Genome-wide association analysis for stripe rust resistance in spring wheat(Triticum aestivum L.) germplasm

Stripe rust is a continuous threat to wheat crop all over the world. It causes considerable yield losses in wheat crop every year. Continuous deployment of adult plant resistance(APR) genes in newly developing wheat cultivars is the most judicious strategy to combat this disease. Herein, we dissected the genetics underpinning stripe rust resistance in Pakistani wheat germplasm. An association panel of 94 spring wheat genotypes was phenotyped for two years to score the infestation of stripe rust on each accession and was scanned with 203 polymorphic SSRs. Based on D' measure, linkage disequilibrium(LD) exhibited between loci distant up to 45 c M. Marker-trait associations(MTAs) were determined using mixed linear model(MLM). Total 31 quantitative trait loci(QTLs) were observed on all 21 wheat chromosomes. Twelve QTLs were newly discovered as well as 19 QTLs and 35 previously reported Yr genes were validated in Pakistani wheat germplasm. The major QTLs were QYr.uaf.2 AL and QYr.uaf.3 BS(PVE, 11.9%). Dissection of genes from the newly observed QTLs can provide new APR genes to improve genetic resources for APR resistance in wheat crop.     

Silicon-mediated genotoxic alterations in Brassica juncea under arsenic stress: A comparative study of biochemical and molecular markers

Arsenic (As), one of the most harmful toxicant at the global level, severely affects plant metabolism when taken up. Interestingly, the presence of silicon (Si) as a fertilizer in As-contaminated soil is an effective strategy to decrease As accumulation in plants. Brassica juncea (var. Varuna) were grown hydroponically to investigate the role of Si at biochemical and molecular levels under arsenite (As³⁺) stress. Seedlings of B. juncea were exposed to As³⁺, Si, and a combination of both elements. Our data demonstrated that seedlings exposed to As³⁺ showed an inhibition in shoot length, chlorophyll, carotenoid, and protein, while co-application of Si improved these growth parameters. Silicon supplementation reduced As accumulation in shoot. Increase/decrease was observed in stress-related parameters (cysteine and proline), antioxidant enzymes (superoxide dismutase, ascorbate peroxidase, and catalase), and oxidative stress markers (malondialdehyde and H₂O₂), which were improved upon co-application of Si as compared to As³⁺ alone treatment. Random amplified polymorphic DNA (RAPD) is a suitable biomarker assay for plants for assessing the genotoxicity. Seven RAPD primers produced a total of 39 and 48 bands in the leaves of the untreated and treated seedlings, respectively. The RAPD band-profiles and genomic template stability were consistent with other growth and physiological parameters. In conclusion, the genotoxic alterations along with the biochemical parameters indicate that the exposure to Si mitigates As³⁺-induced oxidative stress by improving the stress-related parameters and antioxidant system in B. juncea.     

RNAi-mediated knockdown of INHBB increases apoptosis and inhibits steroidogenesis in mouse granulosa cells

Inhibins are members of the TGFβ superfamily and act as suppressors of follicle stimulating hormone (FSH) secretion from pituitary glands via a negative feedback mechanism to regulate folliculogenesis. In this study, the INHBB gene was knocked down by three RNAi-Ready pSIREN-RetroQ-ZsGreen vector- mediated recombinant plasmids to explore the effects of INHBB silencing on granulosa cell (GC) cell cycle, apoptosis and steroid production in vitro. Quantitative real-time polymerase chain reaction, Western blot, flow cytometry and ELISA were performed to evaluate the role of INHBB in the mouse GC cell cycle, apoptosis and steroid production in vitro. The results showed that the relative mRNA and protein expression of INHBB in mouse GCs can be significantly reduced by RNAi with pshRNA-B1, pshRNA-B2 and pshRNA-B3 plasmids, with pshRNA-B3 having the best knockdown efficiency. Downregulation of the expression of INHBB significantly arrests cells in the G1 phase of the cell cycle and increases the apoptosis rate in GCs. This was further confirmed by downregulation of the protein expressions of Cyclin D1, Cyclin E and Bcl2, while the protein expression of Bax was upregulated. In addition, specific downregulation of INHBB markedly decreased the concentration of estradiol and progesterone, which was further validated by the decrease in the mRNA levels of CYP19A1and CYP11A1. These findings suggest that inhibin βB is important in the regulation of apoptosis and cell cycle progression in granulosa cells. Furthermore, the inhibin βB subunit has a role in the regulation of steroid hormone biosynthesis. Evidence is accumulating to support the concept that inhibin βB is physiologically essential for early folliculogenesis in the mouse.     

Production of aromatic amino acids and related compounds from p-hydroxyphenylacetic acid by rumen microorganisms in vitro

Suspensions of mixed rumen bacteria (B), protozoa (P), and mixed rumen microorganisms (BP) prepared from rumen contents of fistulated goats were anaerobically incubated with 1 mM p‐hydroxyphenylacetic acid (HPA) at 39°C for 24 h. Tyrosine (Tyr), phenylalanine (Phe), tryptophan (Trp) and other related compounds in both supernatants and hydrolyzates of microbial cells in all incubations were analyzed by HPLC. Large amounts of Tyr (32.1, 42.7 and 36.1% of disappeared HPA in B, P and BP, respectively) were produced from HPA during a 12 h incubation period. The formation of Tyr in P (178.6 µmol/g MN) was 1.5 and 2 times higher than in B and BP, respectively. Phe (7–11% of the disappeared HPA) and Trp (3–6% of the disappeared HPA) were also synthesized from HPA in B, P, and BP. Phe synthesis in P (46.3 µmol/g MN) was 1.7 times higher than in B but, in contrast, Trp synthesis in B, was 1.6 times higher than in P. The metabolites p‐hydroxyphenylpyruvic acid (in the range of 5–14% of disappeared HPA), phenylacetic acid (1–11%), p‐hydroxybenzoic acid (3–7%) and benzoic acid (1–6%) were produced from HPA in B, P and BP. Phenylpropionic acid (6% of the disappeared HPA) was produced only in B and BP.     

Apparent digestibility coefficients for common feed ingredients in formulated diets for tropical catfish, Mystus nemurus (Cuvier & Valenciennes)

Abstract Apparent digestibility coefficients were determined for six feedstuffs, subsequent to their processing into pelleted feed for tropical freshwater catfish, Mystus nemurus (Cuvier & Valenciennes), using a reference diet and test diets composed of 70% reference diet and 30% test ingredient. It was found that most ingredients were fairly well digested except maize and chicken viscera. Digestibility coefficient for protein (P), dry matter (DM) and energy (E) were: (1) fish meal: 97·8% P,97% DM. 77·88% E: (2) soybean meal: 86% P,95·55% DM, 67·89% E: (3) rice bran: 81% P,85·81% DM, 67·93% E: (4) copra meal: 79·90% P, 86% DM, 71·89% E: (5) maize: 51·93% P, 49·0% DM, 63·40% E and (6) chicken viscera: 37% P,29% DM, 52% E. Digestibility coefficients for feedstuffs indicated that animal protein product, such as fish meal and grain by-product, such as rice bran were more digestible than starchy and fibrous feed, e.g. maize. Chicken viscera was poorly digested largely due to inefficient processing. As most feedstuffs were well digested, ingredient selection should, therefore, be based primarily on their protein content, amino acid composition and palatability.     

Nitrogen management improves lodging resistance and production in maize (Zea mays L.) at a high plant density

Lodging in maize leads to yield losses worldwide.  In this study, we determined the effects of traditional and optimized nitrogen management strategies on culm morphological characteristics, culm mechanical strength, lignin content, root growth, lodging percentage and production in maize at a high plant density.  We compared a traditional nitrogen (N) application rate of 300 kg ha^–1 (R) and an optimized N application rate of 225 kg ha^–1 (O) under four N application modes: 50% of N applied at sowing and 50% at the 10th-leaf stage (N1); 100% of N applied at sowing (N2); 40% of N applied at sowing, 40% at the 10th-leaf stage and 20% at tasseling stage (N3); and 30% of N applied at sowing, 30% at the 10th-leaf stage, 20% at the tasseling stage, and 20% at the silking stage (N4).  The optimized N rate (225 kg ha^–1) significantly reduced internode lengths, plant height, ear height, center of gravity height and lodging percentage.  The optimized N rate significantly increased internode diameters, filling degrees, culm mechanical strength, root growth and lignin content.  The application of N in four split doses (N4) significantly improved culm morphological characteristics, culm mechanical strength, lignin content, and root growth, while it reduced internode lengths, plant height, ear height, center of gravity height and lodging percentage.  Internode diameters, filling degrees, culm mechanical strength, lignin content, number and diameter of brace roots, root volume, root dry weight, bleeding safe and grain yield were significantly negatively correlated with plant height, ear height, center of gravity height, internode lengths and lodging percentage.  In conclusion, treatment ON4 significantly reduced the lodging percentage by improving the culm morphological characteristics, culm mechanical strength, lignin content, and root growth, so it improved the production of the maize crop at a high plant density.