Nuclear genome constitution and other characteristics of somatic hybrids between dihaploid Solanum acaule and tetraploid S. tuberosum

Eleven somatic hybrids (2n = 68 to 74) obtained between S. tuberosum ssp. tuberosum cv. Dejima (2n = 48) and ATDH-1 (2n = 24), an anther-culture-derived dihaploid of S. acaule (Yamada et al., 1997), were characterized by nuclear RFLP markers using 49 single-copy DNA probes distributed throughout the potato genome (2 to 6 probes per chromosome). One of the somatic hybrids, DA8-2, had 72 chromosomes and all the Dejima- and ATDH-1-specific markers (124 and 103 bands, respectively), suggesting the presence of a whole set of both parental chromosomes. The other somatic hybrids lost varying numbers of markers up to seventeen. The pattern of the loss of markers indicated the elimination of five chromosomes among four somatic hybrids. A nucleolar organizer region of chromosome 2 was often eliminated in the somatic hybrids. The somatic hybrids studied here had higher frequencies of multivalent formation than the S. tuberosum parent. They had reasonably good seed set when pollinated with S. tuberosum pollen. Hence, homoeologous recombination between S. acaule and S. tuberosum chromosomes is possible and useful traits from S. acaule may be transferred to the S. tuberosum gene pool. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic resources of salt tolerance in wild Hordeum species

Salt tolerance was evaluated in 340 accessions of Hordeum, consisting of 41 brittle-rachis forms of Hordeum vulgare L. subsp. vulgare (H. agriocrithon) accessions, 154 H. vulgare L. subsp. spontaneum (H. spontaneum) accessions, and 145 accessions of ten other species or subspecies of wild Hordeum. Germination was carried out at concentrations of 171, 257, and 342 mM NaCl. The levels of salt tolerance for seed germination in wild Hordeum species were generally lower than those found by Mano et al. (1996) in cultivated barley; the NaCl tolerance level of the different species were as follows: H. agriocrithon > H. spontaneum > other wild Hordeum species. In addition, leaf injury index was used to assess tolerance at the seedling stage after treatment with 500 mM NaCl solution for four weeks. The levels of salt tolerance at the seedling stage in wild Hordeum species were generally higher than those found by Mano & Takeda (1995) in cultivated barley. Most wild Hordeum species showed high NaCl tolerance at the seedling stage and are considered good sources of germplasm for salt tolerance breeding. This revised version was published online in August 2006 with corrections to the Cover Date.     

The influence of Pueraria mirifica herb containing phytoestrogens on the urinary gonadotropin and estradiol levels in aged menopausal monkeys

We investigated a non‐invasive method of specimen collection for determining the changes of reproductive hormones in aged menopausal monkeys after a long‐term feeding of the Thai herb Pueraria mirifica (PM) containing phytoestrogens. Three groups of aged menopausal monkeys (n = three in each group) were fed daily with 10, 100, or 1000 mg of PM for a 90 day treatment period, and fed with distilled water for 30 and 60 days of the pre‐ and post‐treatment periods, respectively. Urine samples were collected for 14 h daily every 5 days and assayed for follicle stimulating hormone (FSH), luteinizing hormone (LH) and estradiol levels. The result showed that monkeys fed with PM 10, PM 100, and PM 1000 had a decrease in urinary FSH levels during the treatment period, followed by a rebound increase during the post‐treatment period. Urinary estradiol levels tended to decrease and fluctuated between 4.28 and 266.71, 2.85–42.27, and 6.24–203.50% of the pre‐treatment levels in those three groups, respectively. Decreases in urinary LH levels could not be observed in all the three groups. These results suggest that FSH could be a candidate marker to detect the estrogenic effects of phytoestrogens in aged menopausal monkeys when changes of urinary hormones need to be used as an indicator.     

Evaluation of several wild tuber-bearingSolanum species for scab resistance

One hundred accessions of 18 wild diploid potato species were screened for resistance to scab through three cycles of selection, from which a total of 322 clones were selected as resistant. The putative ancestral species of the cultivated potatoes (Solanum bukasovii, S. canasense andS. multidissectum) produced resistant clones with significantly higher rates. Thus, these are primary gene sources for conferring scab resistance to cultivated potatoes.     

Databases for technical aspects of immunohistochemistry: 2021 update

With the aim of sharing information about the technical aspects of immunohistochemistry (IHC) and facilitating the selection of suitable antibodies for histopathological examination, this technical report describes the results of a questionnaire distributed during the period of 2018 to 2019 among members of the Conference on Experimental Animal Histopathology. Additionally, it describes the immunological properties and supplier details (clone, supplier, catalog number, species reactivity, etc.) as well as the IHC staining conditions (fixing solution, fixing time, embedding, antigen retrieval method, antibody dilution, incubation time, incubation temperature, positive control tissue, blocking condition, secondary antibody information, etc.) for a total of 509 primary antibodies (comprising 220 different types). These survey results were an update on the contents reported by CEAH in 2017.     

Chloroplast DNA Variation in the Most Primitive Cultivated Diploid Potato Species Solanum stenotomum Juz. et Buk. and its Putative Wild Ancestral Species using High-Resolution Markers

Solanum stenotomum Juz. et Buk. (2n = 2x = 24) is considered to be the most primitive diploid cultivated species from which all the other Andean cultivated potatoes were originated (Hawkes 1990). To disclose chloroplast DNA (ctDNA) variability and the maternal origin of S. stenotomum, 36 accessions of S. stenotomum and 86 accessions of putative wild ancestral species were determined for ctDNA types and analyzed by high-resolution markers (seven ctDNA microsatellites and an H3 marker). High-resolution markers discriminated 57 different ctDNAs (haplotypes), which were classified into the W-type ctDNA group and C-, S- and A-type ctDNA group, and within the latter group S- and A-type ctDNAs were distinct from each other among many different haplotypes mostly having C-type ctDNA. This ctDNA relationship supported our previous findings obtained for mostly Andean cultivated species (Sukhotu et al. 2004). Compared with other putative ancestral wild species, S. stenotomum showed somewhat limited ctDNA diversity, having two major haplotypes 1 and 2 also found in different wild species in different places. Therefore, the ctDNA in S. stenotomum was of at least dual origins either by successive domestication from different species or else by introgression after initial S. stenotomum arose.     

Net blotch resistance in wild species of Hordeum

Resistance to net blotch was evaluated in 175 Hordeum vulgare subsp. spontaneum (H. spontaneum) accessions and 149 accessions of thirteen species or subspecies of wild Hordeum. Most H. spontaneum accessions showed resistance to each of the four Pyrenophora teres f. teres (P. teres) isolates tested. However, H. spontaneum accessions showed different resistance reactions, depending upon their origin. In particular, some accessions from Afghanistan and Russia showed a high level of resistance, and accessions from Morocco were susceptible. Among the four P. teres isolates, the virulence spectra on the H. spontaneum accessions were more different between isolates from different countries than between those from the same country. Hordeum spontaneum accessions susceptible to the Canadian isolate WRS102, but resistant to the other three isolates were found in Iraq suggesting the geographical differentiation of resistance genes in H. spontaneum. All accessions of the other wild Hordeum species, especially some accessions of H. marinum subsp. gussoneanum, showed high levels of resistance. These resistance genes may be useful candidates for incorporation into cultivated barley. This revised version was published online in July 2006 with corrections to the Cover Date.     

Artificial incubation of Japanese crayfish (Cambaroides japonicus) eggs by using a simple, easy method with a microplate

We developed a simple, easy method with a microplate to artificially incubate Japanese crayfish (Cambaroides japonicus) eggs for their cultivation. We prepared 6-, 12-, 24-, 48-, and 96-well microplates containing sterile water heated to 5, 10, 15, and 20 °C. Fourteen experimental groups for each water temperature were prepared for each of different water volumes (0.125–10 ml) in each well. One embryonic egg was placed in each well. Experiments were also conducted with water collected from the lake where the eggs were harvested from and held at 15 °C. In the microplates with sterile water, high proportions of eggs hatched (60–100%) at 15 °C in all volumes of water, although the proportions of hatching were low (0–20%) at 5, 10, and 20 °C. All eggs died in the experiments that used lake water. We conclude that the 96-well would be the most effective size to hatch crayfish eggs in, because of its convenience. This method using a microplate is simpler and easier compared with methods of previous studies to artificially incubate crayfish eggs, and therefore it might be useful to incubate eggs of other freshwater crayfish species.