A plastidic glucose-6-phosphate dehydrogenase is responsible for hypersensitive response cell death and reactive oxygen species production

Glucose-6-phosphate dehydrogenase (G6PDH) has been implicated in the supply of reduced nicotine amide cofactors for resistance to biotic and abiotic stresses. Here, we show participation of the plastidic P2 isoform of G6PDH in plant immunity. A cytosolic isoform (NbG6PDH-Cyto) and two plastidic isoforms (NbG6PDH-P1 and NbG6PDH-P2) cloned from Nicotiana benthamiana were localized in cytosol and chloroplasts, respectively. Hypersensitive response (HR) cell death and NADPH oxidase (RBOH; respiratory burst oxidase homolog)-dependent reactive oxygen species (ROS) production after recognition of INF1 elicitin, secreted by oomycete Phytophthora infestans, decreased in NbG6PDH-P2-silenced plants, but not in NbG6PDH-Cyto- and NbG6PDH-P1-silenced plants. Silencing of the cytosolic NAD kinase NbNADK1, which phosphorylates NADH to form NADPH, compromised HR cell death and ROS production, and concomitant silencing with NbG6PDH-P2 reduced HR cell death and ROS to levels near those in NbG6PDH-P2-silenced plants. Similarly, silencing NbG6PDH-P2 and NbNADK1 resulted in high susceptibility to P. infestans. These results suggest that NADPH produced by the P2 isoform of G6PDH in chloroplasts is responsible for HR cell death and ROS production mediated by RBOH and that NbNADK1 is involved in this pathway.     

Multiplex PCR and Genescan analysis to detect immunoglobulin heavy chain gene rearrangement in feline B-cell neoplasms

Lymphoid neoplasms are usually diagnosed on the basis of cytological and histopathological findings. However, in some cases, discrimination of lymphoid neoplasms from reactive lymphoid proliferation is difficult. PCR amplification of complementarity-determining region 3 (CDR3) of the immunoglobulin heavy-chain variable region (IGHV) gene can be used to assess clonality of B-cell populations as a supportive diagnostic tool for B-cell neoplasms. Because of the sequence variation and possible somatic hypermutation of the IGHV gene, sensitivity of the PCR-based assay to detect clonal IGHV gene rearrangement largely depends on the sequences and numbers of primer sets. Prior to the development of an efficient assay, we cloned and sequenced 97 IGHV complementary DNAs (48 IGHV-1 and 49 IGHV-3 clones) from normal cat spleens. On the basis of these sequences, we designed 6 forward primers at the variable region and 5 reverse primers at the joining region. Using each of 6 forward primers and a mixture of 5 reverse primers, we amplified CDR3 of IGHV genes and analyzed the PCR products by conventional PAGE and Genescan analyses using fluorescence-labeled primers. Twenty-six feline B-cell neoplasms diagnosed by histopathological and immunohistochemical examinations were subjected to the newly developed analysis of IGHV gene rearrangement. Clonal IGHV gene rearrangement was detected in 22 of 26 (84%) samples by both PAGE and Genescan analyses. To reduce the number of PCR reactions, we constructed a multiplex PCR analysis system using a mixture of IGHV-1- and IGHV-3-specific primers as forward primers and a mixture of 5 joining region reverse primers. Results of the multiplex PCR were 100% concordant with those obtained by each of the singleplex PCRs. The multiplex PCR-based assay and Genescan analysis developed in the present study would be useful and practical tools to detect clonal IGHV gene rearrangement in feline B-cell neoplasms.     

Hemolytic suppression roles of taurine in yellowtail Seriola quinqueradiata fed non-fishmeal diet based on soybean protein

ABSTRACT:   To elucidate the hemolytic suppression roles of taurine and the necessity of dietary taurine supplementation in yellowtail Seriola quinqueradiata fed a diet without fishmeal, juvenile fish with an initial body weight of 250 g were fed for 40 weeks in floating net cages on soybean protein diets supplemented with 0, 3.0, 4.5 and 6.0% taurine. Taurine concentration of the experimental diets were 0.03, 33.9, 52.8 and 71.6 mg/g, respectively. On the 21st week, fish fed the taurine unsupplemented diet had inferior growth and feed performances, higher death, and there were incidences of green liver and hemolytic anemia. In this group, hepatic and plasma taurine concentrations, serum osmolality and osmotic tolerance of erythrocytes (EC50 value) were significantly lower, and plasma hydroperoxide concentration was markedly higher than in the taurine supplemented groups. These conditions markedly improved corresponding with the increase of dietary taurine concentration. These results indicate that taurine plays a role in hemolytic suppression through osmoregulation and biomembrane stabilization in fish. In addition, it is suggested that yellowtail requires dietary taurine as an essential nutrition for maintaining physiological condition normally.     

Muscle fibre distribution in forelimb,hindlimb and trunk muscles in three bat species: The little Japanese horseshoe,greater horseshoe and Egyptian fruit

This study characterised muscle fibres in trunk, forelimb and hindlimb muscles of three bat species: little Japanese horseshoe (Rhinolophus cornutus), greater horseshoe (Rhinolophus ferrumequinum) and Egyptian fruit (Rousettus aegyptiacus). Twenty-seven muscles from trunk, forelimb and hindlimb were dissected, weighed and analysed by immunohistochemistry and sodium didecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and determined their cross-sectional areas (CSA). Results showed that Type IIa and Type IIa/x made the highest proportion of total muscle mass. Moderate proportion was formed by Type IIb. Type I and IIx appeared at very low levels in all bats. Type IIb was the only fibre type detected in patagial muscles in wing membrane of greater horseshoe while other fibre types were not observed. Type I muscle fibres were very few and appeared infrequently in fifteen muscles of Egyptian fruit and in only one muscle in each, greater horseshoe and little Japanese horseshoe. Type IIx was also detected in three muscles in greater horseshoe and only one muscle in Egyptian fruit but none in little Japanese horseshoe. The highest average CSA μm² was detected in Type IIb and values were 734.2μm² for LHB; 1537.9μm² for GHB and 1,720.9μm² for EFB. Lowest and undetermined values were observed for Type I and IIx. These data demonstrate that Type IIa, IIa/x and IIb form significant proportion of adult bat muscle mass and Type IIb is the largest fibre type. The distribution pattern is suggestive of specialised functions of the fibres in relation to orientation and speed of bats during flight.     

Effect of salinity on growth and accumulation of organic and inorganic solutes in the leguminous plants Alhagi pseudoalhagi and Vigna radiata

The response of two leguminous plants Alhagi pseudoalhagi and Vigna radiata to seawater salinity was studied over a period of 30 d. The growth of Vigna radiata was markedly and gradually reduced by increasing salinity levels, whereas that of Alhagi pseudoalhagi was promoted at 9.1 and 16.2 dS m^-1 salinity but then was slightly reduced at 28.2 dS m^-1 salinity. These results indicate that Alhagi pseudoalhagi belongs to the group of halophytic plants. Seawater salinity caused changes in the membrane permeability measured as electrolyte leakage in both plants. Alhagi pseudoalhagi maintained a lower membrane permeability than Vigna radiata. With increasing salinity levels, the membrane permeability decreased in Alhagi pseudoalhagi, whereas, in Vigna radiata it slightly increased at 9.1 dS m^-1. The leaf water potential and the osmotic potential decreased in both plants along with the seawater salinity levels. However, the turgor potential and osmotic adjustment in Alhagi pseudoalhagi were maintained at a higher level than in Vigna radiata. The contributions of organic and inorganic solutes to the osmotic adjustment differed: Alhagi pseudoalhagi achieved osmotic adjustment through Cl^- and Na⁺ uptake from the substrate, while the contribution of K⁺, Ca²⁺, and organic solutes to the osmotic adjustment was limited. These results suggest that the differences in salt tolerance between Alhagi pseudoalhagi and Vigna radiata can not be due to differences in specific-ion effects, but may be related to some factors involved in membrane permeability and osmotic adjustment.     

Effect of soil microorganisms on the growth of roots in rice seedlings

Abstract

Root system of rice seedlings grown on nutrient solution inoculated with soil microorganisms were examined morphologically in comparison with those obtained under sterile condition. In the presence of soil microorganisms, primary roots increased in their number and decreased in the total length. Inoculated plants had more secondary roots equipped with tertiary roots. In addition, longer root hairs developed densely on primary and secondary roots of the inoculated seedlings.

Anatomical examination of the primary roots revealed that the number and width of cortical layers, as well as the length and width of the cortical cells, were increased by the effect of microorganisms. Microbial effect on outer morphology of rice roots, consequently, was estimated to have been induced from the alteration in histological and cytological activities including the activation of the periclinal divisions of the epidermal cells, the inactivation of the transverse divisions of the cortical cells and the activation of the elongation of cortical cells.     

Influence of dietary soy protein and peptide products on bile acid status and distal intestinal morphology of rainbow trout Oncorhynchus mykiss

To investigate the influence of dietary soy protein size on the bile acid status and the distal intestinal morphology in rainbow trout Oncorhynchus mykiss, diets based on soybean meal (SBM), soy protein isolate (SPI) and 4 soy peptide products having different molecular sizes were fed to trout (initial BW, 19?g) for 10?weeks. A diet based on fish meal (Cont) and a diet including the smallest-peptide and low-saponin product (Hi-Nute AM) and supplemented with soya saponin (AMS) were also fed. In fish fed diets SBM and SPI, the gallbladder was atrophied, the proportion of biliary cholyltaurine decreased, and highly vacuolated epithelial cells of mucosal folds and proliferation of connective tissues in the submucosa of the distal intestine were observed. These parameters tended to improve in fish fed soy peptide diets and especially those of fish fed diet AM were similar to fish fed diet Cont. Although the gallbladder of fish fed diet AMS was smaller than fish fed the unsupplemented diet AM, no morphological abnormalities were observed in the distal intestine. These results suggest that soy proteins/peptides with relatively large sizes and with soya saponin affect the bile acid status and distal intestinal morphology of rainbow trout.     

Robust reconstitution of active cell-cycle control complexes from co-expressed proteins in bacteria

ABSTRACT: BACKGROUND: Cell proliferation is an important determinant of plant growth and development. In addition,modulation of cell-division rate is an important mechanism of plant plasticity and is key inadapting of plants to environmental conditions. One of the greatest challenges inunderstanding the cell cycle of flowering plants is the large families of CDKs and cyclins thathave the potential to form many different complexes. However, it is largely unclear whichcomplexes are active. In addition, there are many CDK- and cyclin-related proteins whosebiological role is still unclear, i.e. whether they have indeed enzymatic activity. Thus, abiochemical characterization of these proteins is of key importance for the understanding oftheir function. RESULTS: Here we present a straightforward system to systematically express and purify active CDKcyclincomplexes from E. coli extracts. Our method relies on the concomitant production of aCDK activating kinase, which catalyzes the T-loop phosphorylation necessary for kinaseactivity. Taking the examples of the G1-phase cyclin CYCLIN D3;1 (CYCD3;1), the mitoticcyclin CYCLIN B1;2 (CYCB1;2) and the atypical meiotic cyclin SOLO DANCERS (SDS) inconjunction with A-, B1- and B2-type CDKs, we show that different CDKs can interact withvarious cyclins in vitro but only a few specific complexes have high levels of kinase activity. CONCLUSIONS: Our work shows that both the cyclin as well as the CDK partner contribute to substratespecificity in plants. These findings refine the interaction networks in cell-cycle control andpinpoint to particular complexes for modulating cell proliferation activity in breeding.     

Chromosomal Location of HWA1 and HWA2, Complementary Hybrid Weakness Genes in Rice

Hybrid weakness phenomena in rice reportedly have two causes: those of HWC1 and HWC2 genes and those of HWA1 and HWA2 genes. No detailed study of the latter has been reported. For this study, we first produced crosses among cultivars carrying the weakness-causing allele on the HWA1 and HWA2 loci to confirm the phenotype of the hybrid weakness and the genotypes of the cultivars on the two loci, as reported earlier. We then confirmed that these cultivars belong to Indica. Subsequent linkage analysis of HWA1 and HWA2 genes conducted using DNA markers revealed that both genes are located in the 1,637-kb region, surrounded by the same DNA markers on the long arm of chromosome 11. The possibility of allelic interaction inducing hybrid weakness is discussed.     

Mechanical properties assessment of Cunninghamia lanceolata plantation wood with three acoustic-based nondestructive methods

The objectives of this study were to establish the method of evaluating wood mechanical properties by acoustic nondestructive testing at standing trees and at logs of a Chinese fir (Cunninghamia lanceolata (Lamb.) Hook.) plantation, and to compare three acoustic nondestructive methods for evaluating the static bending modulus of elasticity (MOE), modulus of rupture (MOR), and compressive strength parallel-to-grain (σ_c) of plantation wood as well. Fifteen Chinese fir plantation trees at 36 years of age were selected. Each tree was cut into four logs, for which three values of dynamic modulus of elasticity, i.e., E _sw, of the north and south face based on stress waves to assume the measuring state of the standing tree, E _fr, longitudinal vibration, and E _us, ultrasonic wave, were measured in the green condition. After log measurements, small specimens were cut and air-dried to 12% moisture content (MC). Static bending tests were then performed to determine the bending MOE and MOR, and compressive tests parallel-to-grain were made to determine σ_c. The bending MOE of small clear specimens was about 7.1% and 15.4% less than E _sw and E _us, respectively, and 11.3% greater than E _fr. The differences between the bending MOE and dynamic MOE of logs as determined by the three acoustic methods were statistically significant (P < 0.001). Good correlation (R = 0.77, 0.57, and 0.45) between E _sw, E _fr, and E _us and static MOE, respectively, were obtained (P < 0.001). It can be concluded that longitudinal vibration may be the most precise and reliable technique to evaluate the mechanical properties of logs among these three acoustic nondestructive methods. Moreover, the results indicate that stress wave technology would be effective to evaluate wood mechanical properties both from logs and from the standing tree.