Marker assisted selection (MAS) for downy mildew resistance in grapevines using Rpv3.1 associated markers

Powdery mildew and downy mildew are primary fungal diseases that cause significant damage in viticulture. Therefore, breeding powdery and/or downy mildew resistance is one of the priority subjects in grapevine breeding programs. This study aims to conduct early-selection by marker assisted selection (MAS) method among 869 genotypes obtained through crossbreeding 'Alphonse Lavallee' x 'Regent' cultivars using the markers (GF18-06 and GF18-08) associated with downy mildew resistance gene region Rpv3.1 to develop new grapevine cultivars resistant to downy mildew caused by Plasmopara viticola. A total of 869 hybrid plants which were obtained after crossing 'Alphonse Lavallee' x 'Regent' in a 3-year breeding program were used in the study. The hybrid plants were scored for the resistance level based on their sporulation intensity after artificial inoculation of P. viticola. DNA samples of the hybrid plants were amplified with GF18-06 and GF1808 markers in Polymerase Chain Reaction (PCR) for MAS. The alleles which were associated to Rpv3.1 resistance locus and the results of resistance scoring were compared, and the applicability of the markers in MAS was verified. It was determined that the GF18-08/410 bp marker can be used successfully for MAS. Gf 18-06 marker 385 bp, 390 bp and 407 bp gave false positive results in our population, respectively 8.86%, 9.02% and 37.94%. Therefore, this may limit its use for MAS.National Scientific and Technological Research Council of Turkey [1150176]This work was supported by the National Scientific and Technological Research Council of Turkey (grant number 1150176)

Germinating in Box: An Effective and Practical Method for Grape Seed Germination

Asma ıslahında en önemli engellerden biri, iki üzüm çeşidinin melezlenmesi sonucu elde edilen melezçekirdeklerin çimlenme oranının düşük olmasıdır. Çimlenmeyi etkileyen en önemli faktörler arasında;ortam ve tohum nemi gelir. Ayrıca büyümeyi düzenleyici maddeler (BDM)de, çimlenmeyi olumluetkilemektedir. Bu çalışmada, melez üzüm çekirdeklerine BDM uygulamak ve kapalı ortamlarda kararlı birnem düzeyinde çimlendirilmek suretiyle çimlenme oranını arttırmak amaçlanmıştır. Alphonse Lavalle ×Regent melezlemesinden elde edilen F? melez çekirdekleri, nemlendirilmiş perlitte 4 ay süreyle, +5?’dakatlamayı takiben; torf: perlit (1:1) harç ortamı içeren 5 cm çapındaki torf saksılara ekilmiştir. Çimlenmeyiteşvik etmek için, çekirdekler ekimden önce 24 saat gibberellik asit (GA?, 1000 ppm, benzil aminopurin(BAP, 1000 ppm), GA?+BAP (1000 ppm+1000 ppm), hidrojen peroksit (H?O?, 1 M) çözeltisinde ve suda(kontrol) tutulmuştur. Saksılar, bağıl nem miktarı sürekli %99 düzeyinde olan kapaklı plastik kutularakonulmuş ve bu kutular sıcaklığı 27? olan çimlendirme odalarına alınmıştır. Çimlenen çekirdekler yineplastik kutulara konularak sıcaklığı 25? olan ve kutuların hemen üstü LED lambalar ile aydınlatılmış(PAR=135–317 ?mol m?² s?¹) bitki gelişme odalarına yerleştirilmiştir. Kutuların kapakları her iki odada dakapalı tutulmuştur. Bitkicikler 3–5 gerçek yaprak oluşunca, plastik torbalara alınmış ve daha sonra ısıtmalıseraya alınmıştır. Uygulamalar sonucunda çimlenen çekirdek ve elde edilen bitki yüzdesi kontrolde %60.39iken, 1000 ppm GA? uygulanmış çekirdeklerde %78.32’ye kadar yükselmiştir. Üzüm çekirdeklerinin 24saat GA? çözeltisine batırılmasından sonra plastik kutularda çimlendirilmesi ve bunu takiben yine plastikkutularda geliştirilmesi, asma ıslahı çalışmalarında başarıyı arttıracak etkin bir yöntemdir.One of the main problems of grape breeding is low germination rate of hybrid seeds obtained by crossing of grape varieties. Humidity in grape seeds and germination media have extremely important for germination. In addition, some plant growth regulators (PGR) have positive effects on germination. Hybrid seeds obtained by Alphonse Lavallee and Regent grape cultivars were straficated at 5? for 4 months in damp perlite. Seeds were immersed in PGR solutions such as Gibberellic Acid (GA?, 1000 ppm), Benzyl aminopurine (BAP, 1000 ppm), Hydrogen peroxide (H?O?, 1 M), GA? + BAP (1000 ppm + 1000 ppm) and water as a control. Then, they were sown in torf pots with 5cm diameter containing perlite: torf (v/v, 1:1) mixture. Pots were put in plastic boxes with lid containing humidity 99%. All boxes were placed to germination room with temperature 27? and in dark condition. Germinated seeds were transferred another plastic box and put in growth room illuminated by LED lamps (PAR=135–317 ?mol m?² s?¹) and heated to 25?. Lids of boxes were closed position in both rooms. Seedlings with 3–5 true leaves were transferred to plastic seedling bags containing perlite: torf (v/v, 1:1) and then they were moved to greenhouse. Germination rate and obtained seedling rate were increased up to 78.32%at 1000 ppm GA? application when compared to control (60.39%). Germinating grape seeds after immersing in GA? for 24 hours and thereinafter growing seedlings in plastic boxes are an effective method for hybrid seed germination in grape breeding

S100A8 and S100A9 Positive Cells in Colorectal Carcinoma: Clinicopathological Analysis

Introduction. In colorectal carcinoma, tumoral tissues infiltrate with various immune/inflammatory cells along their invasive margins and the increased S100A8/A9 expression in these immune cells infiltrating the tumor has recently been demonstrated. We examined S100A8/A9 as a potential therapeutic target in the treatment of colorectal carcinoma. Materials and Methods. The current study included a sample of 80 patients diagnosed with CRC (30 cases with distant metastasis, 30 cases with lymph node metastasis, and 20 cases with no metastasis). Peritumoral and intratumoral S100A8 and S100A9 expressing inflammatory cells were counted in primary tumors and their metastasis and correlated with clinicopathological parameters. Results. The peritumoral and intratumoral S100A8/A9 positive cells showed no correlation with age, gender, or depth of tumor invasion. However higher counts of peritumoral and intratumoral S100A8/A9 positive cells were associated with larger tumor size, higher grade, and the presence of metastasis (P<0.05). Conclusion. Our study also found significantly higher number of S100A8/A9 positive cells in the tumor microenvironment among patients with large tumor size, high grade, and metastatic disease. Moreover, in our study, we observed that the expression in the tumor metastasis appeared similar to that of primary tumor

Acinetobacterspp in Intensive Care Unit: Risk Factors Associated with Infection and Mortality

Purpose: Acinetobacter was known as a saprophytic colonizer, yet it is considered an important etiologic agent which is isolated from hospital-acquired infections. In this study, we aimed to investigate fatality rates, factors for fatality of Acinetobacter infections and relationship between combination therapies and survival.Patients and Methods: We evaluated patients with Acinetobacter infection in ICU for demographic and clinical characteristics, combination therapies and adverse effects of these therapeutic options, retrospectively.Results: Ninety-one patients with proved Acinetobacter infection were included in this study. We found that the fatality rate of Acinetobacter infection in ICU was 56%. Advanced age, carbapenem resistance and Acinetobacter isolation from respiratory secretions were significantly related with fatality.  We found that carbapenem resistance increased steadily during a 5 year period, fatality rates significantly decreased with combination therapies including aminoglycosides. There wasn’t significant relation between colistin therapy and survival in patients with Acinetobacter infections.Conclusion: The patients should be well-assessed in terms of fatality risk factors in case of Acinetobacter infections that occur in the intensive care units. The empirical antibiotic treatment started earlier based on such risk factors may decrease fatality. The antimicrobial combinations including aminoglycoside may be considered more in order to the local resistance data

Plasmopara viticola infection process and accumulation of important stilbenes in some grapevine varieties grown in Turkey and in individuals for brined leaves

The detached leaves of three Turkish grapevine varieties (cv. Karasakız, cv. Müşküle and cv. Tekirdağ Sultani) and three hybrid individuals for brined leaves (119, 154, 200) with different resistance levels to downy mildew (Plasmopara viticola) based on sporulation area (mm2) were examined for sporangia number at 7 days, progress of the pathogen and stilbene production at different times after inoculation. A significant positive correlation was determined between number of sporangia on the lesion and sporulation area. The lowest number of stomata infected by the pathogen at 48 h after inoculation (hpi) was determined in extremely resistant 200, followed by highly resistant cv. Müşküle and 154. Vesicles of the pathogen were observed in leaves of all tested plants at 15 hpi; however, in 200 the vesicle primary hypha, which developed at 72 hpi, did not elongate into intercellular spaces of the mesophyll after that time. Elongation occurred in 119 (resistant) at 48 hpi and in cv. Müşküle and 154 at 72 hpi, but development of the pathogen was restricted to 72 hpi. Pathogen mycelia covered the intercostal fields in sensitive (cv. Karasakız) and highly sensitive (cv. Tekirdağ Sultani) varieties at 72 hpi. The amount of resveratrol was found to be high in resistant types, while the highest amount of ?-viniferin was determined in 200 at 72 hpi. Cv. Müşküle and 154 contained higher amounts of pterostilbene than 119. These new results underline the role of ?-viniferin in 200 and pterostilbene in cv. Müşküle and 154 in high resistance toward P. viticola and the importance of these grapevine types in downy mildew management. © 2021, The Author(s) under exclusive licence to Deutsche Phytomedizinische Gesellschaft.NKUBAP.03; 1180226; Central Research Laboratory, CRLThe authors acknowledge the Research Fund of Namık Kemal University (Project Number NKUBAP.03.YL.19.224) for their support, the Turkish Scientific and Technical Research Council (TUBİTAK) (Project No.: 1180226) for obtaining the individuals for brined leaves and Central Research Laboratory (NABILTEM-NKU) for using HPLC. The authors are also grateful to Martha Rowe (University of Nebraska–Lincoln) for improving the language and for useful remarks