Genetic diversity of the `peruvianum-complex' (Lycopersicon peruvianum (L.) Mill. and L. chilense Dun.) revealed by RAPD analysis

Two lines of hexaploid wheat were crossed and the basic generations of parent, F1, F2 and back-cross were sown in a controlled-environment chamber. FreshF1 and back-cross grains were generated, so the material could be handled either as the standard set of basic generations on a whole-plant basis, or as an extended set on an embryo or endosperm basis. The experiment was repeated. Mature grains were harvested and the starch particle size distribution was analysed in 3284 grains from 111 plants. Means and variances were partitioned into additive, dominance and interaction components. Grains from cross-pollinations had B-granule contents between parental values, rather than of the maternal parent, indicating an involvement of the grain genotype. Quantitative models based on endosperm genotype gave a better fit to the data than those based on embryo genotype. The difference in starch B-granule content between the parents was largely due to additive genes. Dominant genes were also indicated, with the first dose in the triploid endosperm having a large effect while the second dose had little or none. Non-allelic interactions were significant in the second experiment where the use of more types of backcross made them more detectable. There were also small and significant residual effects of the maternal plant in the first experiment, attributed to the vigour of the F1 mother plant and to the cytoplasm of Sunco. Narrow-sense heritability was low, between 0.05and 0.18 depending on the generation. Transgressive segregation was not found, suggesting that all alleles tending to increase the B-granule content were found in the Sunco parent and none in ME71. There was also no detectable heterosis in this character. The results show that breeding and selection for a low B-granule content should be possible but a further reduction will require new and complementary genes. This revised version was published online in July 2006 with corrections to the Cover Date.     

Nodulation during vegetative growth of soybean stage does not affect the susceptibility to red crown rot caused by Calonectria ilicicola

Soybean cultivar Enrei and its derivative lines, non-nodulating En1282 and supernodulating Sakukei 4, were grown in an experiment field naturally infested with the red crown rot fungus, Calonectria ilicicola, to determine whether nodulation elicits the disease. Infection frequency by the fungus invariably increased with time and reached about 100% 6 weeks after sowing. Disease severity was invariably slight until 8 weeks after sowing. At harvest, however, disease severity was lowest in En1282 and highest in Sakukei 4. Enrei was intermediately susceptible. Sakukei 4 produced many more nodules than Enrei, while En1282 completely failed to nodulate. For three other pairs of nodulating and non-nodulating isogenic lines, all non-nodulating lines had lower severity than their nodulating counterparts at harvest. Pot experiments with Enrei and its derivatives revealed that inoculation with Bradyrhizobium sp. did not affect disease severity during the vegetative stage of plant growth, i.e., 6 weeks after sowing. These results suggested that the production of nodules in the vegetative stage is not, at least directly, related to susceptibility to the disease, but physiologic changes after anthesis vary depending on the nodulation traits in the respective soybean lines, resulting in different levels of susceptibility to red crown rot.     

Burkholderia gladioli associated with symptoms of bacterial grain rot and leaf-sheath browning of rice plants

Rice plants with bacterial leaf-sheath browning and grain rot were observed in Fukuoka Prefecture in Japan during the autumn seasons of 1995 and 1996. Burkholderia spp. were consistently isolated from the infected leaf sheaths and grains. These isolates were pathogenic and induced symptoms of seedling rot, grain rot, and leaf-sheath browning in rice plants, as well as in some orchidaceous plants (cymbidium, dendrobium, and oncidium leaves), gladiolus leaves, and onion bulbs. On the basis of morphological, physiological and pathological tests, and species-specific polymerase chain reaction, the isolates were identified as belonging to either Burkholderia glumae or Burkholderia gladioli. B. gladioli, as well as B. glumae, attacked rice plants after artificial inoculation and reproduced the symptoms similar to those after natural infections. We confirmed that rice is an additional natural host of B. gladioli. It is clarified that bacterial grain rot of rice is caused not only by B. glumae but also by B. gladioli.     

Involvement of carbon catabolite repression on regulation of endopolygalacturonase gene expression in citrus fruit

 The Acpg1 gene of Alternaria citri encodes an extracellular endopolygalacturonase that is important for virulence in citrus fruits. Expression of Acpg1 is regulated by substrate induction and carbon catabolite repression. A green fluorescent protein (GFP) gene was employed as a reporter gene to define 813 bases upstream of the translation start site comprising the Acpg1 promoter. This upstream sequence contains five putative binding sequences of catabolite repressive element A (CreA), a cis-acting zinc finger repressor involved in carbon catabolite repression. We constructed each CreA-binding site-deleted Acpg1 promoters with GFP reporter gene and transformed them to A. citri. The construct PGPDL4 deleted from −401 to −813 showed both substrate induction and catabolite repression, whereas PGPDL5 additionally deleted from −1 to −84, including one putative CreA-binding site, resulted in a loss of catabolite repression function. Green fluorescence of PGPDL4 was induced by pectin in the peel but was repressed completely in the juice sac area of citrus fruit. However, green fluorescence of PGPDL5 was induced in both the peel and juice sac area, indicating that repression of Acpg1 in the juice sac area is likely accomplished by carbon catabolite repression. Received: October 4, 2002 / Accepted: October 31, 2002 Acknowledgments The authors thank Drs. D. Cullen, A. Van den Wymelenberg, and J. Andrews, University of Wisconsin, for providing pTEFEGFP containing GFP and Dr. T. Tsuge, Nagoya University, for providing transformation vector pSH75. The nucleotide sequence data of Acpg1 promoter region in the DDBJ, EMBL, and GenBank sequence databases is under accession number AB047543. This research was supported in part by grants to K.A. from the Ministry of Education, Culture, Sports, Science, and Technology of Japan.     

Addition of <Emphasis Type="Italic">Pestalotiopsis </Emphasis>spp. to leaf spot pathogens of Japanese persimmon

 In June 1996, a leaf spot disease widely occurred in Japanese persimmon (Diospyros kaki) orchards in Tottori Prefecture, Japan. The main diagnostic symptom was ring spot on the leaves and calyxes of young fruits; in severe cases, lesions developed on more than half of the area of the leaf, resulting in early defoliation. Based on morphological and pathological studies of the isolated fungi, it was shown that Pestalotiopsis longiseta, P. glandicola, P. acaciae, and P. crassiuscula were responsible for the diseases. These fungi, except P. longiseta, were found to be new pathogens of the disease. Received: May 20, 2002 / Accepted: July 25, 2002     

Aldosterone-, corticosterone- and cortisol-secreting adrenocortical carcinoma in a dog: case report

A 12-year-old, intact female beagle exhibited symptoms of polyuria-polydipsia and hyperorexia for two months. Blood tests showed elevated asparate aminotransferase, alanine aminotransferase, alkaline phosphatase and creatine kinase levels, as well as marked hypokalemia. The results of adrenocorticotropic hormone stimulation test showed elevated cortisol, aldosterone and corticosterone concentrations. Abdominal ultrasonography confirmed a mass in the left adrenal gland. Masses were also seen in the liver and caudal vena cava. Diagnosis was a tumor of the adrenal cortex with metastases. Trilostane administration was initiated. The dog initially showed improved demeanor as a result of regulating hormone secretion. However, after 88 days, the dog weakened rapidly, before dying on the 117th day. Pathological findings confirmed a diagnosis of adrenocortical carcinoma.     

Effect of multiple oral dosing of fluconazole on the pharmacokinetics of cyclosporine in healthy beagles

Fluconazole (Fcz) is successfully used in human organ transplant patients as an antifungal therapy. However, Fcz can increase the cyclosporine (CsA) trough level and lead to CsA nephrotoxicity. In canine renal transplantation, CsA has been used as a major immunosuppressant, and it is important to control its trough level. However, the interaction of Fcz with CsA has not yet been reported in dogs. In this study, the effect of Fcz treatment on the pharmacokinetics of CsA in four healthy beagles was investigated using a four-period crossover design. The treatments included CsA alone (A), CsA + multiple-dose Fcz 50 mg (B), CsA + multiple-dose Fcz 25 mg (C) and CsA + single-dose Fcz 50 mg (D). Blood CsA concentrations were measured at 0.5, 1, 2, 4, 6, 8, 10, 12 and 24 hr after CsA administration. The AUC(0-12) and C(max) values for treatment B were significantly higher than those for the other treatments. In particular, the AUC(0-12) of treatment B was about two times higher than that of treatment A. Fcz administration did not significantly prolong the half-life or mean residence time of CsA. The results of our study show that administration of multiple therapeutic doses of Fcz can significantly increase the CsA blood concentration, which might partially depend upon the Fcz blood concentration. When Fcz is used in CsA-based canine renal transplantation, it may be necessary to adjust the CsA trough level by decreasing the dose.     

Histological analysis of implanted embryos in large Japanese field mouse (Apodemus speciosus) and estimation of developmental stage

The large Japanese field mouse (Apodemus speciosus) is a small rodent species endemic to Japan. The genetic characteristics of A. speciosus include different chromosome numbers within the same species. Furthermore, A. speciosus has been used in radiation and genetic research. In the present study, a pregnant A. speciosus was obtained, and histochemical analysis of the implanted embryos was performed and compared with the developmental stages of the mouse (Mus musculus). Although there were some differences, the structures of the implanted embryos, including the primitive streak and placenta of A. speciosus were similar to those of mouse. Our study will be important for the construction of a developmental atlas of A. speciosus.     

Rapid genotyping with DNA micro-arrays for high-density linkage mapping and QTL mapping in common buckwheat (Fagopyrum esculentum Moench)

For genetic studies and genomics-assisted breeding, particularly of minor crops, a genotyping system that does not require a priori genomic information is preferable. Here, we demonstrated the potential of a novel array-based genotyping system for the rapid construction of high-density linkage map and quantitative trait loci (QTL) mapping. By using the system, we successfully constructed an accurate, high-density linkage map for common buckwheat (Fagopyrum esculentum Moench); the map was composed of 756 loci and included 8,884 markers. The number of linkage groups converged to eight, which is the basic number of chromosomes in common buckwheat. The sizes of the linkage groups of the P1 and P2 maps were 773.8 and 800.4 cM, respectively. The average interval between adjacent loci was 2.13 cM. The linkage map constructed here will be useful for the analysis of other common buckwheat populations. We also performed QTL mapping for main stem length and detected four QTL. It took 37 days to process 178 samples from DNA extraction to genotyping, indicating the system enables genotyping of genome-wide markers for a few hundred buckwheat plants before the plants mature. The novel system will be useful for genomics-assisted breeding in minor crops without a priori genomic information.