Development of genomic and EST-SSR markers in radish (Raphanus sativus L.)

Radish (Raphanus sativus L.) belongs to Brassicaceae family and is a close relative of Brassica. This species shows a wide morphological diversity, and is an important vegetable especially in Asia. However, molecular research of radish is behind compared to that of Brassica. For example, reports on SSR (simple sequence repeat) markers are limited. Here, we designed 417 radish SSR markers from SSR-enriched genomic libraries and the cDNA data. Of the 256 SSR markers succeeded in PCR, 130 showed clear polymorphisms between two radish lines; a rat-tail radish and a Japanese cultivar, ‘Harufuku’. As a test case for evaluation of the present SSRs, we conducted two studies. First, we selected 16 SSRs to calculate polymorphism information contents (PICs) using 16 radish cultivars and four other Brassicaceae species. These markers detected 3–15 alleles (average = 9.6). PIC values ranged from 0.54 to 0.92 (average = 0.78). Second, part of the present SSRs were tested for mapping using our previously-examined mapping population. The map spanned 672.7 cM with nine linkage groups (LGs). The 21 radish SSR markers were distributed throughout the LGs. The SSR markers developed here would be informative and useful for genetic analysis in radish and its related species.     

Effect of soil exchangeable potassium content on cesium absorption and partitioning in buckwheat grown in a radioactive cesium-contaminated field

The effect of soil exchangeable (plant-available) potassium (ExK) content on cesium (Cs) absorption and translocation in buckwheat was evaluated in a field contaminated with radioactive Cs (¹³⁴Cs and ¹³⁷Cs, RCs) in 2013. The RCs concentration in buckwheat was significantly positively correlated with the naturally occurring stable Cs (¹³³Cs, SCs) concentration, and was lower at higher soil ExK content. The RCs and SCs were actively absorbed by buckwheat until the flowering stage. The soil ExK content was significantly negatively correlated with soil exchangeable RCs and SCs (ExRCs and ExSCs) concentrations. Greater RCs and SCs absorption by buckwheat in soils with low ExK contents was mainly due to higher soil ExRCs and ExSCs concentrations. Reproductive organs showed the largest differences in SCs concentration between low-ExK and high-ExK plots. The root–shoot and shoot–reproductive organs translocations of SCs markedly decreased with increasing soil ExK content. In the root–shoot and shoot–reproductive organs translocations, the discrimination of SCs and K decreased with decreasing soil ExK content. Our main findings were as follows: (1) because RCs are mainly taken up at the earlier growth stage, potassium should be applied as a basal fertilizer to decrease the RCs concentration in buckwheat; (2) lower soil ExK content led to higher soil ExRCs concentrations, resulting in greater RCs absorption by buckwheat; (3) the high Cs absorption and translocation and weaker discrimination between Cs and K in low ExK content soil may be due to the expression of K transporter(s) with weak discrimination between Cs and K.     

Effect of heat on cambial reactivation during winter dormancy in evergreen and deciduous conifers

Responses of cambium to warming were recorded three times (December 14-27, 1990, January 18-February 3 and February 27-March 13, 1991) on 14-year-old Cryptomeria japonica D. Don and four times (December 12-26, 1990, January 18-February 2, February 26-March 12 and March 28-April 13, 1991) on 27-year-old Larix leptolepis Gord., during a period of winter cambial dormancy. Stem surfaces at breast height, mid-tree height and the crown base were warmed to 25-30 degrees C for 2 weeks. After heat treatment, cambia in the treated regions and in untreated regions 1 m above each treated area were examined by optical and transmission electron microscopy (TEM). In C. japonica, heat treatment often resulted in cambial reactivation in the treated regions, and this response to heat gradually increased as the dormant season passed from winter to spring. Conversely, in L. leptolepis, no cell division was observed in the cambial region of warmed stems until natural resumption of cambial activity, which occurred after bud break.     

Flavonols from saffron flower: tyrosinase inhibitory activity and inhibition mechanism.

A common flavonol, kaempferol, isolated from the fresh flower petals of Crocus sativus L. (Iridaceae) was found to inhibit the oxidation of L-3,4-dihydroxyphenylalanine (L-DOPA) catalyzed by mushroom tyrosinase with an ID(50) of 67 microgram/mL (0.23 mM). Interestingly, its 3-O-glycoside derivatives did not inhibit this oxidation. The inhibition kinetics analyzed by a Lineweaver-Burk plot found kaempferol to be a competitive inhibitor, and this inhibitory activity presumably comes from its ability to chelate copper in the enzyme. This copper chelation mechanism can be applicable for all of the flavonols as long as their 3-hydroxyl group is free. However, quercetin, kaempferol, and galangin each affect the oxidation of L-tyrosine in somewhat different ways.     

Retrospective serological analysis of spontaneous CDV infection in 192 dogs

Spontaneous cases of canine distemper virus (CDV) infection were serologically evaluated. The 192 dogs in which CDV antigen was confirmed from tonsil by immunohistological examination were 2- to 4-months old, of various breeds, and unvaccinated. The prognoses were good in 74 dogs with significantly high levels of anti-CDV passive hemolytic aggregation (PHA) titer. In the other 118 dogs with poor prognoses, anti-CDV PHA titer was not detected. Anti-CDV PHA titer had the most significant association with the prognoses of CDV infection, and could be the most reliable and useful indicator for evaluating such prognoses.     

Granulomatous nephritis and meningoencephalomyelitis caused by Halicephalobus gingivalis in a pony gelding

A ten-year-old Shetland pony gelding showed low appetite, ataxia, peculiar swaying, clouding of consciousness, and ultimately died. At necropsy, multiple coalescing granulomatous foci were detected in the kidneys, and small necrotic lesions were found in the cerebellum. Histologic examination of the renal tissue sections revealed extensive granuloma, and Halicephalobus gingivalis-like nematodes were seen. Similar nematodes were found in the granulomatous or necrotic lesions of the renal lymph nodes and cerebellum, and were also frequently detected in cerebrospinal meningovascular lesions. Morphologic features together with partial ribosomal RNA gene sequences of the nematodes in the lesions revealed that they were H. gingivalis. The present results indicated that H. gingivalis caused granulomatous nephritis and meningoencephalomyelitis in this pony gelding.     

Flow cytometric analysis of thymocyte subpopulations in mice after whole-body X-irradiation.

To determined the cellular kinetics of thymocyte subpopulations in DBA1 mice after whole-body 6.8 Gy X-irradiation, they were analyzed for the expression of several cell surface antigens using flow cytometry. The results show that i) The majority of thymocytes rapidly depleted by irradiation was CD4+8+ cells. ii) radioresistant CD4+8- and CD4-8+ survived 18-48 hr after X-irradiation were considered to be relatively mature type, since they expressed high levels of CD3 and LECAM-1. iii) CD3-positive cells were detected in CD4-8- cells at 72 hr after irradiation.     

Role of sorbitol in manufacturing dried seafood from heated squid meat

SUMMARY: Japanese common squid meat was heat-treated at 80°C for 1 min, cured with 1.0 M sorbitol solution (pH 7.0) at 4°C for 18 h, and dried further at 30°C (60% relative humidity) for 16 h. Osmotic dehydration during the sorbitol curing process and slow moisture vaporization at the initial drying period were observed regardless of the heat denaturation of muscle protein. Simultaneously, lowering the amount of moisture vaporized in the falling rate of the drying period caused a shortening of the total drying time. Furthermore, the shear force of the dried product from heat-treated meat was kept at a lower value by sorbitol curing, although the suppression effect of sorbitol on the hardening of dried meat was lost by protein denaturation. These results are useful for understanding the role of sorbitol in reducing drying time and in eliminating excess hardening of dried squid products.