Excessive Branching Research Articles

The kinetochore protein KNL-1 regulates the actin cytoskeleton to control dendrite branching.

The function of the nervous system is intimately tied to its complex and highly interconnected architecture. Precise control of dendritic branching in individual neurons is central to building the complex structure of the nervous system. Here, we show that the kinetochore protein KNL-1 and its associated KMN (Knl1/Mis12/Ndc80 complex) network partners, typically known for their role in chromosome-microtubule coupling during mitosis, control dendrite branching in the Caenorhabditis elegans mechanosensory PVD neuron. KNL-1 restrains excess dendritic branching and promotes contact-dependent repulsion events, ensuring robust sensory behavior and preventing premature neurodegeneration. Unexpectedly, KNL-1 loss resulted in significant alterations of the actin cytoskeleton alongside changes in microtubule dynamics within dendrites. We show that KNL-1 modulates F-actin dynamics to generate proper dendrite architecture and that its N-terminus can initiate F-actin assembly. These findings reveal that the postmitotic neuronal KMN network acts to shape the developing nervous system by regulating the actin cytoskeleton and provide new insight into the mechanisms controlling dendrite architecture.

Antifungal Activity of Novel Isoindoline-2-Yl Putrescines as Potential Autophagy-Activated Fungicide.

The exacerbation of plant fungal diseases necessitates the development of new fungicides to prevent outbreaks. In this study, five novel isoindoline-2-yl putrescines (ISPs) were synthesized, and their synthetic procedures and gram-scale preparation were explored. When tested at 50 μg mL-1, ISPs did not significantly inhibit mycelial growth on agar plates. However, at 100 μg mL-1, they demonstrated remarkable in vivo efficacy in mitigating Botrytis cinerea infection, especially ISP3 showed curative and protective activities of 91.9 % and 92.6 %, respectively. Moreover, ISP3 also effectively halted lesion expansion of gray mold, Sclerotic rot, and Fusarium scabs, while inducing excessive malformed top mycelial branches of B. cinerea and Sclerotinia sclerotiorum, suppressing sclerotia formation in S. sclerotiorum, and triggering autophagic vacuolization with numerous autophagosomes in the mycelia of these fungi. Molecular docking revealed that ISP3 effectively bound to the active site of BcAtg3, forming hydrogen bonds with Ser279, Gly343, Asp370, and Asp13, along with establishing a stable salt bridge with Asp13. Furthermore, ISP3 possessed favorable ADMET properties. These findings highlight ISP3 as a promising antifungal candidate through autophagy activation.

GLABRA3-mediated trichome branching requires transcriptional repression of MICROTUBULE-DESTABILIZING PROTEIN25.

Microtubules play pivotal roles in establishing trichome branching patterns, which is a model system for studying cell-shape control in Arabidopsis (Arabidopsis thaliana). However, the signaling pathway that regulates microtubule reorganization during trichome branching remains poorly understood. In this study, we report that MICROTUBULE-DESTABILIZING PROTEIN25 (MDP25) is involved in GLABRA3 (GL3)-mediated trichome branching by regulating microtubule stability. Loss of MDP25 function led to excessive trichome branching, and this phenotype in mdp25 could not be rescued by the MDP25 K7A or MDP25 K18A mutated variants. Pharmacological treatment and live-cell imaging revealed increased microtubule stability in the mdp25 mutant. Furthermore, the microtubule collar observed during trichome branching remained more intact in mdp25 compared to the WT under oryzalin treatment. Results of genetic assays further demonstrated that knocking out MDP25 rescued the reduced branching phenotype of gl3 trichomes. In gl3 trichomes, normal microtubule organization was disrupted, and microtubule stability was significantly compromised. Moreover, GL3 physically bound to the MDP25 promoter, thereby inhibiting its expression. Overexpression of GL3 negated the effects of PMDP25-driven MDP25 or its mutant proteins on trichome branching and microtubules in the mdp25 background. Overall, our study uncovers a mechanism by which GL3 inhibits MDP25 transcription, thereby influencing microtubule stability and regulating trichome branching. This mechanism provides a connection between early regulatory components and microtubules during trichome development.

Study of the fracture propagation and the corresponding heat mining performance of multi-branch radial wells geothermal system based on the THM-D coupling model

Due to the multi-well enhanced geothermal systems can significantly improve the heat transfer efficiency and the multi-branch radial well geothermal system can reduce the well construction cost, it is necessary to investigate the fracture propagation and the heat mining performance of the multi-branch radial well geothermal system. Based on the constructed THM-D coupling model, the fracture propagation rules of multi-branch radial wells under the co-function of low-temperature induced thermal stress and radial well induced stress is explored. Then the heat mining prediction study during the heat extraction stage is conducted based on the generated fracture. Furthermore, the influence of radial well characteristic parameters on fracture propagation and the corresponding heat mining performance of multi-branch radial well geothermal system is discussed. The results show that the smaller radial well angle in multi-branch radial well system can generate a larger scope of fracture network between the included angle under the co-function of radial well induced stress and low-temperature induced thermal stress, which will correspond to a larger heat extraction scope in heat mining stage. When the radial well included angle along the direction of maximum horizontal in-situ stress is 120°, the fracture network obtained by fracturing is the most complex. Excessive branching of radial wells can lead to significant induced stress in the plane of radial wells, which will affect the propagation of the fracture network in the direction of longitudinal connection between the doublet wells, thereby affecting the corresponding heat mining performance. With the increase of radial well length, the ability to induce the formation of branching fractures raises, resulting in more complex fracture network. In the heat mining stage of 10-year development, the outlet temperature and heat mining rate respectively increased by 5.83% and 6.9% when the radial well length is 75 m compared to 50 m.

Agro Active Potential of Bacillus subtilis PE7 against Didymella bryoniae (Auersw.), the Causal Agent of Gummy Stem Blight of Cucumis melo.

Microbial agents such as the Bacillus species are recognized for their role as biocontrol agents against various phytopathogens through the production of diverse bioactive compounds. This study evaluates the effectiveness of Bacillus subtilis PE7 in inhibiting the growth of Didymella bryoniae, the pathogen responsible for gummy stem blight (GSB) in cucurbits. Dual culture assays demonstrate significant antifungal activity of strain PE7 against D. bryoniae. Volatile organic compounds (VOCs) produced by strain PE7 effectively impede mycelial formation in D. bryoniae, resulting in a high inhibition rate. Light microscopy revealed that D. bryoniae hyphae exposed to VOCs exhibited abnormal morphology, including swelling and excessive branching. Supplementing a potato dextrose agar (PDA) medium with a 30% B. subtilis PE7 culture filtrate significantly decreased mycelial growth. Moreover, combining a 30% culture filtrate with half the recommended concentration of a chemical fungicide yielded a more potent antifungal effect than using the full fungicide concentration alone, inducing dense mycelial formation and irregular hyphal morphology in D. bryoniae. Strain PE7 was highly resilient and was able to survive in fungicide solutions. Additionally, B. subtilis PE7 enhanced the nutrient content, growth, and development of melon plants while mitigating the severity of GSB compared to fungicide and fertilizer treatments. These findings highlight B. subtilis PE7 as a promising biocontrol candidate for integrated disease management in crop production.

The tomato WRKY-B transcription factor modulates lateral branching by targeting BLIND, PIN4, and IAA15.

Lateral branching is a crucial agronomic trait that impacts crop yield. In tomato ( Solanum lycopersicum ), excessive lateral branching is unfavorable and results in substantial labor and management costs. Therefore, optimizing lateral branching is a primary objective in tomato breeding. Although many genes related to lateral branching have been reported in tomato, the molecular mechanism underlying their network remains elusive. In this study, we found that the expression profile of a WRKY gene, WRKY-B (for WRKY-BRANCING), was associated with the auxin-dependent axillary bud development process. Wrky-b mutants generated by the CRISPR/Cas9 editing system presented fewer lateral branches, while WRKY-B overexpression lines presented more lateral branches than did wild-type plants. Furthermore, WRKY-B can directly target the well-known branching gene BLIND (BL) and the auxin efflux carrier gene PIN4 to activate their expression. Both the bl and pin4 mutants exhibited reduced lateral branching, similar to the wrky-b mutant. The IAA contents in the axillary buds of the wrky-b, bl, and pin4 mutant plants were significantly higher than those in the wild-type plants. In addition, WRKY-B can also directly target the AUX/IAA gene IAA15 and repress its expression. In summary, WRKY-B works upstream of BL, PIN4, and IAA15 to regulate the development of lateral branches in tomato.

Mapping the tree damage index: A case study on the green lane in the urban area of Banda Aceh City

Climate change mitigation efforts implemented by the Government of Banda Aceh by building green lanes with trees. The level of the tree’s ability to absorb carbon is highly correlated with the vitality of the tree itself. The health status of a tree can be known by the tree’s damage level. The aim of the research is to provide information on how much damage to trees and the degree of severity. The research was carried out on green lanes on urban streets in Banda Aceh City as many as 8 lines. The object of research is vegetation with the tree growth phase category, and tree damage index assessment using the forest health monitoring method with indicators of tree damage conditions with parameters of damage location code, damage type code, and damage level code. Based on the results, most of the trees suffered light damage. Of the total number of trees observed, there were 1797 individuals, 252 trees were in good health, 1494 were lightly damaged, 40 trees were medium damaged and 11 trees were heavily damaged. The type of damage experienced was dominated by 46% open wound, 32% cancer, 11% brum (excessive branching) and 11% the rest with other kinds of damage.

Transcription factor WOX11 modulates tolerance to cyst nematodes via adventitious lateral root formation.

The transcription factor WUSCHEL-RELATED HOMEOBOX 11 (WOX11) in Arabidopsis (Arabidopsis thaliana) initiates the formation of adventitious lateral roots upon mechanical injury in primary roots. Root-invading nematodes also induce de novo root organogenesis leading to excessive root branching, but it is not known if this symptom of disease involves mediation by WOX11 and if it benefits the plant. Here, we show with targeted transcriptional repression and reporter gene analyses in Arabidopsis that the beet cyst nematode Heterodera schachtii activates WOX11-mediated adventitious lateral rooting from primary roots close to infection sites. The activation of WOX11 in nematode-infected roots occurs downstream of jasmonic acid-dependent damage signaling via ETHYLENE RESPONSE FACTOR109, linking adventitious lateral root formation to nematode damage to host tissues. By measuring different root system components, we found that WOX11-mediated formation of adventitious lateral roots compensates for nematode-induced inhibition of primary root growth. Our observations further demonstrate that WOX11-mediated rooting reduces the impact of nematode infections on aboveground plant development and growth. Altogether, we conclude that the transcriptional regulation by WOX11 modulates root system plasticity under biotic stress, which is one of the key mechanisms underlying the tolerance of Arabidopsis to cyst nematode infections.

Antagonistic activity of secondary metabolites from rhizofunctional bacteria extracts against <i>Fusarium</i>species

Background: Fusarium species remain important fugal pathogens that produce several mycotoxins with adverse effects on both plant and animals. This work aimed to identify biocontrol agent from rhizofunctional bacteria and assess its antagonistic activity against Fusarium sp. using dual culture technique.
 Methodology: Briefly a circular disc of the Fusarium sp. was inoculated at the center of Potato Dextrose Agar (PDA) plate and incubated for three days. The bacterial isolates were then inoculated about 2cm from the Fusarium hyphal tips and incubated for three days, and zone of inhibition was examined. Isolates that showed antagonistic activities against the fungi were subculture in nutrient broth for three days and the metabolites were extracted using ethyl acetate. The metabolic extracts were tested against the fungi using the agar disc diffusion method.
 Results: Of the 20 rhizofunctional bacterial isolates screened for antagonistic activities against Fusarium sp., 5 showed active antagonism against the fungi with observed clear zone of inhibition in the dual culture, and microscopic examination of the fungal hyphae showed excessive and diffused hyphal branching with hyphal swelling. Ethyl acetate extracts from nutrient broth cultures did not show any zone of inhibition in dual culture against the Fusarium sp. All the 5 bacterial isolates were Gram positive strains but only 2 isolates (2a and 3K) were lipase positive, which may indicate that the mechanisms of antagonism could be due to the production of enzymes that have the ability to hydrolyze the cell wall and membrane lipids of the fungi.
 Conclusion: The rhizoplane and rhizosphere of plants could be great sources of biocontrol agents and that bacterial isolates 2a and 3K have the potential to be used as antifungal agents against Fusarium sp. Molecular identification of 2a and 3K bacterial isolates to the species level is recommended.

Ocular features associated with mutations in ATOH7 gene overlap those with familial exudative vitreoretinopathy.

To report the ocular findings in 3 patients with a mutation in the ATOH7 gene. The clinical findings were collected from the medical records including those for magnetic resonance imaging (MRI). Genetic testing of the ATOH7 gene was performed. Three patients of two families who had poor vision since infancy were studied. The 3 patients had varying degrees of intraocular vascular proliferation associated with advanced retinal detachments as falciform retinal folds or total retinal detachments. This state is referred to as congenital retinal nonattachment. One eye of a sibling had fluorescein angiographic findings of excessive branching of the retinal vessels and fluorescent dye leakage that were consistent with those of familial exudative vitreoretinopathy (FEVR). Bilateral hypoplasia of the optic nerve was found in all three patients by MRI. Genetic analysis showed a known in-frame deletion of the ATOH7 gene in all three patients. This is the first report of a patient with a mutation in the ATOH7 gene that had typical vascular patterns of FEVR in the peripheral retina. The ocular features associated with mutations in the ATOH7 gene overlap those with FEVR at both the early and advanced stages.

Hemp Abnormal Growth Is Attributed to Mono-, Di-, or Tri-Infections of Spiroplasma citri, ‘Candidatus Phytoplasma trifolii’, and Beet Curly Top Virus

As hemp ( Cannabis sativa) emerges as a commercial crop in Nevada, mild to severe abnormal growth has been observed from many plants in commercial fields throughout the growing season. Affected plants exhibited a group of symptoms typically including stunting, leaf yellowing, excessive apical branching, clusters of witches’ broom, leaf rolling upwards, and leaf mottling and mosaic. These symptoms mostly showed up together in a plant or a crop and are defined here as abnormal growth syndrome (AGS). Between 2017 and 2019, the Nevada State Plant Pathology and Molecular Diagnostic Lab received 67 symptomatic hemp samples from Nevada for diagnosis, and ‘ Candidatus Phytoplasma trifolii’ was detected in 14 samples (21%). To investigate additional biotic agents associated with AGS, PCR products generated by primers P1/P7 were cloned into a pGEM-T vector and sequenced, and Spiroplasma citri DNA was found in two samples (3%). All 67 DNA samples were further tested for beet curly top virus (BCTV), and 57 samples (85%) were found to be infected by BCTV. Twelve samples (18%) were coinfected by both ‘ Ca. Phytoplasma trifolii’ and BCTV, and two (3%) were coinfected by S. citri, ‘ Ca. Phytoplasma trifolii’, and BCTV. The findings suggest that BCTV is the most prevalent pathogen causing the hemp abnormal growth in Nevada, but ‘ Ca. Phytoplasma trifolii’ and S. citri may also contribute to the severity and complexity of symptoms. Thus, hemp abnormal growth can be attributed to single, dual, or triple infections of these three leafhopper-vectored mollicutes and virus. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

MBL-1 and EEL-1 affect the splicing and protein levels of MEC-3 to control dendrite complexity.

Transcription factors (TFs) play critical roles in specifying many aspects of neuronal cell fate including dendritic morphology. How TFs are accurately regulated during neuronal morphogenesis is not fully understood. Here, we show that LIM homeodomain protein MEC-3, the key TF for C. elegans PVD dendrite morphogenesis, is regulated by both alternative splicing and an E3 ubiquitin ligase. The mec-3 gene generates several transcripts by alternative splicing. We find that mbl-1, the orthologue of the muscular dystrophy disease gene muscleblind-like (MBNL), is required for PVD dendrite arbor formation. Our data suggest mbl-1 regulates the alternative splicing of mec-3 to produce its long isoform. Deleting the long isoform of mec-3(deExon2) causes reduction of dendrite complexity. Through a genetic modifier screen, we find that mutation in the E3 ubiquitin ligase EEL-1 suppresses mbl-1 phenotype. eel-1 mutants also suppress mec-3(deExon2) mutant but not the mec-3 null phenotype. Loss of EEL-1 alone leads to excessive dendrite branches. Together, these results indicate that MEC-3 is fine-tuned by alternative splicing and the ubiquitin system to produce the optimal level of dendrite branches.

First report of 'Candidatus Phytoplasma asteris' associated with witches'-broom disease of Pinus yunnanensis in China.

Pinus yunnanensis is an evergreen tree belonging to Pinaceae. The species is distributed in the east of Tibet, southwest of Sichuan, southwest of Yunnan, southwest of Guizhou and northwest of Guangxi. It is an indigenous and pioneer tree species for barren mountain afforestation in southwest China. P. yunnanensis has important value to both the building and medicine industries (Liu et al. 2022). In May 2022, P. yunnanensis showing witches'-broom symptom were found in Panzhihua City, Sichuan Province, China. The symptomatic plants had yellow or red needle, and exhibited plexus bud and needle wither. The lateral buds of infected pines developed into twigs. Some lateral buds grew in clusters and a few sprouted needles (Fig.1). The disease was named the P. yunnanensis witches'-broom disease (PYWB) and was found in some areas of Miyi, Renhe, and Dongqu. More than 9% of the pines showed these symptoms in the three areas surveyed, and the disease was spreading. A total of 39 samples were collected from three areas, including 25 symptomatic plants and 14 asymptomatic plants, respectively. The lateral stem tissues of 18 samples were observed under a scanning electron microscope (Hitachi S-3000N). Spherical bodies were found in the phloem sieve cells of symptomatic pines (Fig.1). Total DNA was extracted from 18 plant samples using the CTAB method (Porebski et al. 1997) and subjected to nested-PCR testing. Double-distilled water and DNA extracted from asymptomatic plants were used as negative controls, and DNA extracted from Dodonaea viscosa affected by the D. viscosa witches'-broom disease was used as positive control. Nested PCR was employed to amplify the pathogen's 16S rRNA gene (Lee et al. 1993; Schneider et al. 1993) and 1.2 kb segment were produced (GenBank accessions OP646619; OP646620; OP646621). PCR specific to the ribosomal protein (rp) gene yielded segment of approximately 1.2 kb (Lee et al. 2003)(GenBank accessions OP649589; OP649590; OP649591). The fragment size from 15 samples was consistent with the positive control, confirming the association of phytoplasma with the disease. A BLAST analysis of the 16S rRNA sequences of P. yunnanensis witches'-broom phytoplasma showed that it shared 99.12% ~99.76% identity with that of Trema laevigata witches'-broom phytoplasma (GenBank accession MG755412). The rp sequence shared 99.84%~99.92% identity with that of Cinnamomum camphora witches'-broom phytoplasma (GenBank accession OP649594). An analysis with iPhyClassifier (Zhao et. 2013) showed that the virtual RFLP pattern derived from OP646621 16S rDNA fragment of PYWB phytoplasma is identical (similarity coefficient 1.00) to the reference pattern of 16Sr group I, subgroup B (OY-M, GenBank accession AP006628). The phytoplasma is identified as a 'Candidatus Phytoplasma asteris'-related strain belonging to sub-group 16SrI-B. Interestingly, compared to AP006628, the virtual RFLP pattern derived from OP646619 and OP646620 fragments exhibit differences in three and one cleavage site, with a similarity coefficient of 0.92 and 0.97, respectively (Fig.2). These strains may represent a new subgroup within the 16Sr group I. The phylogenetic tree was reconstructed based on 16S rRNA and rp gene sequences using MEGA versio6.0 (Tamura et al. 2013). The analysis was conducted using the neighbor-joining (NJ) method with 1,000 replicates of bootstrap analysis. The results indicated that the PYWB phytoplasmas grouped into clades including phytoplasmas belonging to 16SrI-B and rpI-B, respectively (Fig.3). In addition, 2-year-old P. yunnanensis were used for grafting assays in nursery, and the twigs from infected pine under natural conditions were used as a scion, and the phytoplasma were detected using nested PCR after grafting for 40 d (Fig.4). In 2008-2014, P. sylvestris and P. mugo in Lituania had excessive branching symptoms that were attributed to 'Ca. Phtyoplasma Pini' (16SrXXI-A) or asteris' (16SrI-A) strains (Valiunas et al. 2015). In 2015, P. pungens with abnormal shoot branching in Maryland were found to be infected by 'Ca. Phytoplasma pini' strain (16SrXXI-B) (Costanzo et al. 2016). To the best of our knowledge, P. yunnanensis is a new host of 'Ca. Phytoplasma asteris'-related strain (16SrI-B) in China. The newly emerged disease is a threat to pines.

A Novel Constitutively Active c . 98 G > C , p.(R33P) Variant in RAB11A Associated with Intellectual Disability Promotes Neuritogenesis and Affects Oligodendroglial Arborization

Whole exome sequencing/whole genome sequencing has accelerated the identification of novel genes associated with intellectual disabilities (ID), and RAB11A which encodes an endosomal small GTPase is among them. However, consequent neural abnormalities have not been studied, and pathophysiological mechanisms underlying the ID and other clinical features in patients harboring RAB11A variants remain to be clarified. In this study, we report a novel de novo missense variant in RAB11A, NM_004663.5: c . 98 G > C , which would result in NP_004654.1: p.(R33P) substitution, in a Japanese boy with severe ID and hypomyelination. Biochemical analyses indicated that the RAB11A-R33P is a gain-of-function, constitutively active variant. Accordingly, the introduction of the RAB11A-R33P promoted neurite extension in neurons like a known constitutively active variant Rab11A-Q70L. In addition, the RAB11A-R33P induced excessive branching with thinner processes in oligodendrocytes. These results indicate that the gain-of-function RAB11A-R33P variant in association with ID and hypomyelination affects neural cells and can be deleterious to them, especially to oligodendrocytes, and strongly suggest the pathogenic role of the RAB11A-R33P variant in neurodevelopmental impairments, especially in the hypomyelination.

StHAB1, a negative regulatory factor in abscisic acid signaling, plays crucial roles in potato drought tolerance and shoot branching.

Abscisic acid (ABA) is critical in drought tolerance and plant growth. Group A protein type 2C phosphatases (PP2Cs) are negative regulators of ABA signaling and plant adaptation to stress. Knowledge about the functions of potato group A PP2Cs is limited. Here, we report that the potato group A PP2C StHAB1 is broadly expressed in potato plants and strongly induced by ABA and drought. Suppression of StHAB1 enhanced potato ABA sensitivity and drought tolerance, whereas overexpression of the dominant mutant StHAB1G276D compromised ABA sensitivity and drought tolerance. StHAB1 interacts with almost all ABA receptors and the Snf1-Related Kinase OST1. Suppressing StHAB1 and overexpressing StHAB1G276D alter potato growth morphology; notably, overexpression of StHAB1G276D causes excessive shoot branching. RNA-sequencing analyses identified that the auxin efflux carrier genes StPIN3, StPIN5, and StPIN8 were up-regulated in StHAB1G276D-overexpressing axillary buds. Correspondingly, the auxin concentration was reduced in StHAB1G276D-overexpressing axillary buds, consistent with the role of auxin in repressing lateral branch outgrowth. The expression of BRANCHED1s (StBRC1a and StBRC1b) was unchanged in StHAB1G276D-overexpressing axillary buds, suggesting that StHAB1G276D overexpression does not cause axillary bud outgrowth via regulation of BRC1 expression. Our findings demonstrate that StHAB1 is vital in potato drought tolerance and shoot branching.

Effects of Electrical Stimulation on Facial Paralysis Recovery after Facial Nerve Injury: A Review on Preclinical and Clinical Studies.

Various methods have been used to improve function and manage facial nerve injury. Although electrical stimulation therapy is frequently used to treat facial paralysis, its effects have been found to vary and no clear standards have been developed. The current review describes the results of preclinical and clinical studies evaluating the effectiveness of electrical stimulation therapy in promoting the recovery of a peripheral facial nerve injury. Evidence is presented showing the efficacy of electrical stimulation in promoting nerve regeneration after peripheral nerve injuries in both animal models and human patients. The ability of electrical stimulation to promote the recovery of facial paralysis was found to depend on the type of injury (compression or transection), the species of animal tested, the type of disease, the frequency and method of electrical stimulation, and the duration of the follow-up. Electrical stimulation, however, can also have potential negative outcomes, such as reinforcing synkinesis, including mistargeted axonal regrowth via inappropriate routes; excessive collateral axonal branching at the lesion site; and multiple innervations at neuromuscular junctions. Because of the inconsistencies among studies and the low quality of evidence, electrical stimulation therapy is not currently regarded as a primary treatment of facial paralysis in patients. However, understanding the effects of electrical stimulation, as determined in preclinical and clinical studies, is important for the potential validity of future research on electrical stimulation.

Ptf1a expression is necessary for correct targeting of spiral ganglion neurons within the cochlear nuclei

Two transcription factors, Atoh1 and Ptf1a, are essential for cochlear nuclei development. Atoh1 is needed to develop glutamatergic neurons, while Ptf1a is required to generate glycinergic and GABAergic neurons that migrate into the cochlear nucleus. While central projections of inner ear afferents are normal following loss of Atoh1, we wanted to know whether the loss of Ptf1a affects central projections. We found that in Ptf1a mutants, initially, afferents show a normal projection; however, a transient posterior expansion of projections to the dorsal cochlear nucleus occurs at a later stage. In addition, in older (E18.5) Ptf1a mutant mice, excessive neuronal branches form beyond the normal projection to the anterior and posterior ventral cochlear nuclei. Our results on Ptf1a null mice are comparable to that observed in loss of function Prickel1, Npr2, or Fzd3 mouse mutants. The disorganized tonotopic projections that we report in Ptf1a mutant embryos might be functionally relevant, but testing this hypothesis requires Ptf1a KO mice at postnatal stages that unfortunately cannot be performed due to their early death.

GTPase-activating protein TBC1D5 coordinates with retromer to constrain synaptic growth by inhibiting BMP signaling

Formation and plasticity of neural circuits rely on precise regulation of synaptic growth. At Drosophila neuromuscular junction (NMJ), Bone Morphogenetic Protein (BMP) signaling is critical for many aspects of synapse formation and function. The evolutionarily conserved retromer complex and its associated GTPase-activating protein TBC1D5 are critical regulators of membrane trafficking and cellular signaling. However, their functions in regulating the formation of NMJ are less understood. Here, we report that TBC1D5 is required for inhibition of synaptic growth, and loss of TBC1D5 leads to abnormal presynaptic terminal development, including excessive satellite boutons and branch formation. Ultrastructure analysis reveals that the size of synaptic vesicles and the density of subsynaptic reticulum are increased in TBC1D5 mutant boutons. Disruption of interactions of TBC1D5 with Rab7 and retromer phenocopies the loss of TBC1D5. Unexpectedly, we find that TBC1D5 is functionally linked to Rab6, in addition to Rab7, to regulate synaptic growth. Mechanistically, we show that loss of TBC1D5 leads to upregulated BMP signaling by increasing the protein level of BMP type II receptor Wishful Thinking (Wit) at NMJ. Overall, our data establish that TBC1D5 in coordination with retromer constrains synaptic growth by regulating Rab7 activity, which negatively regulates BMP signaling through inhibiting Wit level.

Trigeminal Sensory Supply Is Essential for Motor Recovery after Facial Nerve Injury

Recovery of mimic function after facial nerve transection is poor. The successful regrowth of regenerating motor nerve fibers to reinnervate their targets is compromised by (i) poor axonal navigation and excessive collateral branching, (ii) abnormal exchange of nerve impulses between adjacent regrowing axons, namely axonal crosstalk, and (iii) insufficient synaptic input to the axotomized facial motoneurons. As a result, axotomized motoneurons become hyperexcitable but unable to discharge. We review our findings, which have addressed the poor return of mimic function after facial nerve injuries, by testing the hypothesized detrimental component, and we propose that intensifying the trigeminal sensory input to axotomized and electrophysiologically silent facial motoneurons improves the specificity of the reinnervation of appropriate targets. We compared behavioral, functional, and morphological parameters after single reconstructive surgery of the facial nerve (or its buccal branch) with those obtained after identical facial nerve surgery, but combined with direct or indirect stimulation of the ipsilateral infraorbital nerve. We found that both methods of trigeminal sensory stimulation, i.e., stimulation of the vibrissal hairs and manual stimulation of the whisker pad, were beneficial for the outcome through improvement of the quality of target reinnervation and recovery of vibrissal motor performance.

Evidence of mixed infection of phytoplasma and begomovirus associated with Withania somnifera and Capsicum annum plants from Uttar Pradesh, India

Symptoms of yellow discoloration, curling of leaves, and little leaves with excessive branching were observed on Withania somnifera and Capsicum annum plants, with an average incidence of 5 and 8%, respectively, in Hardoi District of Uttar Pradesh, India during 2016–17 and 2017–18. The little leaf and excessive branching suggested the possibility of phytoplasma association. However, the presence of whiteflies in the vicinity and leaf curling suggested begomovirus infection. Three leaf samples from Withania ­somnifera and Capsicum annum symptomatic plants and one from non-symptomatic leaf were used for DNA isolation and were subjected to PCR using P1/P6 primers and nested PCR R16F2n/R16r2 primers, respectively, for the detection of phytoplasma. The begomovirus coat protein-specific primer AV1F/AV1R was used to detect begomovirus infection. Nested PCR amplified the ∼1.2 kb amplicon in all six symptomatic leaves and no amplification was observed in non-symptomatic leaves. The CP region primer yielded ∼800 bp amplicons in all three symptomatic samples of each plant. Amplified products from both primers were eluted, purified, and sequenced. The phytoplasma sequence obtained from Withania somnifera (MH789552) shared the highest sequence identity (99.92%) with other isolates of a clover proliferation group (16SrVI-D) group of phytoplasmas. However, Capsicum annum plants shared the highest identity (99%) with the Ca. P. asteris-related (16SrI-B) group of phytoplasma. In silico RFLP analysis of the 1.2 kbp product of the 16S rRNA sequence of the W. somnifera and C. annum phytoplasma strains submitted to the pDRAW32 tool (https://www.acaclone.com/) and phylogenetic analysis through the MEGA 6.0 tool confirmed that it as a member of the 16SrVI-D subgroup and 16SrI-B subgroup, respectively. The CP gene sequence of the W. somnifera isolate (MW176071) showed maximum identity (99%) with several isolates of the Tomato leaf curl virus reported from various places in India. However, the C. annum isolate (MW420480) shared maximum identity with the Ageratum enation virus. The investigation confirmed the mixed infection of ToLCV and 16SrVI-D group phytoplasma in the W. somnifera plant and 16SrI-B alongwith AEV in C. annum plants.