Identification of Putative Parasitism Genes Expressed in the Esophageal Gland Cells of the Soybean Cyst Nematode Heterodera glycines

Cloning parasitism genes encoding secretory proteins expressed in the esophageal gland cells is the key to understanding the molecular basis of nematode parasitism of plants. Suppression subtractive hybridization (SSH) with the microaspirated contents from Heterodera glycines esophageal gland cells and intestinal region was used to isolate genes expressed preferentially in the gland cells of parasitic stages. Twenty-three unique cDNA sequences from a SSH cDNA library were identified and hybridized to the genomic DNA of H. glycines in Southern blots. Full-length cDNAs of 21 clones were obtained by screening a gland-cell long-distance polymerase chain reaction cDNA library. Deduced proteins of 10 clones were preceded by a signal peptide for secretion, and PSORT II computer analysis predicted eight proteins as extracellular, one as nuclear, and one as plasmalemma localized. In situ hybridization showed that four of the predicted extracellular clones were expressed specifically in the dorsal gland cell, one in the subventral gland cells, and three in the intestine in H. glycines. The predicted nuclear clone and the plasmalemma-localized clone were expressed in the subventral gland cells and the dorsal gland cell, respectively. SSH is an efficient method for cloning putative parasitism genes encoding esophageal gland cell secretory proteins that may have a role in H. glycines parasitism of soybean

The Parasitome of the Phytonematode Heterodera glycines

Parasitism genes expressed in the esophageal gland cells of phytonematodes encode secretions that control the complex process of plant parasitism. In the soybean cyst nematode, Heterodera glycines, the parasitome, i.e., the secreted products of parasitism genes, facilitate nematode migration in soybean roots and mediate the modification of root cells into elaborate feeding cells required to support the growth and development of the nematode. With very few exceptions, the identities of these secretions are unknown, and the mechanisms of cyst nematode parasitism, therefore, remain obscure. The most direct and efficient approach for cloning parasitism genes and rapidly advancing our understanding of the molecular interactions during nematode parasitism of plants is to create gland cell-specific cDNA libraries using cytoplasm microaspirated from the esophageal gland cells of various parasitic stages. By combining expressed sequence tag analysis of a gland cell cDNA library with high throughput in situ expression localization of clones encoding secretory proteins, we obtained the first comprehensive parasitome profile for a parasitic nematode. We identified 51 new H. glycines gland-expressed candidate parasitism genes, of which 38 genes constitute completely novel sequences. Individual parasitome members showed distinct gland cell expression patterns throughout the parasitic cycle. The parasitome complexity discovered paints a more elaborate picture of host cellular events under specific control by the nematode parasite than previously hypothesized

A Profile of Putative Parasitism Genes Expressed in the Esophageal Gland Cells of the Root-knot Nematode Meloidogyne incognita

Identifying parasitism genes encoding proteins secreted from a nematode\u27s esophageal gland cells and injected through its stylet into plant tissue is the key to understanding the molecular basis of nematode parasitism of plants. Meloidogyne incognita parasitism genes were cloned by microaspirating the cytoplasm from the esophageal gland cells of different parasitic stages to provide mRNA to create a gland cell-specific cDNA library by long-distance reverse-transcriptase polymerase chain reaction. Of 2,452 cDNA clones sequenced, deduced protein sequences of 185 cDNAs had a signal peptide for secretion and, thus, could have a role in root-knot nematode parasitism of plants. High-throughput in situ hybridization with cDNA clones encoding signal peptides resulted in probes of 37 unique clones specifically hybridizing to transcripts accumulating within the subventral (13 clones) or dorsal (24 clones) esophageal gland cells of M. incognita. In BLASTP analyses, 73% of the predicted proteins were novel proteins. Those with similarities to known proteins included a pectate lyase, acid phosphatase, and hypothetical proteins from other organisms. Our cell-specific analysis of genes encoding secretory proteins provided, for the first time, a profile of putative parasitism genes expressed in the M. incognita esophageal gland cells throughout the parasitic cycle

A parasitism gene from a plant-parasitic nematode with function similar to CLAVATA3/ESR (CLE) of Arabidopsis thaliana

The Hg-SYV46 parasitism gene is expressed exclusively in the dorsal oesophageal gland cell of parasitic stages of the soybean cyst nematode, Heterodera glycines, and it encodes a secretory protein that contains a C-terminal motif of the CLAVATA3/ESR-related (CLE) family in Arabidopsis thaliana. In shoot and floral meristems of Arabidopsis, the stem cells secret CLV3, a founding member of the CLE protein family, that activates the CLV1/CLV2 receptor complex and negatively regulates WUSCHEL expression to restrict the size of the stem cell population. Mis-expression of Hg-SYV46 in Arabidopsis (ecotype Columbia-0) under control of the CaMV35S promoter resulted in a wus-like phenotype including premature termination of the shoot apical meristem and the development of flowers lacking the central gynoecium. The wus-like phenotype observed was similar to reports of over-expression of CLV3 and CLE40 in Arabidopsis, as was down-regulation of WUS expression in the shoot apices of 35S::Hg-SYV46/Col-0 plants. Expression of 35S::Hg-SYV46 in a clv3-1 mutant of Arabidopsis was able partially or fully to rescue the mutant phenotype, probably dependent upon localization and level of transgene expression. A short root phenotype, as reported for over-expression of CLV3, CLE40 and CLE19 in roots, was also produced in primary 35S::Hg-SYV46/Col-0 transgenic plants. The results suggest a functional similarity of HG-SYV46 to plant-secreted CLE ligands that may play a role in the differentiation or division of feeding cells induced in plant roots by parasitic nematodes

The predictive value of neutrophil-to-lymphocyte ratio for overall survival and pathological complete response in breast cancer patients receiving neoadjuvant chemotherapy

PurposePrevious studies have reported that neutrophil-to-lymphocyte ratio (NLR) at pre-treatment was predictive for overall survival (OS) and pathologic complete response (pCR) in breast cancer (BC) patients receiving neoadjuvant chemotherapy (NAC). This study aims to explore the predictive role of both pre- and post-NLR for OS as well as longitudinal NLR kinetics towards pCR in BC patients undergoing NAC.MethodsWe retrospectively included 501 BC patients who received NAC from 2009 to 2018. NLR at pre-, mid (every two cycles of NAC)-, and post-treatment were collected. Overall, 421 patients were included in the survival analysis. These patients were randomly divided into a training cohort (n = 224) and a validation cohort (n = 197). A multivariable Cox model was built using all significant factors in the multivariable analysis from the training cohort. The performance of the model was verified in the validation cohort by the concordance index (C-index). Longitudinal analysis for pCR prediction of NLR was performed using a mixed-effects regression model among 176 patients who finished eight cycles of NAC.ResultsThe median follow-up time was 43.2 months for 421 patients. In the training cohort, multivariable analysis revealed that ER status, clinical node stage, pCR, pre-NLR, and post-NLR (all p < 0.05) were independent predictors of OS. The OS nomogram was established based on these parameters. The C-indexes of the nomogram were 0.764 and 0.605 in the training and validation cohorts, respectively. In the longitudinal analysis, patients who failed to achieve pCR experienced an augment of NLR during NAC while NLR remained stable among patients with pCR. Pre-NLR tended to be significantly associated with OS in patients of HER2 overexpressing and TNBC subtypes (all p < 0.05), but not in Luminal A and Luminal B subtypes.ConclusionsThis study demonstrated the prognostic value of both pre-NLR and post-NLR on clinical outcomes in BC patients receiving NAC. A novel nomogram was established to predict OS. Non-pCR patients developed increased NLRs during NAC. Routine assessment of NLR may be a simple and affordable tool to predict prognosis for BC patients receiving NAC

Effects of Kevlar® 29 yarn twist on tensile and tribological properties of self-lubricating fabric liner

Yarn twist in textile technology is an important characteristic since it considerably affects the properties of knitted or woven fabrics. Many researchers have investigated the effect of staple-spun yarn twist on the properties of the yarns and fabrics. However, the effects of twist level of Kevlar® 29 filament yarn on the properties of yarn and its resin-impregnated self-lubricating fabric liner are not fully known yet. In this study, we have investigated the effects of Kevlar® 29 twist level on the tensile and tribological properties of the fabric liner (Kevlar® 29/polytetrafluoroethylene fabric-resin composite). Two unexpected findings about the effect of yarn twist have been observed, namely (1) asynchronous twist effect on the yarn’s and the liner’s tensile strength and (2) dissimilar yarn twist effect on the liner’s performance. These findings are mainly attributed to the synergic contributions of the yarn twist and strength and the interaction of the resin with the yarn orientation in the woven fabric structure of the liner

An ECG Monitoring and Alarming System Based On Android Smart Phone

ECG monitoring in daily life is an important means of treating heart disease. To make it easier for the medical to monitor the ECG of their patients outside the hospital, we designed and developed an ECG monitoring and alarming system based on Android smart phone. In our system, an ECG device collec...武汉大学、中国传媒大学、广东工业大学、中国计量学院、Engineering Information Institute、Scientific Research

Engineering the bacterial endophyte Burkholderia pyrrocinia JK-SH007 for the control of lepidoptera larvae by introducing the cry218 genes of Bacillus thuringiensis

To expand the usage of endophytes in agriculture and in forestry, the insecticidal gene cry218 of Bacillus thuringiensis was introduced into a poplar bacterial endophyte Burkholderia pyrrocinia JK-SH007. The cry218 gene was cloned by polymerase chain reaction (PCR) and was inserted into a PHKT2 expression vector that was introduced into the bacterial endophyte JK-SH007. By using sodium dodecyl sulphate polyacryl amide gel electrophoresis (SDS-PAGE) and western blotting, we confirmed that the engineered bacterial endophyte was successfully constructed, and it harboured insecticidal function after the bioassay in planta. The toxicity of the expressed insecticidal protein was analysed on second instar silkworm. The regression equation showed that the median lethal concentration (LC50) of the insecticidal protein was 0.77 (0.57–1.04) g/L at 72 h. The insecticidal bacteria genetically modified in this study have laid the foundation for further exploitation of biocontrol bacteria

First Report of Field Resistance to Afidopyropen, the Novel Pyropene Insecticide, on Bemisia tabaci Mediterranean (Q Biotype) from China

Afidopyropen, a novel biopesticide, is derived from Aspergillus fumigatus, a fungus, and shows promise as a novel insecticidal agent for the management of the whitefly pest Bemisia tabaci in horticultural and economical crop production. In the present work, we monitored the susceptibilities of B. tabaci to afidopyropen in 18 field populations, sampled from 9 provinces of China, and found that, in comparison with the susceptible strain (MED-S), B. tabaci from most field populations were highly susceptible, except for the Haidian population (HD) which exhibited an approximately 40-fold increase in resistance. The HD population also displayed significant cross-resistance to sulfoxaflor (14.5-fold) but little cross-resistance to cyantraniliprole, flonicamid, imidacloprid, pymetrozine, and thiamethoxam. Afidopyropen resistance of the HD population was determined to be incomplete dominant and autosomal, and synergism assays demonstrated that P450 monooxygenases could contribute to the field-evolved afidopyropen resistance observed in the HD population. These results will further our understanding of the molecular underpinnings of insecticide resistance in B. tabaci and can inform the development of field-based pest control tactics to slow the development of afidopyropen resistance and to control whiteflies more sustainably

Resistance Monitoring for Six Insecticides in Vegetable Field-Collected Populations of Spodoptera litura from China

The common cutworm, Spodoptera litura (Fabricius), is a notorious and damaging insect pest of horticultural crops in China, the management of which largely relies on chemical agents that are limited by the development of chemical resistance in target populations. As such, resistance monitoring of S. litura populations is a necessary part of management strategies of insecticide resistance. In the current work, we monitored resistance to six insecticides in field-collected populations of S. litura sampled from eleven provinces across China in 2021. The results show that S. litura populations developed significant resistance against chlorantraniliprole, cyantraniliprole, metaflumizone, and pyridalyl and low levels of resistance to chromafenozide. However, S. litura populations were susceptible or exhibited minimal resistance to tetraniliprole. Possible cross-resistances between chlorantraniliprole, cyantraniliprole, metaflumizone, pyridalyl, and chromafenozide were found by pairwise correlation, which also revealed that tetraniliprole lacked cross-resistance with all insecticides tested. Our results suggest suspending the use of chemical agents against which S. litura displayed significant field-evolved resistance, such as chlorantraniliprole, metaflumizone, and pyridalyl, in favor of pesticides against which S. litura was susceptible or exhibited minimal resistance, such as tetraniliprole and chromafenozide, which may help slow the development of insecticide resistance, and in which field management programs aimed at controlling S. litura in China would benefit from the integration of such survey-informed insecticide application strategies. Moreover, the baseline susceptibility confirmed for the six tested insecticides can contribute to design strategies of resistance management for S. litura