Telomere-to-telomere and gap-free genome assembly of a susceptible grapevine species (Thompson Seedless) to facilitate grape functional genomics.

Grapes are globally recognized as economically significant fruit trees. Among grape varieties, Thompson Seedless holds paramount influence for fresh consumption and for extensive applications in winemaking, drying, and juicing. This variety is one of the most efficient genotypes for grape genetic modification. However, the lack of a high-quality genome has impeded effective breeding efforts. Here, we present the high-quality reference genome of Thompson Seedless with all 19 chromosomes represented as 19 contiguous sequences (N50 = 27.1 Mb) with zero gaps and prediction of all telomeres and centromeres. Compared with the previous assembly (TSv1 version), the new assembly incorporates an additional 31.5 Mb of high-quality sequenced data with annotation of a total of 30 397 protein-coding genes. We also performed a meticulous analysis to identify nucleotide-binding leucine-rich repeat genes (NLRs) in Thompson Seedless and two wild grape varieties renowned for their disease resistance. Our analysis revealed a significant reduction in the number of two types of NLRs, TIR-NB-LRR (TNL) and CC-NB-LRR (CNL), in Thompson Seedless, which may have led to its sensitivity to many fungal diseases, such as powdery mildew, and an increase in the number of a third type, RPW8 (resistance to powdery mildew 8)-NB-LRR (RNL). Subsequently, transcriptome analysis showed significant enrichment of NLRs during powdery mildew infection, emphasizing the pivotal role of these elements in grapevine's defense against powdery mildew. The successful assembly of a high-quality Thompson Seedless reference genome significantly contributes to grape genomics research, providing insight into the importance of seedlessness, disease resistance, and color traits, and these data can be used to facilitate grape molecular breeding efforts.

Urban-rural differences in associations among perceived stress, resilience and self-care in Chinese older adults with multiple chronic conditions.

BACKGROUND: Social determinants of health (SDH) are mainly comprised of structural and intermediary domains. Emerging evidence suggests that the burden of multiple chronic conditions (MCCs) in older adults is exacerbated by structural determinants (e.g. low income and low education). However, less attention was paid to the intermediary determinants (i.e. material circumstances, psychosocial factors and behavioural factors) of MCCs. OBJECTIVES: To investigate the associations among perceived stress, resilience and self-care in Chinese older adults with MCCs by comparing urban and rural groups. METHODS: A convenience sample (125 and 115 participants from urban and rural settings, respectively) of Chinese older adults with MCCs was enrolled between January and April 2022. Hierarchical multiple regression analyses and propensity score weights were used to determine the associations among perceived stress, resilience and self-care. RESULTS: Hypothesis 1 regarding the negative associations between perceived stress and self-care was fully supported in the rural group. However, for the urban group, the negative association was only supported for the relationship between MCCs-related perceived stress and self-care maintenance. Hypothesis 2 was fully supported regarding the positive associations between resilience and the three components of self-care in both groups, although the relationship between resilience and self-care monitoring was marginally significant in the urban group. Hypothesis 3 regarding the moderating effect of resilience was only supported in the relationship between general perceived stress and self-care monitoring in the rural group. After adding the propensity score weights, the moderating effect was no longer statistically significant in the rural group. CONCLUSIONS: The urban-rural disparities in the Chinese context might largely be attributed to the complex interactions of the structural determinants and intermediary determinants. Findings can inform the development of culturally tailored interventions to promote self-care and reduce urban-rural disparities for Chinese older adults with MCCs. IMPLICATIONS FOR PRACTICE: With the increasing number of older adults in China who are living with multiple chronic conditions and the call for effective interventions to improve their health outcomes, current findings can inform the development and implementation of nurse-led culturally tailored interventions to promote self-care and reduce urban-rural disparities for Chinese older adults with MCCs.

Tumor-associated macrophages: Potential therapeutic targets and diagnostic markers in cancer.

Macrophages are plastic and functionally diverse, present in all tissues, and play a key role in organisms from development, homeostasis and repair, to immune responses to pathogens. They are central to many disease states and have emerged as important therapeutic targets for many diseases. Tumor-associated macrophages (TAMs) are the most abundant immune cells in the tumor microenvironment (TME) and are key factors influencing cancer progression, metastasis and tumor recurrence. TAMs can be derived from different sources and exert different pro- or anti-tumor effects based on the type, stage and immune composition of the tumor. TAMs are highly heterogeneous and diverse, and have multiple functional phenotypes. There is still a great deal of controversy regarding the relationship between TAMs and prognosis of cancer patients. In this review, we summarize the characteristics of common markers of TAMs as well as explore the prognostic role of TAMs in different cancers including lung, breast, gastric, colorectal, esophageal and ovarian cancers.

BTG2 suppresses the growth and metastasis of cervical squamous cell carcinoma.

BACKGROUND: Cervical cancer is the fourth most common malignancy in women, of which cervical squamous cell carcinoma (CESC) is the main pathological type of cervical cancer. B-cell translocation gene 2 (BTG2) protein has been recognized as a tumor suppressor in several cancer types. However, BTG2 expression and molecular function in CESC are unknown. METHODS: In this study, we first assessed the expression of BTG2 in tumor tissue specimens from CESC patients using immunohistochemical staining and real-time quantitative PCR, and explored the relationship between BTG2 expression status and clinical manifestations. Next, we constructed BTG2 knockdown and overexpression CESC cell lines to observe the effects of BTG2 on CESC proliferation and metastasis at the cellular level. Finally, we employed a nude mouse xenograft tumor model in an in vivo experiment to observe the effect of BTG2 on tumorigenesis in vivo. RESULTS: The results showed that the expression of BTG2 protein was lower in CESC tissues than in normal tissues, and high BTG2 expression was associated with better survival in CESC patients versus CESC patients. The results of cellular assays confirm that overexpression of BTG2 inhibits the proliferation, migration and metastasis of CESC cells. Nude mouse xenograft tumor model showed that overexpression of BTG2 inhibited tumor growth in vivo, and conversely knockdown of BTG2 promoted tumor growth. CONCLUSION: In summary, our data suggest that BTG2 acts as a tumor suppressor in CESC and inhibits the growth and metastasis of CESC. BTG2 may serve as a potential prognostic marker in CESC and is expected to provide a therapeutic strategy for patients with CESC.

MAPK9 is Correlated with a Poor Prognosis and Tumor Progression in Glioma.

BACKGROUND: Glioma has a high incidence in young and middle-aged adults and a poor prognosis. Because of late diagnosis and uncontrollable recurrence of the primary tumor after failure of existing treatments, glioma patients tend to have a poor prognosis. Recent advances in research have revealed that gliomas exhibit unique genetic features. Mitogen-activated protein kinase 9 (MAPK9) is significantly upregulated in mesenchymal glioma spheres and may be a new target for glioma diagnosis. This study aimed to investigate the potential diagnostic significance and predictive value of MAPK9 in glioma. METHODS: Paraffin-embedded tumor tissues and paracancerous tissues were collected from 150 glioma patients seen at the General Hospital of Northern Theater Command. Immunohistochemistry and western blot assays were used to detect the expression levels of MAPK9. Prognosis and survival analyses were performed using SPSS 26 software for univariate/multivariate analysis and log-rank analysis. Cellular models were used to assess the effect of MAPK9 overexpression and knockdown in vitro. RESULTS: MAPK9 expression was higher in glioma tissues than in paraneoplastic tissues. Prognostic and survival analyses revealed that the MAPK9 expression level is an independent prognostic factor in glioma patients. In addition, overexpression of MAPK9 significantly promoted the proliferation and migration of primary glioma cells, possibly via the Wnt/ß-catenin-regulated EMT pathway. CONCLUSIONS: MAPK9 is an independent prognostic factor in glioma and is involved in tumor progression.

ZIC5 promotes aggressiveness and cancer stemness in cervical squamous cell carcinoma.

BACKGROUND: Cervical cancer is one of the major malignancies causing morbidity and mortality in women in developing countries. ZIC5 has been found to be associated with a variety of cancers, yet the expression and molecular function of ZIC5 in cervical squamous cell carcinoma (CESC) is unknown. METHODS: We examined the expression of ZIC5 in tumors and normal tissues of CESC patients using immunohistochemistry, immunoblotting and fluorescent quantitative PCR, and used statistical methods to explore its relationship with clinical manifestations. Next, we constructed ZIC5 knockdown and overexpression CESC cell lines to observe the effect of ZIC5 on the proliferation and metastasis of CESC cells. Finally, we applied a nude mouse xenograft tumor model to observe the effect of ZIC5 on tumorigenesis in vivo. RESULTS: Our results showed that the expression of ZIC5 was higher in cancer tissues than in normal tissues. Prognostic analysis showed that ZIC5 expression level was an independent prognostic factor in CESC patients, and the results of Transwell, CCK-8 and wound healing assays confirmed that overexpression of ZIC5 could promote the proliferation and migration of CESC cells. A nude mouse xenograft tumor model showed that knockdown of ZIC5 inhibited tumor growth in vivo. Database, immunoblotting assay and in vitro sphere-forming assay confirmed that ZIC5 could promote the stemness of CESC cells. CONCLUSION: ZIC5 is a factor that indicates a poor prognosis of CESC patients and promotes stemness in CESC cells. ZIC5 may be a potential biomarker and therapeutic target for CESC patients.

Traducción y adaptación transcultural al castellano de la escala Perinatal Bereavement Care Confidence Scale (PBCCS) / Translation and cross-cultural adaptation into Spanish of the Perinatal Bereavement Care Confidence Scale (PBCCS)

Objetivo: Realizar la adaptación transcultural de la escala Perinatal Bereavement Care Confidence Scale (PBCCS) al contexto español, así como evaluar la validez de contenido. Método: La traducción y la adaptación transcultural se realizaron mediante la técnica de traducción y retrotraducción, con la participación de un panel de diez expertas. La evaluación de la validez de contenido se llevó a cabo por el mismo grupo de expertas; se calcularon el índice de validez de contenido (CVI), el coeficiente estadístico Kappa (K) y la razón de validez de contenido (CVR) a partir de los resultados de la evaluación. El proceso se realizó en comunicación constante con los autores de la escala original. Resultados: La versión castellana obtenida, la Escala de Confianza frente al Cuidado del Duelo Perinatal (ECCDP), obtuvo un S-CVI/Ave de 0,93 y consta de un total de 43 ítems, clasificados en cuatro dimensiones, manteniendo la misma estructura que la escala original. De los 43 ítems, 37 mostraron un I-CVI con valores por encima de 0,78, y 42 ítems tuvieron un valor de CVR positivo. Conclusiones: La escala ECCDP mantiene la equivalencia con la versión original y es un instrumento con suficiente validez de contenido para medir el nivel de confianza de las matronas y enfermeras en la atención al duelo perinatal en España. Se recomienda un estudio multidisciplinar de validación para evaluar el resto de las propiedades psicométricas del instrumento. (AU)

Objective: To carry out the cross-cultural adaptation of the scale Perinatal Bereavement Care Confidence Scale (PBCCS) to the Spanish context, as well as to evaluate its content validity. Methodology: The translation and cross-cultural adaptation were carried out using the translation and back-translation technique with the participation of a panel of ten experts. The evaluation of content validity was carried out by the same group of experts; the content validity index (CVI), the Kappa statistical coefficient (K) and the content validity ratio (CVR) were calculated. The process was carried out in constant communication with the original scale authors. Results: The Spanish version obtained (ECCDP, Escala de Confianza frente al Cuidado del Duelo Perinatal) had an S-CVI/Ave of 0.93 and consisted of 43 total items, classified into four dimensions, maintaining the same structure as the original scale. 37 of 43 items showed an I-CVI with values above 0.78. Forty-two items had a positive CVR value. Conclusions: The ECCDP scale preserves the equivalence with the original version and is an instrument with sufficient content validity to measure the level of confidence of midwives and nurses in perinatal bereavement care in Spain. A multidisciplinary validation study is recommended to evaluate the remaining psychometric properties of this scale. (AU)

Pterygium Screening and Lesion Area Segmentation Based on Deep Learning.

A two-category model and a segmentation model of pterygium were proposed to assist ophthalmologists in establishing the diagnosis of ophthalmic diseases. A total of 367 normal anterior segment images and 367 pterygium anterior segment images were collected at the Affiliated Eye Hospital of Nanjing Medical University. AlexNet, VGG16, ResNet18, and ResNet50 models were used to train the two-category pterygium models. A total of 150 normal and 150 pterygium anterior segment images were used to test the models, and the results were compared. The main evaluation indicators, including sensitivity, specificity, area under the curve, kappa value, and receiver operator characteristic curves of the four models, were compared. Simultaneously, 367 pterygium anterior segment images were used to train two improved pterygium segmentation models based on PSPNet. A total of 150 pterygium images were used to test the models, and the results were compared with those of the other four segmentation models. The main evaluation indicators included mean intersection over union (MIOU), IOU, mean average precision (MPA), and PA. Among the two-category models of pterygium, the best diagnostic result was obtained using the VGG16 model. The diagnostic accuracy, kappa value, diagnostic sensitivity of pterygium, diagnostic specificity of pterygium, and F1-score were 99%, 98%, 98.67%, 99.33%, and 99%, respectively. Among the pterygium segmentation models, the double phase-fusion PSPNet model had the best results, with MIOU, IOU, MPA, and PA of 86.57%, 78.1%, 92.3%, and 86.96%, respectively. This study designed a pterygium two-category model and a pterygium segmentation model for the images of the normal anterior and pterygium anterior segments, which could help patients self-screen easily and assist ophthalmologists in establishing the diagnosis of ophthalmic diseases and marking the actual scope of surgery.

UCHL1 promotes cancer stemness in triple-negative breast cancer.

BACKGROUND: Triple-negative breast cancer (TNBC) is a special kind of breast cancer with strong ability of invasion and metastasis. UCHL1 belongs to the ubiquitin carboxy-terminal hydrolase family and is found to be increased in a variety of malignancies, but its expression in TNBC is unknown. METHODS: First, we analyzed the expression of UCHL1 in 128 TNBC specimens and paired adjacent normal tissues from 17 TNBC patients undergoing curative resection by immunohistochemistry. Then, the relationship between UCHL1 and cancer stemness was investigated by cell flow cytometry, spheroid formation assays and western blot. Moreover, cell scratch assay and Transwell assays were performed to explore whether UCHL1 promotes the migration and invasion of TNBC cells. Finally, we constructed a xenografts model of TNBC cell lines to observe the effect of UCHL1 on tumorigenesis in vivo. RESULTS: UCHL1 was overexpressed in TNBC tissues and associated with poor prognosis. UCHL1 promoted stem cancer cells properties, including the percentage of CD44+/CD24- cells, sphere-forming ability and CSCs related markers. Furthermore, Scratch assay and Transwell assay proved that UCHL1 enhanced the migration and invasion of TNBC cells. The experimental results of xenografts model in nude mice showed that UCHL1 promoted tumorigenesis of TNBC in vivo. CONCLUSION: UCHL1 may play a role in the malignant progression of TNBC by maintaining the stemness and promoting cell invasion and is expected to become a potential therapeutic target for TNBC patients.

The cancer-testis antigen FBXO39 predicts poor prognosis and is associated with stemness and aggressiveness in glioma.

Gliomas are common brain tumors with poor prognosis. F-box protein39 (FBXO39) is found to be increased in a variety of malignancies, but its expression in gliomas is unknown. Here, we report that FBXO39 as an oncogene is upregulated in glioma and associated with poor prognosis. The degree of FBXO39 positive expression is doubtlessly concomitant with the grade of glioma. The data analyses show that patients with high FBXO39 expression have poor overall survival. What's more, FBXO39 accelerates the invasion and migration abilities of glioma cells and promotes glioma stem cell growth and stemness. In summary, these results suggest that FBXO39 is expected to become a new biological target for judging the prognosis and treatment of glioma.

Prediction of corneal astigmatism based on corneal tomography after femtosecond laser arcuate keratotomy using a pix2pix conditional generative adversarial network.

Purpose: This study aimed to develop a deep learning model to generate a postoperative corneal axial curvature map of femtosecond laser arcuate keratotomy (FLAK) based on corneal tomography using a pix2pix conditional generative adversarial network (pix2pix cGAN) for surgical planning. Methods: A total of 451 eyes of 318 nonconsecutive patients were subjected to FLAK for corneal astigmatism correction during cataract surgery. Paired or single anterior penetrating FLAKs were performed at an 8.0-mm optical zone with a depth of 90% using a femtosecond laser (LenSx laser, Alcon Laboratories, Inc.). Corneal tomography images were acquired from Oculus Pentacam HR (Optikgeräte GmbH, Wetzlar, Germany) before and 3 months after the surgery. The raw data required for analysis consisted of the anterior corneal curvature for a range of ± 3.5 mm around the corneal apex in 0.1-mm steps, which the pseudo-color corneal curvature map synthesized was based on. The deep learning model used was a pix2pix conditional generative adversarial network. The prediction accuracy of synthetic postoperative corneal astigmatism in zones of different diameters centered on the corneal apex was assessed using vector analysis. The synthetic postoperative corneal axial curvature maps were compared with the real postoperative corneal axial curvature maps using the structural similarity index measure (SSIM) and peak signal-to-noise ratio (PSNR). Results: A total of 386 pairs of preoperative and postoperative corneal tomography data were included in the training set, whereas 65 preoperative data were retrospectively included in the test set. The correlation coefficient between synthetic and real postoperative astigmatism (difference vector) in the 3-mm zone was 0.89, and that between surgically induced astigmatism (SIA) was 0.93. The mean absolute errors of SIA for real and synthetic postoperative corneal axial curvature maps in the 1-, 3-, and 5-mm zone were 0.20 ± 0.25, 0.12 ± 0.17, and 0.09 ± 0.13 diopters, respectively. The average SSIM and PSNR of the 3-mm zone were 0.86 ± 0.04 and 18.24 ± 5.78, respectively. Conclusion: Our results showed that the application of pix2pix cGAN can synthesize plausible postoperative corneal tomography for FLAK, showing the possibility of using GAN to predict corneal tomography, with the potential of applying artificial intelligence to construct surgical planning models.

Analysis of the shape characteristics and nutritional components of Akebia trifoliata in Qinba Mountains.

Akebia trifoliata (A. trifoliata) is a widely distributed wild vine that has attracted much attention in recent years due to the edible fruit of food and medicinal value. In this paper, the fruits of A. trifoliata, which are derived from Qinling Mountains (12 genotypes) and Bashan Mountains (4 genotypes) and have been artificially domesticated and cultivated for many years, are taken as the research object to study the fruit characteristics and pulp components of 16 genotypes of A. trifoliata. The results show that the pulp of the 16 genotypes contain a variety of nutrients, among which the average contents of total sugar, total acid, vitamin C, soluble solids and starch are 14.68g/100g, 0.14g/100g, 26.40mg/100g, 17.95% and 5.29g/100g. The fruit contains 17 amino acids, including 7 essential amino acids and 4 organic acids. The latter refers to malic acid, lactic acid, citric acid and fumaric acid, the average contents of which are 1.03g/kg, 3.38g/kg, 0.33g/kg and 0.0149g/kg. Besides, 8 mineral elements in the fruit include 4 macro elements and 4 micro elements. The average contents of the former are potassium (1.83g/kg), calcium (0.23g/kg), phosphorus (0.28g/kg) and magnesium (0.21g/kg), and the average contents of the latter are iron (2.29mg/kg), zinc (2.23mg/kg), copper (1.37mg/kg) and manganese (5.52mg/kg). During the ripening process of A. trifoliata fruit (using HY-9 as the material), the main nutrients in the pulp such as total sugar, soluble solids, starch, amino acids and various mineral elements reach the maximum in stage 3, indicating that stage 3 is the best edible period of A. trifoliata fruit. Through the assignment analysis and comprehensive evaluation of 9 quality indicators (3 apparent characters and 6 main chemical components) of the fruits of the 16 A. trifoliata genotypes from Qinba Mountains, HY-1, HY-2 and HY-9 were finally screened out as the three superior genotypes. This study aims to provide reference for the development and utilization of A. trifoliata wild germplasm resources and the selection of new varieties.

FBXO39 predicts poor prognosis and correlates with tumor progression in cervical squamous cell carcinoma.

BACKGROUND: Cancer/testis antigen (CTA) is a class of antigen molecules mainly expressed in the germinal epithelium of testis and some tumor tissues. FBXO39, also known as F-box protein 39, is a crucial CTA molecule. F-box protein 39 (FBXO39) is overexpressed in cervical squamous cell carcinomas (CESCs), however its function in cancer development and clinical significance are still unknown. METHODS: We used paraffin-embedded tumor tissues from 124 patients and fresh-harvested and paired adjacent normal esophageal tissues from 15 CESC patients who underwent primary surgical resection in Xijing Hospital between 2015 and 2020. The expression level of FBXO39 was evaluated through immunohistochemistry, Western Blot and q-PCR. Prognostic and survival analyses were conducted using univariate/multivariate analysis and log-rank analysis with SPSS 23.0. CCK-8, wound-healing and Transwell assays were applied to demonstrate that FBXO39 promoted the proliferation, migration and invasion. Finally, we constructed a xenografts model of the C-33A cell lines to observe the effect of FBXO39 on tumorigenesis in vivo. RESULTS: Immunohistochemical results showed that FBXO39 was highly expressed in cancer tissues than in corresponding non-cancer tissues. Similarly, we proved this result at protein and mRNA level by Western-Blotting and q-PCR. Prognostic and OS analyses showed that the FBXO39 expression level was an individual prognostic factor in CESC patients. CCK-8, wound-healing and Transwell assays proved that the overexpression of FBXO39 in Si-Ha cells promoted the proliferation, migration and invasion of the cells. Knocking down FBXO39 in C-33A cells inhibited the proliferation, migration and invasion of cells. The experimental results of xenografts model in nude mice showed that the knockdown of FBXO39 in C-33A cells slowed down the growth of tumor. CONCLUSION: FBXO39 is a poor prognostic factor of cervical squamous cell carcinoma, which may provide a novel therapeutic target for CESC.

Grapevine VlbZIP30 improves drought resistance by directly activating VvNAC17 and promoting lignin biosynthesis through the regulation of three peroxidase genes.

Drought stress severely affects grapevine quality and yield, and recent reports have revealed that lignin plays an important role in protection from drought stress. Since little is known about lignin-mediated drought resistance in grapevine, we investigated its significance. Herein, we show that VlbZIP30 mediates drought resistance by activating the expression of lignin biosynthetic genes and increasing lignin deposition. Transgenic grapevine plants overexpressing VlbZIP30 exhibited lignin deposition (mainly G and S monomers) in the stem secondary xylem under control conditions, which resulted from the upregulated expression of VvPRX4 and VvPRX72. Overexpression of VlbZIP30 improves drought tolerance, characterized by a reduction in the water loss rate, maintenance of an effective photosynthesis rate, and increased lignin content (mainly G monomer) in leaves under drought conditions. Electrophoretic mobility shift assay, luciferase reporter assays, and chromatin immunoprecipitation-qPCR assays indicated that VlbZIP30 directly binds to the G-box cis-element in the promoters of lignin biosynthetic (VvPRX N1) and drought-responsive (VvNAC17) genes to regulate their expression. In summary, we report a novel VlbZIP30-mediated mechanism linking lignification and drought tolerance in grapevine. The results of this study may be of value for the development of molecular breeding strategies to produce drought-resistant fruit crops.

Evolution and expression analysis of the grape (Vitis vinifera L.) WRKY gene family.

WRKY proteins comprise a large family of transcription factors that play important roles in plant defence regulatory networks, including responses to various biotic and abiotic stresses. To date, no large-scale study of WRKY genes has been undertaken in grape (Vitis vinifera L.). In this study, a total of 59 putative grape WRKY genes (VvWRKY) were identified and renamed on the basis of their respective chromosome distribution. A multiple sequence alignment analysis using all predicted grape WRKY genes coding sequences, together with those from Arabidopsis thaliana and tomato (Solanum lycopersicum), indicated that the 59 VvWRKY genes can be classified into three main groups (I-III). An evaluation of the duplication events suggested that several WRKY genes arose before the divergence of the grape and Arabidopsis lineages. Moreover, expression profiles derived from semiquantitative PCR and real-time quantitative PCR analyses showed distinct expression patterns in various tissues and in response to different treatments. Four VvWRKY genes showed a significantly higher expression in roots or leaves, 55 responded to varying degrees to at least one abiotic stress treatment, and the expression of 38 were altered following powdery mildew (Erysiphe necator) infection. Most VvWRKY genes were downregulated in response to abscisic acid or salicylic acid treatments, while the expression of a subset was upregulated by methyl jasmonate or ethylene treatments.

Genome-wide analysis of respiratory burst oxidase homologs in grape (Vitis vinifera L.).

Plant respiratory burst oxidase homolog (rboh) genes appear to play crucial roles in plant development, defense reactions and hormone signaling. In this study, a total of seven rboh genes from grape were identified and characterized. Genomic structure and predicted protein sequence analysis indicated that the sequences of plant rboh genes are highly conserved. Synteny analysis demonstrated that several Vvrboh genes were found in corresponding syntenic blocks of Arabidopsis, suggesting that these genes arose before the divergence of the respective lineages. The expression pattern of Vvrboh genes in different tissues was assessed by qRT-PCR and two were constitutively expressed in all tissues tested. The expression profiles were similarly analyzed following exposure to various stresses and hormone treatments. It was shown that the expression levels of VvrbohA, VvrbohB and VvrbohC1 were significantly increased by salt and drought treatments. VvrbohB, VvrbohC2, and VvrbohD exhibited a dramatic up-regulation after powdery mildew (Uncinula necator (Schw.) Burr.) inoculation, while VvrbohH was down-regulated. Finally, salicylic acid treatment strongly stimulated the expression of VvrbohD and VvrbohH, while abscisic acid treatment induced the expression of VvrbohB and VvrbohH. These results demonstrate that the expression patterns of grape rboh genes exhibit diverse and complex stress-response expression signatures.

Effect of GA3 treatment on seed development and seed-related gene expression in grape.

BACKGROUND: The phytohormone gibberellic acid (GA3) is widely used in the table grape industry to induce seedlessness in seeded varieties. However, there is a paucity of information concerning the mechanisms by which GAs induce seedlessness in grapes. METHODOLOGY/PRINCIPAL FINDINGS: In an effort to systematically analyze the cause of this GA3-induced seed abortion, we conducted an in depth characterization of two seeded grape cultivars ('Kyoho' and 'Red Globe'), along with a seedless cultivar ('Thompson Seedless'), following treatment with GA3. In a similar fashion to the seedless control, which exhibited GA3-induced abortion of the seeds 9 days after full bloom (DAF), both 'Kyoho' and 'Red Globe' seeded varieties exhibited complete abortion of the seeds 15 DAF when treated with GA3. Morphological analyses indicated that while fertilization appeared to occur normally following GA3 treatment, as well as in the untreated seedless control cultivar, seed growth eventually ceased. In addition, we found that GA3 application had an effect on redox homeostasis, which could potentially cause cell damage and subsequent seed abortion. Furthermore, we carried out an analysis of antioxidant enzyme activities, as well as transcript levels from various genes believed to be involved in seed development, and found several differences between GA3-treated and untreated controls. CONCLUSION: Therefore, it seems that the mechanisms driving GA3-induced seedlessness are similar in both seeded and seedless cultivars, and that the observed abortion of seeds may result at least in part from a GA3-induced increase in cell damage caused by reactive oxygen species, a decrease in antioxidant enzymatic activities, and an alteration of the expression of genes related to seed development.