Soaking grapevine cuttings in water: a potential source of cross contamination by micro-organisms

Grapevine nurseries soak cuttings in water during propagation to compensate for dehydration and promote root initiation. However, trunk disease pathogens have been isolated from soaking water, indicating cross contamination. Cuttings of Vitis vinifera cv. Sunmuscat and V. berlandieri x V. rupestris rootstock cv. 140 Ruggeri were immersed in sterilized, deionised water for 1, 2, 4, 8 and 16 h. The soaking water was cultured (25°C for 3 days) on non-specific and specific media for fungi and bacteria. The base of each cutting was debarked and trimmed and three 3 mm thick, contiguous, transverse slices of wood cultured at 25°C for 3 days. The soaking water for both cultivars became contaminated with microorganisms within the first hour. Numbers of fungi iso-lated from the wood slices soaked for one hour were significantly greater than those from non-soaked cuttings. The number of bacterial colonies growing from the wood slices increased after soaking for 2‒4 h in Sunmuscat. In a second experiment Shiraz cuttings were soaked for 1, 2, 4, 8 and 24 h. The soaking water became contaminated within the first hour but only the bacterial count increased significantly over time. Microorganisms also established on the container surfaces within the first hour although there were no significant increases over 24 h. These results confirm that soaking cuttings is a potential cause of cross contamination and demonstrate contamination of cuttings occurs after relatively short periods of soaking. Avoiding exposing cuttings to water will reduce the transmission of trunk diseases in propagation

Case Reports1. A Late Presentation of Loeys-Dietz Syndrome: Beware of TGFβ Receptor Mutations in Benign Joint Hypermobility

Background: Thoracic aortic aneurysms (TAA) and dissections are not uncommon causes of sudden death in young adults. Loeys-Dietz syndrome (LDS) is a rare, recently described, autosomal dominant, connective tissue disease characterized by aggressive arterial aneurysms, resulting from mutations in the transforming growth factor beta (TGFβ) receptor genes TGFBR1 and TGFBR2. Mean age at death is 26.1 years, most often due to aortic dissection. We report an unusually late presentation of LDS, diagnosed following elective surgery in a female with a long history of joint hypermobility. Methods: A 51-year-old Caucasian lady complained of chest pain and headache following a dural leak from spinal anaesthesia for an elective ankle arthroscopy. CT scan and echocardiography demonstrated a dilated aortic root and significant aortic regurgitation. MRA demonstrated aortic tortuosity, an infrarenal aortic aneurysm and aneurysms in the left renal and right internal mammary arteries. She underwent aortic root repair and aortic valve replacement. She had a background of long-standing joint pains secondary to hypermobility, easy bruising, unusual fracture susceptibility and mild bronchiectasis. She had one healthy child age 32, after which she suffered a uterine prolapse. Examination revealed mild Marfanoid features. Uvula, skin and ophthalmological examination was normal. Results: Fibrillin-1 testing for Marfan syndrome (MFS) was negative. Detection of a c.1270G > C (p.Gly424Arg) TGFBR2 mutation confirmed the diagnosis of LDS. Losartan was started for vascular protection. Conclusions: LDS is a severe inherited vasculopathy that usually presents in childhood. It is characterized by aortic root dilatation and ascending aneurysms. There is a higher risk of aortic dissection compared with MFS. Clinical features overlap with MFS and Ehlers Danlos syndrome Type IV, but differentiating dysmorphogenic features include ocular hypertelorism, bifid uvula and cleft palate. Echocardiography and MRA or CT scanning from head to pelvis is recommended to establish the extent of vascular involvement. Management involves early surgical intervention, including early valve-sparing aortic root replacement, genetic counselling and close monitoring in pregnancy. Despite being caused by loss of function mutations in either TGFβ receptor, paradoxical activation of TGFβ signalling is seen, suggesting that TGFβ antagonism may confer disease modifying effects similar to those observed in MFS. TGFβ antagonism can be achieved with angiotensin antagonists, such as Losartan, which is able to delay aortic aneurysm development in preclinical models and in patients with MFS. Our case emphasizes the importance of timely recognition of vasculopathy syndromes in patients with hypermobility and the need for early surgical intervention. It also highlights their heterogeneity and the potential for late presentation. Disclosures: The authors have declared no conflicts of interes

Proton Nuclear Magnetic Resonance-Spectroscopic Discrimination of Wines Reflects Genetic Homology of Several Different Grape (V. vinifera L.) Cultivars.

Proton nuclear magnetic resonance spectroscopy coupled multivariate analysis (1H NMR-PCA/PLS-DA) is an important tool for the discrimination of wine products. Although 1H NMR has been shown to discriminate wines of different cultivars, a grape genetic component of the discrimination has been inferred only from discrimination of cultivars of undefined genetic homology and in the presence of many confounding environmental factors. We aimed to confirm the influence of grape genotypes in the absence of those factors.We applied 1H NMR-PCA/PLS-DA and hierarchical cluster analysis (HCA) to wines from five, variously genetically-related grapevine (V. vinifera) cultivars; all grown similarly on the same site and vinified similarly. We also compared the semi-quantitative profiles of the discriminant metabolites of each cultivar with previously reported chemical analyses. The cultivars were clearly distinguishable and there was a general correlation between their grouping and their genetic homology as revealed by recent genomic studies. Between cultivars, the relative amounts of several of the cultivar-related discriminant metabolites conformed closely with reported chemical analyses.Differences in grape-derived metabolites associated with genetic differences alone are a major source of 1H NMR-based discrimination of wines and 1H NMR has the capacity to discriminate between very closely related cultivars.The study confirms that genetic variation among grape cultivars alone can account for the discrimination of wine by 1H NMR-PCA/PLS and indicates that 1H NMR spectra of wine of single grape cultivars may in future be used in tandem with hierarchical cluster analysis to elucidate genetic lineages and metabolomic relations of grapevine cultivars. In the absence of genetic information, for example, where predecessor varieties are no longer extant, this may be a particularly useful approach

Comparison of wine discrimination derived from the ¹H NMR spectra.

<p>(A) PCA Scores Plot (t1/t2), R²X = 0.959, Q² = 0.872. PC1/PC2 accounted for 65.9% of the total variance. (B) PLS-DA Scores Plot (t1/t2), R²X = 0.959, R²Y = 0.991, Q² = 0.954. PC1/PC2 accounted for 65.5% of the total variance. Grape cultivars: m, Merlot; s, Syrah; z, Zinfandel; r, Ruby Cabernet; c, Cabernet Sauvignon.</p

¹H NMR spectrum of Cabernet Sauvignon wine with NOESYPRESAT water peak suppression, (A) sample not lyophilized, (B) sample lyophilized.

<p>¹H NMR spectrum of Cabernet Sauvignon wine with NOESYPRESAT water peak suppression, (A) sample not lyophilized, (B) sample lyophilized.</p

PLS-DA score plots, loading plots and correlation parameters derived from the ¹H NMR spectra of wines as pair wise comparisons (A) Merlot (m) and Syrah (s), R²X = 0.737, R²Y = 0.991, Q² = 0.958.

<p>PC1/PC2 variance accounted for 72.1%. (B) Merlot (m) and Zinfandel (z), R²X = 0.791, R²Y = 0.992, Q² = 0.973. PC1/PC2 accounted for 79.1%. (C) Merlot (m) and Ruby Cabernet (r), R²X = 0.679, R²Y = 0.982, Q² = 0.888. PC1/PC2 variance accounted for 67.9%. (D) Merlot (m) and Cabernet Sauvignon (c), R²X = 0.511, R²Y = 0.991, Q² = 0.891. PC1/PC2 variance accounted for 51.4% (E) Syrah (s) and Ruby Cabernet (r), R²X = 0.705, R²Y = 0.998, Q² = 0.968. PC1/PC2 variance accounted for 70.5%. (F) Syrah (s) and Zinfandel (z), R²X = 0.807, R²Y = 0.992, Q² = 0.989. PC1/PC2 accounted for; 80.7%. (G) Syrah (s) and Cabernet Sauvignon (c), R²X = 0.586, R²Y = 0.985, Q² = 0.945. PC1/PC2 variance accounted for 56.7%. (H) Ruby Cabernet (r) and Zinfandel (z), R²X = 0.822, R²Y = 0.998, Q² = 0.994. PC1/PC2 variance accounted for 80.2%. (I) Cabernet Sauvignon (c) and Zinfandel (z), R²X = 0.897, R²Y = 0.994, Q² = 0.983. PC1/PC2 variance accounted for 89.7%. (J) Cabernet Sauvignon (c) and Ruby Cabernet (r), R²X = 0.775, R²Y = 0.995, Q² = 0.976. PC1/PC2 variance accounted for 74.8%.</p

Dendrogram of wines of five cultivars (M, Merlot; C, Cabernet Sauvignon; R, Ruby Cabernet; Z, Zinfandel; S, Syrah), (4 replicates of each), based on multidimensional analysis of metabolites detected by ¹H NMR spectroscopy.

<p>Dendrogram of wines of five cultivars (M, Merlot; C, Cabernet Sauvignon; R, Ruby Cabernet; Z, Zinfandel; S, Syrah), (4 replicates of each), based on multidimensional analysis of metabolites detected by ¹H NMR spectroscopy.</p

The ¹H-¹H COSY NMR spectrum of lyophilized Cabernet Sauvignon dry red wine.

<p>The ¹H-¹H COSY NMR spectrum of lyophilized Cabernet Sauvignon dry red wine.</p

Identification of volatile compounds used in host location by the black bean aphid, Aphis fabae

Behavioral and electrophysiological responses of winged Aphis fabae to volatiles of faba bean, Vicia faba (var. Sutton dwarf), plants were studied and semiochemicals used in host location were identified. In olfactometer bioassays, aphids spent significantly more time in the region of the olfactometer where V. faba volatiles from an intact plant were present than in control regions with clean air. This response also occurred when an air entrainment sample of a V. faba plant was used as the odor source. Coupled gas chromatography-electroantennography revealed the presence of 16 electrophysiologically active compounds in the air entrainment sample. Fifteen of these were identified as (Z)-3-hexen-1-ol, 1-hexanol, (E)-2-hexenal, benzaldehyde, 6-methyl-5-hepten-2-one, octanal, (Z)-3-hexen-1-yl acetate, (R)-(-)-linalool, methyl salicylate, decanal, undecanal, (E)-caryophyllene, (E)-beta-farnesene, (S)-(-)-germacrene D, and (E,E,)-4,8,12-trimethyl-1,3,7,11-tridecatetraene. An olfactometer response was observed to a 15-component synthetic blend that comprised all identified compounds at the same concentration and ratio as in the natural sample, with the aphids spending significantly more time in the treated regions of the olfactometer where volatiles were present than in the control regions. These data are discussed in the context of insect host location and crop protectio