Samples harvested independently in the same time point as these collected for RNAseq analysis.5 genes wereFrontiers in Plant Apratastat Data Sheet Science www.frontiersin.orgMay Volume ArticleZhang et al.PollenStigma Interactions in Brassica napus L.FIGURE Annotation of stigmaenriched genes.(A) The most extremely represented GO terms in each category (biological procedure, cellular components and molecular functions).(B) Identification from the genes within the SadenosylLmethionine (SAM) cycle and Sadenosylmethioninedependent methyltransferases.Frontiers in Plant Science www.frontiersin.orgMay Volume ArticleZhang et al.PollenStigma Interactions in Brassica napus L.FIGURE Validation of eight randomly chosen genes by qRTPCR.(A) One of the early stage DEGs.(B,C) Two DEGs at late stage.(D,E) Two DEGs at all stages.(F) Stigmaenriched genes.BnaAgD and BnaAgD are genes involved in PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/21543622 SAM cycle.mRNA expression levels were normalized to the expression of ACTIN, and implies from 3 biological replicates are shown.Error bars indicate SE.r represents the correlation coefficient.Frontiers in Plant Science www.frontiersin.orgMay Volume ArticleZhang et al.PollenStigma Interactions in Brassica napus L.selfcompatibility of “Westar” (Okamoto et al Tochigi et al).”W” shows sturdy selfincompatibility and has the identical genetic background with “Westar” except for the induced functional BnSP.As a result, the transgenic B.napus line “W” was perfect to study compatible and incompatible pollenstigma interactions.By observation of “Westar” stigmas min after pollination utilizing TEM, all “W” pollen grains had been identified to become intact (i.e showed no change in morphology), while some “Westar” pollen grains germinated and began to invade the cell wall of the stigma papilla cell (Figure A).A timecourse transcriptome analysis was employed to investigate compatible and incompatible pollenstigma interactions, a moderate transform in gene expression level was observed at , , and min just after pollination (varying from to DEGs), and a drastic modify was located at and min just after pollination (varying from to DEGs) (Figure B; Supplemental File S).A moderate number of DEGs ( in compatible interaction and in incompatible interaction) appeared during all stages of pollination and they were all upregulated; the majority of DEGs have been detected at time points of and min, such as in compatible interaction ( upregulated, downregulated) and in incompatible interaction ( upregulated, downregulated).In the above final results, it could possibly be deduced that pollenstigma interaction would complete min just after pollination, and downstream components were activated in signaling pathways of each compatible and incompatible responses, whilst the signal transduction networks in incompatible response may well be more complicated than that in compatible response.Enriched genes in all stigma samples like unpollinated stigmas have been firstly analyzed in our present study.We discovered the reported pollenstigma interaction genes, the stigma determinant gene BnSRK (Stein et al Takasaki et al Okamoto et al), pollen adhesion associated genes SLG and SLR (Luu et al ,), have been expressed extremely in unpollinated stigma and all pollinated stigmas, which can be in accordance using the demonstration by Nasrallah that the SI response is regulated for the duration of stigma maturation stigmas are initially compatible with selfpollen and acquire the ability to reject selfpollen in conjunction with anther dehiscence days before flower opening or anthesis.Based.
