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Morphology observations were carried under a stereomicroscope (SZX16, Olympus, Tokyo). Samples were fixed in 2.5% glutaraldehyde solution. Fixed samples were dehydrated with gradual ethanol series, dried by critical-point drying method using liquid carbon dioxide (Model HCP-2, Hitachi, Tokyo), gold-coated with an Edwards E-1010 ion sputter coater (Hitachi, Tokyo), and then observed using a S-3000N variable pressure scanning electron microscope (Hitachi, Tokyo). The dp1 locus was mapped by using an F2 population of dp1�C2 and Sheng47 (spp. indica). The locus was mapped selleck chemicals llc to a region between cleaved-amplified polymorphic sequence (CAPS) markers M4 and M6 on the short arm of chromosome 6 ( Luo et al., 2005). We further developed two new CAPS markers M9 and dM1 in this region (Table S2) to narrow the locus to a 10?kb region between M4 and dM1 ( Fig.?6A). The sdp1 locus was mapped by using an F2 population of sdp1 and Minghui 63 (spp. indica). The locus was mapped to a 92?kb region between two sequence-tagged site markers, S14533 and S14625 (Fig. S6). Primer sequences used for vector construction diglyceride are listed in Table S2 in the Supplementary data. For complementation of the dp1�C2 mutant, a 9292?bp genomic fragment containing the entire DP1 coding sequence, 5728?bp of the 5�� upstream region and 2577?bp of the 3�� downstream region was digested form PAC clone P0548D03, and cloned into the pCAMBIA1300 vector to generate plasmid p1300-DP1. For RNA interference analysis, a fragment of the DP1 cDNA (864?bp�C1276?bp of the coding region) was amplified and cloned into pUCCRNAi vector by forward and reverse insertions. The entire fragment was subcloned into pCAMBIA1300A under selleck products the rice ACTIN1 promoter. For pDP1::GUS and pDP1::DP1-GFP (green fluorescent protein), specific primers with suitable adaptors were designed to amplify relative sequences (Table S2), and cloned into pCAMBIA1301 or CAMV35S-sGFP(S65T)-Nos ( Niwa et al., 1999), respectively. These vectors were subsequently transformed into calli derived from mature rice seeds through Agrobacterium-mediated methods ( Hiei et al., 1994). Total RNA was extracted using TRIZOL (Invitrogen, Carlsbad). The RNA was pre-treated with RNase-free DNase I (Takara, Shiga), and first-strand cDNA was synthesized from 2?��g of total RNA using reverse transcriptase (M-MLV, Promega, Madison). The reverse transcription product was used for PCR with gene specific primers (Table S2). Rice ACTIN1 was used as the internal reference. For qRT-PCR, SYBR Green I was added to the reaction system and run on a Chromo 4 real-time PCR detection system (Bio-Rad, Hercules) according to the manufacturer's instructions. Three replicates were carried out for each gene, and each analysis was biologically repeated at least twice. Student's t-test was used to determine significant changes (P?