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A WD40 Repeat Protein from Medicago truncatula Is Necessary for Tissue-Specific

A WD40Repeat Protein from Medicago truncatula Is Necessary for Tissue-Speciﬁc Anthocyanin and Proanthocyanidin Biosynthesis But Not forTrichome Development1[W][OA]Yongzhen Pang,Jonathan P.Wenger,Katie Saathoff,Gregory J.Peel2,Jiangqi Wen,David Huhman,Stacy N.Allen,Yuhong Tang,Xiaofei Cheng,Million Tadege,Pascal Ratet,Kirankumar S.Mysore,Lloyd W.Sumner,M.David Marks,and Richard A.Dixon*Plant Biology Division,Samuel Roberts Noble Foundation,Ardmore,Oklahoma73401(Y.P.,G.J.P.,J.W.,D.H., S.N.A.,Y.T.,X.C.,M.T.,K.S.M.,L.W.S.,R.A.D);Department of Genetics and Cell Biology,University of Minnesota,St.Paul,Minnesota55108(J.P.W.,K.S.,M.D.M.);and Institut des Sciences du Vegetale,CNRS, 91198Gif sur Yvette,France(P.R.)WD40repeat proteins regulate biosynthesis of anthocyanins,proanthocyanidins(PAs),and mucilage in the seed and the development of trichomes and root hairs.We have cloned and characterized a WD40repeat protein gene from Medicago truncatula(MtWD40-1)via a retrotransposon-tagging approach.Deﬁciency of MtWD40-1expression blocks accumulation of mucilage and a range of phenolic compounds,including PAs,epicatechin,otherﬂavonoids,and benzoic acids,in the seed, reduces epicatechin levels without corresponding effects on otherﬂavonoids inﬂowers,reduces isoﬂavone levels in roots,but does not impair trichome or root hair development.MtWD40-1is expressed constitutively,with highest expression in the seed coat,where its transcript proﬁle temporally parallels those of PA biosynthetic genes.Transcript proﬁle analysis revealed that many genes ofﬂavonoid biosynthesis were down-regulated in a tissue-speciﬁc manner in M.truncatula lines harboring retrotransposon insertions in the MtWD40-1gene.MtWD40-1complemented the anthocyanin,PA,and trichome phenotypes of the Arabidopsis(Arabidopsis thaliana)transparent testa glabrous1mutant.We discuss the function of MtWD40-1in natural product formation in M.truncatula and the potential use of the gene for engineering PAs in the forage legume alfalfa(Medicago sativa).Anthocyanins and proanthocyanidins(PAs;also called condensed tannins)areﬂavonoids that beneﬁt both plant and human health.Anthocyanins attract pollinators,protect plant tissues from UV light dam-age,and defend plants against predators(Stapleton and Walbot,1994;Sullivan,1998).PAs are abundant in beverages such as tea,wine,and fruit juice and exhibit antioxidant activity and cardiovascular protective ef-fects(Bagchi et al.,2000;Cos et al.,2004;Dixon et al., 2005).Moreover,a moderate PA level is an important quality trait in forage crops,because PAs can protect ruminant animals from lethal pasture bloat by binding proteins and thereby slowing down their fermentation in the rumen(Li et al.,1996;Aerts et al.,1999;Barry and McNabb,1999;Dixon et al.,2005).The PA biosynthetic pathway in Arabidopsis(Arab-idopsis thaliana)has been studied primarily through the analysis of transparent testa(tt)or transparent testa glabrous(ttg)mutants,which exhibit seed coat(tt)or seed coat and trichome(ttg)phenotypes(Shirley et al., 1995;Lepiniec et al.,2006).The mutated genes have been found to encode either pathway enzymes or transcriptional regulators that function alone or in complexes to control the whole or branches of the pathway(Lepiniec et al.,2006).Anthocyanins and PAs share the same upstream phenylpropanoid/ﬂavonoid pathway,and anthocyanidin is the immediate sub-strate for both glycosylation to anthocyanin or reduc-tion to epicatechin in the biosynthesis of PAs in Arabidopsis(Fig.1).We are studying the formation of PAs in the model legume Medicago truncatula(Xie et al.,2003,2006;Pang et al.,2007,2008).Four structural genes encoding anthocyanidin synthase(ANS),leucoanthocyanidin reductase(LAR),anthocyanidin reductase(ANR), and epicatechin3#-O-glucosyltransferase(UGT72L1) were characterized biochemically and/or genetically1This work was supported by the National Science Foundation Plant Genome Program(grant nos.DBI–0605033and DBI–0703285to R.A.D.and K.S.M.,respectively),by Forage Genetics International, and by the Samuel Roberts Noble Foundation.2Present address:Calgene/Monsanto,19205th Street,Davis,CA 95616.*Corresponding author;e-mail radixon@.The author responsible for distribution of materials integral to the ﬁndings presented in this article in accordance with the policy described in the Instructions for Authors()is: Richard A.Dixon(radixon@).[W]The online version of this article contains Web-only data.[OA]Open Access articles can be viewed online without a sub-scription./cgi/doi/10.1104/pp.109.144022from this species (Xie et al.,2003;Pang et al.,2007,2008).However,little is known about the regulatory network involved in anthocyanin/PA biosynthesis in M.truncatula .A regulatory complex,comprising an R2R3-MYB transcription factor,a basic helix-loop-helix (bHLH)domain protein,and a WD40repeat protein,regulates production of anthocyanins in foliar tissues and PAs and mucilage in seed coats;this complex also controls the formation of root hairs and trichomes on aerial tissues in some but not all plants (Baudry et al.,2004;Broun,2005;Lepiniec et al.,2006;Morita et al.,2006;Serna and Martin,2006;Gonzalez et al.,2008;Zhao et al.,2008).In Arabidopsis,these proteins are encoded by Transparent Testa2(TT2,Myb ),Transparent Testa8(TT8,HLH ),and Transparent Testa Glabrous1(TTG1,WD40repeat ),which together regulate the late ﬂavonoid pathway genes and the PA-speciﬁc pathway gene ANR (Baudry et al.,2004).Loss of function of either TT2or TT8leads to a lack of anthocyanin pigmentation in foliar tissue and a loss of PAs in the seed coat (Nesi et al.,2000,2001).The presence of TTG1is essential and irreplaceable in this complex for anthocyanin/PA biosynthesis,trichome formation,seed mucilage production,and root hair formation (Koornneef,1981;Walker et al.,1999).Several other WD40repeat proteins functionally orthologous to TTG1have been described from other species such as petunia (Petunia hybrida ),Perilla frutescens ,cotton (Gossypium hirsutum ),and maize (Zea mays );mutationof some affects both anthocyanin/PA and trichome phenotypes,whereas mutation of others only affects the anthocyanin/PA phenotype (Lloyd et al.,1992;de Vetten et al.,1997;Sompornpailin et al.,2002;Carey et al.,2004;Humphries et al.,2005).In an attempt to identify genes involved in the regulation of anthocyanin and PA biosynthesis in M.truncatula ,we have screened a Tnt1retrotransposon insertion population for altered leaf (lack of red pig-ment)and seed (transparent testa)phenotypes.This led to the cloning and functional characterization of a gene,MtWD40-1,with high sequence identity to known WD40repeat proteins.MtWD40-1can comple-ment the Arabidopsis ttg1PA and trichome pheno-types,although the Medicago wd40-1mutant retained normal trichomes.Loss of MtWD40-1function has profound and differential effects on ﬂavonoid biosyn-thesis in different plant organs.The potential of MtWD40-1for engineering the PA pathway in alfalfa (Medicago sativa )was also investigated.RESULTSPhenotypic and Genotypic Characterization of M.truncatula Retrotransposon Insertion MutantsOne mutant line (NF0977)drew our attention when screening the M.truncatula Tnt1insertion population for visible anthocyanin phenotypes.This line lacked the typical red pigmentation in the stem,the anthocyanin-rich circle at the base of the axial side of the leaﬂet,and the small red spots on the adaxial side of the leaﬂet,all of which are seen in wild-type ecotype R108(Fig.2,A and B).The seed coat of this mutant line was transpar-ent with a yellowish color that contrasted with the brown pigmentation of the wild type that arises from the presence of oxidized PAs (Fig.2C).To further conﬁrm the PA phenotype,seeds were stained with dimethylaminocinnamaldehyde (DMACA),a reagent that is speciﬁc for PAs and their ﬂavan 3-ol precursors.Mature seeds from the mutant line did not exhibit the typical blue staining characteristic of the reaction of PAs with DMACA (Fig.2C).The seeds from the mutant also produced less mucilage than those of the wild type,as seen by the reduced staining of the seed coat with ruthenium red (Fig.2C).No other obvious phenotypes,such as altered density of glandular or nonglandular trichomes (Fig.2D)or root hairs (Fig.2E),were ob-served in the NF0977mutant.Root hair density ap-peared to be unaffected on both young (4d after germination;Fig.2E)and mature (Supplemental Fig.S1)roots.One of 12plants from the NF0977R2generation exhibiting the lack of pigmentation phenotype was allowed to undergo self-pollination.All 29plants from the R3generation were homozygous,as con-ﬁrmed by PCR with gene-speciﬁc primers and a primer for the Tnt1insert,and retained the visible mutant phenotypes as characterized in Figure 2,AtoFigure 1.The ﬂavonoid pathway leading to anthocyanins and PAs.Enzymes are as follows:PAL,L -Phenylalanine ammonia-lyase;C4H,cinnamate 4-hydroxylase;4CL,4-coumarate:CoA ligase;CHS,chal-cone synthase;CHI,chalcone isomerase;CHR,chalcone reductase;F3H,ﬂavanone 3-hydroxylase;DFR,dihydroﬂavonol reductase;FS,ﬂavone synthase;IFS,isoﬂavone synthase;LAR,leucoanthocyanidin reductase;ANS,anthocyanidin synthase;ANR,anthocyanidin reduc-tase;GT,glucosyltransferase.A WD40Repeat Protein from Medicago truncatulae of thermal asymmetric interlaced (TAIL)-PCR revealed that all individuals possessed a retrotrans-poson insertion in a WD40gene with similarity to the TTG1gene from Arabidopsis.After sequencing and alignment using the available M.truncatula genome database,this Tnt1insertion was found to be be-tween the ﬁrst and second nucleotides of amino acid residue Ser-31of the WD40protein in the NF0977mutant (Fig.2F).A further 20insertion sites in different regions of the genome were also recovered from NF0977(Supplemental Table S1),typical for Tnt1insertional mutagenesis in Medicago (Tadege et al.,2008).None of these insertions was in a gene that would be expected to affect ﬂavonoid biosyn-thesis,although this does not rule out the possibility that the lack of pigmentation phenotype could have been the result of an insertion in one or more of these genes.Therefore,a reverse genetic approach was employed to screen the Tnt1insertion mutant pop-ulation for additional lines with insertions in the WD40gene,and another mutant line,NF2745,wasobtained.The insertion site in line NF2745was between amino acid residues Ser-46and Ile-47(Fig.2F).Homozygous NF2745plants exhibited the same phenotype as NF0977(Fig.2,A–E),strongly suggest-ing that the loss of function of the WD40gene is responsible for the pigmentation phenotypes in the two mutants.Molecular Cloning and Characterization of MtWD40-1BLASTX analysis of the partial WD40sequence against the GenBank database showed that this gene was located on the M.truncatula bacterial artiﬁcial chromosome clone CR940305.Its full-length sequence was predicted to be 1,363bp in length with a 49-bp 5#untranslated region and a 285-bp 3#untranslated region (designated as MtWD40-1;GenBank accession no.EU040206).MtWD40-1is a single-copy gene lack-ing introns,as conﬁrmed by DNA gel-blot analysis and ampliﬁcation of the MtWD40-1open reading frame (ORF)with genomic DNA as template(dataFigure 2.Visible phenotypes result-ing from insertional mutagenesis of MtWD40-1.A,Top,4-d-old seedlings of M.truncatula R108(wild type)and the two insertional mutant lines,show-ing pigmentation below the cotyle-dons.Bottom,aerial parts of older seedlings,shown in the same order.B,Axial side (bottom)and adaxial side (top)of leaves from a wild-type plant (left),NF0977(center),and NF2745(right).C,Mature seeds of the wild type (left),NF0977(center),and NF2745(right),either unstained (top),stained with DMACA reagent to detect PAs (center),or stained with ruthenium red to detect mucilage (bottom).D,Scan-ning electron microscopy analysis of trichomes on young petioles and leaves of the wild type (left),NF0977(center),and NF2745(right).The top panels show nonglandular petiole tri-chomes,the center panels show non-glandular leaf trichomes,and the bottom panels show glandular and nonglandular petiole trichomes.Bars =1mm in top and center panels and 200m m in bottom panels.E,Root hair phenotypes of the wild type (left),NF0977(center),and NF2745(right).Bars =2mm in top panels and 1mm in bottom panels (showing closeups of the hairs just behind the root tip).F,A diagram of the MtWD40-1gene (1,364bp)showing the positions of the inde-pendent Tnt1insertions and the two probe sets on the Medicago Affymetrix GeneChip.Pang et al.not shown).MtWD40-1encodes a predicted protein ORF of343amino acids,with a calculated pI of4.99 and a molecular mass of38kD.The deduced amino acid sequence of MtWD40-1 showed77%to79%identity to other known WD40 repeat proteins from different plant species,such as TTG1from Arabidopsis and AN11from petunia(Fig.3).The four WD40repeat domains are highly con-served among all the WD40repeat proteins,including MtWD40-1,and the last two amino acids in each WD40repeat are identical.Phylogenetic analysis(Fig.4)showed that MtWD40-1is most closely related to TTG1from Arabidopsis.Another Medicago WD40-like protein,MtWD40-2,is less than60%identical to MtWD40-1at the amino acid level and somewhat closer to PAC1frommaize.Figure3.Alignment of deduced amino acid se-quences of plant WD40repeat proteins.The WD40 repeat domains are marked with horizontal bars above the sequences,and the last two amino acids of each repeat domain are marked with stars.Identical residues are highlighted on a black background,and similar residues are highlighted on a gray background.The GenBank accession numbers are as follows:BAE94398, InWDR1from Ipomoea nil;BAE94396,IpWDR1from Ipomoea purpurea;AAC18914,AN11from Petunia hybrida;BAB58883,PFDS from Perilla frutescens; AAM95645,GhTTG3from Gossypium hirsutum; AAK19614,GhTTG1;ABW08112,MtWD40-1; Q9XGN1,AtTTG1;AAM76742,PAC1from Zea mays; AC136505_16.4,MtWD40-2.A WD40Repeat Protein from Medicago truncatulaMtWD40-1Complements the Arabidopsis ttg1andMedicago NF0977Mutants by Interacting with Glabrous3Hairy roots of M.truncatula R108exhibit red antho-cyanin pigmentation (Pang et al.,2008),but this was lacking in the NF0977line.Hairy root transformation,therefore,was used as a rapid method to conﬁrm that MtWD40-1could complement the lack-of-pigment phenotype of the NF0977Tnt1insertion mutant.Red pigmentation was seen in all 101phosphinothricin (ppt)-resistant hairy root lines transformed with MtWD40-1but in none of the 30ppt-resistant NF0977lines transformed with the GUS gene (Fig.5A).Quan-titative reverse transcription (qRT)-PCR conﬁrmed that MtWD40-1,ANS ,and the anthocyanin-speciﬁc gluco-syltransferase UGT78G1(Modolo et al.,2007;Peel et al.,2009)were expressed at higher levels in hairy roots of the MtWD40-1-transformed lines than in the GUS transformants (Fig.5,B–D),thus accounting for the high levels of extractable anthocyanins in the MtWD40-1-expressing lines (Fig.5E).No signiﬁcant differences were observed in the levels of insoluble PAs (PAs that bind to the cell wall and cannot be extracted by organic solvents such as 70%acetone)between the MtWD40-1-expressing NF0977lines and the GUS control lines (Fig.5G)or in the levels of transcripts encoding the PA pathway-speciﬁc genes ANR and UGT72L1(data not shown).In contrast,soluble PA levels decreased slightly in the mutant line complemented with MtWD40-1(Fig.5F),possibly as a result of ﬂux into soluble PAs being diverted back into the anthocyanin pathway.To determine whether MtWD40-1is a functional ortholog of TTG1,the MtWD40-1ORF under the control of the 35S promoter was transformed into the Arabidopsis ttg1-9mutant,and expression of the foreign MtWD40-1gene was conﬁrmed by qRT-PCR (Supplemental Fig.S2).35S :MtWD40-1fully comple-mented the anthocyanin pigmentation,trichome deﬁ-ciency,and seed coat PA phenotypes (Fig.6,A–C).We also tested the ability of MtWD40-1to complement the Arabidopsis ttg1-9mutant when expressed under the control of the Arabidopsis Glabrous2(GL2)pro-moter,which is active in the shoots of ttg1mutants(Szymanski et al.,1998).Again,the phenotype was fully rescued (Fig.6,D–F).To further determine how MtWD40-1might func-tion to restore the trichome phenotype in ttg1-9Arab-idopsis,the yeast two-hybrid system was used to test the interaction of MtWD40-1with GL3,a bHLH pro-tein that regulates trichome development in Arabi-dopsis through interaction with GL1and TTG1(Payne et al.,2000).GL3was fused to the activation domain (AD)of GAL4,and MtWD40-1was fused to the binding domain (BD)of GAL4.Yeast containing empty pGAD424(AD)and pBridge (BD)vectors in conjunc-tion with MtWD40-1did not exhibit b -galactosidase activity (Fig.6G,top),whereas yeast containing GL3-AD and MtWD40-1BD exhibited strong activity (Fig.6G,bottom),suggesting that MtWD40-1can interact with GL3for trichome formation in Arabidopsis,even though it is not necessary for trichome formation in M.truncatula .Tissue-and Development-Speciﬁc Expression of MtWD40-1To determine the developmental expression pattern of MtWD40-1,normalized data were retrieved from the M.truncatula gene expression atlas (Benedito et al.,2008)together with seed coat microarray data (Pang et al.,2008).The expression patterns of two probe sets for MtWD40-1(TC105711and AL372205;probe set locations are shown in Fig.2F)were essentially the same,conﬁrming that,as is also the case for TTG1in Arabidopsis (Walker et al.,1999),MtWD40-1is ex-pressed in all organs,with highest expression in the seed coat (Fig.7A).During seed development,MtWD40-1showed its highest expression level at or before 10d after pollination (dap;Fig.7B),with a subsequent de-cline toward seed maturity.This expression pattern parallels the expression of MtANR and UGT72L1during seed development (Pang et al.,2008).We also analyzed the expression pattern of MtWD40-2in the M.truncatula gene expression atlas (Benedito et al.,2008),where it is represented by probe set Mtr.22605.1.S1_at (/gene-atlas/v2/).The highest expression levelisFigure 4.Unrooted phylogram comparison of the amino acid sequences of MtWD40-1and other func-tionally characterized plant WD40repeat proteins.The sequences used are the same as in Figure 3.The phylogenetic tree was constructed by PAUP*4.0b10,after alignment using MAFFT software.Node support was estimated using neighbor-joining bootstrap anal-ysis (1,000bootstrap replicates).Pang et al.in roots 24h after salt stress and in developing root nodules,but the expression level in these tissues is nearly 2orders of magnitude lower than the maximum expression level of MtWD40-1(in developing seeds).MtWD40-2is expressed around 15-fold lower than MtWD40-1in trichome-containing leaf and petiole tissues and is only expressed very weakly if at all in isolated nonglandular trichomes from M.truncatula (only called present in one out of three Affymetrix data sets;M.David Marks,unpublished data).Further-more,unlike MtWD40-1,MtWD40-2is not induced in M.truncatula hairy roots expressing Arabidopsis TT2.Tissue-Speciﬁc Effects of Loss of MtWD40-1Function on Phenylpropanoid/Flavonoid Pathway Gene Transcripts and MetabolitesTo determine the impacts of the loss of MtWD40-1function on gene expression in seeds,we dissected seeds at 16dap from both the NF0977mutant line and the corresponding wild-type control (ecotype R108)for microarray analysis using the Affymetrix Medicago GeneChip.We have previously shown that phenyl-propanoid/ﬂavonoid biosynthetic pathway genes are highly expressed at 16dap (Pang et al.,2007).The microarray data showed that 152probe sets were down-regulated more than 2-fold in the MtWD40-1mutant line;among these,three probe sets were down-regulated by more than 100-fold,25by more than 5-fold,with the remainder between 2-to 5-fold (Sup-plemental Table S2E).Classiﬁcation using the Gene-Bins ontology tool (.au/utils/GeneBins/index.php)showed that a high percentage (43.5%)of the down-regulated genes were “unclassiﬁed with homology”followed by “bio-synthesis of secondary metabolites”(25.9%;Supple-mental Fig.S3).This latter class consisted primarily of phenylpropanoid/ﬂavonoid pathway genes.Among the 28probe sets that exhibited a more than 5-fold reduction in expression level in the MtWD40-1mutant (Table I),17were associated with the phenylpropanoid/ﬂavonoid pathway and one had no homology to any known gene.The early phenyl-propanoid pathway genes PAL ,4CL ,CHS ,F3#H ,and F3#5#H were all down-regulated,almost 200-fold in the case of one CHS probe set (Table I;Supplemental Table S2).CHS is encoded by a large gene family in Medicago ,and nine different CHS probe setswereFigure 5.Genetic complementation of the anthocyanin and PA phe-notypes of the NF0977retrotransposon insertion line.A,Pigmentation of hairy roots of the NF0977line expressing GUS (left)and MtWD40-1(right).B,qRT-PCR analysis of MtWD40-1transcript levels in hairy roots of NF0977expressing GUS or MtWD40-1.C,As above,showing ANS transcript levels.D,As above,showing UGT78G1transcript levels.E,Anthocyanin levels from NF0977expressing GUS or MtWD40-1(three independent lines of each).F,As above,showing insoluble PA levels.G,As above,showing soluble PA levels.FW,Fresh weight.A WD40Repeat Protein from Medicago truncatuladown-regulated more than5-fold(Supplemental Table S2).The two later anthocyanin pathway genes,DFR and ANS,were down-regulated by2.6-fold and9.2/ 10.0-fold,respectively(Table I;Supplemental Table S2),suggesting that MtWD40-1regulates both early and later anthocyanin pathway genes in seeds.Three genes speciﬁc for the PA pathway,LAR,ANR,and UGT72L1,were down-regulated3.9-,34.6-,and14.7-fold,respectively,highlighting the speciﬁc involve-ment of MtWD40-1in the regulation of PA biosynthesis. MtWD40-1might also regulate additional branches of theﬂavonoid pathway,as seen by the40.6-fold and 2.1-fold reductions in expression ofﬂavonol synthase and a putative isoﬂavone O-glycosyltransferase in the MtWD40-1mutant.Another271probe sets were up-regulated in seeds of the mutant,most of them associated with primary metabolism or stress responses,but no phenylpropa-noid/ﬂavonoid pathway genes were up-regulated (data not shown).The large number of changes observed in nonphen-ylpropanoid/ﬂavonoid pathway genes in the above experiment could potentially occur as a result of the additional retrotransposon insertions in line NF0977. Therefore,we reexamined changes in keyﬂavonoid pathway gene transcripts in seeds and other organs,in both NF0977and the independent retrotransposon insertion line NF2745,using qRT-PCR(Table II). MtWD40-1transcript levels were more strongly down-regulated in tissues of NF2745than in NF0977 (Table II;Supplemental Tables S3and S4).Compared with wild-type R108,P AL and CHI transcript levels were least affected in the two MtWD401retrotranspo-son insertion mutants.The most consistent changes observed as a result of loss of MtWD40-1function were strong reductions of CHS expression inﬂower(but only determined for one probe set corresponding to TC138581)and seed,DFR1expression in leaf and ﬂower,ANS expression in stem,leaf,and seed,LAR Figure6.Genetic complementation of the Arabidopsis ttg1-9mutant.A,Leaves of the ttg1-9mutant line.B,Leaves of the ttg1-9mutantexpressing35S::MtWD40-1.C,Seed coat pigmentation of the wildtype,ttg1-9,and ttg1-9expressing35S::MtWD40-1.D,A single leaf ofttg1-9showing the glabrous phenotype.E,A single leaf of the ttg1-9mutant expressing GL2::MtWD40-1showing the restoration of thetrichome phenotype.F,Seed coat pigmentation of the wild type,ttg1-9,and ttg1-9expressing GL2::MtWD40-1.G,Yeast two-hybrid analysis ofthe interaction between MtWD40-1and Arabidopsis GL3(see text fordetails).Figure7.MtWD40-1transcript levels in M.truncatula ecotype Jema-long A17as determined by microarray analysis.A,Tissue-speciﬁcexpression.B,MtWD40-1transcript levels during seed development.The data were retrieved from the M.truncatula gene expression atlas(Benedito et al.,2008)and the seed coat microarray data set(Panget al.,2008).Pang et al.and ANR expression inﬂower and seed,and UGT72L1 expression in seed(Table II;Supplemental Tables S3 and S4).Thus,although MtWD40-1is most strongly expressed in the seed(coat),its loss of function can affectﬂavonoid pathway gene expression in multiple tissues.To further investigate the impact of loss of WD40-1 expression onﬂavonoid biosynthesis,levels of phen-ylpropanoid-derived secondary metabolites were measured by ultra-high-performance liquid chroma-tography coupled to electrospray ionization quadrupole time-of-ﬂight mass spectrometry(UPLC-ESI-QTOF-MS)in various tissues of wild-type R108 and the two independent retrotransposon insertion lines(Table III).The greatest effects were seen in developing seed,where levels of epicatechin and its glucoside(Fig.8)as well as cyanidin3-O-glucoside, kaempferol3-O-rutinoside,and two benzoic acid de-rivatives were reduced to undetectable levels in the insertion lines.In contrast,although epicatechin and its conjugate were likewise undetectable inﬂowers of the two mutant lines,levels of cyanidin3-O-glucoside and otherﬂavonoids were increased(Table III),in spite of the apparently strong reduction in CHS expression in these lines.Loss of function of MtWD40-1had little effect on the levels of threeﬂavonoids in leaves but resulted in reduced isoﬂavone(biochanin A)and aurone levels in roots(Table III).Flavonol(kaempferol 3-O-rutinoside)levels were reduced in developing seed of the mutant lines,consistent with the reduction inﬂavonol synthase expression(Table I).The less consistent results of MtWD40-1down-regulation in nonseed tissue could either be because natural product levels are more variable as a result of environmental factors in nonseed tissues or because of effects of different additional retrotransposon inserts in the two mutant lines.Overexpression of MtWD40-1in Medicago Hairy Roots Ectopic expression of the Arabidopsis MYB tran-scription factor TT2in M.truncatula hairy rootsTable I.The gene probe sets that were down-regulated more than5-fold in developing seed of the M.truncatula NF0977mutant compared with the wild-type controlExpression values were obtained from RMA(Irizarry et al.,2003).Probe Sets Target DescriptionRatio(R108/NF0977)P a Q bMtr.20567.1.S1_at Type III polyketide synthase;naringenin-chalcone synthase(CHS)198.050.0000170.060224 Mtr.20185.1.S1_x_at Naringenin-chalcone synthase;type III polyketide synthase(CHS)105.580.0001470.089928 Mtr.39897.1.S1_at Similar to CPRD12protein,partial(61%)104.580.0000010.0219 Mtr.20185.1.S1_at Naringenin-chalcone synthase;type III polyketide synthase(CHS)95.970.0006660.121225 Mtr.36333.1.S1_at Similar toﬂavonoid3#-hydroxylase(fragment),partial(21%;F3#H)85.310.0000020.032849 Mtr.49421.1.S1_at2OG-Fe(II)oxygenase79.120.0000050.043799 Mtr.14017.1.S1_at Flavonol synthase(based on similarity;FLS)40.620.0000180.060224 Mtr.6517.1.S1_at Similar to gray pubescenceﬂavonoid3#-hydroxylase,partial(49%;F3#H)36.770.0002640.105576 Mtr.44985.1.S1_at Anthocyanidin reductase,complete(ANR)34.550.0000380.089283 Mtr.14428.1.S1_x_at Naringenin-chalcone synthase;type III polyketide synthase(CHS)26.530.0004140.115755 Mtr.51818.1.S1_at Predicted protein23.290.000860.124286 Mtr.16432.1.S1_at Myb,DNA-binding;homeodomain-like23.040.0033350.151484 Mtr.14428.1.S1_at Naringenin-chalcone synthase;type III polyketide synthase(CHS)22.590.0007740.124286 Mtr.20187.1.S1_at Naringenin-chalcone synthase;type III polyketide synthase(CHS)16.740.0000150.060224 Mtr.20187.1.S1_x_at Naringenin-chalcone synthase;type III polyketide synthase(CHS)15.910.0000490.089283 Mtr.21996.1.S1_x_at Weakly similar to glucosyltransferase-13(fragment;UGT72L1)14.720.0006390.121225 Mtr.49572.1.S1_s_at Naringenin-chalcone synthase;type III polyketide synthase(CHS)14.190.0001910.096689 Mtr.47287.1.S1_at Weakly similar to myosin heavy chain-related temporary automatedfunctional11.910.0003970.115149Mtr.3858.1.S1_at Leucoanthocyanidin dioxygenase,anthocyanidin synthase,partial(24%;ANS)9.980.0157260.183253Mtr.28774.1.S1_at Anthocyanidin synthase,partial(53%;ANS)9.230.0000690.089283 Mtr.28714.1.S1_at Homolog to chalcone synthase3(Sinapis alba),partial(12%;CHS)7.920.0144030.182863 Mtr.48474.1.S1_at Weakly similar to nodulin N21family protein integral membraneprotein domain,partial(91%)7.650.0029090.147343Mtr.6511.1.S1_at Similar to GTP-binding protein,partial(47%) 6.750.0001890.096689 Mtr.25305.1.S1_at Weakly similar to The Arabidopsis Information Resource gene2827885-GOpep0.168409.m01848;expressed protein6.000.000220.10046Mtr.18797.1.S1_at Proteinase inhibitor I3,Kunitz legume;Kunitz inhibitor ST1-like 5.970.005590.164484 Mtr.32965.1.S1_at Similar to cytochrome b5DIF-F,partial(36%) 5.790.0089920.176801 Mtr.7095.1.S1_at Similar to Na+/H+antiporter NHX6,partial(28%) 5.690.0011180.127332 Mtr.41031.1.S1_at Homolog to4-coumarate-CoA ligase(4CL) 5.260.0001570.089928a The P values were obtained using associative analysis(Dozmorov and Centola,2003).b The Q values were obtained using extraction of differential gene expression(Leek et al.,2006).A WD40Repeat Protein from Medicago truncatula。

trimAl Phylogenetics Alignment Trimming Tool说明书

trimAl: a tool for automated alignment trimming in large-scale phylogenetics analyses Salvador Capella-Gutiérrez, Jose M. Silla-Martínez and Toni GabaldónTutorialVersion 1.2trimAl tutorialtrimAl is a tool for the automated trimming of Multiple Sequence Alignments. A format inter-conversion tool, called readAl, is included in the package. You can use the program either in the command line or webserver versions. The command line version is faster and has more possibilities,so it is recommended if you are going to use trimAl extensively.The trimAl webserver included in Phylemon 2.0 provides a friendly user interface and the opportunity to perform many different downstream phylogenetic analyses on your trimmed alignment. This document is a short tutorial that will guide you through the different possibilities of the program.Additional information can be obtained from where a more comprehensive documentation is available.If you use trimAl or readAl please cite our paper:trimAl: a tool for automated alignment trimming in large-scale phylogenetic analyses.Salvador Capella-Gutierrez;Jose M.Silla-Martinez;Toni Gabaldon.Bioinformatics 2009 25: 1972-1973.If you use the online webserver phylemon or phylemon2, please cite also this reference:Phylemon:a suite of web tools for molecular evolution,phylogenetics and phylogenomics.Tárraga J, Medina I, Arbiza L, Huerta-Cepas J, Gabaldón T, Dopazo J, Dopazo H. Nucleic Acids Res. 2007 Jul;35 (Web Server issue):W38-42.1. Program Installation.If you have chosen the trimAl command line version you can download the source code from the Download Section in trimAl's wikipage.For Windows OS users, we have prepared a pre-compiled trimAl version to use in this OS. Once the user has uncompressed the package, the user can find a directory,called trimAl/bin, where trimAl and readAl pre-compiled version can be found.Meanwhile for the OS based on Unix platform, e.g. GNU/Linux or MAC OS X, the user should compile the source code before to use these programs. To compile the source code, you have to change your current directory to trimAl/source and just execute "make".Once you have the trimAl and readAl binaries program, you should check if trimAl is running in appropriate way executing trimal program before starting this tutorial.2. trimAl. Multiple Sequence Alignment dataset.In order to follow this tutorial, we have prepared some examples. These examples have been taken from and you can use the codes from these files to get more information about it in this database.You can find three different directories called Api0000038, Api0000040 and Api0000080 with different files. The directory contains these files:A file .seqs with all the unaligned sequences.A file .tce with the Multiple Sequence Alignment produced by T-Coffee1.A file .msl with the Multiple Sequence Alignment produced by Muscle2.A file .mft with the Multiple Sequence Alignment produced by Mafft3.A file .clw with the Multiple Sequence Alignment produced by Clustalw4.A file .cmp with the different names of the MSAs in the directory. This file would be used by trimAl to get the most consistent MSA among the different alignments.You can use any directory to follow the present tutorial.3. Useful trimAl's features.Among the different trimAl parameters, there are some features that can be useful to interpret your alignment results:-htmlout filename. Use this parameter to have the trimAl output in an html file. In this way you can see the columns/sequences that trimAl maintains in the new alignment in grey color while the columns/sequences that have been deleted from the original alignment are in white color.-colnumbering. This parameter will provide you the relationship between the column numbers in the trimmed and the original alignment.-complementary. This parameter lets the user get the complementary alignment, in other words,when the user uses this parameter trimAl will render the columns/sequences that would be deleted from the original alignment.-w number. The user can change the windows size, by default 1, to take into account the surrounding columns in the trimAl's manual methods. When this parameter is fixed, trimAl take into account number columns to the right and to the left from the current position to compute any value, e.g. gap score, similarity score, etc. If the user wants to change a specific windows size value should use the correspond parameter-gw to change window size applied only a gap score assessments, -sc to change window size applied only to similiraty score calculations or -cw to change window size applied only to consistency part.4. Useful trimAl's/readAl's features.Both programs, trimAl and readAl, share common features related to the MSA conversion. It is possible to change the output format for a given alignment, by default the output format is the same than the input one, you can produce an output in different format with these options: -clustal. Output in CLUSTAL format.-fasta. Output in FASTA format.-nbrf. Output in PIR/NBRF format.-nexus. Output in NEXUS format.-mega. Output in MEGA format.-phylip3.2. Output in Phylip NonInterleaved format.-phylip. Output in Phylip Interleaved format.5. Getting Information from Multiple Sequence Alignment.trimAl computes different scores, such as gap score or similarity score distribution, from a given MSA. In order to obtain this information, we can use different parameters through the command line version.To do this part,we are going to use the MSA called Api0000038.msl.This file is in the Api0000038 directory.$ cd Api0000038$ trimal -in Api0000038.msl -sgt$ trimal -in Api0000038.msl -sgc$ trimal -in Api0000038.msl -sct$ trimal -in Api0000038.msl -scc$ trimal -in Api0000038.msl -sidentYou can redirect the trimAl output to a file. This file can be used in subsequent steps as input of other programs, e.g.gnuplot,,microsoft excel,etc,to do plots of this information.$ trimal -in Api0000038.msl -scc > SimilarityColumnsFor instance, in the lines below you can see how to plot the information generated by trimAl using the GNUPLOT program.$ gnuplotplot 'SimilarityColumns' u 1:2 w lp notitleset yrange [-0.05:1.05]set xrange [-1:1210]set xlabel 'Columns'set ylabel 'Residue Similarity Score'plot 'SimilarityColumns' u 1:2 w lp notitleexitIn this other example you can see the gaps distribution from the alignment. This plot also was generated using GNUPLOT$ trimal -in Api0000038.msl -sgt > gapsDistribution$ gnuplotset xlabel '% Alignment'set ylabel 'Gaps Score'plot 'gapsDistribution' u 7:4 w lp notitleexit6. Using user-defined thresholds.If you do not want to use any of the automated procedures included in trimAl (see sections 7 and 8) you can set your own thresholds to trim your alignment. We will use the parameter -htmlout filename for each example so differences can be visualized. In this example, we will use the Api0000038.msl file from the Api0000038 directory.Firstly, we are going to trim the alignment only using the -gt value which is defined in the [0 - 1] range. In this specific example, those columns that do not achieve a gap score, at least, equal to 0.190, meaning that the fraction of gaps on these columns are smaller than this value, will be deleted from the input alignment.$ trimal -in Api0000038.msl -gt 0.190 -htmlout ex01.htmlYou can see different parts of the alignment in the image below.This figure has been generated from the trimAl's HTML file for the previous example.In this other example, we can see the effect to be more strict with our threshold. An usual consequence of higher stringency is that the trimmed MSA has fewer columns. Be careful so you do not remove too much signal$ trimal -in Api0000038.msl -gt 0.8 -htmlout ex02.htmlTo be on the safe side, you can set a minimal fraction of your alignment to be conserved. In this example,we have reproduced the previous example with the difference that here we required to the program that, at least, conserve the 80% of the columns from the original alignment. This will remove the most gappy 20% of the columns or stop at the gap threshold set.$ trimal -in Api0000038.msl -gt 0.8 -cons 80 -htmlout ex03.htmlSecondly,we are going to introduce other manual threshold-st value.In this case,this threshold,also defined in the[0-1]range,is related to the similarity score.This score measures the similarity value for each column from the alignment using the Mean Distance method, by default we use Blosum62 similarity matrix but you can introduce any other matrix (see the manual). In the example below, we have used a smaller threshold to know its effect over the example.$ trimal -in Api0000038.msl -st 0.003 -htmlout ex04.htmlIn this example, similar to the previous example, we have required to conserve a minimum percentage of the original alignment in a independent way to fixed by the similarity threshold.A given threshold maintains a larger number of columns than the cons threshold, trimAl selects this first one.$ trimal -in Api0000038.msl -st 0.003 -cons 30 -htmlout ex05.htmlThirdly, we are going to see the effect of combining two different thresholds. In this case, trimAl only maintains those columns that achieve or pass both thresholds.$ trimal -in Api0000038.msl -st 0.003 -gt 0.19 -htmlout ex06.htmlFinally, we are going to see the effect of combining two different thresholds with the cons parameter. In this case, if the number of columns that achieve or pass both thresholds is equal or greater than the percentage fixed by cons parameter, trimAl chose these columns. However, if the number of columns that achieve or pass both thresholds is less than the number of columns fixed by cons parameter, trimAl relaxes both to thresholds in order to retrieve those columns that lets to achieve this minimum percentage.$ trimal -in Api0000038.msl -st 0.003 -gt 0.19 -cons 60 -htmlout ex07.html7. Selection of the most consistent alignment.trimAl can select the most consistent alignment when more than one alignment is provided for the same sequences (and in the same order) using the -compareset filename parameter. To do this part, we are going to move to Api0000040 directory, we can find there a file calledApi0000040.cmp listing the alignment paths. Using this file, we execute the instruction below to select the most consistent alignment among the alignment provided$ trimal -compareset Api0000040.cmpAs in previous section, once trimAl has selected the most consistent alignment, we can get information about the alignment selected using the appropriate parameters. For example, we can use the follow instructions to know the consistency value for each column in the alignment or its consistency values distribution$ trimal -compareset Api0000040.cmp -sct$ trimal -compareset Api0000040.cmp -sccAlso, we can trim the selected alignment using a specific threshold related to the consistency value. To do that, we should use the -ct value where the value is a number defined in the [0 - 1] range. This number refers to the average conservation of residue pars in that column with respect to the other alignments.$ trimal -compareset Api0000040.cmp -ct 0.6 -htmlout ex08.htmlOn the same way than the previous section, we can define a minimum percentage of columns that should be conserve in the new alignment. For this purpose, we have to use the cons parameter as we explained before.$ trimal -compareset Api0000040.cmp -ct 0.6 -cons 50 -htmlout ex09.htmlFinally, we can combine different thresholds, in fact, we can use all of them as well as we can define a minimum percentage of columns that should be conserve in the output alignment. In the line below, you can see an example of this situation.$ trimal -compareset Api0000040.cmp -ct 0.6 -cons 50 -gt 0.8 -st 0.01-htmlout ex10.html8. Applying automated methods.One of the most powerful aspects of trimAl is that it provides you with several automated options.This option will automatically select the most appropriate thresholds for your alignment after examining the distribution of various parameters along your alignment. Among the alignment features that trimAl takes into account to compute these optimal cut-off are the gap distribution, the similarity distribution, the identity score, etc.You can find a complete explanation about all of these methods in the trimAl's Publications Section.Here,we provide some examples on how to use these methods.The automated methods, gappyout, strict and strictpus, can be used independently if you are working with one or more than one alignment, in the last case, for the same sequences.In the lines below, you can see how to use the gappyout method in both ways. This method will eliminate the most gappy fraction of the columns from your alignment. For this, we are going to continue using the same directory than the previous section.$ trimal -compareset Api0000040.cmp -gappyout -htmlout ex11.html$ trimal -in Api0000040.mft -gappyout -htmlout ex12.htmlIn this case, we are going to use the same files than in the example before but we have changed the method to trim the alignmnet. Now, we are using strict and strictplus methods. These two methods combine the information on the fraction of gaps in a column and their similarity scores, being strictplus for more stringent than strict method.$ trimal -compareset Api0000040.cmp -strict -htmlout ex13.html$ trimal -in Api0000040.clw -strictplus -htmlout ex14.htmling an heuristic method to decide which is the best automated method for a given MSA.Finally, we implemented an heuristic method to decide which is the best automated method to trim a given alignment. The heuristic method takes into account alignment features such as the number of sequences in the alignment as well as some measures about the identity score among the sequences in the alignment or among the best pairwise sequences in that MSA. According to these characteristics trimAl will decide upon one of the two automated methods (gappyout or strictplus).To illustrate how to use this method, we provide a couple of example using the same directory than the section before. First, we used trimAl to selecte the most consistent alignment and then we trimmed that alignmnet using our heuristic method.$ trimal -compareset Api0000040.cmp -automated1 -htmlout ex15.htmlThen, we trim a single MSA using the previously mentioned method.$ trimal -in Api0000040.msl -automated1 -htmlout ex16.html10. Getting more information.We hope that this short introduction to trimAl's features has been useful to you.We advise you to visit periodically the trimAl's wikipage()where you could get the latest news about the program as well as more information, examples, etc, about trimAl's package. You can also subscribe to the mailing list if you want to be updated in new trimAl developing.11. References.1.T-Coffee: A novel method for fast and accurate multiple sequence alignment.Notredame C, Higgins DG, Heringa J. J Mol Biol. 2000 Sep 8;302(1):205-17.2.MUSCLE:multiple sequence alignment with high accuracy and highthroughput. Edgar RC.Nucleic Acids Res. 2004 Mar 19;32(5):1792-7.3.MAFFT: a novel method for rapid multiple sequence alignment based on fastFourier transform. Katoh K, Misawa K, Kuma K, Miyata T. Nucleic Acids Res. 2002 Jul 15;30(14):3059-66.4.CLUSTAL W:improving the sensitivity of progressive multiple sequencealignment through sequence weighting,position-specific gap penalties and weight matrix choice. Thompson JD, Higgins DG, Gibson TJ. Nucleic Acids Res. 1994 Nov 11;22(22):4673-80.。

电路保护器产品说明书

-55 to +15050Maximum repetitive peak reverse voltage250 C/10 seconds at terminalsSingle phase half-wave 60Hz,resistive or inductive load,V V V SYMBOLS UNITSAA V V RRMV RMSV DC I (AV)I FSMV F 1.030.0Operating junction temperature range Maximum RMS voltage Maximum DC blocking voltageMaximum average forward rectified currentat T L =100 CPeak forward surge current, 8.3ms single half sine-wave superimposed on rated load Maximum instantaneous forward voltage at 1.0A Maximum DC reverse current T A =25 C at rated DC blocking voltage T A =125 C I R 0.5R q JA T J 80.0C Typical thermal resistanceC/W Ratings at 25 C ambient temperature unless otherwise specified.SS18SS110SS11560426080568010070100150105150200140200Case : Molded plastic bodyTerminals : Solder plated, solderable per MIL-STD-750,Method 2026Polarity : Polarity symbol marking on body Mounting Position : AnyWeight : 0.0023 ounce, 0.07 grams for capacitive load current derate by 20%.FeaturesThe plastic package carries Underwriters Laboratory Flammability Classification 94V-0For surface mounted applicationsBuilt-in strain relief,ideal for automated placement Low reverse leakageHigh forward surge current capability High temperature soldering guaranteed Mechanical DataMaximum Ratings And Electrical Characteristics SS12~SS1200.55SS1201.0Amp Surface Mounted Schottky Barrier RectifiersSS16402840SS14201420SS12Parameter 0.700.850.95100.05A m -55 to +150Storage temperature rangeT STGC-55 to +125DO-214AC/SMADimensions in inches and (millimeters)0.067 (1.70)0.051 (1.30)0.012(0.305)0.006(0.152)4531.545SS145FIG. 3-TYPICAL FORWARD VOLTAGE CHARACTERISTICSNumber of cyclesFIG. 2-MAXIMUM NON-REPETITIVE PEAK FORWARDSURGE CURRENT PERLEGFIG. 1- DERATING CURVE OUTPUT RECTIFIED CURRENTA v e ra g e F o r w a r d R e c t i f i e d C u r r e n tA m p e r e sP e a k F o r w a r d S u r a g e C u r r e n t FIG. 4-TYPICAL REVERSE LEAKAGE CHARACTERISTICSLead Temperature1.00.90.80.70.60.50.40.30.2A m p e r e sRatings And Characteristic CurvesSuggested Pad LayoutI N S T A N T A N E O U S F O R W A R D C U R R E N T ,A M P E R E SINSTANTANEOUS FORWARD VOLTAGE,VOLTS5010.010.10.01PERCENT OF PEAK REVERSE VOLTAGE,%I N S T A N T A N E O U S R E V E R S E C U R R E N T ,M I C R O A M P E R E S1001010.10.0950.215Symbol Unit (mm)A C 3.90D 0.0605.452.41EB Unit (inch)0.1541.520.0661.68Suggested Soldering Temperature ProfileR ecom m ended reflow m ethods:IR ,vapor phase oven,hot air oven,w ave solder.The device can be exposed to a m axim um tem perature of 265°C for 10seconds.D evices can be cleaned using standard industry m ethods and solvents.If reflow temperatures exceed the re co m m e nde d p ro file,d evice s m a y n ot m e et th e p erfo rm ance re q uire m ents.NotePackage InformationCarrier Dimension(mm)A0B0K0D02.804.05.304.02.362.01.550.251.7512E F 5.50P0P1P2T W 0.1TolerancePackage SpecificationsPackage Reel SizeQ'TY/Reel Box Size 11'528510QTY/Box Carton Size355*310*310Reel DIA.278(mm)(Kpcs)Q'TY/Carton 80(mm)(mm)(Kpcs)(Kpcs)SMA13'7.534015360*360*360330120。

华为GPON数据查看

华为GPON数据查看MA5680T> enable 进⼊特权模式MA5680T # config 进⼊终端配置MA5680T (config)# switch language-mode 中英⽂切换MA5608T(config)#display ont info 0 6 0 all 查看pon⼝下所有的光猫MA5608T(config)#display ont optical-info 11 42 查看单个光衰MA5608T(config)#display ont optical-info 5 all 查看pon下所有的光衰MA5608T(config)#interface gpon 0/1 进⼊端⼝MA5608T(config-if-gpon-0/0)#ont deactivate 0 32 关闭32个光猫(config-if-gpon-0/0)#display ont info 0 3 查看第32个光猫MA5608T(config-if-gpon-0/0)#ont activate 4 2 打开第32个光猫MA5608T(config-if-gpon-0/7)#display ont optical-info 1 39 查看单个光猫的光MA5608T(config)#display ont wan-info 0/2 32 连接状态MA5608T(config)#display service-port vlan 91 查看第⼏个端⼝粮可以使⽤MA5680T (config)#display current-configuration port 0/2/3 查看光猫所⽤个数MA5680T (config)#display ont info by-loid HET002778643 ⽤loid找光猫MA5680T (config)#display ont info by-sn 485754435696EA9A⽤sn找光猫MA5680T (config)#display mac-address vlan 1001 查询1001的⽤户MA5680T (config)#display service-port 0/0/14 ont 5 查看序号MA5680T (config)#display log all 查询历史操作MA5680T (config)#display ont port state 1 2 eth-port all 查看是否插⽹线MA5680T (config)#display ont autofind all 全局搜光猫(config)#display patch all 查看版本和补丁(config)#display board 0 查看槽位信息和单板状态：(config)#display board 0/1 查看0/1槽位单板下的所有ONU的状态：(config-epon-0/1)#display ont info 2 3 查看0/1槽的2号PON⼝的3号 ONU的详细信息(config-epon-0/1)#ont version 2 3 查看0/1槽的2号PON⼝的3号ONU的版本(config-epon-0/1)#display ont optical-info 2 3 查看查看0/1槽的2号PON⼝的3号ONU的收光功率(config-epon-0/1)#display statistics ont-line-qualtiy 2 3 查看查看0/1槽的2号PON⼝的3号ONU的光路误码(config)#display ont-line-profile epon profile-id 在全局模式下查看模板信息，查看EPON ONU 的线路模板(config)#display ont-srv-profile epon profile-id 查看EPON ONU的业务模板(config)#display dba-profile 查看ONU的dba模板(config)#display traffic table ip from-index 0 查看ONU的流量模板(config)#display service-port port 0/1/2 ont 3 查看0/1/2的ONT3的业务流(config)#display mac-address port 0/1/2 ont 3 查看0/1/2的ONT3学到的MAC 地址(config)#display mac-address vlan 100 查看VLAN100中学到的MAC 地址。

LGO处理GPS静态数据的方法与基线超限时的处理方法

LGO 里F 检验超限怎么办笼统的来说，此时可以重新处理基线，看看是浮点解还是固定解，如果还是超限那么改为手动处理基线，查看基线残差图，剔除残差较大的卫星或时段，然后再平差，看看是否通过。

详细来讲的话。

可一分为以下几个步骤：1，导入数据后基线处理时，进行自动处理，处理完保存结果；2，找到误差超限的点，删除与之相关的基线进行重新处理，这时，改为手动处理，观测时间长的作为参考点，短的作为流动站，进行逐个处理，并保存结果；3，如误差还是超限，则找到对应的点观测值，开窗进行卫星的时间处理，剔除掉信号不好的卫星时段，但也要注意观测时间必须保证不低于规范规定的时间；然后进行手动处理基线，并保存结果；4，处理完所有的基线后，误差不超限，就进行网平差，再输出报告，查看是否F检验超限，若仍旧超限，则重复第三步。

注意：在查看F 检验时，值太大（100以上）应该是点的初始坐标不对，在处理基线前，先进行单点定位；若值小于100，应该是卫星的时段不好，通过剔除不好的卫星或者卫星时段，即可，但需要有耐心，得反复的进行卫星处理。

同时也可以在网平差的结果中查看残差，对着残差信息进行处理会更直观些！LGO 的功能及应用概述Leica Geo Office （LGO ）内业处理软件包支持Leica所有仪器类型，以统一的方式管理TPS 、GPS 和水准数据，功能强大，易于使用，是适合用户自定义和智能向导的工具。

图1 LGO内业处理软机包LGO包括标准软件、其它选项以及工具三大部分，不同的安装和配置选项支持用户的不同需求。

标准软件主要包括数据输入/输出、项目管理和查看、报表、编码列表管理等功能；其它选项主要包括水准处理、GPS处理、网平差以及GIS/CAD输出等功能；常用工具则包括数据管理器、软件上传以及格式编辑器。

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3GPP TS 36.331 V13.2.0 (2016-06)

3GPP TS 36.331 V13.2.0 (2016-06)Technical Specification3rd Generation Partnership Project;Technical Specification Group Radio Access Network;Evolved Universal Terrestrial Radio Access (E-UTRA);Radio Resource Control (RRC);Protocol specification(Release 13)The present document has been developed within the 3rd Generation Partnership Project (3GPP TM) and may be further elaborated for the purposes of 3GPP. The present document has not been subject to any approval process by the 3GPP Organizational Partners and shall not be implemented.This Specification is provided for future development work within 3GPP only. The Organizational Partners accept no liability for any use of this Specification. Specifications and reports for implementation of the 3GPP TM system should be obtained via the 3GPP Organizational Partners' Publications Offices.KeywordsUMTS, radio3GPPPostal address3GPP support office address650 Route des Lucioles - Sophia AntipolisValbonne - FRANCETel.: +33 4 92 94 42 00 Fax: +33 4 93 65 47 16InternetCopyright NotificationNo part may be reproduced except as authorized by written permission.The copyright and the foregoing restriction extend to reproduction in all media.© 2016, 3GPP Organizational Partners (ARIB, ATIS, CCSA, ETSI, TSDSI, TTA, TTC).All rights reserved.UMTS™ is a Trade Mark of ETSI registered for the benefit of its members3GPP™ is a Trade Mark of ETSI registered for the benefit of its Members and of the 3GPP Organizational PartnersLTE™ is a Trade Mark of ETSI currently being registered for the benefit of its Members and of the 3GPP Organizational Partners GSM® and the GSM logo are registered and owned by the GSM AssociationBluetooth® is a Trade Mark of the Bluetooth SIG registered for the benefit of its membersContentsForeword (18)1Scope (19)2References (19)3Definitions, symbols and abbreviations (22)3.1Definitions (22)3.2Abbreviations (24)4General (27)4.1Introduction (27)4.2Architecture (28)4.2.1UE states and state transitions including inter RAT (28)4.2.2Signalling radio bearers (29)4.3Services (30)4.3.1Services provided to upper layers (30)4.3.2Services expected from lower layers (30)4.4Functions (30)5Procedures (32)5.1General (32)5.1.1Introduction (32)5.1.2General requirements (32)5.2System information (33)5.2.1Introduction (33)5.2.1.1General (33)5.2.1.2Scheduling (34)5.2.1.2a Scheduling for NB-IoT (34)5.2.1.3System information validity and notification of changes (35)5.2.1.4Indication of ETWS notification (36)5.2.1.5Indication of CMAS notification (37)5.2.1.6Notification of EAB parameters change (37)5.2.1.7Access Barring parameters change in NB-IoT (37)5.2.2System information acquisition (38)5.2.2.1General (38)5.2.2.2Initiation (38)5.2.2.3System information required by the UE (38)5.2.2.4System information acquisition by the UE (39)5.2.2.5Essential system information missing (42)5.2.2.6Actions upon reception of the MasterInformationBlock message (42)5.2.2.7Actions upon reception of the SystemInformationBlockType1 message (42)5.2.2.8Actions upon reception of SystemInformation messages (44)5.2.2.9Actions upon reception of SystemInformationBlockType2 (44)5.2.2.10Actions upon reception of SystemInformationBlockType3 (45)5.2.2.11Actions upon reception of SystemInformationBlockType4 (45)5.2.2.12Actions upon reception of SystemInformationBlockType5 (45)5.2.2.13Actions upon reception of SystemInformationBlockType6 (45)5.2.2.14Actions upon reception of SystemInformationBlockType7 (45)5.2.2.15Actions upon reception of SystemInformationBlockType8 (45)5.2.2.16Actions upon reception of SystemInformationBlockType9 (46)5.2.2.17Actions upon reception of SystemInformationBlockType10 (46)5.2.2.18Actions upon reception of SystemInformationBlockType11 (46)5.2.2.19Actions upon reception of SystemInformationBlockType12 (47)5.2.2.20Actions upon reception of SystemInformationBlockType13 (48)5.2.2.21Actions upon reception of SystemInformationBlockType14 (48)5.2.2.22Actions upon reception of SystemInformationBlockType15 (48)5.2.2.23Actions upon reception of SystemInformationBlockType16 (48)5.2.2.24Actions upon reception of SystemInformationBlockType17 (48)5.2.2.25Actions upon reception of SystemInformationBlockType18 (48)5.2.2.26Actions upon reception of SystemInformationBlockType19 (49)5.2.3Acquisition of an SI message (49)5.2.3a Acquisition of an SI message by BL UE or UE in CE or a NB-IoT UE (50)5.3Connection control (50)5.3.1Introduction (50)5.3.1.1RRC connection control (50)5.3.1.2Security (52)5.3.1.2a RN security (53)5.3.1.3Connected mode mobility (53)5.3.1.4Connection control in NB-IoT (54)5.3.2Paging (55)5.3.2.1General (55)5.3.2.2Initiation (55)5.3.2.3Reception of the Paging message by the UE (55)5.3.3RRC connection establishment (56)5.3.3.1General (56)5.3.3.1a Conditions for establishing RRC Connection for sidelink communication/ discovery (58)5.3.3.2Initiation (59)5.3.3.3Actions related to transmission of RRCConnectionRequest message (63)5.3.3.3a Actions related to transmission of RRCConnectionResumeRequest message (64)5.3.3.4Reception of the RRCConnectionSetup by the UE (64)5.3.3.4a Reception of the RRCConnectionResume by the UE (66)5.3.3.5Cell re-selection while T300, T302, T303, T305, T306, or T308 is running (68)5.3.3.6T300 expiry (68)5.3.3.7T302, T303, T305, T306, or T308 expiry or stop (69)5.3.3.8Reception of the RRCConnectionReject by the UE (70)5.3.3.9Abortion of RRC connection establishment (71)5.3.3.10Handling of SSAC related parameters (71)5.3.3.11Access barring check (72)5.3.3.12EAB check (73)5.3.3.13Access barring check for ACDC (73)5.3.3.14Access Barring check for NB-IoT (74)5.3.4Initial security activation (75)5.3.4.1General (75)5.3.4.2Initiation (76)5.3.4.3Reception of the SecurityModeCommand by the UE (76)5.3.5RRC connection reconfiguration (77)5.3.5.1General (77)5.3.5.2Initiation (77)5.3.5.3Reception of an RRCConnectionReconfiguration not including the mobilityControlInfo by theUE (77)5.3.5.4Reception of an RRCConnectionReconfiguration including the mobilityControlInfo by the UE(handover) (79)5.3.5.5Reconfiguration failure (83)5.3.5.6T304 expiry (handover failure) (83)5.3.5.7Void (84)5.3.5.7a T307 expiry (SCG change failure) (84)5.3.5.8Radio Configuration involving full configuration option (84)5.3.6Counter check (86)5.3.6.1General (86)5.3.6.2Initiation (86)5.3.6.3Reception of the CounterCheck message by the UE (86)5.3.7RRC connection re-establishment (87)5.3.7.1General (87)5.3.7.2Initiation (87)5.3.7.3Actions following cell selection while T311 is running (88)5.3.7.4Actions related to transmission of RRCConnectionReestablishmentRequest message (89)5.3.7.5Reception of the RRCConnectionReestablishment by the UE (89)5.3.7.6T311 expiry (91)5.3.7.7T301 expiry or selected cell no longer suitable (91)5.3.7.8Reception of RRCConnectionReestablishmentReject by the UE (91)5.3.8RRC connection release (92)5.3.8.1General (92)5.3.8.2Initiation (92)5.3.8.3Reception of the RRCConnectionRelease by the UE (92)5.3.8.4T320 expiry (93)5.3.9RRC connection release requested by upper layers (93)5.3.9.1General (93)5.3.9.2Initiation (93)5.3.10Radio resource configuration (93)5.3.10.0General (93)5.3.10.1SRB addition/ modification (94)5.3.10.2DRB release (95)5.3.10.3DRB addition/ modification (95)5.3.10.3a1DC specific DRB addition or reconfiguration (96)5.3.10.3a2LWA specific DRB addition or reconfiguration (98)5.3.10.3a3LWIP specific DRB addition or reconfiguration (98)5.3.10.3a SCell release (99)5.3.10.3b SCell addition/ modification (99)5.3.10.3c PSCell addition or modification (99)5.3.10.4MAC main reconfiguration (99)5.3.10.5Semi-persistent scheduling reconfiguration (100)5.3.10.6Physical channel reconfiguration (100)5.3.10.7Radio Link Failure Timers and Constants reconfiguration (101)5.3.10.8Time domain measurement resource restriction for serving cell (101)5.3.10.9Other configuration (102)5.3.10.10SCG reconfiguration (103)5.3.10.11SCG dedicated resource configuration (104)5.3.10.12Reconfiguration SCG or split DRB by drb-ToAddModList (105)5.3.10.13Neighbour cell information reconfiguration (105)5.3.10.14Void (105)5.3.10.15Sidelink dedicated configuration (105)5.3.10.16T370 expiry (106)5.3.11Radio link failure related actions (107)5.3.11.1Detection of physical layer problems in RRC_CONNECTED (107)5.3.11.2Recovery of physical layer problems (107)5.3.11.3Detection of radio link failure (107)5.3.12UE actions upon leaving RRC_CONNECTED (109)5.3.13UE actions upon PUCCH/ SRS release request (110)5.3.14Proximity indication (110)5.3.14.1General (110)5.3.14.2Initiation (111)5.3.14.3Actions related to transmission of ProximityIndication message (111)5.3.15Void (111)5.4Inter-RAT mobility (111)5.4.1Introduction (111)5.4.2Handover to E-UTRA (112)5.4.2.1General (112)5.4.2.2Initiation (112)5.4.2.3Reception of the RRCConnectionReconfiguration by the UE (112)5.4.2.4Reconfiguration failure (114)5.4.2.5T304 expiry (handover to E-UTRA failure) (114)5.4.3Mobility from E-UTRA (114)5.4.3.1General (114)5.4.3.2Initiation (115)5.4.3.3Reception of the MobilityFromEUTRACommand by the UE (115)5.4.3.4Successful completion of the mobility from E-UTRA (116)5.4.3.5Mobility from E-UTRA failure (117)5.4.4Handover from E-UTRA preparation request (CDMA2000) (117)5.4.4.1General (117)5.4.4.2Initiation (118)5.4.4.3Reception of the HandoverFromEUTRAPreparationRequest by the UE (118)5.4.5UL handover preparation transfer (CDMA2000) (118)5.4.5.1General (118)5.4.5.2Initiation (118)5.4.5.3Actions related to transmission of the ULHandoverPreparationTransfer message (119)5.4.5.4Failure to deliver the ULHandoverPreparationTransfer message (119)5.4.6Inter-RAT cell change order to E-UTRAN (119)5.4.6.1General (119)5.4.6.2Initiation (119)5.4.6.3UE fails to complete an inter-RAT cell change order (119)5.5Measurements (120)5.5.1Introduction (120)5.5.2Measurement configuration (121)5.5.2.1General (121)5.5.2.2Measurement identity removal (122)5.5.2.2a Measurement identity autonomous removal (122)5.5.2.3Measurement identity addition/ modification (123)5.5.2.4Measurement object removal (124)5.5.2.5Measurement object addition/ modification (124)5.5.2.6Reporting configuration removal (126)5.5.2.7Reporting configuration addition/ modification (127)5.5.2.8Quantity configuration (127)5.5.2.9Measurement gap configuration (127)5.5.2.10Discovery signals measurement timing configuration (128)5.5.2.11RSSI measurement timing configuration (128)5.5.3Performing measurements (128)5.5.3.1General (128)5.5.3.2Layer 3 filtering (131)5.5.4Measurement report triggering (131)5.5.4.1General (131)5.5.4.2Event A1 (Serving becomes better than threshold) (135)5.5.4.3Event A2 (Serving becomes worse than threshold) (136)5.5.4.4Event A3 (Neighbour becomes offset better than PCell/ PSCell) (136)5.5.4.5Event A4 (Neighbour becomes better than threshold) (137)5.5.4.6Event A5 (PCell/ PSCell becomes worse than threshold1 and neighbour becomes better thanthreshold2) (138)5.5.4.6a Event A6 (Neighbour becomes offset better than SCell) (139)5.5.4.7Event B1 (Inter RAT neighbour becomes better than threshold) (139)5.5.4.8Event B2 (PCell becomes worse than threshold1 and inter RAT neighbour becomes better thanthreshold2) (140)5.5.4.9Event C1 (CSI-RS resource becomes better than threshold) (141)5.5.4.10Event C2 (CSI-RS resource becomes offset better than reference CSI-RS resource) (141)5.5.4.11Event W1 (WLAN becomes better than a threshold) (142)5.5.4.12Event W2 (All WLAN inside WLAN mobility set becomes worse than threshold1 and a WLANoutside WLAN mobility set becomes better than threshold2) (142)5.5.4.13Event W3 (All WLAN inside WLAN mobility set becomes worse than a threshold) (143)5.5.5Measurement reporting (144)5.5.6Measurement related actions (148)5.5.6.1Actions upon handover and re-establishment (148)5.5.6.2Speed dependant scaling of measurement related parameters (149)5.5.7Inter-frequency RSTD measurement indication (149)5.5.7.1General (149)5.5.7.2Initiation (150)5.5.7.3Actions related to transmission of InterFreqRSTDMeasurementIndication message (150)5.6Other (150)5.6.0General (150)5.6.1DL information transfer (151)5.6.1.1General (151)5.6.1.2Initiation (151)5.6.1.3Reception of the DLInformationTransfer by the UE (151)5.6.2UL information transfer (151)5.6.2.1General (151)5.6.2.2Initiation (151)5.6.2.3Actions related to transmission of ULInformationTransfer message (152)5.6.2.4Failure to deliver ULInformationTransfer message (152)5.6.3UE capability transfer (152)5.6.3.1General (152)5.6.3.2Initiation (153)5.6.3.3Reception of the UECapabilityEnquiry by the UE (153)5.6.4CSFB to 1x Parameter transfer (157)5.6.4.1General (157)5.6.4.2Initiation (157)5.6.4.3Actions related to transmission of CSFBParametersRequestCDMA2000 message (157)5.6.4.4Reception of the CSFBParametersResponseCDMA2000 message (157)5.6.5UE Information (158)5.6.5.1General (158)5.6.5.2Initiation (158)5.6.5.3Reception of the UEInformationRequest message (158)5.6.6 Logged Measurement Configuration (159)5.6.6.1General (159)5.6.6.2Initiation (160)5.6.6.3Reception of the LoggedMeasurementConfiguration by the UE (160)5.6.6.4T330 expiry (160)5.6.7 Release of Logged Measurement Configuration (160)5.6.7.1General (160)5.6.7.2Initiation (160)5.6.8 Measurements logging (161)5.6.8.1General (161)5.6.8.2Initiation (161)5.6.9In-device coexistence indication (163)5.6.9.1General (163)5.6.9.2Initiation (164)5.6.9.3Actions related to transmission of InDeviceCoexIndication message (164)5.6.10UE Assistance Information (165)5.6.10.1General (165)5.6.10.2Initiation (166)5.6.10.3Actions related to transmission of UEAssistanceInformation message (166)5.6.11 Mobility history information (166)5.6.11.1General (166)5.6.11.2Initiation (166)5.6.12RAN-assisted WLAN interworking (167)5.6.12.1General (167)5.6.12.2Dedicated WLAN offload configuration (167)5.6.12.3WLAN offload RAN evaluation (167)5.6.12.4T350 expiry or stop (167)5.6.12.5Cell selection/ re-selection while T350 is running (168)5.6.13SCG failure information (168)5.6.13.1General (168)5.6.13.2Initiation (168)5.6.13.3Actions related to transmission of SCGFailureInformation message (168)5.6.14LTE-WLAN Aggregation (169)5.6.14.1Introduction (169)5.6.14.2Reception of LWA configuration (169)5.6.14.3Release of LWA configuration (170)5.6.15WLAN connection management (170)5.6.15.1Introduction (170)5.6.15.2WLAN connection status reporting (170)5.6.15.2.1General (170)5.6.15.2.2Initiation (171)5.6.15.2.3Actions related to transmission of WLANConnectionStatusReport message (171)5.6.15.3T351 Expiry (WLAN connection attempt timeout) (171)5.6.15.4WLAN status monitoring (171)5.6.16RAN controlled LTE-WLAN interworking (172)5.6.16.1General (172)5.6.16.2WLAN traffic steering command (172)5.6.17LTE-WLAN aggregation with IPsec tunnel (173)5.6.17.1General (173)5.7Generic error handling (174)5.7.1General (174)5.7.2ASN.1 violation or encoding error (174)5.7.3Field set to a not comprehended value (174)5.7.4Mandatory field missing (174)5.7.5Not comprehended field (176)5.8MBMS (176)5.8.1Introduction (176)5.8.1.1General (176)5.8.1.2Scheduling (176)5.8.1.3MCCH information validity and notification of changes (176)5.8.2MCCH information acquisition (178)5.8.2.1General (178)5.8.2.2Initiation (178)5.8.2.3MCCH information acquisition by the UE (178)5.8.2.4Actions upon reception of the MBSFNAreaConfiguration message (178)5.8.2.5Actions upon reception of the MBMSCountingRequest message (179)5.8.3MBMS PTM radio bearer configuration (179)5.8.3.1General (179)5.8.3.2Initiation (179)5.8.3.3MRB establishment (179)5.8.3.4MRB release (179)5.8.4MBMS Counting Procedure (179)5.8.4.1General (179)5.8.4.2Initiation (180)5.8.4.3Reception of the MBMSCountingRequest message by the UE (180)5.8.5MBMS interest indication (181)5.8.5.1General (181)5.8.5.2Initiation (181)5.8.5.3Determine MBMS frequencies of interest (182)5.8.5.4Actions related to transmission of MBMSInterestIndication message (183)5.8a SC-PTM (183)5.8a.1Introduction (183)5.8a.1.1General (183)5.8a.1.2SC-MCCH scheduling (183)5.8a.1.3SC-MCCH information validity and notification of changes (183)5.8a.1.4Procedures (184)5.8a.2SC-MCCH information acquisition (184)5.8a.2.1General (184)5.8a.2.2Initiation (184)5.8a.2.3SC-MCCH information acquisition by the UE (184)5.8a.2.4Actions upon reception of the SCPTMConfiguration message (185)5.8a.3SC-PTM radio bearer configuration (185)5.8a.3.1General (185)5.8a.3.2Initiation (185)5.8a.3.3SC-MRB establishment (185)5.8a.3.4SC-MRB release (185)5.9RN procedures (186)5.9.1RN reconfiguration (186)5.9.1.1General (186)5.9.1.2Initiation (186)5.9.1.3Reception of the RNReconfiguration by the RN (186)5.10Sidelink (186)5.10.1Introduction (186)5.10.1a Conditions for sidelink communication operation (187)5.10.2Sidelink UE information (188)5.10.2.1General (188)5.10.2.2Initiation (189)5.10.2.3Actions related to transmission of SidelinkUEInformation message (193)5.10.3Sidelink communication monitoring (195)5.10.6Sidelink discovery announcement (198)5.10.6a Sidelink discovery announcement pool selection (201)5.10.6b Sidelink discovery announcement reference carrier selection (201)5.10.7Sidelink synchronisation information transmission (202)5.10.7.1General (202)5.10.7.2Initiation (203)5.10.7.3Transmission of SLSS (204)5.10.7.4Transmission of MasterInformationBlock-SL message (205)5.10.7.5Void (206)5.10.8Sidelink synchronisation reference (206)5.10.8.1General (206)5.10.8.2Selection and reselection of synchronisation reference UE (SyncRef UE) (206)5.10.9Sidelink common control information (207)5.10.9.1General (207)5.10.9.2Actions related to reception of MasterInformationBlock-SL message (207)5.10.10Sidelink relay UE operation (207)5.10.10.1General (207)5.10.10.2AS-conditions for relay related sidelink communication transmission by sidelink relay UE (207)5.10.10.3AS-conditions for relay PS related sidelink discovery transmission by sidelink relay UE (208)5.10.10.4Sidelink relay UE threshold conditions (208)5.10.11Sidelink remote UE operation (208)5.10.11.1General (208)5.10.11.2AS-conditions for relay related sidelink communication transmission by sidelink remote UE (208)5.10.11.3AS-conditions for relay PS related sidelink discovery transmission by sidelink remote UE (209)5.10.11.4Selection and reselection of sidelink relay UE (209)5.10.11.5Sidelink remote UE threshold conditions (210)6Protocol data units, formats and parameters (tabular & ASN.1) (210)6.1General (210)6.2RRC messages (212)6.2.1General message structure (212)–EUTRA-RRC-Definitions (212)–BCCH-BCH-Message (212)–BCCH-DL-SCH-Message (212)–BCCH-DL-SCH-Message-BR (213)–MCCH-Message (213)–PCCH-Message (213)–DL-CCCH-Message (214)–DL-DCCH-Message (214)–UL-CCCH-Message (214)–UL-DCCH-Message (215)–SC-MCCH-Message (215)6.2.2Message definitions (216)–CounterCheck (216)–CounterCheckResponse (217)–CSFBParametersRequestCDMA2000 (217)–CSFBParametersResponseCDMA2000 (218)–DLInformationTransfer (218)–HandoverFromEUTRAPreparationRequest (CDMA2000) (219)–InDeviceCoexIndication (220)–InterFreqRSTDMeasurementIndication (222)–LoggedMeasurementConfiguration (223)–MasterInformationBlock (225)–MBMSCountingRequest (226)–MBMSCountingResponse (226)–MBMSInterestIndication (227)–MBSFNAreaConfiguration (228)–MeasurementReport (228)–MobilityFromEUTRACommand (229)–Paging (232)–ProximityIndication (233)–RNReconfiguration (234)–RNReconfigurationComplete (234)–RRCConnectionReconfiguration (235)–RRCConnectionReconfigurationComplete (240)–RRCConnectionReestablishment (241)–RRCConnectionReestablishmentComplete (241)–RRCConnectionReestablishmentReject (242)–RRCConnectionReestablishmentRequest (243)–RRCConnectionReject (243)–RRCConnectionRelease (244)–RRCConnectionResume (248)–RRCConnectionResumeComplete (249)–RRCConnectionResumeRequest (250)–RRCConnectionRequest (250)–RRCConnectionSetup (251)–RRCConnectionSetupComplete (252)–SCGFailureInformation (253)–SCPTMConfiguration (254)–SecurityModeCommand (255)–SecurityModeComplete (255)–SecurityModeFailure (256)–SidelinkUEInformation (256)–SystemInformation (258)–SystemInformationBlockType1 (259)–UEAssistanceInformation (264)–UECapabilityEnquiry (265)–UECapabilityInformation (266)–UEInformationRequest (267)–UEInformationResponse (267)–ULHandoverPreparationTransfer (CDMA2000) (273)–ULInformationTransfer (274)–WLANConnectionStatusReport (274)6.3RRC information elements (275)6.3.1System information blocks (275)–SystemInformationBlockType2 (275)–SystemInformationBlockType3 (279)–SystemInformationBlockType4 (282)–SystemInformationBlockType5 (283)–SystemInformationBlockType6 (287)–SystemInformationBlockType7 (289)–SystemInformationBlockType8 (290)–SystemInformationBlockType9 (295)–SystemInformationBlockType10 (295)–SystemInformationBlockType11 (296)–SystemInformationBlockType12 (297)–SystemInformationBlockType13 (297)–SystemInformationBlockType14 (298)–SystemInformationBlockType15 (298)–SystemInformationBlockType16 (299)–SystemInformationBlockType17 (300)–SystemInformationBlockType18 (301)–SystemInformationBlockType19 (301)–SystemInformationBlockType20 (304)6.3.2Radio resource control information elements (304)–AntennaInfo (304)–AntennaInfoUL (306)–CQI-ReportConfig (307)–CQI-ReportPeriodicProcExtId (314)–CrossCarrierSchedulingConfig (314)–CSI-IM-Config (315)–CSI-IM-ConfigId (315)–CSI-RS-Config (317)–CSI-RS-ConfigEMIMO (318)–CSI-RS-ConfigNZP (319)–CSI-RS-ConfigNZPId (320)–CSI-RS-ConfigZP (321)–CSI-RS-ConfigZPId (321)–DMRS-Config (321)–DRB-Identity (322)–EPDCCH-Config (322)–EIMTA-MainConfig (324)–LogicalChannelConfig (325)–LWA-Configuration (326)–LWIP-Configuration (326)–RCLWI-Configuration (327)–MAC-MainConfig (327)–P-C-AndCBSR (332)–PDCCH-ConfigSCell (333)–PDCP-Config (334)–PDSCH-Config (337)–PDSCH-RE-MappingQCL-ConfigId (339)–PHICH-Config (339)–PhysicalConfigDedicated (339)–P-Max (344)–PRACH-Config (344)–PresenceAntennaPort1 (346)–PUCCH-Config (347)–PUSCH-Config (351)–RACH-ConfigCommon (355)–RACH-ConfigDedicated (357)–RadioResourceConfigCommon (358)–RadioResourceConfigDedicated (362)–RLC-Config (367)–RLF-TimersAndConstants (369)–RN-SubframeConfig (370)–SchedulingRequestConfig (371)–SoundingRS-UL-Config (372)–SPS-Config (375)–TDD-Config (376)–TimeAlignmentTimer (377)–TPC-PDCCH-Config (377)–TunnelConfigLWIP (378)–UplinkPowerControl (379)–WLAN-Id-List (382)–WLAN-MobilityConfig (382)6.3.3Security control information elements (382)–NextHopChainingCount (382)–SecurityAlgorithmConfig (383)–ShortMAC-I (383)6.3.4Mobility control information elements (383)–AdditionalSpectrumEmission (383)–ARFCN-ValueCDMA2000 (383)–ARFCN-ValueEUTRA (384)–ARFCN-ValueGERAN (384)–ARFCN-ValueUTRA (384)–BandclassCDMA2000 (384)–BandIndicatorGERAN (385)–CarrierFreqCDMA2000 (385)–CarrierFreqGERAN (385)–CellIndexList (387)–CellReselectionPriority (387)–CellSelectionInfoCE (387)–CellReselectionSubPriority (388)–CSFB-RegistrationParam1XRTT (388)–CellGlobalIdEUTRA (389)–CellGlobalIdUTRA (389)–CellGlobalIdGERAN (390)–CellGlobalIdCDMA2000 (390)–CellSelectionInfoNFreq (391)–CSG-Identity (391)–FreqBandIndicator (391)–MobilityControlInfo (391)–MobilityParametersCDMA2000 (1xRTT) (393)–MobilityStateParameters (394)–MultiBandInfoList (394)–NS-PmaxList (394)–PhysCellId (395)–PhysCellIdRange (395)–PhysCellIdRangeUTRA-FDDList (395)–PhysCellIdCDMA2000 (396)–PhysCellIdGERAN (396)–PhysCellIdUTRA-FDD (396)–PhysCellIdUTRA-TDD (396)–PLMN-Identity (397)–PLMN-IdentityList3 (397)–PreRegistrationInfoHRPD (397)–Q-QualMin (398)–Q-RxLevMin (398)–Q-OffsetRange (398)–Q-OffsetRangeInterRAT (399)–ReselectionThreshold (399)–ReselectionThresholdQ (399)–SCellIndex (399)–ServCellIndex (400)–SpeedStateScaleFactors (400)–SystemInfoListGERAN (400)–SystemTimeInfoCDMA2000 (401)–TrackingAreaCode (401)–T-Reselection (402)–T-ReselectionEUTRA-CE (402)6.3.5Measurement information elements (402)–AllowedMeasBandwidth (402)–CSI-RSRP-Range (402)–Hysteresis (402)–LocationInfo (403)–MBSFN-RSRQ-Range (403)–MeasConfig (404)–MeasDS-Config (405)–MeasGapConfig (406)–MeasId (407)–MeasIdToAddModList (407)–MeasObjectCDMA2000 (408)–MeasObjectEUTRA (408)–MeasObjectGERAN (412)–MeasObjectId (412)–MeasObjectToAddModList (412)–MeasObjectUTRA (413)–ReportConfigEUTRA (422)–ReportConfigId (425)–ReportConfigInterRAT (425)–ReportConfigToAddModList (428)–ReportInterval (429)–RSRP-Range (429)–RSRQ-Range (430)–RSRQ-Type (430)–RS-SINR-Range (430)–RSSI-Range-r13 (431)–TimeToTrigger (431)–UL-DelayConfig (431)–WLAN-CarrierInfo (431)–WLAN-RSSI-Range (432)–WLAN-Status (432)6.3.6Other information elements (433)–AbsoluteTimeInfo (433)–AreaConfiguration (433)–C-RNTI (433)–DedicatedInfoCDMA2000 (434)–DedicatedInfoNAS (434)–FilterCoefficient (434)–LoggingDuration (434)–LoggingInterval (435)–MeasSubframePattern (435)–MMEC (435)–NeighCellConfig (435)–OtherConfig (436)–RAND-CDMA2000 (1xRTT) (437)–RAT-Type (437)–ResumeIdentity (437)–RRC-TransactionIdentifier (438)–S-TMSI (438)–TraceReference (438)–UE-CapabilityRAT-ContainerList (438)–UE-EUTRA-Capability (439)–UE-RadioPagingInfo (469)–UE-TimersAndConstants (469)–VisitedCellInfoList (470)–WLAN-OffloadConfig (470)6.3.7MBMS information elements (472)–MBMS-NotificationConfig (472)–MBMS-ServiceList (473)–MBSFN-AreaId (473)–MBSFN-AreaInfoList (473)–MBSFN-SubframeConfig (474)–PMCH-InfoList (475)6.3.7a SC-PTM information elements (476)–SC-MTCH-InfoList (476)–SCPTM-NeighbourCellList (478)6.3.8Sidelink information elements (478)–SL-CommConfig (478)–SL-CommResourcePool (479)–SL-CP-Len (480)–SL-DiscConfig (481)–SL-DiscResourcePool (483)–SL-DiscTxPowerInfo (485)–SL-GapConfig (485)。

P6450-Logic-Analyzer-Probe-with-D-Max-Probing-Technology-Instructionsl

*P071247801*
071-2478-01
Copyright © Tektronix. All rights reserved. Licensed software products are owned by Tektronix or its subsidiaries or suppliers, and are protected by national copyright laws and international treaty provisions.
Tektronix products are covered by U.S. and foreign patents, issued and pending. Information in this publication supersedes that in all previously published material. Speciﬁcations and price change privileges reserved.
TEKTRONIX and TEK are registered trademarks of Tektronix, Inc.
D-Max is a registered trademark of Tektronix, Inc. MagniVu is a trademark of Tektronix, Inc.
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Contacting Tektronix

New England Biolabs Pulse Marker

Pulse Marker ™50−1,000 kbCatalog Number D2416Storage Temperature 2−8 °C. Do not freeze.Product DescriptionPulse Marker ™50−1,000 kb consists of ~21successive concatemers of lambda DNA embedded in 1% low melting point agarose with 10mM Tris-HCl, pH 8.0, 1mM EDTA, and 50% glycerol as the storage buffer. Supplied in a convenient graduated syringe, the recommended plug size is ~20µl or two small graduations on the syringe. One syringe contains 25plugs which is sufficient for 25 uses. The concentration of DNA is ~1µg/20µl plug.Suitable for use as molecular weight marker in pulsed field gel electrophoresis.Figure 1.Gel imageof Pulse Marker ™Precautions and DisclaimerThis product is for R&D use only, not for drug,household, or other uses. Please consult the Material Safety Data Sheet for information regarding hazards and safe handling practices. Storage/StabilityStore at 2−8 °C. Do not freeze.ProcedurePosition syringe cap facing up and remove the cap. Remove storage buffer using the plunger and extrude the agarose from the syringe. Slice plugs from the end with a clean sharp blade. Retract the agarose into the syringe and replace the cap. Each plug is sufficient for one lane. Place the plug against the side of the well facing in the direction of electrophoretic migration.ResultsCHEF (Contour-clamped Homogeneous Electric Field) Gel Analysis - A 1% agarose gel prepared in 0.5X TBE was subjected to pre-electrophoresis using running conditions for 1hour. Samples were added and run in 0.5X TBE at 14 °C for 45.5 hours at 150 V. Pulse times were ramped from 5−120 seconds. After staining in 0.5µg/ml ethidium bromide and analysis with UV (301nm) transillumination, at least15(48.5−776kb, see Figure 1) of 21 possible ladder bands (see Table 1) were resolved.References1.Birren, B. and Lai, E. Pulsed Field GelElectrophoresis: A Practical Guide. Academic Press, San Diego, 1993.2.Chu, G. Pulsed field electrophoresis in contour-clamped homogeneous electric fields for the resolution of DNA by size or topology. Electrophoresis, 10, 290-295 (1989).Pulse Marker is a trademark of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich CoRS,PHC 08/10-1Sigma brand products are sold through Sigma-Aldrich, Inc.Sigma-Aldrich, Inc. warrants that its products conform to the information contained in this and other Sigma-Aldrich publications. Purchaser must determine the suitability of the product(s) for their particular use. Additional terms and conditions may apply.Please see reverse side of the invoice or packing slip.。

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Lead Time 30 30 30 30 30 30 30 30 30 30 30 30
Machine Size 200T 200T 200T 200T 200T 200T 200T 120T 120T 120T 120T 200T
Machine Rate/Hr-USD 19.3 19.3 19.3 19.3 19.3 19.3 19.3 15 15 15 15 19.3
FACTORY SIGNATURE
1 1 1 1 1 1 1 1 1 1
DAILY CAPACITY 1494.00 1494.00 1494.00 8337.00 8337.00 8800.00 8800.00 8800.00 8800.00 8800.00 8800.00 8800.00
TOOL PRICE USD US$
PROJECT NAME ITEM# REV # Quote Date PDP PO# TOOLING FACTORY RESPONSIBLE PM PAYMENT TERMS INVOICE# OTHER Grand Total Tooling Cost #600232 PL3793&PL6452
Mold Material 母模仁：S136热处理公模仁：S136热处理母模仁：S136热处理公模仁：S136热处理母模仁：S136热处理公模仁：S136热处理母模仁：S136热处理公模仁：S136热处理母模仁：S136热处理公模仁：S136热处理母模仁：S136热处理公模仁：S136热处理母模仁：S136热处理公模仁：S136热处理母模仁：S136热处理公模仁：S136热处理母模仁：S136热处理公模仁：S136热处理母模仁：S136热处理公模仁：S136热处理母模仁：S136热处理公模仁：S136热处理母模仁：S136热处理公模仁：S136热处理
DISCLAIMER / OWNERSHIP
Total Molding Labor :
All tooling referenced herein shall be the sole and exclusive property of Performance Designed Products LLC ("PDP"). PDP shall have the right, with reasonable advanced notice, at any time to remove, collect and/or move the tooling from the factory location to any other location of PDP's choosing without any interference from the above-referenced factory. The above-referenced factory at no point has the right to inject parts for any company other than PDP, or use the tooling for any other purpose, without PDP's express prior written consent. Acceptance of this tooling plan by the above-referenced factory indicates acceptance of the foregoing terms in their entirety, and PDP reserves all its rights to enforce these terms, and any violation thereof, by the above-referenced factory.
Performance Designed Products Tool Plan Confidential
Currency Conversion Rate:
Finish/ TOOL NUMBER DESCRIPTION Texture
Plastic Material PC PC PC ABS ABS ABS ABS PC PC PC PC ABS
Cycle Time 53.00 53.00 53.00 38.00 38.00 36.00 36.00 36.00 36.00 36.00 36.00 36.00
Lifetime Cycles 500K 500K 500K 500K 500K 500K 500K 500K 500K 500K 500K 500K CAVITY 1 1 1 1 1 1 1 1 1 1 1 1 X X X X X X X X X X X X 1 1 1 4 4 4 4 4 4 4 4 4
Unit Cost-US$
頭帶 (HEADBAND ）頭帶 PDP_3793T1202_060513 (HEADBAND ）頭帶 PDP_3793T1203_060513 (HEADBAND ） PDP_3793T1201_060513 PDP_3793T1204_060513 左耳殼上蓋（earcup-L-top) PDP_3793T1205_060513 右耳殼上蓋（earcup-R-top) PDP_3793T1206_060513 左耳殼下蓋（earcup-L-bottom) PDP_3793T1207_060513 右耳殼下蓋（earcup-R-bottom) PDP_3793T1208_060513 左耳殼中蓋（earcup-L-mid) PDP_3793T1209_060513 右耳殼中蓋（earcup-R-mid) PDP_3793T1210_060513 左導光板（LIGHT GUIDE plate-L) PDP_3793T1211_060513 右导光板（LIGHT GUIDE plate-R) PDP_3793T1212_060513 耳套蓋 (speaker_plate-r-top)