Photoassimilate partitioning in hypernodulation mutant of soybean (Glycine max (L.) Merr.) NOD1-3 and its parent williams in relation to nitrate inhibition of nodule growth

Nodule growth of a hypernodulating soybean (Glycine max (L.) Merr.) mutant line NOD1-3 was compared to that of its wild-type parent cv. Williams from 14 to 18 days after planting (DAP) in the absence of nitrate treatment (hereafter referred to as “0 mM treatment”) or with 5 mM nitrate treatment. The growth rate determined by increase in the diameter of the nodules was relatively lower in the mutant NOD1-3 than that of the parent Williams under nitrogen-free conditions (0 mM nitrate). The inhibition of nodule growth by 5 mM nitrate started at 1 d after the onset of the nitrate treatment in Williams, while the inhibition did not occur before the application of the nitrate treatment for 2 d in NOD1-3. The nodule growth was completely inhibited after 2 d in Williams and after 3 d in NOD1-3 during the 5 mM nitrate treatment period. After 4 d of 5 mM nitrate treatment, the nodule dry weight decreased by 22% in NOD1-3 and by 58% in Williams, respectively. The treatment with 5 mM nitrate decreased the acetylene reduction activity (ARA) in NOD1-3 by 60% per plant and by 50% per nodule g DW and these parameters were less sensitive to the treatment than those in Williams in which the inhibition rate was 90% per plant and 80% per nodule g DW. These results indicate that NOD1-3 is partially nitrate-tolerant in terms of individual nodule growth as well as total nodule dry weight and Nz fixation activity. A whole shoot of Williams and NOD1-3 plants was exposed to ¹⁴CO₂ for 120 min followed by 0 or 5 mM nitrate treatment for 2 d, and the partitioning of the photoassimilates among the organs was analyzed. Under 0 mM nitrate treatment, the percentages of the distribution of ¹⁴C radioactivity between the nodules and roots were 63 and 37% in Williams and 89 and 11% in NOD1-3. Under the 5 mM nitrate conditions, the percentages of the distribution of ¹⁴C between the nodules and roots changed to 14 and 86% in Williams and 39 and 61% in NOD1-3, respectively. These results indicated that the hypernodulating mutant NOD1-3 supplied a larger amount of photoassimilates to the nodules than to the roots under nitrogen-free conditions, and that the nitrate depression of photoassimilate transport to the nodules was less sensitive than that of the parent line.     

Rapid and reversible nitrate inhibition of nodule growth and N2 fixation activity in soybean (Glycine max (L.) Merr.)

Nodulated soybean (Glycine max. (L) Merr. cv. Williams) plants were hydroponically cultured, and various combinations of 1-week culture with 5 or 0 mm nitrate were applied using 13-d-old soybean seedlings during three successive weeks. The treatments were designated as 0-0-0, 5-5-5, 5-5-0, 5-0-0, 5-0-5, 0-5-5, and 0-0-5, where the three sequential numbers denote the nitrate concentration (mm) applied in the first-second-third weeks. The size of the individual nodule was measured periodically using a slide caliper. All the plants were harvested after measurement of the acetylene reduction activity (ARA) at the end of the treatments. In the 0-0-0 treatment, the nodules grew continuously during the treatment period. Individual nodule growth was immediately suppressed after 5 mm nitrate supply. However, the nodule growth rapidly recovered by changing the 5 mm nitrate solution to a 0 mm nitrate solution in the 5-0-0 and 5-5-0 treatments. In the 5-0-5 treatment, nodule growth was completely inhibited in the first and the third weeks with 5 mm nitrate, but the nodule growth was enhanced in the second week with 0 mm nitrate. The nodule growth response to 5 mm nitrate was similar between small and large size nodules. After the 5-5-5, 5-0-5, 0-0-5, and 0-5-5 treatments, where the plants were cultured with 5 mm nitrate in the last third week, the ARA per plant was significantly lower compared with the 0-0-0 treatment. On the other hand, the ARA after the 5-0-0 and 5-5-0 treatments was relatively higher than that after the 0-0-0 treatment, possibly due to the higher photosynthate supply associated with the vigorous vegetative growth of the plants supplemented with nitrate nitrogen. It is concluded that both soybean nodule growth and N₂ fixation activity sensitively responded to the external nitrate level, and that these parameters were reversibly regulated by the current status of nitrate in the culture solution, possibly through sensing of the nitrate concentration in roots and / or nodules.     

The genetic organization of the capsular polysaccharide biosynthesis region of Actinobacillus pleuropneumoniae serotype 15

Nucleotide sequence determination and analysis of the cps gene involved in the capsular polysaccharide biosynthesis of Actinobacillus pleuropneumoniae serotype 15 revealed the presence of three open reading frames, designated as cps15ABC genes. At the protein level, Cps15A and Cps15B showed considerably high homology to CpsA (67.0 to 68.7%) and CpsB (31.7 to 36.8%), respectively, of A. pleuropneumoniae serotypes 1, 4 and 12, revealing the common genetic organization of the cps among serotypes 1, 4, 12 and 15. However, Cps15C showed no homology to any proteins of A. pleuropneumoniae serotypes, indicating that cps15C may be specific to serotype 15. This study will provide the basic molecular knowledge necessary for the development of diagnostics and a vaccine for A. pleuropneumoniae serotype 15.     

Prevalence of equine proliferative enteropathy in Hidaka district,Hokkaido, over five seasons

Equine proliferative enteropathy (EPE) is an equine infectious disease that can lead to severe weight loss and hyperplasia of the intestinal mucosa due to infection with Lawsonia intracellularis. In this study, we investigated the prevalence of EPE in a major Thoroughbred breeding area: Hidaka district, Hokkaido, Japan. Of the 252 symptomatic horses that we tested, 192 EPE cases (76.2%), including 8 fatal cases, were confirmed from April 2015 to March 2020 by etiological and/or serological investigation. Most of the EPE cases were observed in foals (88.5%), with fewer cases in yearlings (7.3%) and adults (4.2%). Asymptomatic infection was observed in 62.9% of the horses kept with affected horses. These results suggest that EPE is an enzootic disease in Hidaka district.     

Effect of deep placement of calcium cyanamide,coated urea,and urea on soybean (Glycine max (L.) Merr.) seed yield in relation to different inoculation methods

Abstract

The main objective of this study was to increase the productivity of soybean [Glycine max (L.) Merr. cv. Enrei] seed by deep placement of controlled release nitrogen fertilizers and by the application of different methods of inoculation of bradyrhizobia. Ten days old seedlings in an inoculated paper pot (IPP), in a non-inoculated paper pot (NIPP), and those grown in a vermiculite bed without paper pot (DT) were transplanted to an upland field converted from a drained paddy field in Nagaoka. In addition to the basal application of 16 kg N ha^?1 in the surface layer (Control), deep placement of 100 kg N ha^?1 of urea (Urea), 100-day type coated urea (CU-100), and calcium cyanamide (CaCN₂) treatments were applied at the depth of 20 cm. In the IPP method, a significantly higher seed yield was obtained with the deep placement of CaCN₂ and CU-100 compared with the Urea and Control treatments. A similar tendency was observed for the DT and NIPP methods. Among the same N fertilizer treatments, the seed yield for IPP and DT tended to exceed that for NIPP, although the NIPP roots also showed nodulation probably due to infection with indigenous bradyrhizobia. The percentage of nitrogen derived from atmospheric N₂ estimated by the simple relative ureide method was higher in the plants with CU-100 and CaCN₂ compared with those with the Urea and Control treatments at the RI stage, suggesting that the basal deep placement of CaCN₂ or CU-100 for soybean cultivation enabled the supply of N without concomitant depression of N₂ fixation. Thus the deep placement of cheaper CaCN₂ was found to be as effective as that of CU-100 for enhancing the soybean seed yield.     

Effects of inhibitors for signaling components on the expression of the genes for nitrate reductase and nitrite reductase in excised barley leaves

Nitrate-induced accumulation of mRNAs for nitrate reductase and nitrite reductase was strongly inhibited by treatment with inhibitors of Ca²⁺ channels, protein phosphatases, and tyrosine protein kinases in excised barley leaves. It was shown that these components may be involved in the nitrate-response signaling pathways for the expression of nitrate and nitrite reductase genes in barley leaves.     

Aspen regeneration in the Colorado Front Range: differences at local and landscape scales

Elk (Cervus elaphus) populations in Rocky Mountain National Park are higher than at any time in the past century, and heavy browsing by elk may interfere with aspen (Populus tremuloides Michx.) regneration. We used aerial photographs to identify all aspen stands within Rocky Mountain National Park, and all aspen stands within the elk winter range range (defined as 2400 to 2800 m elevation) in three portions of the adjacent Roosevelt National Forest. From this population of aspen stands, we randomly selected 57 stands for evaluation of aspen regeneration. Stands that contained stems younger than 30 years and taller than 2.5 m tall were classified as regenerating successfully. Only 20% of the aspen stands in Estes Valley contained a cohort of regenerating aspen stems, whereas 45-to-75% of aspen stands across the larger landscape of the Front Range had regenerating cohorts of aspen. Within the elk winter range of the Roosevelt National Forest, 13 of 17 aspen stands were regenerating. In the elk winter range on the east side of the Park but outside of Estes Valley, 11 of 15 aspen stands were regenerating successfully. Only a few aspen stands exist in the elk winter range on the western side of the Park, and none of the five aspen stands sampled in Kawuneeche Valley had a regenerating cohort. The lack of regeneration in Kawuneeche Valley may result from locally heavy elk use in both winter and summer. In the summer elk range at higher elevations in the Park (2800 to 3200 m), 16 of 23 stands had regenerated. At landscape scales, all locations outside of the heavily impacted Estes Valley averaged about two cohorts/stand that regenerated after the mid-1960s. All stands that lacked a regenerating cohort showed evidence of moderate-to-severe damage from elk browsing of stems. No regenerating stands showed evidence of severe browsing. We conclude that at landscape scales, regeneration within aspen stands is very common across the Front Range, except in local areas of the highest elk use where little regeneration has occurred in the past 30 years.