Pathogenic and genetic diversity of Xanthomonas translucens pv. undulosa in North Dakota

Bacterial leaf streak (BLS), caused by Xanthomonas translucens pv. undulosa, has become more prevalent recently in North Dakota and neighboring states. From five locations in North Dakota, 226 strains of X. translucens pv. undulosa were collected and evaluated for pathogenicity and then selected strains were inoculated on a set of 12 wheat cultivars and other cereal hosts. The genetic diversity of all strains was determined using repetitive sequence-based polymerase chain reaction (rep-PCR) and insertion sequence-based (IS)-PCR. Bacterial strains were pathogenic on wheat and barley but symptom severity was greatest on wheat. Strains varied greatly in aggressiveness, and wheat cultivars also showed differential responses to several strains. The 16S ribosomal DNA sequences of the strains were identical, and distinct from those of the other Xanthomonas pathovars. Combined rep-PCR and IS-PCR data produced 213 haplotypes. Similar haplotypes were detected in more than one location. Although diversity was greatest (≈92%) among individuals within a location, statistically significant (P ≤ 0.001 or 0.05) genetic differentiation among locations was estimated, indicating geographic differentiation between pathogen populations. The results of this study provide information on the pathogen diversity in North Dakota, which will be useful to better identify and characterize resistant germplasm.     

Effect of Cadmium and Iron on Rice (Oryza Sativa L.) Plant in Chelator-Buffered Nutrient Solution

ABSTRACT

To better understand the mechanisms responsible for differences in uptake and distribution of cadmium (Cd), nutrient-solution experiments were conducted with different varieties of rice (Oryza sativa), ‘Khitish’ and ‘CNRH3’. The plants were grown in a complete nutrient solution with different levels of pCd (-log free Cd⁺² activity) and pFe [-log free iron (Fe⁺²) activity]. The required concentrations of chelating agent and metals were determined using a computerized chemical equilibrium model such as Geochem-PC. Experimental treatments included a combination of four pCd activity levels (0, 7.9, 8.2, and 8.5) applied as Cd (NO₃)₂ 4H₂O, and two pFe activity levels (17.0 and 17.8) applied as FeCl₃. The application of both Cd and Fe in solution culture significantly affected plant growth, yield, and Cd accumulation in plant tissue. In general, yield of rice was decreased by an increase in amount of solution Cd; however, yield response varied among the cultivars. At the 7.9 pCd level, yields of rice cultivars ‘Khitish’ and ‘CNRH3’ were reduced to 69% and 65%, respectively, compared with control plants. Root Cd concentrations ranged from 2.6 mg kg^?1 (control plants) to 505.7 mg kg^?1 and were directly related to solution Cd concentrations. In rice plants, Cd toxicity symptoms resembled Fe chlorosis. Differential tolerance of varieties to phytotoxicity was not readily visible, but a significant interaction of substrate Cd and variety was obtained from dry-matter yields. Significant interactions indicated that response of tissue Cd concentration, plant Cd uptake, and translocation of Cd to the aerial parts were dependent on variety as well as substrate Cd. Uptake of Cd by roots was significantly higher than by shoots. Higher Cd uptake by rice plants decreased the uptake of other beneficial metals.

The effect of Cd and Fe on the rate of phytometallophore release was also studied in the nutrient solution. Among the rice genotypes, ‘Khitish’ was the most sensitive to Cd toxicity. In both genotypes, with the onset of visual Cd-toxicity symptoms, the release of phytometallophore (PM) was enhanced. Among the rice varieties, ‘Khitish’ had the highest rate of PM release. Treatments with the metal ions studied produced a decrease in chlorophyll and enzyme activity. A decrease in concentrations of chlorophyll pigments in the third leaf was observed due to the highest activity level of Cd (pCd 7.9). Activities of enzymes such as peroxidase (POD) and superoxide dismutase (SOD) are altered by toxic amounts of Cd. Changes in enzyme activities occurred at the lowest activity of Cd (pCd 8.5) in solution. Peroxidase activity increased in the third leaf. Results showed that in contrast with growth parameters, the measurements of enzyme activities may be included as early biomarkers in a plant bioassay to assess the phytotoxicity of Cd-contaminated solution on rice plants. Evidence that Cd uptake and translocation are genetically controlled warrants the selection of varieties that assimilate the least Cd and that translocate the least metal to the plant part to be used for human and animal consumption.     

Carbon, nitrogen and phosphorus budget in shrimp (Penaeus monodon) culture ponds in eastern India

Nutrient budget for shrimp (Penaeus monodon) culture was performed in ten culture ponds (0.4–1.0 ha) in Orissa, India, at stocking density of 10.0–22.10/m². The average initial body weight of shrimps was 0.02 g. The culture period was for 4 months. Feed was the main input of nitrogen (N), phosphorus (P) and organic carbon (OC). The FCR varied from 1.35 to 1.75. At harvest time, the average weight of shrimps varied from 28 to 32 g. Total N, P and OC applied to these ponds through all the inputs ranged from 106.64 to 292.63, 23.17 to 57.55 kg and 763.10 to 1,831.20 kg per crop, respectively. Feed alone accounted for 94.43–95.03 % total N, 91.92–95.36 % total P and 80.33–92.48 % total OC, respectively. The harvest of shrimp accounted for recovery of 25.45–36.88 (av. 30.12) %, 10.07–12.94 (av. 11.16) % and 15.80–20.81 (av. 18.01) % of added N, P and OC, respectively. N, P and OC accumulated in sediment were 49.11, 64.07 and 64.82 %, respectively, of total nutrient retention in the culture system. Unaccounted N, P and OC were 13.29, 21.83 and 18.14 %, respectively, of the total nutrient retention in the system. N, P and OC outputs in the discharged water during harvest were 7.48, 2.94 and 2.03 %, respectively.     

Growth and enzymatic activity of maize (Zea mays L.) plant: Solution culture test for copper dioxide nano particles

Copper (Cu) is an essential micronutrient for plants, which acts either as the metal component of enzymes or as a functional structural or a regulatory co-factor of a large number of enzymes. To understand the possible benefits of applying nanotechnology to agriculture, the first step should be to analyze penetration and transport of nano-particles in plants. The present study was conducted to test the hypothesis that copper nanoparticle would enter into the plant cell and govern the growth of maize plant.A solution culture experiment was conducted to investigate the effect of Cu nano-particles (<50nm) on the growth and enzymatic activity of maize (Zea mays L.) plant. Bioaccumulation of Cu nano-particles in plant was also investigated. Results showed that Cu nano-particles can enter into the plant cell through roots and leaves. Bioaccumulation increased with increasing concentration of Cu nano-particles (NPs), and agglomeration of particles was observed in the cells using transmission-electron microscopy. Application of Cu nano-particles through solution culture as well as spray enhanced the growth (51%) of maize plant in comparison to control. The different enzymatic activities like glucose-6-phosphate dehydrogenase, succinate dehydrogenase, superoxide dismutase, catalase, andguaiacol peroxidase were studied to find a possible pathway through which NPs may affect the enzymatic activity of plant. Amongst the enzymes, the activity of glucose-6-phosphate dehydrogenase was highly influenced by copper oxide (CuO)nano-particles application by spray as well as in solution. Experimental results revealed that CuOnano-particles affected the pentose phosphate pathway of maize plant. The obtained experimental results provided conclusive evidence to indicate that the nano-particles considered under this study could enter into the plant cell, easily be assimilated by plants and also enhanced its growth by regulating the different enzyme activities.     

Common occurrence of a unique Cryptosporidium ryanae variant in zebu cattle and water buffaloes in the buffer zone of the Chitwan National Park, Nepal

There are very few studies on the diversity and public health significance of Cryptosporidium species in zebu cattle and water buffaloes in developing countries. In this study, PCR-restriction fragment length polymorphism and DNA sequence analyses of the small-subunit (SSU) rRNA gene were used to genotype Cryptosporidium specimens from 12 zebu cattle calves, 16 water buffalo calves, and four swamp deer (Cervus duvaucelii) collected from the buffer zone of the Chitwan National Park, Nepal. All Cryptosporidium specimens from cattle and buffaloes belonged to Cryptosporidium ryanae, whereas those from deer belonged to Cryptosporidium ubiquitum. Comparison of the SSU rRNA gene sequences obtained with those from earlier studies has identified a nucleotide substitution unique to all C. ryanae isolates from Nepal, in addition to some sequence heterogeneity among different copies of the gene. The finding of the dominance of a unique C. ryanae variant in both zebu cattle and water buffaloes in Nepal indicates that there is unique cryptosporidiosis transmission in bovine animals in the study area, and cross-species transmission of some Cryptosporidium spp. can occur between related animal species sharing the same habitats.     

Effect of Waterborne Zinc on Survival, Growth, and Feed Intake of Indian Major Carp, Cirrhinus mrigala (Hamilton)

The effect of waterborne zinc on survival, growth, and feed intake of Indian major carp, Cirrhinus mrigala (Hamilton), advanced fry was studied under laboratory condition. Survival rates of C. mrigala advanced fry (2.71?±?0.49 g) after 30 days exposure to control (0.01), 0.03, 0.06, 0.10, and 0.15 mg/L zinc using the static renewal method in freshwater at pH 7.3?±?0.2, temperature 26?±?2°C, and total hardness 114?±?16 mg/L as CaCO₃ were 100%. Growth of the fish exposed to 0.10 and 0.15 mg/L of zinc was significantly lower (P?<?0.05) than in control (0.01), 0.03, and 0.06 mg/L of zinc after 30 days of exposure. However, there were no significant differences (P?>?0.05) in fish growth between 0.03 and 0.06 mg/L zinc concentrations. Feed intake rates were significantly (P?<?0.05) reduced in the fish exposed to 0.10 mg/L and higher levels of zinc. The zinc accumulation in the whole body of the fish increased with increasing concentrations of the metal.     

Nodulation response to molybdenum supplementation in alfalfa and its correlation with root and shoot growth in low pH soil

Molybdenum (Mo) is a critical micronutrient for nitrogen (N) fixation in legumes. Low pH limits the availability of Mo, thereby reducing nodulation and N fixation. This study investigates the effect of Mo supplementation on alfalfa nodulation and its correlation with root and shoot biomass in low-pH soil. Three experiments were conducted in the greenhouse, involving 14 genotypes of alfalfa subjected to four different treatments, unlimed low-pH soil (5.2) with Mo applied (lithium (Li)^?Mo⁺), low-pH soil (5.2) without Mo (Li^?Mo^?), limed soil (pH 7.3) with Mo applied (Li⁺Mo⁺), and limed soil (pH 7.3) without Mo (Li⁺Mo^?). Foliar application of Mo resulted in a significant increase in nodule counts in the 14 alfalfa cultivars grown in low-pH soil (5.2) even though to a lesser extent than in limed soil with neutral pH (7.3). The increase in number of nodules correlated positively with plant root weight and upper plant biomass.     

Optimization of lipase-catalyzed synthesis of ginsenoside Rb1 esters using response surface methodology

In the lipase (Novozyme 435)-catalyzed synthesis of ginsenoside Rb1 esters, different acyl donors were found to affect not only the degree of conversion but also the regioselectivity. The reaction of acyl donors with short carbon chain was more effective, showing higher conversion than those with long carbon chain. Among the three solvent systems, the reaction in tert-amyl alcohol showed the highest conversion rate, while the reaction in the mixed solvent of t-BuOH and pyridine (1:1) had the lowest conversion rate. To allow the increase of GRb1 lipophilicity, we decided to further study the optimal condition of synthesis of GRb1 with vinyl decanoate with 10 carbon chain fatty acids in tert-amyl alcohol. Response surface methodology (RSM) was employed to optimize the synthesis condition. From the ridge analysis with maximum responses, the maximum GRb1 conversion was predicted to be 61.51% in a combination of factors (40.2 h, 52.95 degrees C, substrate mole ratio 275.57, and enzyme amount 39.81 mg/mL). Further, the adequacy of the predicted model was examined by additional independent experiments at the predicted maximum synthesis conditions. Results showed that the RSM was effective to optimize a combination of factors for lipase-catalyzed synthesis of ginsenoside Rb1 with vinyl decanoate.     

Resistance of wheat to Mycosphaerella graminicola involves early and late peaks of gene expression

Large-scale cDNA-AFLP profiling identified numerous genes with increased expression during the resistance response of wheat to the Septoria tritici blotch fungus, Mycosphaerella graminicola. To test whether these genes were associated with resistance responses, primers were designed for the 14 that were most strongly up-regulated, and their levels of expression were measured at 12 time points from 0 to 27 days after inoculation (DAI) in two resistant and two susceptible cultivars of wheat by real-time quantitative polymerase chain reaction. None of these genes was expressed constitutively in the resistant wheat cultivars. Instead, infection of wheat by M. graminicola induced changes in expression of each gene in both resistant and susceptible cultivars over time. The four genes chitinase, phenylalanine ammonia lyase, pathogenesis-related protein PR-1, and peroxidase were induced from about 10- to 60-fold at early stages (3 h–1 DAI) during the incompatible interactions but were not expressed at later time points. Nine other genes (ATPase, brassinosteroid-6-oxidase, peptidylprolyl isomerase, peroxidase 2, 40S ribosomal protein, ADP-glucose pyrophosphorylase, putative protease inhibitor, methionine sulfoxide reductase, and an RNase S-like protein precursor) had bimodal patterns with both early (1–3 DAI) and late (12–24 DAI) peaks of expression in at least one of the resistant cultivars, but low if any induction in the two susceptible cultivars. The remaining gene (a serine carboxypeptidase) had a trimodal pattern of expression in the resistant cultivar Tadinia. These results indicate that the resistance response of wheat to M. graminicola is not completed during the first 24 h after contact with the pathogen, as thought previously, but instead can extend into the period from 18 to 24 DAI when fungal growth increases dramatically in compatible interactions. Many of these genes have a possible function in signal transduction or possibly as regulatory elements. Expression of the PR-1 gene at 12 h after inoculation was much higher in resistant compared to susceptible recombinant-inbred lines (RILs) segregating for the Stb4 and Stb8 genes for resistance. Therefore, analysis of gene expression could provide a faster method for separating resistant from susceptible lines in research programs. Significant differential expression patterns of the defense-related genes between the resistant and susceptible wheat cultivars and RILs after inoculation with M. graminicola suggest that these genes may play a major role in the resistance mechanisms of wheat.