Joel J. Sohlberg

Auxin can act independently of CRC, LUG, SEU, SPT and STY1 in style development but not apical‐basal patterning of the Arabidopsis gynoecium

Patterning of the Arabidopsis thaliana gynoecium is dependent on the localization and concentration of the plant hormone auxin and it has been previously reported that STYLISH1 (STY1) activates transcription of the auxin biosynthesis gene YUCCA4 (YUC4) and affects gynoecium development. Here, the relationship between auxin, STY1 and other regulators of gynoecium development was examined. Exogenous auxin in droplets of lanolin paste were applied to young gynoecia; auxin biosynthesis rate was measured and STY1 overexpression or chemically mediated polar auxin transport (PAT) inhibition were induced in various mutants. The style phenotype of sty1-1sty2-1 mutants was restored by exogenous application of auxin, and STY1 over-activation resulted in an elevated auxin biosynthesis rate. Both over-activation of STY1 and inhibition of PAT restored the stylar defects of several unrelated mutants, but with regard to gynoecium apical-basal patterning the mutants responded differently to inhibition of PAT. These results suggest that reduced auxin concentrations cause the sty1-1 sty2-1 phenotype, that STY1 induces auxin biosynthesis, that elevated apical auxin concentrations can compensate for the loss of several style-promoting factors, and that auxin may act downstream of, or in parallel with these during style development but is dependent on their action in apical-basal patterning.

Analysis of the WUSCHEL-RELATED HOMEOBOX gene family in the conifer picea abies reveals extensive conservation as well as dynamic patterns.

BackgroundMembers of the WUSCHEL-RELATED HOMEOBOX (WOX) gene family have important functions during all stages of plant development and have been implicated in the development of morphological novelties during evolution. Most studies have examined the function of these genes in angiosperms and very little is known from other plant species.ResultsIn this study we examine the presence and expression of WOX genes in the conifer Picea abies. We have cloned 11 WOX genes from both mRNA and genomic DNA and examined their phylogenetic relationship to WOX genes from other species as well as their expression during somatic embryogenesis and in adult tissues.ConclusionsOur study shows that all major radiations within the WOX gene family took place before the angiosperm-gymnosperm split and that there has been a recent expansion within the intermediate clade in the Pinaceae family. Furthermore, we show that the genes from the intermediate clade are preferentially expressed during embryo development in Picea abies. Our data also indicates that there are clear orthologs of both WUS and WOX5 present in the P. abies genome.

WUSCHEL-RELATED HOMEOBOX 8/9 is important for proper embryo patterning in the gymnosperm Norway spruce

Summary The WUSCHEL-RELATED HOMEOBOX 8/9 is important for correct orientation of the cell division plane and for cell fate determination during embryo pattern formation in the gymnosperm Norway spruce.

WUSCHEL-RELATED HOMEOBOX 2 is important for protoderm and suspensor development in the gymnosperm Norway spruce

BackgroundDistinct expression domains of WUSCHEL-RELATED HOMEOBOX (WOX) gene family members are involved in patterning and morphogenesis of the early embryo in Arabidopsis. However, the role of WOX genes in other taxa, including gymnosperms, remains elusive. Here, we use somatic embryos and reverse genetics for studying expression and function of PaWOX2, the corresponding homolog of AtWOX2 in the gymnosperm Picea abies (Pa; Norway spruce).ResultsThe mRNA level of PaWOX2 was transiently up-regulated during early and late embryogeny. PaWOX2 mRNA in early and early late embryos was detected both in the embryonal mass and in the upper part of the suspensor. Down-regulation of PaWOX2 during development of early embryos resulted in aberrant early embryos, which failed to form a proper protoderm. Cells on the surface layer of the embryonal mass became vacuolated, and new embryogenic tissue differentiated from the embryonal mass. In addition, the aberrant early embryos lacked a distinct border between the embryonal mass, and the suspensor and the length of the suspensor cells was reduced. Down-regulation of PaWOX2 in the beginning of embryo development, before late embryos were formed, caused a significant decrease in the yield of mature embryos. On the contrary, down-regulation of PaWOX2 after late embryos were formed had no effect on further embryo development and maturation.ConclusionsOur data suggest an evolutionarily conserved function of WOX2 in protoderm formation early during embryo development among seed plants. In addition, PaWOX2 might exert a unique function in suspensor expansion in gymnosperms.

The WUSCHEL‐RELATED HOMEOBOX 3 gene PaWOX3 regulates lateral organ formation in Norway spruce

Summary In angiosperms, WUSCHEL‐RELATED HOMEOBOX 3 (WOX3) genes are required for the recruitment of founder cells from the lateral domains of shoot meristems that form lateral regions of leaves. However, the regulation of the formation of lateral organs in gymnosperms remains unknown. By using somatic embryos of Norway spruce (Picea abies) we have studied the expression and function of PaWOX3 during embryo development. The mRNA abundance of PaWOX3 was determined by quantitative real‐time PCR, and the spatial expression of PaWOX3 was analysed by histochemical β‐glucuronidase (GUS) assays and in situ mRNA hybridization. To investigate the function of PaWOX3, we analysed how downregulation of PaWOX3 in RNA interference lines affected embryo development and morphology. PaWOX3 was highly expressed in mature embryos at the base of each cotyledon close to the junction between the cotyledons, and in the lateral margins of cotyledons and needles, separating them into an adaxial and an abaxial side. Downregulation of the expression of PaWOX3 caused defects in lateral margin outgrowth in cotyledons and needles, and reduced root elongation. Our data suggest that the WOX3 function in margin outgrowth in lateral organs is conserved among the seed plants, whereas its function in root elongation may be unique to gymnosperms.