene-controlled traits of horticultural crops.

Redesigning the tomato fruit shape for mechanized production.

Crop breeding for mechanized harvesting has driven modern agriculture. In tomato, machine harvesting for industrial processing varieties became the norm in the 1970s. However, fresh-market varieties whose fruits are suitable for mechanical harvesting are difficult to breed because of associated reduction in flavour and nutritional qualities. Here we report the cloning and functional characterization of fs8.1, which controls the elongated fruit shape and crush resistance of machine-harvestable processing tomatoes. FS8.1 encodes a non-canonical GT-2 factor that activates the expression of cell-cycle inhibitor genes through the formation of a transcriptional module with the canonical GT-2 factor SlGT-16. The fs8.1 mutation results in a lower inhibitory effect on the cell proliferation of the ovary wall, leading to elongated fruits with enhanced compression resistance. Our study provides a potential route for introducing the beneficial allele into fresh-market tomatoes without reducing quality, thereby facilitating mechanical harvesting.

Effect of Pr/Zn on the anti-humidity and acetone-sensing properties of Co3O4 prepared by electrospray.

Co3O4 is a P-type metal-oxide semiconductor which can realize acetone detection at a lower temperature, but the lower working temperature brings the enhanced humidity effect. In order to solve the problem of a Co3O4 gas sensor being easily affected by humidity, an acetone-sensing material of Co3O4 mixed with Pr/Zn was prepared by electrospray in this work. The optimal working temperature of Pr/Zn-Co3O4 is 160 °C, and the detection limit can reach 1 ppm. The fluctuation of the acetone response is about 7.7% in the relative humidity range of 30-90%. Compared with pure Co3O4, the anti-humidity property of this material is obviously enhanced, but the gas-sensing response deteriorates. Compared with Pr-Co3O4, the anti-humidity and acetone sensing properties of Pr/Zn-Co3O4 were both improved. The morphology, composition, crystal state and energy state of the material were analyzed by SEM, EDS, XRD and XPS. The material of Pr/Zn-Co3O4 is a multi-component mixed material composed of PrCoO3, ZnO, Pr6O11 and Co3O4. The improved anti-humidity and acetone sensing properties exhibited by this material are the result of the synergistic effect of ZnO and Pr3+.

A Transcriptional Network Promotes Anthocyanin Biosynthesis in Tomato Flesh.

Dietary anthocyanins are important health-promoting antioxidants that make a major contribution to the quality of fruits. It is intriguing that most tomato cultivars do not produce anthocyanins in fruit. However, the purple tomato variety Indigo Rose, which has the dominant Aft locus combined with the recessive atv locus from wild tomato species, exhibits light-dependent anthocyanin accumulation in the fruit skin. Here, we report that Aft encodes a functional anthocyanin activator named SlAN2-like, while atv encodes a nonfunctional version of the anthocyanin repressor SlMYBATV. The expression of SlAN2-like is responsive to light, and the functional SlAN2-like can activate the expression of both anthocyanin biosynthetic genes and their regulatory genes, suggesting that SlAN2-like acts as a master regulator in the activation of anthocyanin biosynthesis. We further showed that cultivated tomatoes contain nonfunctional alleles of SlAN2-like and therefore fail to produce anthocyanins. Consistently, expression of a functional SlAN2-like gene driven by the fruit-specific promoter in a tomato cultivar led to the activation of the entire anthocyanin biosynthesis pathway and high-level accumulation of anthocyanins in both the peel and flesh. Taken together, our study exemplifies that efficient engineering of complex metabolic pathways could be achieved through tissue-specific expression of master transcriptional regulators.

[Growth of rifampin-dependent Mycobacterium tuberculosis in conditions without rifampin].

OBJECTIVE: To study the growth of rifampin-dependent Mycobacterium tuberculosis in conditions without rifampin. METHODS: A typical rifampin-dependent strain was identified and a single colony growing in the optimal concentration of rifampin was selected and inoculated (10(-4) mg/ml) to L-J and 12B liquid media, cultured at 37 degrees C. The bacterial growth and characteristics were studied. RESULTS: In the L-J medium containing rifampin, bacterial growth was observed in two weeks, but in the medium without rifampin tiny colonies were not observed until three weeks. In the 12B media with and without rifampin, bacterial growth was evident on day 9 and day 13 respectively. Thick and larger colonies and cauliflower-like growth were observed in media containing rifampin, but smaller colonies and fried egg-like growth were observed in media without rifampin. Microscopic study showed thick and heavily stained bacteria in media containing rifampin, but small and lightly stained bacteria in media without rifampin. Transmission electron microscopy demonstrated that bacteria growing in the presence of rifampin had complete cell walls, but those growing in the absence of rifampin had abnormal cell walls. CONCLUSION: Rifampin-dependent Mycobacterium tuberculosis may be changed into bacterial L-forms in media without rifampin.