Overexpressing GLUTAMINE SYNTHETASE 1;2 maintains carbon and nitrogen balance under high-ammonium conditions and results in increased tolerance to ammonium toxicity in hybrid poplar.

The glutamine synthetase/glutamic acid synthetase (GS/GOGAT) cycle plays important roles in N metabolism, growth, development, and stress resistance in plants. Excess ammonium (NH4+) restricts growth, but GS can help to alleviate its toxicity. In this study, the 84K model clone of hybrid poplar (Populus alba × P. tremula var. glandulosa), which has reduced biomass accumulation and leaf chlorosis under high-NH4+ stress, showed less severe symptoms in transgenic lines overexpressing GLUTAMINE SYNTHETASE 1;2 (GS1;2-OE), and more severe symptoms in RNAi lines (GS1;2-RNAi). Compared with the wild type, the GS1;2-OE lines had increased GS and GOGAT activities and higher contents of free amino acids, soluble proteins, total N, and chlorophyll under high-NH4+ stress, whilst the antioxidant and NH4+ assimilation capacities of the GS1;2-RNAi lines were decreased. The total C content and C/N ratio in roots and leaves of the overexpression lines were higher under stress, and there were increased contents of various amino acids and sugar alcohols, and reduced contents of carbohydrates in the roots. Under high-NH4+ stress, genes related to amino acid biosynthesis, sucrose and starch degradation, galactose metabolism, and the antioxidant system were significantly up-regulated in the roots of the overexpression lines. Thus, overexpression of GS1;2 affected the carbon and amino acid metabolism pathways under high-NH4+ stress to help maintain the balance between C and N metabolism and alleviate the symptoms of toxicity. Modification of the GS/GOGAT cycle by genetic engineering is therefore a potential strategy for improving the NH4+ tolerance of cultivated trees.

Resection and reconstruction for radial polydactyly Type IV-D in 206 cases: a retrospective clinical analysis.

BACKGROUND: Radial Polydactyly Type IV-D deformity is difficult to treat because of the most complex bone and soft tissue anomalies. Resection and reconstruction for one of the two thumbs was an option for treatment. OBJECTIVE: The study was to present our method of resection and reconstruction with a new incision for radial polydactyly Type IV-D and evaluate the clinical efficacy comprehensively using Rotterdam assessment system in a large sample. METHODS: 206 cases of type IV-D thumb duplication underwent resection and reconstruction surgical treatment between 2010 and 2019. Two equal triangle flap incisions were designed around the radial thumb. The radial thumb was resected and the ulnar thumb was reconstructed in aspects of bone, tendons, ligaments and abductor pollicis brevis. The clinical results were evaluated using Rotterdam assessment system. RESULTS: The mean follow-up period was 2.2 years (SD 1.5). The mean age of the patients was 9 months (SD 1.8) at the time of operation. The mean ranges of active IP and MP joint flexion and extension were 110° and 26°. The mean angulations for IP and MP joint instabilities were 3° and 11°, relatively. Angulation for palmar abduction was 58°. The mean appearance domain score was 8.9. The average parental satisfaction score was 2.5 and the average patient-reported pain score was 2.1. The mean functional domain score for all patients was 6.6. The average appearance domain score was 8.9. The mean patient-reported domain score was 4.5. The mean Rotterdam outcome score was 20.0, equivalent to 67% of the full score. The postoperative score of patients over two years old was significantly lower than that of patients under two years old. CONCLUSION: Resection and reconstruction method with two equal triangle flap incisions was a recommended treatment for radial polydactyly Type IV-D. LEVEL OF EVIDENCE: IV.

Genome Identification and Expression Profiles in Response to Nitrogen Treatment Analysis of the Class I CCoAOMT Gene Family in Populus.

Lignin is essential for the characteristics and quality of timber. Nitrogen has significant effects on lignin contents in plants. Nitrogen has been found to affect wood quality in plantations and lignin content in plants. Caffeoyl-CoA 3-O-methyltransferase (CCoAOMT) is an important methyltransferase in lignin biosynthesis. However, the classification of woody plant CCoAOMT gene family members and the regulation mechanism of nitrogen are not clear. Bioinformatics methods were used to predict the members, classification, and transcriptional distribution of the CCoAOMT gene family in Populus trichocarpa. The results showed that there were five PtCCoAOMTs identified, and they could be divided into three sub-groups according to their structural and phylogenetic features. The results of tissue expression specificity analysis showed that: PtCCoAOMT1 was highly expressed in roots and internodes; PtCCoAOMT2 was highly expressed in roots, nodes, and internodes, PtCCoAOMT3 was highly expressed in stems; PtCCoAOMT4 was highly expressed in young leaves, and, PtCCoAOMT5 was highly expressed in roots. Different forms and concentrations of nitrogen had varying effects on the expression patterns of genes in different plant tissue types. The results of real-time PCR showed that the expression levels of PtCCoAOMT1 and PtCCoAOMT2 in stems increased significantly under different forms of nitrogen. PtCCoAOMT3 and PtCCoAOMT4 were induced by nitrate nitrogen in upper stems and lower leaves, respectively. PtCCoAOMT4 and PtCCoAOMT5 were induced by different concentrations of nitrate nitrogen in lower stems and roots, respectively. These results could provide valuable information for revealing the differences between functions and expression patterns of the various CCoAOMT gene family members under different forms and concentrations of exogenous nitrogen in poplar.

Molecular Mechanisms of Diverse Auxin Responses during Plant Growth and Development.

The plant hormone auxin acts as a signaling molecule to regulate numerous developmental processes throughout all stages of plant growth. Understanding how auxin regulates various physiological and developmental processes has been a hot topic and an intriguing field. Recent studies have unveiled more molecular details into how diverse auxin responses function in every aspect of plant growth and development. In this review, we systematically summarized and classified the molecular mechanisms of diverse auxin responses, and comprehensively elaborated the characteristics and multilevel regulation mechanisms of the canonical transcriptional auxin response. On this basis, we described the characteristics and differences between different auxin responses. We also presented some auxin response genes that have been genetically modified in plant species and how their changes impact various traits of interest. Finally, we summarized some important aspects and unsolved questions of auxin responses that need to be focused on or addressed in future research. This review will help to gain an overall understanding of and some insights into the diverse molecular mechanisms of auxin responses in plant growth and development that are instrumental in harnessing genetic resources in molecular breeding of extant plant species.

Genome-wide identification of FRK genes in Populus trichocarpa and their expression under different nitrogen treatments.

Fructokinase (FRK) is the main fructose phosphorylase and plays an important role in catalyzing the irreversible reaction of free fructose phosphorylation. In order to study the regulatory effect of different forms and concentrations of nitrogen on PtFRK genes in Populus trichocarpa, seven genes encoding the hypothetical FRK proteins were identified in Populus trichocarpa genome by bioinformatics method. Phylogenetic analysis revealed that PtFRK family genes can be divided into two subgroups: SI (PtFRK 1, 3, 4, 6) and SII (PtFRK 2, 5, 7). The tissue-specific expression data obtained from PopGenIE indicate that PtFRK2, 3, 4 and 5 are expressed highly in the stem. Quantitative real-time RT-PCR illustrate that PtFRK1-7 showed different expression patterns in different tissues under different concentrations and morphological nitrogen application. Under high nitrate treatment, the expression levels of PtFRK1, 2, 3 and 6 in stem increased significantly, while under low nitrate treatment, only the expression of PtFRK1, 4 in the upper stem and the expression of PtFRK3, 5 in the lower stem increased significantly. In contrast, ammonium tends to inhibit the expression of PtFRKs in lower stems, the expression levels of PtFRK2, 3, 4 and 5 are significantly reduced under ammonium treatment. However, high ammonium had significant effects on PtFRK6 in the apical bud and upper leaves, which were 6 and 8 times of the control, respectively. These results laid the foundation for the study of the PtFRK gene family of poplar and provided a theoretical basis for the molecular mechanism of nitrogen regulating cell wall development. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12298-021-01055-6.

Doppler sonography for ulnar neuropathy at the elbow.

INTRODUCTION: The aim of this study was to determine the value of Doppler sonography for evaluation of ulnar neuropathy at the elbow (UNE). METHODS: A total of 102 patients with a clinical suspicion of UNE and 50 healthy controls were examined by Doppler and gray-scale sonography. RESULTS: Intraneural vascularization was found in 31 (46.3%) of 67 patients with confirmed UNE and in 3 (6.0%) of 50 healthy controls. Combining gray-scale sonography measurement of nerve size and the presence of intraneural vascularization increased sensitivity by 3%, but decreased the specificity. Patients with intraneural vascularization had more severe disease than those without intraneural vascularization (P < 0.05). The variables obtained by Doppler sonography were associated with the severity of UNE (P < 0.05). CONCLUSIONS: Doppler sonography is not helpful for diagnosing UNE but can be used to assess the severity of UNE. Muscle Nerve 54: 258-263, 2016.

An alternative method to reconstruct grade-3 hypoplastic thumb by hemi-metacarpal transfer.

We designed a new method using hemi-longitudinal second metacarpal bone to reconstruct grade-3 hypoplastic thumbs. Seven patients were treated in two stages. In the first stage, the second metacarpal was split longitudinally and transferred to reconstruct the first metacarpal. In the second stage, opponensplasty was performed by transferring the FDS tendon. Bone union was achieved in all cases. All patients could oppose to their middle finger at least. They managed to do daily activities such as writing, eating, using smartphones and so on. This is a useful procedure to preserve a 5-digit hand with good function in treating grade-3 hypoplastic thumbs, with no harm to the foot and no need for vascular anastomosis. LEVEL OF EVIDENCE: IV.

Characterization of DmToll and DmToll7 homologue in Litopenaeus vannamei based on structure analysis.

Toll-like receptors (TLRs) are a family of pattern recognition receptors (PRRs) that recognize invading pathogens and activate downstream signaling pathways. The number of 10 Tolls is found in Litopenaeus vannamei but have not yet been identified as the corresponding Toll homologue of model animal. In this study, we predicted the three-dimensional (3D) structures of 10 LvTolls (LvToll1-10) with AlphaFold2 program. The per-residue local distance difference test (pLDDT) scores of LvTolls showed the predicted structure of LvTolls had high accuracy (pLDDT>70). By structural analysis, 3D structures of LvToll2 and LvToll3 had high similarity with Drosophila melanogaster Toll and Toll7, respectively. 3D structure of LvToll7 and LvToll10 were not similar to that of other LvTolls. Moreover, we also predicted that LvSpätzle4 had high structural similarity to DmSpätzle. There were 9 potential hydrogen bonds in LvToll2-LvSpätzle4 complex. Importantly, co-immunoprecipitation assay showed that LvToll2 could bind with LvSpätzle4. Collectively, this study provides new insight for researching invertebrate immunity by identifying the protein of model animal homologue.

How to prevent injury to the palmar cutaneous branch of median nerve and ulnar nerve in a palmar incision in carpal tunnel release, a cadaveric study.

OBJECTIVE: To identify palmar cutaneous branches of median nerve and ulnar nerve (PCBMN and PCBUN) and try to find a safe path at wrist and forearm in the decompression procedure of carpal tunnel syndrome. MATERIALS AND METHODS: Ten formalin-fixed and five fresh-frozen cadaveric forearms were included in the study. The cross point of longitude of middle finger and distal wrist crease was defined as 0 point. Distal wrist crease (DWC) and 0 point were chosen as references for measurements. Several points on the pathway of PCBMN and PCBUN were measured. RESULTS: The average distance between the origin of the PCBMN and PCBUN to the DWC was 4.95 ± 0.88 cm, 10.12 ± 1.50 cm, separately. The average distance between DWC and the point where PCBMN and PCBUN separated from their trunk was found to be 2.09 ± 0.31 cm, 2.90 ± 0.50 cm, separately. The distances between PCBMN, PCBUN and 0 point at DWC level was found to be 0.61 ± 0.12 cm, 0.47 ± 0.31 cm, separately. The diameters of two cutaneous branches were 0.10 ± 0.02 cm, 0.11 ± 0.04 cm, separately. CONCLUSION: The general longitudinal palmar incision could avoid injuries to recurrent branch of median nerve and distal branches of palmar cutaneous nerve can be avoided macroscopically. The area about 5 mm ulnar and 6 mm radial to 0 point at wrist level was a relatively safe area.

Classification and treatment of congenital central slip hypoplasia.

OBJECTIVES: The aim of this study is to make a diagnosis and a classification for congenital central slip hypoplasia. The surgical treatment was determined according to the classification. METHODS: A retrospective study of 25 treated digits in 13 patients with congenital central slip hypoplasia was carried out. The central slip was classified into two types. Type I: The distance between the insertion of central slip and the proximal interphalangeal joint was shorter than or equal to 5 mm. Type II: The distance between the insertion of central slip and the proximal interphalangeal joint was longer than 5 mm. Tendon advancement or tendon graft was used for type I or II, respectively. RESULTS: The preoperative mean extension lag was 91° (range, 80°-100°), and the mean follow-up duration was 18 months (range, 9-24 months). The postoperative mean extension lag was 19° (range, 0°-50°). No matter whether in type I or II, the postoperative ranges of proximal interphalangeal joint extension had significant improvement compared with the preoperative ones. There was no statistical difference of proximal interphalangeal joint extension lag changes before and after surgery between the two types. CONCLUSION: Congenital central slip hypoplasia could be classified into two types. Either tendon advancement or tendon graft might be effective, which depended on the classification.

Manure properties, soil conditions and managerial factors regulate greenhouse vegetable yield with organic fertilizer application across China.

To better understand the responses of vegetable yields in a greenhouse system to organic fertilizer through a quantitative evaluation based on peer-reviewed journal articles and in consideration of environmental managerial factors. We conducted a meta-analysis of 453 paired observations from 68 peer-reviewed journal articles to assess the response of vegetable yields in greenhouse vegetable systems in China to organic fertilization. Compared with the control (no organic fertilizer), organic fertilization significantly increased the yields of vegetables by 44.11% on average. The response of vegetable yields to organic fertilizer tended to increase with the increasing experimental duration. Organic fertilizer application had the greatest potential for leafy vegetables (+76.44%), in loamy soils (+53.94%), at moderate organic fertilizer carbon input levels (+54.13%), and in soils with moderate initial soil total nitrogen levels (+50.89%). Aggregated boosted tree analysis indicated that organic fertilizer carbon inputs, vegetable type and experimental duration were the predominant factors that manipulated the response of vegetable yields to organic fertilizer application. The rational application of farmyard manure would be a promising strategy for increasing vegetable yields in greenhouse vegetable systems in China. Factoring in vegetable type, carbon and nitrogen inputs of organic fertilizer, and soil texture would benefit vegetable yields with the application of organic fertilizer.

Identification and expression analysis of the PtGATL genes under different nitrogen and carbon dioxide treatments in Populus trichocarpa.

Pectin is one of the most important components of the plant cell wall. Galacturonosyltransferase-like (GATL) is an important enzyme involved in forming pectin in Arabidopsis thaliana. In this study, 12 PtGATL genes were identified and characterized based on the Populus trichocarpa genome using bioinformatics methods. The results showed that the PtGATLs contained four typical motifs, including DXD, LPPF, GLG, and HXXGXXKPW. According to phylogenetic analysis, PtGATLs were divided into six groups. Chromosome distribution and genome synteny analysis showed that there were 11 segmental-duplicated gene pairs with repeated fragments on chromosomes 2, 5, 7, 8, 10, and 14. Tissue-specific expression profiles indicated that these PtGATLs had different expression patterns. The transcription level of PtGATLs was regulated by different carbon dioxide and nitrogen concentrations. In conclusion, the identification and analysis of PtGATL genes in poplar provide important information on the gene function. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-022-03129-y.

Factors shaping soil organic carbon stocks in grass covered orchards across China: A meta-analysis.

Orchard grass coverage has been widely adopted to increase fruit yield by improving soil fertility. However, the impact of the environment on the changes in soil organic carbon (SOC) stocks consecutive to orchard grass coverage remain poorly quantified at a large scale. The present study aimed to examine the responses of SOC stocks to grass coverage at a soil depth of 0-30 cm in orchards compared to clean tillage. A total of 342 observations across China from 139 peer-reviewed publications were subjected to meta-analysis. Aggregated boosted tree analysis was performed, evaluating the determinants of SOC stocks, such as plant traits (e.g., fruit tree type, grass type, orchard age, and grass age), edaphic variables (e.g., initial SOC and nitrogen concentration, soil pH, and soil clay content), climatic factors (e.g., mean annual precipitation (MAP) and mean annual temperature (MAT)), and management practices (e.g., grass source, grass growing mode, fertilization, grass mowing, placement of mowed residues, and irrigation). On average, orchard grass coverage significantly enhanced SOC stocks by 21.47% (percentage change) compared to clean tillage. Biotic and abiotic factors influenced this increase in SOC stocks following grass coverage in orchards to different extents. Grass age and soil clay content were the main determinants driving the variation in the SOC stocks following grass coverage in orchards. Thus, we propose an efficient way to optimize C sequestration in grass covered orchards, regarding plant traits, climatic factors, edaphic variables, and management practices. Longer than 12 months of surface grass coverage with cultivated grass species in mature deciduous fruit orchards (≥5 years) efficiently increased SOC stocks. This is particularly the case for acidic (pH < 6.5) soils with low C content (SOM < 15 g kg-1) in areas with suitable rainfall and temperature conditions (MAP ≥ 400 mm, MAT ≥ 10 °C). Collectively, this meta-analysis identified orchard grass coverage as a promising practice for significantly increasing SOC stocks at 0-30 cm across large geospatial locations in China.

Bioinformatics analysis of PAE family in Populus trichocarpa and responsiveness to carbon and nitrogen treatment.

Plant Pectin acetylesterase (PAE) belongs to family CE13 of carbohydrate esterases in the CAZy database. The ability of PAE to regulate the degree of acetylation of pectin, an important polysaccharide in the cell wall, affects the structure of plant cell wall. In this study, ten PtPAE genes were identified and characterized in Populus trichocarpa genome using bioinformatics methods, and the physiochemical properties such as molecular weight, isoelectric points, and hydrophilicity, as well as the secondary and tertiary structure of the protein were predicted. According to phylogenetic analysis, ten PtPAEs can be divided into three evolutionary clades, each of which had similar gene structure and motifs. Tissue-specific expression profiles indicated that the PtPAEs had different expression patterns. Real-time quantitative PCR (RT-qPCR) analysis showed that transcription level of PtPAEs was regulated by different CO2 and nitrogen concentrations. These results provide important information for the study of the phylogenetic relationship and function of PtPAEs in Populus trichocarpa. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-02918-1.

Genome-wide analysis of UGDH genes in Populus trichocarpa and responsiveness to nitrogen treatment.

Plant UDP-glucose 6-dehydrogenase (UGDH) is an important enzyme for the formation of hemicellulose and pectin. Previous studies on UGDH have primarily focused on the biosynthesis of cell wall polysaccharides, while few studies have focused on their regulation by exogenous nitrogen. In the present study, four genes encoding PtUGDH proteins were analyzed by bioinformatics methods. And, the expression profiles of PtUGDH genes under different nitrogen treatments were evaluated with qRT-PCR. The results showed that PtUGDHs have conserved NAD coenzyme binding motif GAGYVGG and the catalytic motif GFGGSCFQKDIL. According to the phylogenetic analysis, PtUGDHs were divided into two subgroups. PtUGDH3 and PtUGDH4 were closely related to AtUGDH1 (important for normal development of Arabidopsis cell wall structure). Chromosomal distribution and genome synteny analysis revealed four segmental-duplicated gene pairs on chr4, 8, 10 and 17. Tissue-specific data from PlantGenIE showed that PtUGDH3 and PtUGDH4 were highly expressed in stems. The qRT-PCR detection showed that the expression of PtUGDH3 in the lower stem and PtUGDH2 of upper leaves were significantly increased induced by low ammonium or nitrate condition. This comprehensive analysis of the UGDH family in poplar provides new insights into their regulation by nitrogen, and would increase our understanding of the roles of UGDHs in hemicellulose and pectin biosynthesis in the cell wall and during poplar development. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-02697-9.

Genome-wide analysis of the RGP gene family in Populus trichocarpa and their expression under nitrogen treatment.

Reversible glycosylation polypeptide (RGP) is a type of plant-specific protein, primarily involved in the biosynthesis of cell wall polysaccharides, which in turn changes the shape of the cell walls and affects the wood properties of plants. Poplar is a major industrial timber species, and the RGP gene has not been studied. This study uses bioinformatics methods to predict physical and chemical characters such as molecular weight, isoelectric point, and hydrophilicity; and fluorescent quantitative method to determine the effect of different forms of nitrogen on the transcription level of the gene family. The results showed that there are six RGP homologous genes in the Populus trichocarpa genome, which were distributed on the six chromosomes of P. trichocarpa. The family members have a simple gene structure and contain four exons and introns. Phylogenetic tree analysis showed that RGP genes all belong to Class I in P. trichocarpa. Tissue-specific expression analysis showed that PtRGP1 and PtRGP2 were highly expressed in the stems, PtRGP4 and PtRGP5 were highly expressed in the upper leaves, PtRGR3 and PtRGR6 were expressed in stems and internodes, but the relative expression is not high. Quantitative real-time RT-PCR (qRT-PCR) analyses revealed that PtRGP3 and 6 were up-regulated in the upper stem in response to the low ammonium and high nitrate treatments. The influence of nitrogen on the expression of PtRGP3 and 6 genes may affect the formation of the plant secondary cell wall. This study lays a foundation for further study on the function of RGP genes in P. trichocarpa.

Functional Research on Three Presumed Asparagine Synthetase Family Members in Poplar.

Asparagine synthetase (AS), a key enzyme in plant nitrogen metabolism, plays an important role in plant nitrogen assimilation and distribution. Asparagine (Asn), the product of asparagine synthetase, is one of the main compounds responsible for organic nitrogen transport and storage in plants. In this study, we performed complementation experiments using an Asn-deficient Escherichia coli strain to demonstrate that three putative asparagine synthetase family members in poplar (Populussimonii× P.nigra) function in Asn synthesis. Quantitative real-time PCR revealed that the three members had high expression levels in different tissues of poplar and were regulated by exogenous nitrogen. PnAS1 and PnAS2 were also affected by diurnal rhythm. Long-term dark treatment resulted in a significant increase in PnAS1 and PnAS3 expression levels. Under long-term light conditions, however, PnAS2 expression decreased significantly in the intermediate region of leaves. Exogenous application of ammonium nitrogen, glutamine, and a glutamine synthetase inhibitor revealed that PnAS3 was more sensitive to exogenous glutamine, while PnAS1 and PnAS2 were more susceptible to exogenous ammonium nitrogen. Our results suggest that the various members of the PnAS gene family have distinct roles in different tissues and are regulated in different ways.

Proteomic analysis of trans-hemispheric motor cortex reorganization following contralateral C7 nerve transfer.

Nerve transfer is the most common treatment for total brachial plexus avulsion injury. After nerve transfer, the movement of the injured limb may be activated by certain movements of the healthy limb at the early stage of recovery, i.e., trans-hemispheric reorganization. Previous studies have focused on functional magnetic resonance imaging and changes in brain-derived neurotrophic factor and growth associated protein 43, but there have been no proteomics studies. In this study, we designed a rat model of total brachial plexus avulsion injury involving contralateral C7 nerve transfer. Isobaric tags for relative and absolute quantitation and western blot assay were then used to screen differentially expressed proteins in bilateral motor cortices. We found that most differentially expressed proteins in both cortices of upper limb were associated with nervous system development and function (including neuron differentiation and development, axonogenesis, and guidance), microtubule and cytoskeleton organization, synapse plasticity, and transmission of nerve impulses. Two key differentially expressed proteins, neurofilament light (NFL) and Thy-1, were identified. In contralateral cortex, the NFL level was upregulated 2 weeks after transfer and downregulated at 1 and 5 months. The Thy-1 level was upregulated from 1 to 5 months. In the affected cortex, the NFL level increased gradually from 1 to 5 months. Western blot results of key differentially expressed proteins were consistent with the proteomic findings. These results indicate that NFL and Thy-1 play an important role in trans-hemispheric organization following total brachial plexus root avulsion and contralateral C7 nerve transfer.