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The MYB- basic helix-loop-helix (bHLH)-WD40 complexes regulating anthocyanin and proanthocyanidin (PA) biosynthesis in plants are not fully understood. Here Medicago truncatula bHLH MtTT8 was characterized as a central component of these ternary complexes that control anthocyanin and PA biosynthesis. Mttt8 mutant seeds have a transparent testa phenotype with reduced PAs and anthocyanins. MtTT8 restores PA and anthocyanin productions in Arabidopsis tt8 mutant. Ectopic expression of MtTT8 restores anthocyanins and PAs in mttt8 plant and hairy roots and further enhances both productions in wild-type hairy roots. Transcriptomic analyses and metabolite profiling of mttt8 mutant seeds and M. truncatula hairy roots (mttt8 mutant, mttt8 mutant complemented with MtTT8, or MtTT8 overexpression lines) indicate that MtTT8 regulates a subset of genes involved in PA and anthocyanin biosynthesis. MtTT8 is genetically regulated by MtLAP1, MtPAR and MtWD40-1. Combinations of MtPAR, MtLAP1, MtTT8 and MtWD40-1 activate MtTT8 promoter in yeast assay. MtTT8 interacts with these transcription factors to form regulatory complexes. MtTT8, MtWD40-1 and an MYB factor, MtPAR or MtLAP1, interacted and activated promoters of anthocyanidin reductase and anthocyanidin synthase to regulate PA and anthocyanin biosynthesis, respectively. Our results provide new insights into the complex regulation of PA and anthocyanin biosynthesis in M. truncatula.

Silica-filled polydimethylsiloxane (PDMS) composite membranes are prepared on a polytetrafluoroethylene support structure. The structure and the performance of the membranes are characterized by scanning electron microscope, Fourier transform infrared spectroscopy, X-ray diffraction, and thermogravimetry. The pervaporation process for acetic/water separation is performed within the PDMS membranes. The vulcanization temperature was found to have a great influence on the separation performance of the membrane. The addition of silica can significantly improve the pervaporation flux and enhance the thermal stability of the membrane. With an increase in the feed temperature, selectivity decreases and permeation flux increases. Performed with a pure PDMS membrane vulcanized at 30°C, the separation factor at first will increase, then decrease when the feed flow rate was increased from 14 to 38 L · h−1. The maximum separation factor is achieved when the feed flow rate is 26 L · h−1. POLYM. ENG. SCI., 2011. © 2011 Society of Plastics Engineers