Skin color is an important factor in pear breeding programs. The degree of red coloration is determined by the content and composition of anthocyanins. In plants, many MYB transcriptional factors are involved in regulating anthocyanin biosynthesis. In this study, a R2R3-MYB transcription factor gene, PyMYB10, was isolated from Asian pear (Pyrus pyrifolia) cv. 'Aoguan'. Sequence analysis suggested that the PyMYB10 gene was an ortholog of MdMYB10 gene, which regulates anthocyanin biosynthesis in red fleshed apple (Malus x domestica) cv. 'Red Field'. PyMYB10 was identified at the genomic level and had three exons, with its upstream sequence containing core sequences of cis-acting regulatory elements involved in light responsiveness. Fruit bagging showed that light could induce expression of PyMYB10 and anthocyanin biosynthesis. Quantitative real-time PCR revealed that PyMYB10 was predominantly expressed in pear skins, buds, and young leaves, and the level of transcription in buds was higher than in skin and young leaves. In ripening fruits, the transcription of PyMYB10 in the skin was positively correlated with genes in the anthocyanin pathway and with anthocyanin biosynthesis. In addition, the transcription of PyMYB10 and genes of anthocyanin biosynthesis were more abundant in red-skinned pear cultivars compared to blushed cultivars. Transgenic Arabidopsis plants overexpressing PyMYB10 exhibited ectopic pigmentation in immature seeds. The study suggested that PyMYB10 plays a role in regulating anthocyanin biosynthesis and the overexpression of PyMYB10 was sufficient to induce anthocyanin accumulation.
Generation of genetically uniform non-human primates may help to establish animal models for primate biology and biomedical research. In this study, we have successfully cloned cynomolgus monkeys (Macaca fascicularis) by somatic cell nuclear transfer (SCNT). We found that injection of H3K9me3 demethylase Kdm4d mRNA and treatment with histone deacetylase inhibitor trichostatin A at one-cell stage following SCNT greatly improved blastocyst development and pregnancy rate of transplanted SCNT embryos in surrogate monkeys. For SCNT using fetal monkey fibroblasts, 6 pregnancies were confirmed in 21 surrogates and yielded 2 healthy babies. For SCNT using adult monkey cumulus cells, 22 pregnancies were confirmed in 42 surrogates and yielded 2 babies that were short-lived. In both cases, genetic analyses confirmed that the nuclear DNA and mitochondria DNA of the monkey offspring originated from the nucleus donor cell and the oocyte donor monkey, respectively. Thus, cloning macaque monkeys by SCNT is feasible using fetal fibroblasts.
The effects of ultrasound on the molecular weight of apple pectin were investigated. The structure and rheological properties of the degradation products were also tentatively identified by High Performance Liquid Chromatography-Photodiode Array Detector (HPLC-PAD), Infrared spectroscopy (IR), Nuclear Magnetic Resonance spectroscopy (NMR) and Rheometer. The results indicated that the weight-average molecular weight of apple pectin decreased obviously after ultrasound treatment. The molecular weight of degradation products had a uniform and narrow distribution. Ultrasound intensity and temperature play an important role in the degradation reaction. Degradation kinetics model of apple pectin fitted to 1/M(t) - 1/M(0) = kt from 5 to 45 °C. The degree of methylation of apple pectin reduced according to IR analysis when ultrasound was applied. Ultrasound treatment could not alter the primary structure of apple pectin according to the results determined by HPLC, IR and NMR. Meanwhile, the viscosity of apple pectin was 10(3) times as large as that of ultrasound-treated apple pectin. The ultrasound-treated apple pectin showed predominantly viscous responses (G' < G") over the same frequency range. The results suggested that ultrasound provided a viable alternative method for the modification of pectin.
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