Molecular regulation of tomato male reproductive development.

The reproductive success of flowering plants, which directly affects crop yield, is sensitive to environmental changes. A thorough understanding of how crop reproductive development adapts to climate changes is vital for ensuring global food security. In addition to being a high-value vegetable crop, tomato is also a model plant used for research on plant reproductive development. Tomato crops are cultivated under highly diverse climatic conditions worldwide. Targeted crosses of hybrid varieties have resulted in increased yields and abiotic stress resistance; however, tomato reproduction, especially male reproductive development, is sensitive to temperature fluctuations, which can lead to aborted male gametophytes, with detrimental effects on fruit set. We herein review the cytological features as well as genetic and molecular pathways influencing tomato male reproductive organ development and responses to abiotic stress. We also compare the shared features among the associated regulatory mechanisms of tomato and other plants. Collectively, this review highlights the opportunities and challenges related to characterizing and exploiting genic male sterility in tomato hybrid breeding programs.

Intraoperative assessment of bone viability through improved analysis and visualization of dynamic contrast-enhanced fluorescence imaging: technique report.

Bone devitalization is believed to be a critical determinant of complications such as infection or nonunion. However, intraoperative assessment of bone devitalization, particularly in open fractures and infections, remains highly subjective resulting in variation in treatment. Optical imaging tools, particularly dynamic contrast-enhanced fluorescence imaging, can provide real-time, intraoperative assessment of bone and soft tissue perfusion, which informs the tissues' ability to heal and fight infection. We describe a novel technique to apply indocyanine green-based fluorescence imaging, using a device that is frequently used in the operating room to assess skin or flap perfusion in plastic surgery, to assess bone and deep tissue perfusion in three pertinent cases: (1) a chronic infection/nonunion after a Gustilo type 3A tibia fracture (patient 1), (2) an acute Gustilo type 3C tibia open fracture with extensive degloving/soft tissue stripping (patient 2), and (3) an atrophic nonunion of the humerus (patient 3). In all three cases, fluorescence imaging (both time-specific fluorescence and maximum fluorescence) and derived kinetic maps of time-to-peak, ingress slope, and egress slope demonstrated clear spatial variation in perfusion that corresponded to the patient pathogenesis. The impact of this information on patient outcome will need to be evaluated in future clinical trials; however, these cases demonstrate in principle that optical imaging information has the potential to inform surgical practice, reduce the variation in treatment, and improve outcomes observed in these challenging patients.

Heterotypic transcriptional condensates formed by prion-like paralogous proteins canalize flowering transition in tomato.

BACKGROUND: Paralogs that arise from gene duplications during genome evolution enable genetic redundancy and phenotypic robustness. Variation in the coding or regulatory sequence of paralogous transcriptional regulators diversifies their functions and relationships, which provides developmental robustness against genetic or environmental perturbation. The fate transition of plant shoot stem cells for flowering and reproductive success requires a robust transcriptional control. However, how paralogs function and interact to achieve such robustness is unknown. RESULTS: Here, we explore the genetic relationship and protein behavior of ALOG family transcriptional factors with diverse transcriptional abundance in shoot meristems. A mutant spectrum covers single and higher-order mutant combinations of five ALOG paralogs and creates a continuum of flowering transition defects, showing gradually enhanced precocious flowering, along with inflorescence simplification from wild-type-like to progressively fewer flowers until solitary flower with sterile floral organs. Therefore, these paralogs play unequal roles and act together to achieve a robust genetic canalization. All five proteins contain prion-like intrinsically disordered regions (IDRs) and undergo phase separation. Accumulated mutations following gene duplications lead to IDR variations among ALOG paralogs, resulting in divergent phase separation and transcriptional regulation capabilities. Remarkably, they retain the ancestral abilities to assemble into a heterotypic condensate that prevents precocious activation of the floral identity gene ANANTHA. CONCLUSIONS: Our study reveals a novel genetic canalization mechanism enabled by heterotypic transcriptional condensates formed by paralogous protein interactions and phase separation, uncovering the molecular link between gene duplication caused IDR variation and robust transcriptional control of stem cell fate transition.

Dynamic evolution of small signalling peptide compensation in plant stem cell control.

Gene duplications are a hallmark of plant genome evolution and a foundation for genetic interactions that shape phenotypic diversity1-5. Compensation is a major form of paralogue interaction6-8 but how compensation relationships change as allelic variation accumulates is unknown. Here we leveraged genomics and genome editing across the Solanaceae family to capture the evolution of compensating paralogues. Mutations in the stem cell regulator CLV3 cause floral organs to overproliferate in many plants9-11. In tomato, this phenotype is partially suppressed by transcriptional upregulation of a closely related paralogue12. Tobacco lost this paralogue, resulting in no compensation and extreme clv3 phenotypes. Strikingly, the paralogues of petunia and groundcherry nearly completely suppress clv3, indicating a potent ancestral state of compensation. Cross-species transgenic complementation analyses show that this potent compensation partially degenerated in tomato due to a single amino acid change in the paralogue and cis-regulatory variation that limits its transcriptional upregulation. Our findings show how genetic interactions are remodelled following duplications and suggest that dynamic paralogue evolution is widespread over short time scales and impacts phenotypic variation from natural and engineered mutations.

Drugs used in regional block analgesia for thyroidectomy: A network meta-analysis of randomized controlled trials.

BACKGROUND: Postoperative pain has become one of the major obstacles to postoperative recovery and can lead to increasing demand for opioids. So far, a wide variety of drugs is used for regional block anesthesia, but which one is the best remains unclear. Thus, this study aims to obtain the best drug for thyroid surgery regional block based on their efficacy and side effects through Bayesian network meta-analysis. METHODS: Systemically searched MEDLINE, CENTRAL, Embase, and web of science on July 3, 2021 to obtain randomized controlled trials (RCTs) focusing on adults who underwent open thyroidectomy and regional blocks, and took opioids requirement, and the postoperative pain level as outcomes. The "Gemtc" package of R-4.1.1 was used for Bayesian network meta-analysis based on extracted data. RESULTS: We retrieved 398 articles, and finally included 31 RCTs, which included a total of 2221 patients. Patients received levobupivacaine had the lowest requirement rate for opioids and lowest pain score at T6 and T12. Non-steroidal anti-inflammatory drugs show significant superiority neither in the opioids requirement nor the pain score. Clonidine has a better potential to act as a qualified anesthetic adjuvant compared to adrenaline. The ranking results of the subgroup analysis were consistent with the integrated analysis. CONCLUSION: Levobupivacaine has a relatively greater advantage in reducing opioids requirement and decreasing the pain score of patients, and bilateral superficial cervical plexus blockade and pre thyroidectomy intervention have more advantages than local wound infiltration and post thyroidectomy intervention in alleviating patients' pain.

Progress on the Mechanism of Visceral Hypersensitivity in Nonerosive Reflux Disease.

Nonerosive reflux disease (NERD) is the most common type of gastroesophageal reflux disease (GERD). Its clinical symptoms can recur, and clinical treatment is often ineffective, causing patients severe economic and psychological burden. In recent years, studies that have explored in-depth the pathogenesis of NERD have found that visceral hypersensitivity (VH) plays an important role. VH refers to the phenomenon that viscera react strongly to nociceptive stimuli or produce a negative reaction to physiological stimuli due to the decrease of one's visceral pain threshold. Studies have found that the VH mechanism in NERD primarily includes abnormal neurotransmitters, the activation of acid-sensitive receptors, and abnormal psychological factors-all of which we review in this article.

ROS regulated reversible protein phase separation synchronizes plant flowering.

How aerobic organisms exploit inevitably generated but potentially dangerous reactive oxygen species (ROS) to benefit normal life is a fundamental biological question. Locally accumulated ROS have been reported to prime stem cell differentiation. However, the underlying molecular mechanism is unclear. Here, we reveal that developmentally produced H2O2 in plant shoot apical meristem (SAM) triggers reversible protein phase separation of TERMINATING FLOWER (TMF), a transcription factor that times flowering transition in the tomato by repressing pre-maturation of SAM. Cysteine residues within TMF sense cellular redox to form disulfide bonds that concatenate multiple TMF molecules and elevate the amount of intrinsically disordered regions to drive phase separation. Oxidation triggered phase separation enables TMF to bind and sequester the promoter of a floral identity gene ANANTHA to repress its expression. The reversible transcriptional condensation via redox-regulated phase separation endows aerobic organisms with the flexibility of gene control in dealing with developmental cues.

Controlling the thermally activated delayed fluorescence of axially chiral organic emitters and their racemate for information encryption.

A pair of axially chiral organic enantiomers were facilely prepared through a one-pot sequential synthesis. They exhibit circularly polarized luminescence activities and have thermally activated delayed fluorescence (TADF) and aggregation-induced emission enhancement properties. Meanwhile, these two enantiomers present remarkable and reversible thermochromism in the crystalline state, enabling dual-colour TADF switching between orange and red. However, when they form cocrystals, the resulting racemate shows opposite thermochromic behaviors. These intriguing results probably emanate from their different optical activities, leading to distinct molecular packing modes and molecular conformation variations. Moreover, information encryption based on thermochromism of organic enantiomers and their racemate has been presented for the first time. This work may expand the application scope of chiral organic luminogens and pave a new way to construct intelligent luminescent systems.

Author Correction: Genome of Tripterygium wilfordii and identification of cytochrome P450 involved in triptolide biosynthesis.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

[Effect of double-leaf perforator free flap posterolateral calf peroneal artery on reconstruction of oropharyngeal anatomy after ablation of advanced oropharyngeal carcinoma].

OBJECTIVE: To evaluate the effect of using free double- leaf perforator flap posterolateral calf peroneal artery in anatomical reconstruction of the oropharyngeal structure after ablation of advanced oropharyngeal carcinoma. METHODS: Twenty-six patients with oropharyngeal defects after ablation of oropharyngeal malignancies were recruited, including 12 with carcinoma in the tongue base, 5 in the latenral pharyngeal wall and 9 in the soft palate. Between July, 2016 and July, 2018, the patients underwent surgeries for reconstruction of the oropharyngeal defects using flaps. The areas of tissue defects repaired by double-leaf perforator flaps ranged from 40.5 to 72.5 cm2. Reconstruction was performed for oropharyngeal defects in the soft palate, pterygopalate, parapharyngeal, pterygo- mandibular, and tongue base tissues. The patients' outcomes including mouth opening, functions of deglutition, linguistic function, restoration of palatopharyngeal anatomical structure and postoperative survival were evaluated, and their quality of life was assessed using FACT-H&N scale (Chinese Edition). RESULTS: All the 26 patients with transplantation of the free flaps survived. Six months after the operation, the oropharyngeal function and anatomical structure of the patients were basically restored. The questionnaire survey showed that the patients' physical, social/family, emotional and functional conditions, the total score of the core scale, items scores for the head and neck, and the total score of the scale all improved significantly after the operation compared with those before the operation (P < 0.05). CONCLUSIONS: The free peroneal artery bilobate perforator flap in the posterolateral crus, which seldom has anatomical variations of the blood vessels, allows flexible design and contains rich tissue volume to facilitate defect repair with different approaches and ranges. The application of this flap, which is an ideal perforator flap for reconstruction of the oropharyngeal structure and function, can improve the quality of life of patients following operations for advanced oropharyngeal cancer.

Endosteal and periosteal blood flow quantified with dynamic contrast-enhanced fluorescence to guide open orthopaedic surgery.

Due to the lack of objectively measurable or quantifiable methods to assess the bone perfusion, the success of removing devitalized bone is based almost entirely on surgeon's experience and varies widely across surgeons and centers. In this study, an indocyanine green (ICG)-based dynamic contrast-enhanced fluorescence imaging (DCE-FI) has been developed to objectively assess bone perfusion and guide surgical debridement. A porcine trauma model (n = 6 pigs × 2 legs) with up to 5 conditions of severity in loss of flow in each, was imaged by a commercial fluorescence imaging system. By applying the bone-specific hybrid plug-compartment (HyPC) kinetic model to four-minute video sequences, the perfusion-related metrics, such as peak intensity, total bone blood flow (TBBF) and endosteal bone blood flow to TBBF fraction (EFF) were calculated. The results shown that the combination of TBBF and EFF can effectively differentiate injured from normal bone with the accuracy, sensitivity and specificity of 89%, 88% and 90%, respectively. Our subsequent first in human bone blood flow imaging study confirmed DCE-FI can be successfully translated into human orthopaedic trauma patients.

Loss of Function of an RNA Polymerase III Subunit Leads to Impaired Maize Kernel Development.

Kernel size is an important factor determining grain yield. Although a number of genes affecting kernel development in maize (Zea mays) have been identified by analyzing kernel mutants, most of the corresponding mutants cannot be used in maize breeding programs due to low germination or incomplete seed development. Here, we characterized small kernel7, a recessive small-kernel mutant with a mutation in the gene encoding the second-largest subunit of RNA polymerase III (RNAPΙΙΙ; NRPC2). A frame shift in ZmNRPC2 leads to a premature stop codon, resulting in significantly reduced levels of transfer RNAs and 5S ribosomal RNA, which are transcribed by RNAPΙΙΙ. Loss-of-function nrpc2 mutants created by CRISPR/CAS9 showed significantly reduced kernel size due to altered endosperm cell size and number. ZmNRPC2 affects RNAPIII activity and the expression of genes involved in cell proliferation and endoreduplication to control kernel development via physically interacting with RNAPIII subunits RPC53 and AC40, transcription factor class C1 and Floury3. Notably, unlike the semidominant negative mutant floury3, which has defects in starchy endosperm, small kernel7 only affects kernel size but not the composition of kernel storage proteins. Our findings provide novel insights into the molecular network underlying maize kernel size, which could facilitate the genetic improvement of maize in the future.

Genome of Tripterygium wilfordii and identification of cytochrome P450 involved in triptolide biosynthesis.

Triptolide is a trace natural product of Tripterygium wilfordii. It has antitumor activities, particularly against pancreatic cancer cells. Identification of genes and elucidation of the biosynthetic pathway leading to triptolide are the prerequisite for heterologous bioproduction. Here, we report a reference-grade genome of T. wilfordii with a contig N50 of 4.36 Mb. We show that copy numbers of triptolide biosynthetic pathway genes are impacted by a recent whole-genome triplication event. We further integrate genomic, transcriptomic, and metabolomic data to map a gene-to-metabolite network. This leads to the identification of a cytochrome P450 (CYP728B70) that can catalyze oxidation of a methyl to the acid moiety of dehydroabietic acid in triptolide biosynthesis. We think the genomic resource and the candidate genes reported here set the foundation to fully reveal triptolide biosynthetic pathway and consequently the heterologous bioproduction.

Immunogenicity of 987P fimbriae of enterotoxigenic Escherichia coli surface-displayed on Lactobacillus casei.

As most pathogens invade the bodies through the mucosa, it is crucial to develop vaccines that induce mucosal immunity. To this end, we generated a safe and effective vaccine candidate that displayed fimbrial protein 987P of enterotoxigenic Escherichia coli (ETEC) on the surface of Lactobacillus casei (L.casei) CICC 6105 by using poly-γ-glutamate synthetase A (PgsA) as an anchoring matrix. After gavage inoculation of the recombinant strain pLA-987P/L.casei into specific-pathogen-free (SPF) BALB/c mice, high levels of mucosal immunoglobulin A (IgA) were induced in fecal samples, intestine and lung lavage fluids and systemic immunoglobulin G of IgG subclasses (IgG1, IgG2b, and IgG2a) was produced in serum. T-cell proliferation assays showed the stimulation index (SI) of the groups immunized with pLA-987P/L.casei to be significantly higher than that of the control group. The recombinant L.casei promoted T cells to produce both Th1 and Th2 cytokines, while the number of splenic IL-4 Spot forming cells (SFC) exceeded the number of IFN-γ SFC by 2.26-fold (P < .01). >83.3% of the vaccinated mice were protected from challenge with a lethal dose of virulent strain C83916. These results indicate that the recombinant L.casei expressing ETEC 987P fimbrial protein could elicit a protective immune response against ETEC 987P infection effectively.

[De novo domestication to create new crops].

Crop improvement by domestication and traditional breeding often results in fitness penalties and loss of genetic diversity, which greatly threatens crop production and food security under the challenging global climate. De novo domestication has been proposed as a novel strategy for crop breeding. By combining multi-omics, genome editing and synthetic biology approaches, domestication of wild or semi-wild plant species can be accelerated by rapidly introducing desirable traits without causing an associated drag on their inherent traits. In this review, we summarize the history of crop domestication, emphasize the urgency for breeding strategy innovation, and discuss recent progress of de novo crop domestication.

Evolution of buffering in a genetic circuit controlling plant stem cell proliferation.

Precise control of plant stem cell proliferation is necessary for the continuous and reproducible development of plant organs1,2. The peptide ligand CLAVATA3 (CLV3) and its receptor protein kinase CLAVATA1 (CLV1) maintain stem cell homeostasis within a deeply conserved negative feedback circuit1,2. In Arabidopsis, CLV1 paralogs also contribute to homeostasis, by compensating for the loss of CLV1 through transcriptional upregulation3. Here, we show that compensation4,5 operates in diverse lineages for both ligands and receptors, but while the core CLV signaling module is conserved, compensation mechanisms have diversified. Transcriptional compensation between ligand paralogs operates in tomato, facilitated by an ancient gene duplication that impacted the domestication of fruit size. In contrast, we found little evidence for transcriptional compensation between ligands in Arabidopsis and maize, and receptor compensation differs between tomato and Arabidopsis. Our findings show that compensation among ligand and receptor paralogs is critical for stem cell homeostasis, but that diverse genetic mechanisms buffer conserved developmental programs.

Correlation of the oxidative stress indices and Cd exposure using a mathematical model in the earthworm, Eisenia fetida.

With the increase in heavy metal pollution, it is of great significance to evaluate the ecological security and early warning of cadmium (Cd) contaminated soil. In this paper, a mathematical model was established for the first time by combining the advantages of the factor analysis method and the analytic hierarchy process, and was used to screen and analyze the ecological indices of oxidative stress in earthworms under Cd exposure. The experiment lasted for 40 days, removing one earthworm every 10 days. The Cd2+ concentration gradient was set at 0, 1, 10, 20, 100, 200, 400 and 800 mg kg-1. The ecological indices measured were total protein (TP), peroxidase (POD), superoxide dismutase (SOD), glutathione peroxidase (GPX), glutathione-S-transferase (GST), catalase (CAT), acetylcholinesterase (AChE) and malondialdehyde (MDA) levels. The results showed that when the earthworm was exposed to Cd2+ for 10 days and 30 days, in the head tissues, the key indices to focus on for monitoring were both POD. At 20 days and 40 days, the key indices were both TP. For the tail tissue tests, under Cd exposure for 10 days, the key indicator focused on for monitoring was MDA. After 20 days of exposure, the key monitoring indicator was AChE. At 30 days, it was CAT, and at 40 days, it was TP. This study provides a theoretical basis for the prompt, inexpensive, accurate and scientific early warning of metal contaminated soils and establishes a foundation for application of the screening model for other ecological indicators.