The interaction of O3 and CO2 concentration, exposure timing and duration on stem rust severity on winter wheat variety 'Coker 9553'.

Wheat rusts, elevated ozone (O3), and carbon dioxide (CO2) are simultaneously impacting wheat production worldwide, but their interactions are not well understood. This study investigated whether near-ambient O3 is suppressive or conducive to stem rust (Sr) of wheat, considering the interactions with ambient and elevated CO2. Winter wheat variety 'Coker 9553' (Sr-susceptible; O3 sensitive) was inoculated with Sr (race QFCSC) following pre-treatment with four different concentrations of O3 (CF, 50, 70, and 90 ppbv) at ambient CO2 levels. Gas treatments were continued during the development of disease symptoms. Disease severity, measured as percent sporulation area (PSA), significantly increased relative to the CF control only under near-ambient O3 conditions (50 ppbv) in the absence of O3-induced foliar injury. Disease symptoms at higher O3 exposures (70 and 90 ppbv) were similar to or less than the CF control. When Coker 9553 was inoculated with Sr while exposed to CO2 (400; 570 ppmv) and O3 (CF; 50 ppbv) in four different combinations, and seven combinations of exposure timing and duration, PSA significantly increased only under continuous treatment with O3 for six weeks or pre-inoculation treatment for three weeks, suggesting that O3-predisposes wheat to the disease rather than enhancing disease post-inoculation. O3 singly and in combination with CO2 increased PSA on flag leaves of adult Coker 9553 plants while elevated CO2 alone had little effect on PSA. These findings show that sub-symptomatic O3 conditions are conducive to stem rust, contradicting the current consensus that biotrophic pathogens are suppressed by elevated O3. This suggests that sub-symptomatic O3 stress may enhance rust diseases in wheat-growing regions.

Chromosome Location Contributing to Ozone Tolerance in Wheat.

Breeding wheat for higher grain yield can contribute to global food security and sustainable production on less land. Tropospheric ozone can injure wheat plants and subsequently reduce grain yield. Identification of ozone tolerance in the wheat genome can assist plant breeders in developing new sources of tolerant germplasm. Our objective was to use the 'Chinese Spring' monosomic lines to screen for ozone response and identify the chromosomic locations contributing to ozone tolerance based on foliar injury. Two methodologies, Continuous Stirred Tank Reactors and Outdoor Plant Environment Chambers, were used to expose wheat monosomic lines to varying concentrations and durations of ozone. Each wheat monosomic line in 'Chinese Spring' has a missing chromosome in each of the wheat subgenomes (A, B, and D). In both methodologies, we found significant and repeatable data to identify chromosome 7A as a major contributor to tolerance to ozone injury in 'Chinese Spring'. In every experiment, the absence of chromosome 7A resulted in significant injury to wheat due to ozone. This was not the case when any other chromosome was missing.

BrAPI-an application programming interface for plant breeding applications.

MOTIVATION: Modern genomic breeding methods rely heavily on very large amounts of phenotyping and genotyping data, presenting new challenges in effective data management and integration. Recently, the size and complexity of datasets have increased significantly, with the result that data are often stored on multiple systems. As analyses of interest increasingly require aggregation of datasets from diverse sources, data exchange between disparate systems becomes a challenge. RESULTS: To facilitate interoperability among breeding applications, we present the public plant Breeding Application Programming Interface (BrAPI). BrAPI is a standardized web service API specification. The development of BrAPI is a collaborative, community-based initiative involving a growing global community of over a hundred participants representing several dozen institutions and companies. Development of such a standard is recognized as critical to a number of important large breeding system initiatives as a foundational technology. The focus of the first version of the API is on providing services for connecting systems and retrieving basic breeding data including germplasm, study, observation, and marker data. A number of BrAPI-enabled applications, termed BrAPPs, have been written, that take advantage of the emerging support of BrAPI by many databases. AVAILABILITY AND IMPLEMENTATION: More information on BrAPI, including links to the specification, test suites, BrAPPs, and sample implementations is available at https://brapi.org/. The BrAPI specification and the developer tools are provided as free and open source.

Mapping of powdery mildew resistance gene Pm53 introgressed from Aegilops speltoides into soft red winter wheat.

KEY MESSAGE: A powdery mildew resistance gene was introgressed from Aegilops speltoides into winter wheat and mapped to chromosome 5BL. Closely linked markers will permit marker-assisted selection for the resistance gene. Powdery mildew of wheat (Triticum aestivum L.) is a major fungal disease in many areas of the world, caused by Blumeria graminis f. sp. tritici (Bgt). Host plant resistance is the preferred form of disease prevention because it is both economical and environmentally sound. Identification of new resistance sources and closely linked markers enable breeders to utilize these new sources in marker-assisted selection as well as in gene pyramiding. Aegilops speltoides (2n = 2x = 14, genome SS), has been a valuable disease resistance donor. The powdery mildew resistant wheat germplasm line NC09BGTS16 (NC-S16) was developed by backcrossing an Ae. speltoides accession, TAU829, to the susceptible soft red winter wheat cultivar 'Saluda'. NC-S16 was crossed to the susceptible cultivar 'Coker 68-15' to develop F2:3 families for gene mapping. Greenhouse and field evaluations of these F2:3 families indicated that a single gene, designated Pm53, conferred resistance to powdery mildew. Bulked segregant analysis showed that multiple simple sequence repeat (SSR) and single nucleotide polymorphism (SNP) markers specific to chromosome 5BL segregated with the resistance gene. The gene was flanked by markers Xgwm499, Xwmc759, IWA6024 (0.7 cM proximal) and IWA2454 (1.8 cM distal). Pm36, derived from a different wild wheat relative (T. turgidum var. dicoccoides), had previously been mapped to chromosome 5BL in a durum wheat line. Detached leaf tests revealed that NC-S16 and a genotype carrying Pm36 differed in their responses to each of three Bgt isolates. Pm53 therefore appears to be a new source of powdery mildew resistance.

Characterization of Resistance to Stripe Rust in Contemporary Cultivars and Lines of Winter Wheat from the Eastern United States.

Stripe rust, caused by Puccinia striiformis f. sp. tritici, has been an important disease of winter wheat (Triticum aestivum) in the eastern United States since 2000, when a new strain of the pathogen emerged. The new strain overcame the widely used resistance gene, Yr9, and was more aggressive and better adapted to warmer temperatures than the old strain. Host resistance is the most effective approach to manage stripe rust. Winter wheat lines with resistance to the new strain in the field are common, but the genes conferring this resistance are mostly unknown. The objectives of this research were to characterize the all-stage resistance and adult-plant resistance (APR) to stripe rust in a representative group of contemporary winter wheat cultivars and breeding lines and to identify the resistance genes when possible. Of the 50 lines evaluated for all-stage resistance at the seedling stage, nearly all were susceptible to the new strain. Based on a linked molecular marker, seven lines had resistance gene Yr17 that confers resistance to both old and new strains; however, this resistance was difficult to identify in the seedling stage. Of the 19 lines evaluated for APR, all expressed APR compared with a very susceptible check. Nine had race-specific APR to the new strain and nine had APR to both old and new strains. The remaining line, 26R61, had all-stage resistance to the old strain (conferred by resistance gene Yr9) and a high level of APR to the new strain. APR was expressed as low infection type, low percent leaf area diseased, and long latent period at heading stage under both low and high temperature regimes and could be identified as early as jointing stage. Based on tests for linked molecular markers, the most widely used slow-rusting APR genes, Yr18 and Yr29, were not present in any of the lines. The results of this research indicate that effective all-stage resistance was conferred only by Yr17 and that APR was common and likely conferred by unknown race-specific genes rather than genes conferring slow rusting that are more likely to be durable.

Expression of cancer-testis antigens in endometrial carcinomas using a tissue microarray.

Cancer-testis (CT) antigens are expressed in a variety of malignant tumors, but in normal adult tissue, they are only expressed in testicular germ cells. Owing to this tumor-associated expression pattern, these antigens are of major interest as potential targets for immunotherapy and possibly for diagnostic purposes. This study was performed to analyze the expression of four CT antigens, NY-ESO-1, MAGE-A3, MAGE-A4, and CT7/MAGE-C1, in endometrial carcinoma using immunohistochemistry, and to correlate expression with histologic subtypes, grade, and expression of WT1 and p53. Formalin-fixed paraffin-embedded tissues of 130 endometrial carcinomas of the following types and grades were analyzed using a tissue microarray: 85 endometrioid carcinomas (FIGO grade 1, 39; grade 2, 11; and grade 3, 35), 18 papillary serous carcinomas, 12 clear cell carcinomas, 13 malignant mixed mullerian tumors, one mucinous adenocarcinoma, and one undifferentiated carcinoma. The following anti-CT monoclonal antibodies/antigens were studied by immunohistochemistry: monoclonal antibody ES121/NY-ESO-1, monoclonal antibody M3H67/MAGE-A3, monoclonal antibody 57B/MAGE-A4, and monoclonal antibody CT7-33/CT7. The CT expression data were compared to WT1 and p53 protein expression as analyzed in a previous study. Positive staining with anti-CT monoclonal antibodies was graded as follows: focal, <5% positive cells; 1+, 5-25% cells; 2+, 26-50% cells; 3+, 51-75%; and 4+, >75% cells. The 3+ and 4+ staining patterns were considered homogeneous patterns of potential clinical significance and were scored positive for statistical analysis. In low-grade tumors, the most immunoreactivity was seen with mAb M3H67 but little labeling was observed with the other monoclonal antibodies. In high-grade tumors, monoclonal antibodies M3H67 (25%), 57B (23%), and CT7-33 (20%) showed the highest reactivity, while ES121 showed the lowest immunoreactivity (6%). The staining pattern was mostly heterogeneous. Statistical significance was found solely for the correlation of monoclonal antibody 57B staining and p53 expression. No correlation was found for any anti-CT monoclonal antibody staining and clinical stage or for anti-CT staining and WT1 expression. CT antigens CT7, MAGE-A3 and MAGE-A4, but not NY-ESO-1, are expressed in high-grade endometrial carcinomas, and expression of MAGE-A4 is correlated with the presence of overexpressed p53.

Her-2/neu expression and amplification in early stage ovarian surface epithelial neoplasms.

OBJECTIVE: The aim of this study is to examine Her-2/neu gene amplification and protein overexpression in a spectrum of ovarian neoplasms using both immunohistochemical (IHC) and fluorescence in situ hybridization (FISH) techniques that are FDA approved. This study is focused on early stage tumors including both carcinomas and borderline tumors. METHODS: FDA-approved IHC and FISH for Her-2/neu were performed on formalin-fixed, paraffin-embedded tissue from 79 ovarian neoplasms representing a broad spectrum of tumor types as well as four normal ovaries. All tumors were either stage I or stage II. Tumor and normal tissue were studied collectively using a tissue microarray (TMA). HercepTest (DAKO) and PathVysion Her-2/neu probe kit (Vysis Inc.) were used for IHC and FISH analysis. RESULTS: FISH analysis of serous carcinomas demonstrated Her-2/neu gene amplification in 3 (18%) of 17 cases. Two of three cases showing Her-2/neu gene amplification were scored 1+ using IHC, while the remaining case was scored as 0. Analysis of endometrioid carcinomas demonstrated Her-2/neu amplification using FISH in 1 of 10 (10%) cases. IHC in this case was scored 2+ (positive). None of the remaining 44 tumors, including clear cell carcinoma (n = 12), transitional cell carcinoma (n = 1), mixed epithelial carcinoma (n = 7), carcinoma not otherwise specified (n = 1), and 31 borderline tumors (mucinous, n = 17; endometrioid, n = 7; serous, n = 7), showed Her-2/neu gene amplification or protein overexpression. Normal ovaries were negative as well. CONCLUSIONS: Amplification of Her-2/neu in early stage ovarian neoplasms is infrequent, 6.7% overall. Due to the limited number of informative cases, we were unable to determine the clinical significance of Her-2/neu amplification in this study. Her-2/neu amplification was restricted to carcinomas and was not encountered in ovarian borderline tumors.

Wilms Tumor Gene (WT1) and p53 expression in endometrial carcinomas: a study of 130 cases using a tissue microarray.

OBJECTIVE: With the exception of ovarian serous carcinoma, Wilms tumor suppressor gene (WT1) expression in common gynecologic carcinomas has not been described in detail. We studied a large number of endometrial carcinomas to determine the range of tumors that express WT1; this could have prognostic and therapeutic significance. METHODS: We studied the immunohistochemical expression of WT1 and p53 in 130 primary human endometrial carcinomas of various histological subtypes, grades, and stages using a tissue microarray. The clinical data were retrieved from the medical records. RESULTS: WT1 was expressed in a wide variety of endometrial cancers and was most marked in malignant mixed Mullerian tumors (MMMTs) (70% positive). WT1 expression was significantly correlated with high histological grade, and there was a trend toward a worse clinical outcome for patients whose tumors expressed WT1. An association between expression of WT1 and p53 and between these and outcome was noted in a univariate analysis, but only stage and p53 status remained prognostically significant independent variables. CONCLUSION: WT1 is expressed in appreciable numbers of endometrial cancers, particularly MMMTs. These findings support further investigation of WT1 as a possible therapeutic target in gynecologic malignancies.

Differential expression of WT1 and p53 in serous and endometrioid carcinomas of the endometrium.

Wilms' tumor antibody (WT1) has recently been reported to be reactive in most ovarian and peritoneal serous carcinomas, but few studies have looked at WT1 reactivity in endometrial carcinomas. p53, like WT1, is a tumor suppressor gene and in its mutated form is frequently present in endometrial serous carcinoma. Routine immunohistochemical staining for p53 and WT1 was performed in 70 endometrial carcinomas (39 endometrioid and 31 serous) of varying differentiation using tissue microarrays. Only 2 (7.5%) serous carcinomas and none of the endometrioid carcinomas (0%) were reactive for WT1. p53 immunoreactivity was found in 26 (83.9%) serous carcinomas and in 2 (5.1%) endometrioid carcinomas. We conclude that WT1 and p53 expression are not related and that WT1 expression in endometrial serous carcinoma differs from that of its extrauterine counterparts.

Fallopian tube and primary peritoneal carcinomas associated with BRCA mutations.

PURPOSE: The aims of this study were to determine the incidence of BRCA mutations among Ashkenazi Jewish patients with fallopian tube carcinoma (FTC) or primary peritoneal carcinoma (PPC), to study the clinicopathologic features of these cancers, and to estimate the risks of these cancers in association with a BRCA mutation. PATIENTS AND METHODS: A retrospective review at two institutions identified 29 Jewish patients with FTC and 22 Jewish patients with PPC. These patients were genotyped for the three Ashkenazi Jewish BRCA founder mutations (185delAG and 5382insC in BRCA1 and 6174delT in BRCA2). Surgical and pathologic information, family history, and survival data were obtained from hospital records. All statistical tests were two sided. RESULTS: Germline BRCA mutations were identified in five of 29 patients with FTC (17%) and nine of 22 patients with PPC (41%). Mutation carriers had a younger mean age at diagnosis than patients without a mutation (60 v 70 years; P =.002). The overall median survival was 148 months for mutation carriers and 41 months for patients without a mutation (P =.04). For BRCA mutation carriers, the lifetime risks of FTC and PPC were 0.6% and 1.3%, respectively. CONCLUSION: Substantial proportions of Ashkenazi Jewish patients with FTC or PPC are BRCA mutation carriers. Patients with BRCA-associated FTC or PPC are younger at diagnosis and have improved survival compared with patients without a BRCA mutation. Although the lifetime risks of FTC or PPC for patients with BRCA heterozygotes are greater than those for the general population, the absolute risks seem relatively low.

Clinically inapparent invasive vulvar carcinoma in an area of persistent Paget's disease: a case report.

BACKGROUND: Invasive vulvar carcinoma is reported to occur in 5 to 20% of patients with vulvar Paget's disease. We report a case in which a clinically inapparent invasive lesion was discovered on reexcision of microscopically persistent vulvar Paget's disease. CASE: A 58-year-old woman presented with a diagnosis of vulvar Paget's disease. A wide local excision of the lesion was performed and pathologic analysis revealed microscopic Paget's disease at two of the margins. The patient returned for a follow-up 4 months later and a vulvar biopsy revealing persistent Paget's cells was obtained from the area of the prior microscopically positive surgical margin. A reexcision was performed from the normal-appearing vulva and invasive vulvar carcinoma was noted in this specimen. CONCLUSIONS: This case demonstrates several concerning aspects of this disease, most important of which is that the clinically apparent lesion did not contain the clinically significant invasive lesion. Invasive vulvar carcinoma may occur in association with microscopically persistent vulvar Paget's disease, a condition often encountered after primary treatment with wide local excision.