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Effect of hormone treatment on Norway spruce seedlings [Hypocotyl]

ABSTRACT: We analyze the underlying molecular mechanisms for the observed changes to cambium and xylem differentiation by assessing gene expression changes after hormone applications.

ORGANISM(S): Picea abies

PROVIDER: GSE178533 | GEO | 2024/06/20

REPOSITORIES: GEO

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Effect of hormone treatment on Norway spruce seedlings [Hypocotyl]

Project description:Effect of hormone treatment on Norway spruce seedlings [Hypocotyl]

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Project description:The goal is to look at changes in the pattern of expression of the xylem transcriptome through the growth season in two spruces (Picea glauca and Picea abies).

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Transcriptome-wide analysis of active xylem in White Spruce and Norway Spruce during the growth season

Project description:The goal is to look at changes in the pattern of expression of the xylem transcriptome through the growth season in two spruces (Picea glauca and Picea abies). One-color comparison of active xylem collected in June, July, August and September, in two spruce species. Six biological repetitions per time point and specie, for a total of 48 slides.

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The Populus Homeobox Gene ARBORKNOX2 Regulates Secondary Growth in trees

Project description:Currently little is known about the genetic mechanisms regulating the vascular cambium or the secondary growth of stems. We show here that the Populus Class I KNOX homeobox gene ARBORKNOX2 (ARK2) regulates both cell division in the cambium region and the differentiation of daughter cells in secondary xylem and phloem. ARK2 is expressed in the shoot apical meristem, and the vascular cambium region, reflecting some overlap in the regulation of these meristems. ARK2 is expressed broadly in the cambium region and in differentiating lignified cells types before becoming progressively restricted to the cambium. Populus overexpressing ARK2 present stem phenotypes with precocious cambium formation, delayed differentiation of cambium daughter cells, a wider cambium region, and ultimately less phloem fibers and secondary xylem. In contrast, Populus expressing RNAi or amicroRNA that target ARK2 transcripts present precocious differentiation of secondary phloem fibers and xylem, and ultimately more secondary xylem tissue and thicker secondary cell walls in phloem fibers and secondary xylem cells. These phenotypes in turn correlate with changes in the expression of genes affecting cell division, auxin, and cell wall synthesis and lignification that indicate that ARK2 primarily affects woody tissue development by regulation of cell differentiation. Notably, wood properties associated with secondary cell wall synthesis are negatively associated with ARK2 expression, including lignin and cellulose content. Together, our results suggest that ARK2 functions primarily by negatively regulating cell differentiation during secondary growth. We propose that ARK2 may identify a co-evolved regulatory module that influences complex wood properties relevant to ecological, industrial, and biofuels applications.

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A common graft formation pathway in Norway spruce and Arabidopsis thaliana identifies the PAT gene family as conserved regulators of tissue attachment and wound healing [RNAseq: Grafted-samples]

Project description:The widespread use of plant grafting has enabled different plants to join and combine their best properties to improve stress tolerance, growth and yields. Grafting is commonly performed between closely related eudicots or gymnosperms where mechanisms permit tissue fusion yet limit success as plants become unrelated. To investigate these aspects, we developed a micrografting method using young conifer tree seedlings that enabled divergent conifer members to successfully graft. Conifer grafts showed rapid connection of phloem and xylem at the junction, while a genome-wide transcriptome analysis of the Picea abies (Norway spruce) healing junction revealed differential expression of thousands of genes including those related to auxin response and cell wall biogenesis. We compared these genes to those induced during Arabidopsis thaliana graft healing and found a common activation of cambium, cell division, phloem and xylem-related genes. A gene regulatory network analysis revealed that PHYTOCHROME A SIGNAL TRANSDUCTION 1 (PAT1) acted as a central hub during Picea grafting and was also induced during Arabidopsis grafting. Arabidopsis mutants lacking PATs failed to attach tissues or successfully graft, while complementing Arabidopsis PAT mutants with the Picea abies PAT1 homolog could rescue tissue attachment and enhance callus formation. Together, our data demonstrate a competency for young tissues to graft to distantly related species and identifies the PAT gene family as conserved regulators of graft healing and tissue regeneration in eudicots and gymnosperms.

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A common graft formation pathway in Norway spruce and Arabidopsis thaliana identifies the PAT gene family as conserved regulators of tissue attachment and wound healing [RNAseq: Hornome-Treatment-samples]

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Hunting monolignol transporter(s): membrane proteomics and biochemical transport assays with membrane vesicles of Norway spruce

Project description:Monolignol transport during lignification is a partially solved puzzle: both the mechanism(s) and the transported form of monolignols are unknown in developing xylem of trees. We tested a hypothesis of an active, plasma membrane (PM)-localized transport of monolignol monomers, dimers, and/or glucosidic forms with membrane vesicles prepared of developing xylem, phloem, and lignin-forming, tissue-cultured cells of Norway spruce (Picea abies L. Karst.), as well as of a control material, non-lignifying tobacco (Nicotiana tabacum L.) BY-2 cells. Xylem and BY-2 vesicles transported both coniferin and p-coumaryl alcohol glucoside, but inhibitor assays suggested this transport being over the tonoplast. Based on similar inhibitor assays, lignin-forming, tissue42 cultured cells of spruce had coniferin transport putatively localizing on PM. Liquid chromatography-mass spectrometry (LC-MS/MS) analysis of membrane proteins isolated from spruce developing xylem, phloem and tissue-cultured cells revealed multiple transporters. These were compared to a transporter gene set that was gained by a correlation analysis with a selected set of spruce monolignol biosynthesis genes. Biochemical membrane vesicle assays showed no support for the ABC-transporter-mediated monolignol transport but point to secondary active transporters (such as MFS- or MATE-transporters). In contrast, proteomic and co-expression analyses suggest a role for ABC-transporters and MFS-transporters.

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Protein Expression in TW Formation

Project description:Tangential sections of 30 µm thickness were taken across the secondary phloem and developing secondary xylem, sections were then pooled to represent the tissues from specific developmental stages. Phloem (P) comprises 3 sections, the cambium (C) - 2 sections, the radial expansion zone (E) - 4 sections, the first to third xylem development zones (X1-X3) - 3 sections each, and the forth to sixth xylem development zones (X4-X6) - 6 sections each.

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Transcriptome profiles and comparison of Arabidopsis thaliana woody (soc1-ful) and WT (Col-0) stems.

Project description:The douple mutant Arabidopsis thaliana soc1 ful, in contrast with WT, produces an interfascicular cambium and a large wood cylinder is the flowering stem. We present the RNAseq data for polyA mRNA of different developmental stages of cambium and wood formation in Arabidopsis thaliana. We sequenced 7 stages; 4 in the woody mutant soc1-6 ful-7 (herbaceous, cambium initiation, wood initiation and leaf) and 3 stages in the WT Col-0 (herbaceous, cambium and leaf). The corresponding stem anatomy is also presented in the manuscript indicating the stage of cambium development and the production of secondary xylem.

2016-05-21 | GSE69167 | GEO

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