Transgenic cucumber expressing an endogenous class III chitinase gene has reduced symptoms from Botrytis cinerea

A class III chitinase gene (CHI2) is induced in cucumber plants (Cucumis sativa L.) in response to infection by pathogenic microorganisms. Infection of Botrytis cinerea, causal agent of gray mold disease on cucumber, also induces CHI2 expression. To investigate whether CHI2 is involved in resistance to gray mold disease, transgenic cucumber plants were produced to overexpress the CHI2 gene. One line was analyzed in detail in terms of disease resistance. The transgenic cucumber plant (CC2) constitutively expressed CHI2 and reduced the symptoms of B. cinerea for 4 days after inoculation compared with nontransgenic plants. However, this inhibitory effect was not absolute, and CC2 eventually developed serious disease symptoms. Chitinase activity of the crude extract from CC2 leaves was higher than that from nontransgenic plants. A high-molecular-weight fraction containing CHI2 from CC2 leaves had fungistatic activity against B. cinerea. Interestingly, the low-molecular-weight fraction from CC2 leaves with CHI2 removed also had fungistatic activity against B. cinerea. Not only the introduced chitinase activity but also the endogenous defense reactions activated by overexpression of CHI2 may be involved in the enhanced gray mold disease resistance in CC2.     

Inflammation of the cardiac coronary artery in ICR mice

Inflammation of the cardiac coronary artery in ICR mice is occasionally observed in toxicity studies; however, this has not been well explored histologically. Herein, we investigated the detailed histology of the associated lesions in 6–8-week-old ICR mice. Coronary artery inflammation in the right ventricular wall was observed in 10 of 142 mice (7.0%). Histopathological examination revealed hypertrophy of the vascular smooth muscle cells and perivascular infiltration of macrophages in mild cases. In moderate to marked cases, single-cell necrosis of vascular smooth muscle cells, hemorrhage of the tunica media, and fibrinoid necrosis of the vessel wall were observed, in addition to the changes seen in mild cases. Electron microscopic examination of moderate cases revealed a discontinuous internal elastic lamina suggestive of rupture, and vascular smooth muscle cells beneath the elastic lamina showed degeneration and necrosis. These findings suggest that the lesions developed as a rupture of the internal elastic lamina and necrosis of vascular smooth muscle cells, while leaked plasma components caused vascular and perivascular inflammation. In ICR mice, dystrophic calcinosis (DCC) is known to occur rarely in the right ventricle. DCC is defined as focal calcification in necrotic myocardial fibers, the pathogenesis of which is considered to involve ectopic calcification. Since calcification was not observed in any part of the heart, including the inflammation region, the pathophysiology of cardiac arterial inflammation seen in our ICR mice was considered to differ from that of DCC.     

Plasma histamine concentrations in horses administered sodium penicillin,guaifenesin–xylazine–ketamine and isoflurane with morphine or butorphanol

ObjectiveVarious drugs administered to horses undergoing surgical procedures can release histamine. Histamine concentrations were evaluated in horses prepared for surgery and administered butorphanol or morphine intraoperative infusions.Study designProspective studies with one randomized.AnimalsA total of 44 client-owned horses.MethodsIn one study, anesthesia was induced with xylazine followed by ketamine–diazepam. Anesthesia was maintained with guaifenesin–xylazine–ketamine (GXK) during surgical preparation. For surgery, isoflurane was administered with intravenous (IV) morphine (group M: 0.15 mg kg^–1 and 0.1 mg kg^–1 hour^–1; 15 horses) or butorphanol (group B: 0.05 mg kg^–1 and 0.01 mg kg^–1 hour^–1; 15 horses). Histamine and morphine concentrations were measured using enzyme-linked immunoassay before opioid injection (time 0), and after 1, 2, 5, 30, 60 and 90 minutes. In a subsequent study, plasma histamine concentrations were measured in 14 horses before drug administration (baseline), 15 minutes after IV sodium penicillin and 15 minutes after starting GXK IV infusion. Statistical comparison was performed using anova for repeated measures. Pearson correlation compared morphine and histamine concentrations. Data are presented as mean ± standard deviation. Significance was assumed when p ≤ 0.05.ResultsWith histamine, differences occurred between baseline (3.2 ± 2.4 ng mL^–1) and GXK (5.2 ± 7.1 ng mL^–1) and between baseline and time 0 in group B (11.9 ± 13.4 ng mL^–1) and group M (11.1 ± 12.4 ng mL^–1). No differences occurred between baseline and after penicillin or between groups M and B. Morphine concentrations were higher at 1 minute following injection (8.1 ± 5.1 ng mL^–1) than at 30 minutes (4.9 ± 3.1 ng mL^–1) and 60 minutes (4.0 ± 2.5 ng mL^–1). Histamine correlated with morphine at 2, 30 and 60 minutes.Conclusions and clinical relevanceGXK increased histamine concentration, but concentrations were similar with morphine and butorphanol.     

Quality evaluation and prediction of Citrullus lanatus by 1H NMR-based metabolomics and multivariate analysis

(1)H NMR spectrometry in combination with multivariate analysis was considered to provide greater information on quality assessment over an ordinary sensory testing method due to its high reliability and high accuracy. The sensory quality evaluation of watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai) was carried out by means of (1)H NMR-based metabolomics. Multivariate analyses by partial least-squares projections to latent structures-discrimination analysis (PLS-DA) and PLS-regression offered extensive information for quality differentiation and quality evaluation, respectively. The impact of watermelon and rootstock cultivars on the sensory qualities of watermelon was determined on the basis of (1)H NMR metabolic fingerprinting and profiling. The significant metabolites contributing to the discrimination were also identified. A multivariate calibration model was successfully constructed by PLS-regression with extremely high reliability and accuracy. Thus, (1)H NMR-based metabolomics with multivariate analysis was considered to be one of the most suitable complementary techniques that could be applied to assess and predict the sensory quality of watermelons and other horticultural plants.     

A pollen extender medium technology for seedless watermelon production by pollination with soft X-ray irradiated pollen

The inactivation of pollen by soft X-ray irradiation and subsequent artificial pollination are time-intensive practices used in the production of seedless watermelons (Citrullus lanatus L.). Watermelon generally has a lot of staminate flowers; however, they only have a small amount of pollen. Watermelon pollen cannot be used in pollination under the present situation. Therefore, the objective of this study was to determine which is the most effective pollen extender medium for cultivation of watermelon with soft X-ray irradiated pollen. In this experiment, ‘Agar,’ ‘Marriage powder,’ and ‘Sekishoshi’ (Lycopodium dyed red with safflower pigment) were used as extender media with soft X-ray irradiated pollen at equal or twice the weight of the pollen. When ‘Sekishoshi’ was used as an extender medium, fruit set was very low. A lot of deformed fruit was produced when agar was used with the pollen. On the other hand, when ‘Marriage powder’ was used in equal proportions with the pollen, fruit set was about 70% and, moreover, Brix was high. Thus, the mixture of ‘Marriage powder’ with an equal amount of pollen was the best for use in actual cultivation. We conclude, then, that soft X-ray irradiated pollen in an extender medium can be effectively adapted for producing seedless watermelons.     

Variable response of growth and arbuscular mycorrhizal colonization of maize plants to preceding crops in various types of soils

The effects of the preceding crops, sunflower (mycorrhizal host) and mustard (nonhost), on arbuscular mycorrhizal (AM) colonization and growth of succeeding maize were examined in 17 soils in an attempt to clarify the influence of soil characteristics on the effects of preceding crops. Shoot weight and P uptake of maize planted after sunflower were much higher than those after mustard in 14 soils, although the preceding crop had little effect on soil-P availability. AM colonization of maize after sunflower was much higher than that after mustard. The effect of the preceding crop was eliminated by soil sterilization. These results suggested that the differences in maize growth were caused by differences in the AM colonization. Correlation analysis of the effect of the preceding crop and soil properties showed that the difference in the effects on maize growth could not be explained by soil chemical properties, but only by the AM colonization of the preceding sunflower crop. In one of the 17 soils, however, the effect was not evident despite the higher AM colonization of sunflower. This soil was sterilized, and the effect of inoculation by AM fungi (AMF) on maize was examined. However, it was found that the inoculation increased AM colonization but did not improve maize growth at any P level, suggesting that the effect of AMF was unusually inhibited in this soil by unknown soil physicochemical properties. In most soils, however, the preceding mycorrhizal host crop, sunflower, improved the growth and AM colonization of maize depending on the AM colonization of sunflower.     

Effects of epidermal growth factor (EGF) on fetal islet B cells in vitro

ABSTRACT. It has been reported that when the rat fetus is treated with streptozotocin (STZ) in vivo, islet B cells are destroyed but later recover. To investigate the process of the recovery of B cells after in vitro treatment of the fetal pancreas with STZ and the role of epidermal growth factor (EGF) in the recovery of B cells, we measured the level of insulin released from the cultured fetal pancreas and examined it histologically. As a result, we immunohistologically confirmed the regeneration of B cells in the pancreas that had been cultured for 48 hr after destruction of islet B cells by STZ treatment. An immunohistologic study using proliferating cell nuclear antigen (PCNA) showed that without the addition of EGF, the cell division index was significantly higher in the STZ-treated group (STZ group) than in the untreated group (intact group), whereas with the addition of EGF, the cell division index increased in both groups, but EGF did not have a significant cell division-promoting effect on the pancreas in the STZ group. The addition of EGF caused a significant decrease in the concentration of insulin in culture medium in both groups. These results indicate that EGF has a cell growth-promoting effect on intact fetal pancreas in vitro but has the effect of inhibiting the release of insulin, and thus suggest that EGF does not trigger the regeneration of islet B cells.     

Transformation of trifoliate orange with rice chitinase gene and the use of the transformed plant as a rootstock

RCC2, cDNA clone encoding rice class-I chitinase, was introduced into trifoliate orange (Poncirus trifoliata Raf.). Chitinase activity of the transformed plants was higher than that of the non-transformed plants. The introduced gene was not detected in the scions grafted onto the transformed plants. The effect of the introduction of foreign gene into rootstocks on the scions was discussed.     

Calcium ion promotes successful penetration of powdery mildew fungi into barley cells

To determine whether Ca²⁺ promotes powdery mildew penetration, Ca²⁺-treated barley coleoptiles were inoculated with conidia of pathogenic and nonpathogenic fungi. Penetration efficiency of the pathogenic powdery mildew Blumeria graminis was enhanced by Ca²⁺ treatment, but that of the necrotrophic pathogen Helminthosporium sp. remained unaffected. Similarly, when actin-dependent penetration resistance is suppressed with cytochalasin A, Ca²⁺ treatment specifically enhanced penetration of the nonpathogenic powdery mildew Erysiphe pisi but not that of other nonpathogens. Calmodulin inhibitors suppressed the promotive effect of Ca²⁺ on B. graminis penetration. These results suggest that barley powdery mildew specifically requires Ca²⁺ and calmodulin for penetration.