Ovarian activity and pregnancy in the Siberian tiger, Panthera tigris altaica, assessed by fecal gonadal steroid hormones analyses

Feces were collected from two female and one male Siberian tigers, Panthera tigris altaica. Steroid hormones were extracted from lyophilized feces and quantified by enzyme immunoassay. The fecal contents of estradiol-17beta (E(2)) and testosterone in the females and male, respectively, changed markedly throughout the year. The fecal E(2) contents of females Nos. 179 and 238 increased at 26.4 +/- 8.0 and 28.0 +/- 14.2 day intervals, respectively. However, the fecal contents of progesterone (P(4)) in the female kept alone did not change. In contrast, the other female, which was kept with a male, had increased fecal P(4) contents after copulation. The fecal progesterone levels of the pregnant female remained high during her 106-day pregnancy.     

A serosurvey of Borna disease virus infection in wild rats by a capture ELISA

For a serological diagnostic test for Borna disease (BD), we developed a capture ELISA with specificity and sensitivity based on detection of antibodies against BD virus (BDV) p40 protein. Using our capture ELISA system, the antibody response of rats inoculated intracerebrally with BDV at 4 weeks after birth showed a sharp increase from 1 to 4 weeks postinoculation (p.i.) and a steady level after 5 weeks p.i. To investigate prevalence of BDV infection among wild rats, we examined sera of Rattus norvegicus in Kami-iso town, Oshima district, Hokkaido, suggesting that rats in this area had not been infected by BDV.     

Lens formation in the absence of optic cup in rat embryos irradiated with soft X-ray

In order to investigate the effect of soft X-ray irradiation on ocular development, pregnant rats were exposed to a single 12.5 Gy irradiation on embryonic day 9 (ED 9). The embryos obtained by laparotomy on ED 12 and 21 were examined for ocular abnormalities under a binocular stereo-microscope and a light microscope. The ED 12 embryos were stained with osmium tetroxide to facilitate the observation. The stereo-microscopic examination on ED 12 and 21 revealed various types of ocular abnormalities characterized primarily by aplasia or hypoplasia of the optic cup and invaginated lens placode. The light microscopic examination further confirmed these findings histomorphologically, and the hypoplastic abnormalities were classified into three types: (1) hypoplasia of the optic cup and invaginated lens placode, (2) complete malformation of the optic cup and hypoplasia of the invaginated lens placode, and (3) complete malformation of the optic cup and invaginated lens placode. Because the lens was formed in the complete absence of the retina, the development and differentiation of the retina and lens do not seem to be tightly synchronized. Thus, this sequential analysis on ocular abnormalities during the early stage of development supports the notion that the presence of the retina is not always necessary for the development of the lens.     

Need for flagella for complete virulence of Pseudomonas syringae pv. tabaci: genetic analysis with flagella-defective mutants ?fliC and ?fliD in host tobacco plants

The flagellins purified from Pseudomonas syringae pv. tabaci induce a hypersensitive reaction in nonhost tomato cells. To investigate the role of flagella and flagellin in the compatible interaction, we generated two types of flagella-defective mutant. The fliC mutant lost the fliC gene that encodes flagellin protein, whereas the fliD mutant lost the fliD gene that encodes HAP2-capping protein. The two mutants had markedly reduced ability to cause disease symptoms in tobacco leaves. Furthermore, propagation of the mutants in tobacco leaves was less than that in wild-type pv. tabaci. Compared to the inoculation with wild-type pv. tabaci, inoculation with the two mutants did not markedly induce the expression of typical defense response-related genes such as PAL and hsr203J. Complementation of each fliC and fliD gene to the corresponding deficient mutant restored motility and virulence. These results indicate that flagella of P. syringae pv. tabaci are indispensable organelles for complete virulence on host tobacco plants.     

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.     

Effects of D-genome chromosomes on crossability of hexaploid triticale (X Triticosecale Wittmack) with maize

With the aim of producing polyhaploids of hexaploid triticale, 20 genotypes from a CIMMYT breeding programme and eight D-genome chromosome substitution lines of ‘Rhino’ were crossed with maize. In crosses between 20 triticale genotypes and maize, 15 lines produced embryos. Frequencies of embryo formation ranged from 0.0 to 5.4%, with an average of 1.1%. From a total of 200 pollinated spikes, 62 plants were regenerated. Most regenerated plants were polyhaploids with 21 chromosomes, and few aneuhaploids with 22 chromosomes were found. In crosses of triticale substitution lines with maize, all the lines produced embryos, while ‘Rhino’ produced no embryos at all. Higher frequencies of embryo formation were obtained in substitution lines with chromosomes 2D and 4D. These results suggest that D-genome chromosomes in a triticale genetic background have the effect of increasing the frequency of polyhaploid production in triticale x maize crosses.     

Microbial Immobilization and Plant Uptake of Different N Forms in Three Forest Types in Shikoku District, Southern Japan

Soil microbial immobilization and plant uptake of N were evaluated for three forest types in Kochi, Shikoku district. During 196-d laboratory incubation, soil NO₃-N production in the Hinoki cypress forest was negligible for the initial 40 d and then rapidly increased, whereas NO₃-N production was rapid from the beginning in Japanese cedar and deciduous hardwood forests. Microbial immobilization of the labeled ¹⁵N decreased in the order of NH₄-N>glycine-N>NO₃-N. The ¹⁵N immobilization was higher for soil in the Hinoki cypress forest than other two soils. The delayed NO₃-N production in the Hinoki cypress forest was likely related with low availability of NH₄-N due to NH₄-N immobilization and substantial NO₃-N immobilization. In the field experiment, ¹⁵N uptake by roots decreased in the order of NH₄-N>NO₃-N>glycine-N. The absorption of the labeled ¹³C suggested direct uptake of organic N. The preference of N forms by root uptake was not different among forest types. Trees in three forest types can absorb inorganic and organic forms of N, suggesting trees absorb the N form that is the most abundant in the soil.