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Phased small-interfering RNAs (phasiRNAs) are a special class of small RNAs, which are generated in 21- or 24-nt intervals from transcripts of precursor RNAs. Although phasiRNAs have been found in a range of organisms, their biological functions in plants have yet to be uncovered. Here we show that phasiRNAs generated by the photopheriod-sensetive genic male sterility 1 (Pms1) locus were associated with photoperiod-sensitive male sterility (PSMS) in rice, a germplasm that started the two-line hybrid rice breeding. The Pms1 locus encodes a long-noncoding RNA PMS1T that was preferentially expressed in young panicles. PMS1T was targeted by miR2118 to produce 21-nt phasiRNAs that preferentially accumulated in the PSMS line under long-day conditions. A single nucleotide polymorphism in PMS1T nearby the miR2118 recognition site was critical for fertility change, likely leading to differential accumulation of the phasiRNAs. This result suggested possible roles of phasiRNAs in reproductive development of rice, demonstrating the potential importance of this RNA class as regulators in biological processes.

Summary: Post‐transcriptional gene silencing in plants results from independent activities of diverse small RNA types. In anthers of grasses, hundreds of loci yield noncoding RNAs that are processed into 21‐ and 24‐nucleotide (nt) phased small interfering RNAs (phasiRNAs); these are triggered by miR2118 and miR2275.We characterized these 'reproductive phasiRNAs' from rice (Oryza sativa) panicles and anthers across seven developmental stages. Our computational analysis identified characteristics of the 21‐nt reproductive phasiRNAs that impact their biogenesis, stability, and potential functions.We demonstrate that 21‐nt reproductive phasiRNAs can function in cis to target their own precursors. We observed evidence of this cis regulatory activity in both rice and maize (Zea mays). We validated this activity with evidence of cleavage and a resulting shift in the pattern of phasiRNA production.We characterize biases in phasiRNA biogenesis, demonstrating that the Pol II‐derived 'top' strand phasiRNAs are consistently higher in abundance than the bottom strand. The first phasiRNA from each precursor overlaps the miR2118 target site, and this impacts phasiRNA accumulation or stability, evident in the weak accumulation of this phasiRNA position. Additional influences on this first phasiRNA duplex include the sequence composition and length, and we show that these factors impact Argonaute loading. [ABSTRACT FROM AUTHOR]

Chicken egg yolk immunoglobulin (IgY) is a superior alternative to mammalian immunoglobulin. However, the practical application of IgY in research, diagnostics, and functional food is limited due to complex or time-consuming purification procedures. The objective of this study was to develop a simple, safe, large-scale separation method for IgY from egg yolk. Egg yolk was diluted with 6-fold delipidation solutions made of different types (pectin, λ-carrageenan, carboxymethylcellulose, methylcellulose, and dextran sulfate) and concentrations (0.01, 0.05, 0.1, 0.15, and 0.2%) of polysaccharides, respectively. The yolk solution was adjusted to pH 5.0, and then kept overnight at 4°C before being centrifuged at 4°C. The resulting supernatant was added to 35% (w/v) (NH4)2SO4 and then centrifuged. The precipitant, which contained IgY, was dissolved in distilled water and then dialyzed. SDS-PAGE and Western blotting were utilized to conduct qualitative analysis of IgY; high-performance liquid chromatography (HPLC) was used for quantitative analysis. The immunoreactivity of IgY was measured by ELISA. The results showed that yield, purity, and immunoreactivity varied with types and concentrations of polysaccharides. The optimal isolation of IgY for pectin, λ-carrageenan, dextran sulfate, and carboxymethylcellulose was at the concentration of 0.1%; for methylcellulose, optimal isolation was at 0.15%. The best results were obtained in the presence of 0.1% pectin. In this condition, yield and purity can reach 8.36 mg/mL egg yolk and 83.3%, respectively, and the negative effect of IgY on immunoreactivity can be minimized. The procedure of isolation was simplified to 2 steps with a higher yield of IgY, avoiding energy- and time-consuming methods. Therefore, the isolation condition under study has a great potential for food industry production of IgY on a large scale. [ABSTRACT FROM AUTHOR]

An economical, effective and large-scale method for the preparation of immunoglobulin of chicken egg yolk (IgY) was developed. The method was based on an improved water dilution combining with polyethylene glycol 6000 (PEG6000) precipitation and anion-exchange chromatography. The egg yolk was diluted with 8 volumes of aseptic water, the pH of which was adjusted to 5.2 with 0.1 mol/L HCl, then the mixture was centrifuged at 5 000 x g after standing for 8 h at 4 degrees C. The crude IgY was obtained from the supernatant layer with the yield of 93.47%. The purification of IgY was achieved by 6% PEG6000 precipitation and DEAE-Toyo-pearl 650 M anion-exchange chromatographic packing. The optimum purified conditions by ion-exchange chromatography were as follows: equilibration with 0.05 mol/L phosphate buffer (pH 7.0) and elution with 0. 075 mol/L phosphate buffer (pH 7.0). The purity of the IgY fraction was consistently greater than 95% with high activity (73.77%) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). This method is superior to the traditional ion-exchange chromatography that can't simultaneously obtain high purity and high recovery. Moreover, this method is suitable for a large-scale IgY preparation from chicken egg yolk.