Breeding for higher yield, early maturity, wider adaptability and waterlogging tolerance in soybean (Glycine max L.): A case study.

Breeding for higher yield and wider adaptability are major objectives of soybean crop improvement. In the present study, 68 advanced breeding lines along with seven best checks were evaluated for yield and attributing traits by following group balanced block design. Three blocks were constituted based on the maturity duration of the breeding lines. High genetic variability for the twelve quantitative traits was found within and across the three blocks. Several genotypes were found to outperform check varieties for yield and attributing traits. During the same crop season, one of the promising entries, NRC 128,was evaluated across seven locations for its wider adaptability and it has shown stable performance in Northern plain Zone with > 20% higher yield superiority over best check PS 1347. However, it produced 9.8% yield superiority over best check in Eastern Zone. Screening for waterlogging tolerance under artificial conditions revealed that NRC 128 was on par with the tolerant variety JS 97-52. Based on the yield superiority, wider adaptability and waterlogging tolerance, NRC 128 was released and notified by Central Varietal Release Committee (CVRC) of India, for its cultivation across Eastern and Northern Plain Zones of India.

Linkage mapping of Mungbean yellow mosaic India virus (MYMIV) resistance gene in soybean.

Mungbean Yellow Mosaic India Virus (MYMIV) is one of the most prevalent pathogen that limits soybean production in India. In this study RILs derived from JS335, dominant but MYMIV susceptible variety and PI171443, donor of MYMIV resistance gene in most of the MYMIV resistant varieties released in India and F2 population derived from SL525, a resistant variety released for northern India and NRC101, a susceptible genotype were used to study the inheritance of MYMIV resistance and map the gene responsible for MYMIV resistance. F1s were found to be completely susceptible. F2:3 and RILs population segregated to fit a ratio of 1:2:1 and 1:1 indicating that a single recessive gene controlled resistance to MYMIV. BSA was performed using 144 polymorphic SSR markers. MYMIV resistance gene was mapped on chr 6 (LG C2) within a 3.5-cM genome region between two SSR markers GMAC7L and Satt322 whose size was estimated to be 77.115 kb (position of 12,259,594-12,336,709 bp). This is the first report on linkage mapping of MYMIV resistance gene in soybean. This will be helpful in breeding soybean varieties for resistance against MYMIV responsible for wide spread damage to soybean crop in India using Marker Assisted Selection.

In vivo induction of antioxidant response and oxidative stress associated with genotoxicity and histopathological alteration in two commercial fish species due to heavy metals exposure in northern India (Kali) river.

Heavy metals can significantly bioaccumulate in fish tissues. The step wise mechanism of heavy metal toxicities on fish health is still limited. The present study assessed the tissue-specific antioxidant response and oxidative stress biomarkers of commercially important fish species namely, Channa striatus and Heteropneustes fossilis inhabiting Kali River of northern India where heavy-metal load is beyond the World Health Organisation - maximum permissible limits. Heavy metals chromium (Cr), nickel (Ni), lead (Pb) and cadmium (Cd) were elevated in both fish species compared to recommended values of the Federal Environmental Protection Agency (FEPA), 1999 for edible fishes. Reduced glutathione (GSH), superoxide dismutase (SOD) and catalase (CATA) activities in all tissues (brachial, neural, renal and hepatic) were altered. Cellular lipid and protein compromisation in both fishes induced by heavy metals was determined by lipid peroxidation (LPO) and protein carbonylation (PC) assays. Micronucleus (MN) test of erythrocytes and comet assay of liver cells confirmed genotoxicity. Histopathology of the liver, kidney and brain of affected fishes was distorted significantly with its reference fishes thereby affecting the quality and quantity of these fish stocks. This raises a serious concern as these fishes are consumed by the local population which would ultimately affect human health.

Diversity and stability of cultured vaginal lactobacilli in pregnant women from a multi-ethnic urban UK population.

AIM: To determine the diversity and stability of cultured vaginal lactobacilli in a multi-ethnic population of pregnant women. METHODS AND RESULTS: A single-centre, prospective, cohort study was performed in a tertiary perinatal centre in East London, UK. Self-collected vaginal swabs at 13 and 20 weeks gestation were obtained from women attending for routine antenatal care and cultured for lactobacilli. In women who provided both swabs, 37 of 203 (18%) had no lactobacilli cultured at either time. Only 53 (26%) had the same species at both times. Black women were less likely to have lactobacilli cultured at 13 weeks (P = 0·014), and Black and Asian women were less likely to have lactobacilli cultured at 20 weeks (P = 0·002) compared with those in the White and Other groups. CONCLUSIONS: Significant differences exist between ethnic groups in the carriage and stability of vaginal lactobacilli. SIGNIFICANCE AND IMPACT OF THE STUDY: These differences have implications for the design of interventions aimed at normalizing the vaginal microbiota in pregnant women.

Placental-specific Igf2 knockout mice exhibit hypocalcemia and adaptive changes in placental calcium transport.

Evidence is emerging that the ability of the placenta to supply nutrients to the developing fetus adapts according to fetal demand. To examine this adaptation further, we tested the hypothesis that placental maternofetal transport of calcium adapts according to fetal calcium requirements. We used a mouse model of fetal growth restriction, the placental-specific Igf2 knockout (P0) mouse, shown previously to transiently adapt placental System-A amino acid transporter activity relative to fetal growth. Fetal and placental weights in P0 mice were reduced when compared with WT at both embryonic day 17 (E17) and E19. Ionized calcium concentration [Ca(2+)] was significantly lower in P0 fetal blood compared with both WT and maternal blood at E17 and E19, reflecting a reversal of the fetomaternal [Ca(2+)] gradient. Fetal calcium content was reduced in P0 mice at E17 but not at E19. Unidirectional maternofetal calcium clearance ((Ca) K (mf)) was not different between WT and P0 at E17 but increased in P0 at E19. Expression of the intracellular calcium-binding protein calbindin-D(9K), previously shown to be rate-limiting for calcium transport, was increased in P0 relative to WT placentas between E17 and E19. These data show an increased placental transport of calcium from E17 to E19 in P0 compared to WT. We suggest that this is an adaptation in response to the reduced fetal calcium accumulation earlier in gestation and speculate that the ability of the placenta to adapt its supply capacity according to fetal demand may stretch across other essential nutrients.

Increased maternofetal calcium flux in parathyroid hormone-related protein-null mice.

The role of parathyroid hormone-related protein (PTHrP) in fetal calcium homeostasis and placental calcium transport was examined in mice homozygous for the deletion of the PTHrP gene (PTHrP-/- null; NL) compared to PTHrP+/+ (wild-type; WT) and PTHrP+/- (heterozygous; HZ) littermates. Fetal blood ionized calcium was significantly reduced in NL fetuses compared to WT and HZ groups at 18 days of pregnancy (dp) with abolition of the fetomaternal calcium gradient. In situ placental perfusion of the umbilical circulation at 18 dp was used to measure unidirectional clearance of (45)Ca across the placenta in maternofetal ((Ca)K(mf)) and fetoplacental ((Ca)K(fp)) directions; (Ca)K(fp) was < 5% of (Ca)K(mf) for all genotypes. At 18 dp, (Ca)K(mf) across perfused placenta and intact placenta ((Ca)K(mf(intact))) were similar and concordant with net calcium accretion rates in vivo. (Ca)K(mf) was significantly raised in NL fetuses compared to WT and HZ littermates. Calcium accretion was significantly elevated in NL fetuses by 19 dp. Placental calbindin-D(9K) expression in NL fetuses was marginally enhanced (P < 0.07) but expression of TRPV6/ECaC2 and plasma membrane Ca2+-ATPase (PMCA) isoforms 1 and 4 were unaltered. We conclude that PTHrP is an important regulator of fetal calcium homeostasis with its predominant effect being on unidirectional maternofetal transfer, probably mediated by modifying placental calbindin-D(9K) expression. In situ perfusion of mouse placenta is a robust methodology for allowing detailed dissection of placental transfer mechanisms in genetically modified mice.

Identification of SSR markers associated with seed coat permeability and electrolyte leaching in soybean.

Seed coat permeability and electrolyte leaching are the important traits that have been negatively associated with seed longevity in soybean. The objective of this study was to use SSR markers to identify genomic regions significantly associated with QTLs controlling seed coat permeability and electrolyte leaching in a segregating F2 population derived from a cross of Birsa soya-1 x JS 71-05. Parental polymorphism survey using 145 SSR markers identified 21 polymorphic ones, which were used to genotype 153 F2 individuals. Four independent markers (Satt434, Satt538, Satt281 and Satt598) were significantly (P=0.05) associated with seed coat permeability. One of these markers (Satt 281) also showed significant association with electrolyte leaching that partly supported the observed positive correlation (r = 0.425) between the two traits. Markers for seed coat permeability individually explained 3.9% to 4.5% of the total phenotypic variation, while the marker linked with electrolyte leaching explained 5.6% of the total variation.

Isolation, molecular cloning and in vitro expression of rhesus monkey (Macaca mulatta) prominin-1.s1 complementary DNA encoding a potential hematopoietic stem cell antigen.

Human prominin-1 (CD133 or AC133) is an important cell surface marker used to isolate primitive hematopoietic stem cells. The commercially available antibody to human prominin-1 does not recognize rhesus prominin-1. Therefore, we isolated, cloned and characterized the complementary DNA (cDNA) of rhesus prominin-1 gene and determined its coding potential. Following the nomenclature of prominin family of genes, we named this cDNA as rhesus prominin-1.s1. The amino acid sequence data of the putative rhesus prominin-1.s1 could be used in designing antigenic peptides to raise antibodies for use in isolation of pure populations of rhesus prominin-1(+) hematopoietic cells. To the best of our knowledge, there has been no previously published report about the isolation of a prominin-1 cDNA from rhesus monkey (Macaca mulatta).

Artificial perfusion of the fetal circulation of the in situ mouse placenta: methodology and validation.

Here we present methodology and validation (including measurement of unidirectional maternofetal clearance (Kmf) of (45)Ca and (14)C-mannitol) for in situ perfusion of the mouse placenta. On day 18 of gestation (term=19 days) mice were anaesthetised and the uterus delivered into a saline bath (40 degrees C). A fetus was selected, the umbilical artery and vein catheterised and perfused with Krebs Ringer (pH 7.4) at 60 microl/min. (45)Ca/(14)C-mannitol (2 microCi/5 microCi in 50 microl saline) was injected via maternal tail vein. Perfusate samples were collected every 5 min for 45 min. Maternal carotid artery pressure was monitored throughout perfusion. A terminal maternal cardiac blood sample was taken and analysed. Placentas were immersion fixed and processed for electron microscopy. Kmf for (45)Ca and (14)C-mannitol was calculated as perfusate [(45)Ca or (14)C-mannitol] x perfusion rate/maternal plasma [(45)Ca or (14)C-mannitol]xplacental weight. Maternal cardiac blood chemistry at termination (n=8-15, mean+/-SEM) was as follows: pH 7.153+/-0.016, PCO(2) 45.48+/-2.06 mmHg, PO(2) 66.47+/-7.10 mmHg, Na(+) 151.4+/-1.2 mmol/l, K(+) 5.54+/-0.17 mmol/l, Ca(2+) 1.15+/-0.03 mmol/l, glucose 7.2+/-0.5 mmol/l, and lactate 1.76+/-0.77 mmol/l. A successful 45 min perfusion in which perfusate recovery was >95% occurred in >50% of animals. Perfusion did not alter placental morphology or carotid pressure. Kmf (microl/min/g placenta) for (45)Ca (66.0+/-8.4 (n=7)) was significantly higher than Kmf for (14)C-mannitol (20.0+/-2.4 (n=5)) (p<0.01). These data demonstrate physiological perfusion of the mouse placenta in situ and its usefulness for measurement of solute transfer.

Electrical potential difference between mother and conceptus in the mouse.

This study aimed to determine whether there is a maternofetal potential difference (PD) in the mouse. The mean (+/- SEM) in vivo electrical potential difference (PD(sa)) between saline filled catheters in the maternal subcutaneous space (s) and the fetal amniotic sac (a) measured, according to strict criteria, in anaesthetised MF1 mice at a gestational age of 18-20 (term 20) days was 3.9+/-0.5 mV (significantly different from zero P<0.0001) in 16 conceptuses from 11 mice with the amniotic sac positive with respect to the maternal catheter. The PD(sv) between maternal tail vein (v) and maternal subcutaneous space was -0.8+/-0.4 mV (n=3: not significantly different from zero). Measurement of PD between two different maternal subcutaneous catheters (n=4) was < or =0.5 mV. This study shows that there is a maternofetal PD in the mouse and provides the foundation for studies addressing its mechanism of generation in this species.

Effect of diabetes mellitus on rat placenta cellularity.

Transcellular placental maternofetal flux of calcium and magnesium is reduced in diabetic pregnancy in the rat which might be due to changes in placental cellularity. In order to investigate this wet and dry weight, DNA and protein content were measured in placentas from untreated diabetic (D(O)), insulin-treated diabetic (D(I)) and control rats (C) on day 21 of gestation (term=23 days). Wet and dry weights (mg; mean+/-S.E.M.) were 418+/-13, 474+/-19, 416+/-14 and 66+/-3, 75+/-3, 67+/-3 in C, D(O) and D(I) groups, respectively. Total DNA and protein content (mg) was 1.8+/-0.2, 1.7+/-0.1, 1.5+/-0.1 and 50.4+/-2.4, 54.9+/-2.6, 51.9+/-3.3 in C, D(O) and D(I) groups, respectively. The data suggest that placental cellularity is unaffected by maternal diabetes mellitus in the rat and is unlikely to directly affect maternofetal flux of calcium and magnesium.

Antigen expression during murine gamma-herpesvirus infection.

Gammaherpesviruses (gammaHV) establish a life-long latency in the host and are associated with a number of malignant human diseases. It is generally believed that T cells play a major role in controlling the initial acute infection and subsequently maintaining the virus in a quiescent state. However, the nature of the T cell response to gamma-herpesvirus infections is poorly understood. In the current report we took advantage of a mouse model of gammaHV infection (murine herpesvirus-68, MHV-68) to investigate the T cell response to different phases of the infection. Intranasal infection with MHV-68 induces an acute infection in lung epithelial cells and long-term latency in B cells. The kinetics of the CD8+ T cell response to different lytic cycle and latency-associated antigens was highly complex and distinct patterns of response could be identified. These responses were regulated by multiple factors including differences in temporal expression of the relevant antigens, differences in the presentation of antigen in different organs, and differential expression of antigen in different types of antigen presenting cells. For example, some antigens were expressed at distinct phases of the infection and in specific organs or subsets of antigen presenting cells. In addition, recent data suggest that in addition to B cells, both macrophages and dendritic cells harbor latent MHV-68 infection, adding further complexity to their role in controlling the T cell response to this infection.

Latent murine gamma-herpesvirus infection is established in activated B cells, dendritic cells, and macrophages.

Intranasal infection of mice with the murine gamma-herpesvirus MHV-68 results in an acute lytic infection in the lung, followed by the establishment of lifelong latency. Development of an infectious mononucleosis-like syndrome correlates with the establishment of latency and is characterized by splenomegaly and the appearance of activated CD8+ T cells in the peripheral blood. Interestingly, a large population of activated CD8+ T cells in the peripheral blood expresses the V beta 4+ element in their TCR. In this report we show that MHV-68 latency in the spleen after intranasal infection is harbored in three APC types: B cells, macrophages, and dendritic cells. Surprisingly, since latency has not previously been described in dendritic cells, these cells harbored the highest frequency of latent virus. Among B cells, latency was preferentially associated with activated B cells expressing the phenotype of germinal center B cells, thus formally linking the previously reported association of latency gene expression and germinal centers to germinal center B cells. Germinal center formation, however, was not required for the establishment of latency. Significantly, although three cell types were latently infected, the ability to stimulate V beta 4+CD8+ T cell hybridomas was limited to latently infected, activated B cells.

Effects of acute maternal hyperglycaemia and hyperosmolality on maternofetal transfer of calcium and magnesium across the in situ perfused rat placenta.

The effects of acute maternal hyperglycaemia and hyperosmolality on maternofetal placental transfer of Ca, Mg and 51Cr-EDTA were investigated in the rat. On day 21 of gestation (term = 23 d) the fetal circulation of the in situ placenta of anaesthetised rats was perfused with a Mg-free Krebs Ringer solution and the unidirectional maternofetal fluxes of Ca (CaJmf) and Mg (MgJmf), and the unidirectional maternofetal clearance of 51Cr-EDTA ((EDTA)Kmf) were determined before and during maternal hyperglycaemia, hyperosmolality and volume expansion, attained by infusing 30% D-glucose, 25% mannitol and 0.9% saline solutions, respectively, into the maternal circulation. MgJmf was significantly reduced during glucose infusion (23.9 +/- 1.2 v 28.2 +/- 1.4 nmol min(-1) g(-1) placenta during control perfusion (mean +/- SEM); p < 0.005) and during mannitol infusion (28.2 +/- 1.0 v 33.5 +/- 1.5 nmol min(-1) g(-1) placenta; p < 0.001). CaJmf and (EDTA)Kmf were not significantly altered by maternal hyperglycaemia or hyperosmolality. There was no significant change in MgJmf during infusion of saline into the maternal circulation. Maternal plasma Na concentration was significantly reduced in both glucose and mannitol infusion experiments, whereas maternal plasma Mg concentration was significantly reduced only during glucose infusion. We postulate that the reduced maternal plasma Na concentration in the glucose and mannitol experiments might decrease MgJmf via alteration of placental Na+/Mg2+ exchange activity.

Murine gammaherpesvirus M2 gene is latency-associated and its protein a target for CD8(+) T lymphocytes.

Murine gammaherpesvirus 68 (MHV-68) infection of mice is a potential model with which to address fundamental aspects of the pathobiology and host control of gammaherpesvirus latency. Control of MHV-68 infection, like that of Epstein-Barr virus, is strongly dependent on the cellular immune system. However, the molecular biology of MHV-68 latency is largely undefined. A screen of the MHV-68 genome for potential latency-associated mRNAs revealed that the region encompassing and flanking the genomic terminal repeats is transcriptionally active in the latently infected murine B-cell tumor line S11. Transcription of one MHV-68 gene, that encoding the hypothetical M2 protein, was detected in virtually all latently infected S11 cells and in splenocytes of latently infected mice, but not in lytically infected fibroblasts. Furthermore, an epitope was identified in the predicted M2 protein that is recognized by CD8(+) T cells from infected mice and a cytotoxic T lymphocyte line that recognizes this epitope killed S11 cells, indicating that the M2 protein is expressed during latent infection and is a target for the host cytotoxic T lymphocyte response. This work therefore provides essential information for modeling MHV-68 latency and strategies of immunotherapy against gammaherpesvirus-related diseases in a highly tractable animal model.

Urinary excretion of pyridinium crosslinks in healthy 4-10 year olds.

Urinary pyridinoline and deoxypyridinoline, pyridinium crosslinks released during breakdown of mature collagen, might serve as useful markers of bone resorption. Before their role can be identified, reference values must be established. In this study, free pyridinoline (f-Pyr), free deoxypyridinoline (f-DPyr), and creatinine (Cr) were measured in first morning void urine samples from 250 girls and 265 boys between the ages of 4 and 10 years. Overall, there was a decrease in f-Pyr:Cr and f-DPyr:Cr ratios with increasing age in both sexes, but there was a wide range of values for individuals of similar ages. Further studies are required to assess whether urinary pyridinium crosslink excretion is sufficiently deranged in conditions affecting bone metabolism for the measurement of these compounds to be of clinical value.

Detection of non-deletional type of hereditary persistence of fetal hemoglobin (HPFH) condition associated with 619 bp ß(°)-thalassemia deletion.

DNA samples from a family (parents and a son) with hereditary persistence of fetal hemoglobin (HPFH) condition were subjected to amplification of a 1.214 kbp DNA fragment from ß-globin gene using polymerase chain reaction (PCR). The aim of this study was to identify the type of HPFH i.e. deletional or non deletional. Non deletional type of HPFH was identified in two samples and moreover, these samples were found to be associated with 619bp ß(°)-thalassemia deletion. This is the first report on the association of non deletional HPFH with 619bp ß(°)-thalassemia deletion.

Analysis of sickle cell gene using polymerase chain reaction & restriction enzyme Bsu 361.

A 772bp DNA fragment from human beta-globin gene has been amplified by polymerase chain reaction (PCR) and subjected to restriction enzyme analysis using Bsu 361, an isoschizomer of restriction enzyme Mst II. This protocol has been designed basically to enhance the analytical facility for the detection of sickle cell mutation. A 430bp DNA fragment was found to be associated with the mutant locus, whereas 228bp and 202bp DNA fragments were generated from the normal locus. This difference of about 202bp in the resulting fragments from the mutant and normal loci has improved discriminatory power in the genotype analysis of the sickle cell mutation.

The effect of diabetes mellitus on urinary calcium excretion in pregnant rats and their offspring.

The effect of maternal diabetes mellitus on renal calcium excretion in pregnant rats and their offspring has been examined in order to ascertain the role of the kidney in the disturbed calcium homeostasis of infants born to diabetic mothers. Diabetic pregnant (DP) rats exhibited severe hypercalciuria which greatly exceeded the urinary calcium losses (UCaV) in non-diabetic pregnant (CP) or non-pregnant diabetic (D) rats. Means +/- S.E.M. for UCaV at day 21 (mmol/24 h) were: DP = 1.12 +/- 0.09 (n = 7); CP = 0.06 +/- 0.01 (n = 7); D = 0.63 +/- 0.06 (n = 7) (P < 0.001 DP vs CP and DP vs D). The profile for urinary calcium excretion in the three groups was different from that of other measured ions. The degree of natriuresis, for example, was comparable in DP and D rats at all stages studied. Although magnesium output was significantly greater in DP than D rats on days 14 and 21, this appeared to result from an additive effect of the magnesiuresis seen when pregnancy and diabetes were studied separately. The marked renal calcium wasting of diabetic pregnancy will have implications for overall calcium balance in the mother. For example, an enhanced intestinal calcium absorption was seen in DP rats in the second half of gestation. Means +/- S.E.M. for day 21 (mmol/24 h) were: DP = 3.8 +/- 0.8 (n = 7); CP = 1.4 +/- 0.3 (n = 7); D = 1.6 +/- 0.3 (n = 7) (P < 0.05 DP vs CP and DP vs D).(ABSTRACT TRUNCATED AT 250 WORDS)