Prediction of grain protein content in winter wheat through leaf color measurements using a chlorophyll meter

A field trial was conducted over a 3-year period at the Hokkaido Kitami Agricultural Experiment Station to examine whether the grain protein content (GPC) of a winter wheat cultivar (Triticum aestivum L. cv. Chihokukomugi) suitable for Japanese noodle-making could be predicted before harvest. The prediction of the GPC was accurate based on the color of the second leaf (just below the flag leaf) at the end of the emergence of the inflorescence, when nitrogen application was graded. In order to evaluate the reliability of this test, a survey of 95 wheat fields in the eastern part of Hokkaido was also carried out during a 3-year period. The prediction of the GPC for this cultivar based on the color of the second leaf was less accurate across many sites. The results of this survey, however, suggested that the leaf color could be used as an index for ranking the GPC as low or high in relation to processing requirements. When the leaf color value of the second leaf measured with a chlorophyll meter at the end of the emergence of the inflorescence was less than 40, it was predicted that the GPC would be lower than the processing requirement. This index could be applied to the cultivars grown in the eastern part of Hokkaido, except for those grown on peat soils.     

Cloning and characterization of a NADP+-malic enzyme gene from Bradyrhizobium japonicum USDA110

Malic enzymes have been considered to play a key role in energy metabolism for nitrogenase reaction in bacteroids. To elucidate the physiological role of the malic enzymes in Bradyrhizobium japonicum bacteroids, a putative malic enzyme gene Bjtme1 was cloned by polymerase chain reaction (PCR) using degenerated primers from conserved regions of the protein sequences of bacterial malic enzymes and draft sequence data of the Bradyrhizobium japonicum USDA110 genome sequence project. To confirm the characteristics of the Bjtme1 gene, the protein encoded by this gene was over-expressed using a pET32a(+) system and it exhibited a NADP⁺-malic enzyme (EC 1.1.1.40) activity, indicating that Bjtme1 was the gene of the NADP⁺-malic enzyme. This is the first report on the cloning and characterization of the NADP⁺-malic enzyme gene from B. japonicum, and the gene structure was compared with that of NADP⁺-malic enzyme genes of other rhizobia.     

Combining pre-transplanting phosphorus application and green manure incorporation: a trial for reducing fertilizer input

Pre-transplanting phosphorus application is a fertilization method that can reduce phosphorus (P) fertilization of the field and increase P use efficiency. We investigated whether P use efficiency can be increased further by combining pre-transplanting phosphorus application and the green manure practice, using cabbage as a test plant in a field experiment performed from 2006 to 2008. The pre-transplanting phosphorus application successfully reduced P fertilization of the field to half without any yield losses in 2006 and 2008. However, growth and yield could not be maintained with the pre-transplanting phosphorus application when P fertilization was halved in 2007. Differences among seasons and discrepancies in the results compared to previous studies may have arisen from the differences in the field management and the nursery soil used. No benefits of green manure in terms of the subsequent cabbage yield were detected. Consequently, there was no positive effect of the combination of pre-transplanting phosphorus application and incorporation of green manure. Studies with mycorrhizal plants as test plants may be necessary to verify the mechanism of enhanced P supply by the incorporation of green manure, and to examine any potential benefit of combining green manure and pre-transplanting phosphorus application practices.     

Near-isogenic lines of durum wheat: their development and plant characteristics

Summary The effect of specific plant characteristics on the grain and biomass yield of durum wheat can be accurately determined by using isogenic lines, which, however, were not usually available. This study reported the effects of long glume, glaucousness, glume pubescence, black glume and purple culm on the yield and its associated characteristics in near-isogenic lines of durum wheat cv. LD222, which were developed by continuous backcrossing. The long glume trait which resulted in a large photosynthetic area did not enhance yield. Increased glume size associated with the P gene tended to increase the main culm dominance, characterized such as vigorous main culm and weak tillers, and plant height, but to reduce tillering and spike number. Consequently, grain yield and harvest index declined. Under the adequate water supplying condition of the present study, the glaucous trait was beneficial for grain yield. The trait of glume pubescence did not excert any significant effect on the yield related characteristics in the LD222 background. The traits of black glume and purple culm reduced the number of spikes per unit area and the number of kernels per unit area.     

Development of EST-SSR markers of Ipomoea nil

Although Japanese morning glory (Ipomoea nil (L.) Roth.) has been used intensively for genetic studies, DNA markers have not been developed in Ipomoea nil sufficient to cover all chromosomes. Therefore, we conducted microsatellite (simple sequence repeats, SSR) marker development in I. nil for future genetic studies. From 92,662 expressed sequence tag (EST) sequences, 514 unique microsatellite-containing ESTs were identified. Primer pairs were designed automatically in 326 SSRs. Of 150 SSRs examined, 75 showed polymorphisms among strains. A phenogram based on the SSR genotypes revealed the genetic relation among seven Japanese morning glories from five different regions of the world and an ivyleaf morning glory (I. hederacea Jacq.). The developed SSR markers might be applicable for genetic studies of morning glories and their relatives.     

Evaluation of soybean germplasm conserved in NIAS genebank and development of mini core collections

Genetic variation and population structure among 1603 soybean accessions, consisted of 832 Japanese landraces, 109 old and 57 recent Japanese varieties, 341 landrace from 16 Asian countries and 264 wild soybean accessions, were characterized using 191 SNP markers. Although gene diversity of Japanese soybean germplasm was slight lower than that of exotic soybean germplasm, population differentiation and clustering analyses indicated clear genetic differentiation among Japanese cultivated soybeans, exotic cultivated soybeans and wild soybeans. Nine hundred ninety eight Japanese accessions were separated to a certain extent into groups corresponding to their agro-morphologic characteristics such as photosensitivity and seed characteristics rather than their geographical origin. Based on the assessment of the SNP markers and several agro-morphologic traits, accessions that retain gene diversity of the whole collection were selected to develop several soybean sets of different sizes using an heuristic approach; a minimum of 12 accessions can represent the observed gene diversity; a mini-core collection of 96 accession can represent a major proportion of both geographic origin and agro-morphologic trait variation. These selected sets of germplasm will provide an effective platform for enhancing soybean diversity studies and assist in finding novel traits for crop improvement.     

Evaluation of cytokine messenger RNA expression in peripheral blood mononuclear cells from dogs with canine demodicosis

Using RT-PCR and semi-quantitative PCR, mRNA expression for canine interferon (IFN)-gamma, interleukin (IL)-2, IL-4, IL-5, IL-10, tumor necrosis factor (TNF)-alpha and transforming growth factor (TGF)-beta in peripheral blood mononuclear cells (PBMCs) was examined in dogs with or without demodicosis. mRNA expression for IFN-gamma as well as TNF-alpha in dogs with demodicosis (localized (LD) and generalized (GD)) was slightly lower than those in dogs without demodicosis (healthy controls). Expression of IL-5 mRNA in dogs with demodicosis was higher than that in control dogs, but there were no significant differences in IL-4 and IL-10 mRNA expression levels among the three groups. On the other hand, expression levels of TGF-beta mRNA in dogs with GD were higher than those in control dogs and dogs with LD. The expression levels of IL-5 and TGF-beta mRNA decreased in all three dogs with GD which showed resolution of the clinical signs. Taken together, these results suggest that the Th2-like response in PBMCs from dogs with demodicosis is up-regulated, and that subsequent increased expression of IL-5 and TGF-beta mRNA in dogs with GD is reversible after treatment. Therefore, these cytokines, particularly IL-5, might be a useful clinical index of the clinical course in demodicosis. Also, increased TGF-beta mRNA expression might be a key factor for revealing the difference in the mechanism of onset between LD and GD.     

Effect of improvement cutting on the community structure of aphyllophoraceous fungi on Okinawa Island

Improvement cutting, which decreases tree density by cutting and thinning while leaving logging residues, has been practiced on Okinawa Island since the 1970s and changes not only the vegetation composition but also the amount and composition of woody debris on the forest floor. To quantitatively compare the effect of improvement cutting with the effects of woody debris and vegetation on the community structure of aphyllophoraceous fungi, we conducted a field survey of vegetation composition, the amount and composition of woody debris, and the aphyllophoraceous fungi flora on Okinawa Island in December 2006. We established 3 plots in forest stands without logging and 13 plots in a forest that had undergone improvement cutting from 1 to 22 years before our field survey. The number and volume of woody debris tended to be highest soon after improvement cutting. We recorded 1,172 fungal samples belonging to 81 species in four orders. Fungal species richness was correlated with the composition of the woody debris. Fungal species composition in forest plots 1 year after improvement cutting differed from most of the other logged forests. Variation partitioning revealed that the effects of improvement cutting explained 14.1 % of the total variation in fungal species composition, and that differences in woody debris and vegetation not related to improvement cutting explained 40.4 %. Changes in the composition of woody debris caused by improvement cutting explained at most 2.7 % of the variation. This suggests that improvement cutting does not have a strong effect on fungal community structure, but woody debris does.     

Salt tolerance of grain crops in relation to ionic balance ability to absorb microelements

A greenhouse experiment was conducted to investigate the effect of exchangeable Na on the growth and absorption of metal elements in barley, rye, and maize. The plants were cultivated in soils whose exchangeable sodium percentages (ESP) were 6.6 (saline soil: Saline), 17.4 (saline-sodic soil: Sodic 1), and 39.6 (sodic soil: Sodic 2), which were prepared from Tottori sand dune soil (Control). The dry weight (DW) and concentrations of metal elements Ca, Mg, Mn, Zn, and Cu) in shoots were analyzed. The shoot DW was smaller with higher ESP, but in barley the difference between all the treatments was no longer observed with time. In Sodic soils, the growth of barley was vigorous, whereas rye growth was poor, and maize plants died by 5 weeks after planting. The Na concentration in shoots of all the species was higher with higher ESP. The K concentration in shoots was low at the early growth stage, but in barley it was higher in the Saline and both Sodic soils than in the Control at the subsequent stages. The concentrations of Ca and Mg in shoots of barley and maize in the Saline and both Sodic soils were higher than those in the Control, but in rye the concentrations were lowest in Sodic 2. The concentrations of Mn, Zn, and Cu in barley shoots in the Saline and bothSodic soils tended to be higher than those in the Control, whereas in rye they were lower than in the Control in both Sodic soils. Barley showed a higher ability to absorb low available microelements than rye and maize. These results indicate that barley is tolerant to sodicity as well as salinity, maize is tolerant to salinity, but is very sensitive to sodicity, and rye is moderately sensitive to both stresses. We suggest that the tolerance of grain crops to ESP involves a tolerance to a high Na concentration in shoots, the ability to keep suitable concentrations of essential cations in the presence of a high concentration of Na in shoots and the ability to absorb low available microelements.     

Molecular identification of `Candidatus Mycoplasma haemovis' in sheep with hemolytic anemia

We examined the presence of hemoplasmas, hemotropic mycoplasmas, among 11 sheep (Ovis aries) with regenerative and hemolytic anemia and found six of them were positive by real-time PCR. The positive samples were then subjected to conventional PCR for direct sequencing of the 16S rRNA gene. Nucleotide sequences of all the positive samples were identified as the 16S rRNA gene of `Candidatus Mycoplasma haemovis' by phylogenetic analysis, demonstrating the infections with this particular hemoplasma species in Japan.