Development of mutants with varying flowering times by targeted editing of multiple SVP gene copies in Brassica napus L.

Manipulation of flowering time to develop cultivars with desired maturity dates is fundamental in plant breeding.It is desirable to generate polyploid rapeseed(Brassica napus L.)germplasm with varying flowering time controlled by a few genes.In the present study,Bna SVP,a rapeseed homolog of the Arabidopsis SVP(Short Vegetative Phase)gene,was characterized and a set of mutants was developed using a CRISPR/Cas9-based gene-editing tool.A single construct targeting multiple sites was successfully applied to precisely mutate four copies of Bna SVP.The induced mutations in these copies were stably transmitted to subsequent generations.Homozygous mutants with loss-of-function alleles and free transgenic elements were generated across the four Bna SVP homologs.All mutant T₁lines tested in two environments(summer and winter growing seasons)showed early-flowering phenotypes.The decrease in flowering time was correlated with the number of mutated Bna SVP alleles.The quadruple mutants showed the shortest flowering time,with a mean decrease of 40.6%–50.7%in length relative to the wild type under the two growth conditions.Our study demonstrates the quantitative involvement of Bna SVP copies in the regulation of flowering time and provides valuable resources for rapeseed breeding.     

Effect of ginger (Zingiber officinale Roscoe) on growth performance, nutrient digestibility, serum metabolites, gut morphology, and microflora of growing guinea fowl

A 56-day feeding trial was conducted to investigate the effect of dietary supplementation of dried ginger meal (DGM) on the growth performance, nutrient digestibility, serum parameters, gut morphology, and microflora of growing helmeted guinea fowl (Numidia meleagris). One hundred sixty 28-day-old male keets were assigned to four dietary treatments. There were 40 birds per treatment replicated four times with 10 keets each. The experimental diets consisted of the basal diet (control), ginger-supplemented diets containing 20, 40, and 60 g/kg DGM, respectively. Guinea fowls fed diet supplemented with DGM had higher (P <0.05) final body weight, body weight gain and lower (P <0.05) feed intake. Optimum weight gain was obtained with supplementation level of 36.15-g DGM (R ²?=?0.923). Guinea fowls fed diet supplemented with 40 g/kg DGM recorded the highest (P <0.05) coefficient of total tract apparent digestibility of dry matter, ether extract and longest (P <0.05) duodenal and ileal villus heights. The crypt depth of the duodenum and ileum reduced (P <0.05) with increasing level of dietary supplementation of DGM. Dietary supplementation of DGM resulted in increased (P <0.05) total serum protein, serum albumin and low (P <0.05) serum cholesterol concentration. Ileum content of birds fed diet supplemented with 40 g/kg DGM recorded the highest (P <0.05) lactobacillus count. In conclusion, dietary inclusion of 40-g DGM per kilogram diet is hereby recommended for improved growth, apparent nutrient digestibility, gut morphology, serum chemistry, and stimulation of balanced intestinal microflora in growing guinea fowl.     

Molecular characterization of Malaysian fowl adenovirus (FAdV) serotype 8b species E and pathogenicity of the virus in specific-pathogen-free chicken

BackgroundInclusion body hepatitis (IBH) is an economically important viral disease primarily affecting broiler and breeder chickens. All 12 serotypes of fowl adenovirus (FAdV) can cause IBH.ObjectivesTo characterize FAdV isolates based on phylogenetic analysis, and to study the pathogenicity of FAdV-8b in specific-pathogen-free (SPF) chickens following virus inoculation via oral and intramuscular (IM) routes.MethodsSuspected organ samples were subjected to virus isolation and polymerase chain reaction (PCR) for FAdV detection. Hexon gene sequencing and phylogenetic analysis were performed on FAdV-positive samples for serotype identification. One FAdV-8b isolate, UPM/FAdV/420/2017, was selected for fiber gene characterization and pathogenicity study and was inoculated in SPF chickens via oral and IM routes.ResultsThe hexon gene phylogenetic analysis revealed that all isolates belonged to FAdV-8b. The fiber gene-based phylogenetic analysis of isolate UPM/FAdV/420/2017 supported the grouping of that isolate into FAdV species E. Pathogenicity study revealed that, chickens infected with UPM/FAdV/420/2017 via the IM route had higher clinical score values, higher percent mortality, higher degree of the liver lesions, higher antibody response (p < 0.05), and higher virus shedding amounts (p < 0.05) than those infected via the oral route. The highest virus copy numbers were detected in liver and gizzard.ConclusionsFAdV-8b is the dominant FAdV serotype in Malaysia, and pathogenicity study of the FAdV-8b isolate UPM/FAdV/420/2017 indicated its ability to induce IBH in young SPF chickens when infected via oral or IM routes.     

Influence of interposition of pink muscle fiber into dorsal ordinary muscle on 5'-IMP degrading activity in various fish species

ABSTRACT: To clarify the cause of differences in the temporal change of K -value among fish species living in the same habitat water temperature, the influence of the interposition of pink muscle fibers into dorsal ordinary muscle on 5'-inosine monophosphate (5'-IMP) degrading activity was examined. Fourteen fish species from the Sakishima Islands (habitat water temperature 28°C) and from Nagasaki (habitat water temperature 17°C) were used for the sample fishes. Each of 5'-IMP and ρ-nitrophenol phosphate (ρ-NPP) degrading activities showed a peak at near pH 8.0 and near pH 5.0, respectively. These activities were somewhat higher in fish from Nagasaki than in fish from the Sakishima Islands. The interposition percentage of pink muscle fibers into dorsal ordinary muscle correlated significantly ( P < 0.05) with the 5'-IMP degrading activity at pH 7.0. Furthermore, the activity at pH 7.0 correlated significantly ( P < 0.001) with the increasing rate of K -value at 32°C (ΔK₃₂). These results suggest that the difference in ΔK₃₂ among fish species in the same habitat water temperature might be caused by differences in 5'-IMP degrading activity because of the differences in the interposition rates of pink muscle fibers into dorsal ordinary muscle.     

Immunoprotective role of LaSota vaccine under immunosuppressive conditions in chicken challenged with velogenic avian avulavirus-1

The first objective of the present study was to evaluate if the antibodies induced by the live LaSota and killed Newcastle disease (sub-genotype VIIi) vaccines protect the chickens against exposure with pathogenic avian avulavirus-1 (AAvV-1) of chicken and/or pigeon origins. The second objective was to study the effect of vaccines on stressed birds (dexamethasone, aflatoxin, and heat stressed) with respect to antibody production and protection against pathogenic AAvV-1 challenge. Sixty-one-day-old Hubbard chickens were divided into six groups (gA–gF) with ten animals each. All the groups received LaSota (10⁵ EID₅₀, 0.1 ml per chick) on days 7 and 27 via eye drop and one intramuscular injection of a killed vaccine (sub-genotype VIIi) (10^7.5 EID₅₀, 1 ml) on day 18, except the control birds received the PBS only. Moreover, group gC-DEX received dexamethasone intramuscularly at a dose rate of 1-mg/kg body weight daily; gD-AFLA had received aflatoxin as oral gavage at a dose rate of 30 ppb daily, and gE-HEAT was kept under heat stressed (38 °C) till challenged. All the groups were challenged with AAvV-1 strain of chicken origin of sub-genotype VIIi, except the group gA-pigeon was challenged with pigeon-origin strain (sub-genotype VIm). The result showed that the gA-pigeon and gB-chicken vaccinate showed 100% and 80% protection. The immunosuppressive birds produced low pre-challenge HI titer, and protection was observed at 40%, 50%, and 70% in gC-DEX, gD-AFLA, and gE-HEAT, respectively. Our findings suggest the stress factors such as aflatoxin in the feed and dexamethasone are immunosuppressive in nature and suppress the immune response and its associated protective role during infection.

     

Genetic variability in bread wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) of Pakistan based on polymorphism for high molecular weight glutenin subunits

Variation in bread wheat including pre and post green revolutions varieties of Pakistan along with landraces was investigated for high molecular weight Glutenin subunits (HMW Gs) encoded at three genes (Glu-A1, Glu-B1, Glu-D1) with SDS-PAGE. The germplasm was diverse and unique on the basis of HMW Gs compositions and out of 14 alleles detected at all the Glu-1 loci, three belonged to Glu-A1, nine to Glu-B1 and two to Glu-D1 locus. High variation was observed in the landraces and higher gene diversity was observed between the populations as compared to the gene diversity within populations, whereas a reverse pattern of gene diversity was observed when populations were pooled across the region (higher within the regions than between the regions). A lack of relationship between the HMW Gs diversity and the altitude of collection site was observed. A data base has been generated in this study which could be expanded/exploited for cultivar development or management of gene bank.     

Genetic Variation of Glutelin Acidic Subunit Polypeptides in Bangladesh Rice Genetic Resources

Bangladesh rice genetic resources collected from six distinct regions of the country were examined to obtain the diversity in glutelin acidic subunit polypeptides. Seed glutelins from 576 Bangladesh rice cultivars representing seven ecotypes were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), isoelectric focusing (IEF) electrophoresis and two-dimensional electrophoresis (2-DE: SDS-PAGE/IEF) analyses. Glutelin acidic subunit was separated into four bands, α-1, α-2, α-3 and α-4, and the variation in each of bands was detected by SDS-PAGE analysis. A higher molecular size component of α-1(α-1′) was identified in the cultivars tested. In case of α-2, α-2H with high molecular mass, α-2L with low molecular mass and α-2H/L were detected. α-3 band variation showed α-3H, α-3L, and α-3H/L, while for α-4, α-4H, α-4L bands were identified. In IEF analysis, a total of 16 bands with independent pI ranging from pI 6.30 to 7.52 were identified for the glutelin acidic subunit among the cultivars. The maximum and minimum numbers of IEF bands found were 13 and 9, respectively. The α-2L less cultivars were also lacking in pI 6.80 polypeptide in IEF. Result of 2-DE showed that pI 6.80 polypeptide was the main component of α-2L band. Transplanted Aman ecotype was the most diverse with respect to glutelin variation. Geographical distribution of glutelin variation in the Transplanted Aman ecotype differed according to the regions. The above results indicate that Bangladesh rice cultivars possess great genetic diversity in glutelin acidic subunit polypeptides. This study of indigenous rice cultivars from Bangladesh provides useful information regarding their breeding potential.     

The prevalence of <Emphasis Type="Italic">Mycobacterium bovis</Emphasis>-infection and atypical mycobacterioses in cattle in and around Morogoro,Tanzania

A study was conducted to determine the prevalence of Mycobacterium bovis-infection and atypical mycobacterioses in different cattle herd management systems in and around Morogoro, Tanzania. Between April and June 2005, a total of 728 bovines from 49 herds were tested for M. bovis-infection and atypical mycobacterioses. Milk samples were taken from tuberculin positive animals and analysed for the presence of mycobacteria. Total prevalences of 2.5% and 10.1% were found for M. bovis-infection and atypical mycobacterioses respectively, with more M. bovis-infection in cattle in the extensive management system and more atypical mycobacterioses in cattle in the intensive management system. From 8 out of 42 milk samples (19%) atypical mycobacteria were cultured. A higher prevalence of M. bovis-infection in the extensive sector could be due to several factors. In addition, such high prevalence puts herd owners and their families at risk for BTB. Therefore control of BTB, as well as education of cattle owners is important, especially in the extensive sector.     

An aneuploid mouse strain carrying human chromosome 21 with Down syndrome phenotypes

Aneuploidies are common chromosomal defects that result in growth and developmental deficits and high levels of lethality in humans. To gain insight into the biology of aneuploidies, we manipulated mouse embryonic stem cells and generated a trans-species aneuploid mouse line that stably transmits a freely segregating, almost complete human chromosome 21 (Hsa21). This "transchromosomic" mouse line, Tc1, is a model of trisomy 21, which manifests as Down syndrome (DS) in humans, and has phenotypic alterations in behavior, synaptic plasticity, cerebellar neuronal number, heart development, and mandible size that relate to human DS. Transchromosomic mouse lines such as Tc1 may represent useful genetic tools for dissecting other human aneuploidies.