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.     

Underwater sound detection by cephalopod statocyst

Abstract:   The cephalopod receptor of particle motion was identified. In a previous study, it was suggested that statocysts served this function, but there was no direct supporting evidence, and epidermal hair cells had not been conclusively ruled out. Experiments on Octopus ocellatus were conducted using respiratory activity as an indicator of sound perception. Intact animals clearly responded to 141-Hz particle motion at particle accelerations below 1.3 × 10⁻³ m/s², and the mean perception threshold at this frequency was approximately 6.0 × 10⁻⁴ m/s². Specimens in which the statoliths had been surgically removed did not show any response for accelerations up to 3.9 × 10⁻³ m/s² at 141 Hz, which was approximately 16 dB greater than the mean perception threshold at this frequency. Specimens that had undergone a control operation in which the statoliths remained intact showed positive responses at 2.8 × 10⁻³ m/s² for the same frequency stimulus. This indicates that the statocyst, which is morphologically similar to the inner ear system in fish, is responsible for the observed responses to particle motion in O. ocellatus . This is the first direct evidence that cephalopods detect kinetic sound components using statocysts.     

Measurements of auditory sensitivity in common carp <Emphasis Type="Italic">Cyprinus carpio</Emphasis> by the auditory brainstem response technique and cardiac conditioning method

ABSTRACT:   Recently, the non-invasive auditory brainstem response (ABR) technique has been applied to the determination of hearing sensitivity in fish. The technique has some advantages over, and methodological differences from, the classical techniques. The auditory thresholds of common carp were measured by the ABR technique. The classical method was applied to measure cardiac response by electrocardiogram (ECG) after conditioning with electric shock. The most sensitive frequency obtained by ABR was 505 Hz, and by ECG, 1000 Hz. The shapes of the audiogram obtained by ABR and ECG were similar, though threshold levels in both audiograms differed from each other. Furthermore, audiograms obtained by using behavioral techniques were compared with those taken by the ABR and ECG techniques to assess the validity of the ABR technique for testing fish hearing sensitivity. The measured threshold levels increased in the order of behavioral, ABR, and ECG techniques, in the range below 1000 Hz.     

Concentrations of toxic metals and essential minerals in the mane hair of healthy racing horses and their relation to age

Concentrations of trace elements (As, Al, Pb, Cd, Hg, Se, Si, P, Na, K, Ca, Mg, Fe, Cu, Zn, Mn, Cr, Ni and Mn) in the mane hair obtained from 9 female and 15 male healthy racing Thoroughbred horses aged 2-5 years were analyzed by the inductively coupled plasma atomic emission spectrometry (ICP-AES) method. No significant differences between the female and male horses were observed in the mean concentrations of those minerals. Significantly positive correlations with age were observed in Cd (r=0.546, p<0.01) and Mo (r=0.733, p<0.001). Significantly negative correlations with age were observed in Hg (r= -0.726, p<0.001), Mn (r= -0.450, p<0.05) and Fe (r=-0.642, p<0.01). This reference range of trace elements in the mane hair of racing horses should be used to assess disease and the nutritional status in equine practice.     

Pea extracellular Cu/Zn-superoxide dismutase responsive to signal molecules from a fungal pathogen

We previously reported that the release of O₂ ⁻ from isolated pea cell walls was enhanced by a 70-kDa glycoprotein elicitor but was suppressed by mucin-type glycopeptide suppressors (supprescins A and B) prepared from pycnospore germination fluid of Mycosphaerella pinodes, causal agent of Mycosphaerella blight of pea. Here, we show that superoxide dismutase (SOD) in the apoplast fluid/cell wall of pea seedlings responds to the fungal elicitor and suppressor molecules. In a pharmacological study and with internal amino acid sequencing, the apoplastic SOD in a pea cultivar Midoriusui was found to be a Cu/Zn type SOD. We cloned a full-length cDNA of the Cu/Zn-SOD and designated it as PsCu/Zn-SOD1. An increase in PsCu/Zn-SOD1 mRNA and the PsCu/Zn-SOD1 protein was induced by treatment with the elicitor more intensively than by wounding. Such induction by the elicitor or wounding, however, was inhibited by the concomitant presence of supprescins. The SOD activity of recombinant PsCu/Zn-SOD1 was regulated directly by these signal molecules in a manner similar to their effect on the SOD activity in the apoplastic fluid and in the cell wall-bound proteins. Based on these findings, we discuss a role for PsCu/Zn-SOD1 in the pea defense response. The nucleotide sequence data of PsCu/Zn-SOD1 reported are available in the DDBJ/EMBL/GenBank databases under accession number AB189165.     

Management approach using simple indices of deer density and status of understory vegetation for conserving deciduous hardwood forests on a regional scale

We investigated the validity and efficiency of a survey using sight per unit effort (SPUE) of sika deer and shrub-layer decline rank (SDR), which is an index of decline in the physical structure of a whole stand caused by sika deer, based on data collected on a broad scale. This survey was to be used to manage a deer population in order to conserve a forest ecosystem. First, we evaluated the spatial and temporal scales of deer density that are most appropriate for predicting decline in the status of understory vegetation. The model with SPUE calculated in a buffer with a radius of 4.5 km using data for the past 4 years was found to be the best. We showed that our knowledge of the relationship between deer density and status of shrub-layer vegetation is improved by identifying the most suitable spatial and temporal scales of SPUE for predicting SDR. Next, we quantified the effects of SPUE and environmental components on SDR in stands. We found that SPUE had the greatest effect on SDR among all explanatory variables. Moreover, the area under the curve (AUC) was large in a model that only used SPUE (AUC = 0.718). This result suggests that the variation in SDR among stands was explained well by SPUE regardless of differences in the forest environment. Furthermore, we identified the effective values of SPUE for preventing shrub-layer vegetation from declining through deer density control. We conclude that a management system based on SPUE and SDR is a simple and valid method for managing deer populations in order to conserve forest ecosystems.     

Reactivity of secondary hydroxyl groups in methylβ-d-xylopyranoside toward aβ-o-4-type quinone methide

Methyl-d-xylopyranoside was allowed to react with-O-4-type quinone methide without a catalyst to elucidate the reactivities of secondary hydroxyl groups at the C2, C3, and C4 positions. Benzyl ether-type lignin-carbohydrate complex (LCC) compounds linked at the C2 and C4 positions were predominant, at a ratio of 23. However, the reactivity of the hydroxyl group at the C3 position was quite low. These results strongly suggest that the reactivity of the C2 hydroxyl group in xylan toward quinone methide intermediate is higher than that of the C3 hydroxyl group during biosynthesis of LCCs.     

HIV-1 integrase and neuraminidase inhibitors from Alpinia zerumbet

AIDS and influenza are viral pandemics and remain one of the leading causes of human deaths worldwide. The increasing resistance of these diseases to synthetic drugs demands the search for novel compounds from plant-based sources. In this regard, the leaves and rhizomes of Alpinia zerumbet, a traditionally important economic plant in Okinawa, were investigated for activity against HIV-1 integrase (IN) and neuraminidase (NA). The aqueous extracts of leaves and rhizomes had IN inhibitory activity with IC(50) values of 30 and 188 μg/mL, whereas against NA they showed 50% inhibition at concentrations of 43 and 57 μg/mL, respectively. 5,6-Dehydrokawain (DK), dihydro-5,6-dehydrokawain (DDK), and 8(17),12-labdadiene-15,16-dial (labdadiene) were isolated from the rhizomes and were tested for enzyme inhibitions. DK and DDK strongly inhibited IN with IC(50) of 4.4 and 3.6 μg/mL, respectively. Against NA, DK, DDK, and labdadiene exhibited mixed type of inhibition with respective IC(50) values of 25.5, 24.6, and 36.6 μM and K(i) values ranging from 0.3 to 2.8 μM. It was found that DDK is a slow and time-dependent reversible inhibitor of NA, probably with a methoxy group as its functionally active site. These results suggest that alpinia could be used as a source of bioactive compounds against IN and NA and that DK and DDK may have possibilities in the design of drugs against these viral diseases.