Overexpressing the Multiple-Stress Responsive Gene At1g74450 Reduces Plant Height and Male Fertility in Arabidopsis thaliana.
A subset of genes in Arabidopsis thaliana is known to be up-regulated in response to a wide range of different environmental stress factors. However, not all of these genes are characterized as yet with respect to their functions. In this study, we used transgenic knockout, overexpression and reporter gene approaches to try to elucidate the biological roles of five unknown multiple-stress responsive genes in Arabidopsis. The selected genes have the following locus identifiers: At1g18740, At1g74450, At4g27652, At4g29780 and At5g12010. Firstly, T-DNA insertion knockout lines were identified for each locus and screened for altered phenotypes. None of the lines were found to be visually different from wildtype Col-0. Secondly, 35S-driven overexpression lines were generated for each open reading frame. Analysis of these transgenic lines showed altered phenotypes for lines overexpressing the At1g74450 ORF. Plants overexpressing the multiple-stress responsive gene At1g74450 are stunted in height and have reduced male fertility. Alexander staining of anthers from flowers at developmental stage 12-13 showed either an absence or a reduction in viable pollen compared to wildtype Col-0 and At1g74450 knockout lines. Interestingly, the effects of stress on crop productivity are most severe at developmental stages such as male gametophyte development. However, the molecular factors and regulatory networks underlying environmental stress-induced male gametophytic alterations are still largely unknown. Our results indicate that the At1g74450 gene provides a potential link between multiple environmental stresses, plant height and pollen development. In addition, ruthenium red staining analysis showed that At1g74450 may affect the composition of the inner seed coat mucilage layer. Finally, C-terminal GFP fusion proteins for At1g74450 were shown to localise to the cytosol.
Department of Plant Sciences, University of Oxford, Oxford, OX1 3RB, Oxfordshire, United Kingdom.
Anne M
AM
Visscher
Department of Plant Sciences, University of Oxford, Oxford, OX1 3RB, Oxfordshire, United Kingdom.
Eric J
EJ
Belfield
Department of Plant Sciences, University of Oxford, Oxford, OX1 3RB, Oxfordshire, United Kingdom.
Daniela
D
Vlad
Department of Plant Sciences, University of Oxford, Oxford, OX1 3RB, Oxfordshire, United Kingdom.
Niloufer
N
Irani
Department of Plant Sciences, University of Oxford, Oxford, OX1 3RB, Oxfordshire, United Kingdom.
Ian
I
Moore
Department of Plant Sciences, University of Oxford, Oxford, OX1 3RB, Oxfordshire, United Kingdom.
Nicholas P
NP
Harberd
D017360:Arabidopsis / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D005298:Fertility / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D018506:Gene Expression Regulation, Plant; D055786:Gene Knockout Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D056626:Inflorescence / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D018613:Microscopy, Confocal; D009154:Mutation; D010641:Phenotype; D051479:Plant Infertility / Q000235:genetics; D030821:Plants, Genetically Modified; D011058:Pollen / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013312:Stress, Physiological / Q000235:genetics*
D029681:Arabidopsis Proteins; C000608319:At1g74450 protein, Arabidopsis; D049452:Green Fluorescent Proteins