This paper describes a new protocol to develop doubled-haploid (DH) Brassica napus lines with improved resistance to Sclerotinia sclerotiorum. In this protocol, haploid seedlings derived from microspore cultures of B. napus were used to produce haploid calli for in vitro mutation-selection. For routine screening, mutation was induced by EMS (ethylmethane sulfonate) or occurred spontaneously, and screening for resistant mutants occurred on media with added oxalic acid (OA) as a selection agent. In tests with selected lines, the optimal concentration of EMS for mutation was determined to be 0.15%, and the optimal concentration of OA for in vitro screening was 3 mmol/l (half lethal dose was 3.1 mmol/l) for the first cycle of screening. There was an accumulated effect of OA toxicity on calli over two cycles of screening, but the growth and capacity of the surviving calli for regenerating seedlings were not affected by OA. Of the 54 DH lines produced from the in vitro mutation-selection, two DH lines of resistant mutants, named M083 and M004, were selected following seedling and glasshouse tests. The resistance of M083 and M004 to S. sclerotiorum following tests with both mycelial inoculum and OA was greater than that of their donor lines and the resistant control Zhongyou 821. In both glasshouse and field disease nurseries, disease indices on M083 and M004 were less than 50% of those of the control. The time required for M083 and M004 to mature was 14 days and 10 days shorter, respectively, than that of their donor lines. Furthermore, M083 had more pods per inflorescence, a greater 1,000 seed weight and higher yield than its donor line. Random amplified polymorphic DNA characterisation showed that M083 had DNA band patterns that differed from its donor line.
The aim of this study was to compare the effect of DL-selenomethionine (SM) with 2 routinely used Se sources, sodium selenite (SS) and seleno-yeast (SY), on relative bioavailability based on antioxidant activity and tissue Se content. Six hundred thirty 131-day-old brown laying hens were randomly assigned to 7 treatments for 168 d (24 wks) with 6 replicates of 15 hens per replicate. The SS and SY animals were supplemented a cornmeal and soybean diet that supplied a total Se 0.3 mg/kg whereas SM was added at 4 different levels to the total Se at 0.1, 0.3, 0.5 and 0.7 mg/kg. All hens fed the Se-supplemented diet showed higher glutathione peroxidase (GSH-Px) activity (P < 0.01), higher superoxide dismutase (SOD) activity (P < 0.05), lower malondialdehyde (MDA) content (P < 0.05) in plasma, and greater Se contents in egg yolks, albumen, leg muscle, breast muscle, liver, and plasma compared with those fed the control diet (P < 0.01). The organic sources (SY and SM) exhibited a greater ability to increase the GSH-Px activity (P < 0.01) and Se content in albumen (P < 0.01), leg, and breast muscles (P = 0.0099 and P = 0.0014, respectively) than the SS that was added at 0.3 mg Se/kg. The higher SM added levels increased the GSH-Px activity until the dose of 0.5mg Se/kg (P < 0.01).The greater Se concentrations in albumen, muscle and liver appeared in the higher SM-added level, as well as above the dose of 0.1 mg Se/kg (P < 0.01). In addition, hens fed the diet with SM accumulated more Se in albumen, leg, and breast muscle than those fed diets with SY (P < 0.05). These results confirmed the higher ability of organic Se sources to increase the antioxidant activity and Se deposition in egg albumen, leg, and breast muscles compared with SS, and demonstrated a significantly better efficiency of SM compared with SY for albumen and muscle Se enrichment.
Genomes carry millions of noncoding variants, and identifying the tiny fraction with functional consequences is a major challenge for genomics. We assessed the role of selection on long noncoding RNAs (lncRNAs) for domestication-related changes in rice grains. Among 3363 lncRNA transcripts identified in early developing panicles, 95% of those with differential expression (329 lncRNAs) between Oryza sativa ssp. japonica and wild rice were significantly down-regulated in the domestication event. Joint genome and transcriptome analyses reveal that directional selection on lncRNAs altered the expression of energy metabolism genes during domestication. Transgenic experiments and population analyses with three focal lncRNAs illustrate that selection on these loci led to increased starch content and grain weight. Together, our findings indicate that genome-wide selection for lncRNA down-regulation was an important mechanism for the emergence of rice domestication traits.
This study evaluated the effects of long-term Bacillus subtilis CGMCC 1.921 supplementation on the performance, egg quality, and fecal/cecal microbiota of laying hens. A total of 360 28-week-old Hy-Line Brown laying hens were randomly allocated into 5 treatments with 6 replicates of 12 birds each for 24 weeks. The experimental treatments included a basal diet without additions (Con) and the basal diet supplemented with 1.0 × 105 (B1), 1.0 × 106 (B2), 1.0 × 107 (B3), and 1.0 × 108 (B4) cfu/g B. subtilis CGMCC 1.921. The results showed that feed:egg ratio significantly decreased (P < 0.05) in groups B1 (wk 13 to 16, 17 to 20, 21 to 24, and one to 24), B2 (wk 13 to 16, 17 to 20, and 21 to 24), B3 (wk 13 to 16, 17 to 20, 21 to 24, and one to 24), and B4 (wk 13 to 16, 17 to 20, 21 to 24, and one to 24). However, egg production, egg weight, and feed intake were not significantly different (P > 0.05) among treatments. Eggshell strength significantly improved (P < 0.05) in groups B1 (wk 8, 16, 20, and 24), B2 (wk 20 and 24), and B3 (wk 8, 16, 20, and 24). Fecal E. coli counts significantly decreased (P < 0.05) in groups B1 (wk 16), B2 (wk 12, 16, 20, and 24), B3 (wk 12, 20, and 24), and B4 (wk 16, 20, and 24). Lactobacillus in cecal digesta of groups B1, B3, and B4 increased significantly (P < 0.01). Bifidobacterium in cecal digesta of groups B1, B2, B3, and B4 increased significantly (P < 0.05). Bifidobacterium counts increased linearly (P = 0.015) and quadratically (P = 0.004) as B. subtilis CGMCC 1.921 supplementation increased. Compared with Con, E. coli in the cecal digesta of groups B2 and B4 decreased significantly (P < 0.01). C. perfringens in the cecal digesta of groups B3 and B4 decreased significantly (P < 0.05). E. coli:Lactobacillus ratio decreased in group B1 (P < 0.05) and B2, B3, and B4 (P < 0.01). Therefore, the probiotic B. subtilis CGMCC 1.921 effectively improved performance and egg quality via the reduction of fecal E. coli and beneficial modulation of cecal microbiota.
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