Leaf developmental stage affects sulfate depletion and specific sulfate transporter expression during sulfur deprivation in Brassica napus L.

BRASSICA NAPUS was grown under hydroponic conditions and responses to the removal of the external supply of sulfur (S) were analysed in roots and in leaves of different developmental age. The concentrations of sulfate and nitrate were greatest in the older leaves and least in younger leaves, whilst phosphate was greatest in roots and youngest leaves and least in old leaves. S-deprivation resulted in decreases in tissue sulfate concentrations at variable rates in the order: roots and young leaves > middle-aged leaves > oldest leaves. Phosphate concentrations were unaffected and nitrate concentrations were only depleted in the oldest leaves. Expression of representative members of the sulfate transporter gene family was assessed by Northern blotting in the respective tissues. Group 1 transporters (high affinity type) were induced in response to S-deprivation in all tissues except old leaves, where no expression was detected, and to the greatest extent in roots. Groups 2 and 5 (a BRASSICA Group 5 sulfate transporter is reported here, accession number: AJ311389) transporters showed either no or only a small induction by S-deprivation. Group 4 transporters (localised in the tonoplast membrane and thought to be involved in vacuolar sulfate efflux) were induced by S-deprivation with a complex pattern: 4;1 was expressed in root and mature leaves, was strongly induced by sulfur-deprivation in roots, and was also induced in the middle-aged leaves alone; 4;2 was only expressed under S-deprivation in parallel with the observed pattern of tissue sulfate concentrations. Expression patterns indicated that both differences in intracellular sulfate pools and localised aspects of the signal transduction pathway link tissue sulfate-status and sulfur-nutrition regulated gene expression.

[1]  M. Hawkesford,et al.  Sulphur Assimilation and Effects on Yield and Quality of Wheat , 1999 .

[2]  R. Hell,et al.  Regulation of Sulfate Uptake and Expression of Sulfate Transporter Genes in Brassica oleracea as Affected by Atmospheric H2S and Pedospheric Sulfate Nutrition1 , 2004, Plant Physiology.

[3]  Hideki Takahashi,et al.  Vacuolar Sulfate Transporters Are Essential Determinants Controlling Internal Distribution of Sulfate in Arabidopsis , 2004, The Plant Cell Online.

[4]  Peter Buchner,et al.  Plant sulphate transporters: co-ordination of uptake, intracellular and long-distance transport. , 2004, Journal of experimental botany.

[5]  E. J. Evans,et al.  Influence of nitrogen and sulphur on the glucosinolate profile of rapeseed (Brassica napus L.) , 1994 .

[6]  J. Thompson,et al.  The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. , 1997, Nucleic acids research.

[7]  M. Hawkesford,et al.  Diagnosing sulfur deficiency in field-grown oilseed rape (Brassica napus L.) and wheat ( Triticum aestivum L.) , 2000, Plant and Soil.

[8]  E. J. Evans,et al.  Sulphur uptake and distribution in double and single low varieties of oilseed rape (Brassica napus L.) , 1993, Plant and Soil.

[9]  Hideki Takahashi,et al.  Phloem-Localizing Sulfate Transporter, Sultr1;3, Mediates Re-Distribution of Sulfur from Source to Sink Organs in Arabidopsis1 , 2003, Plant Physiology.

[10]  M. Van Montagu,et al.  Regulation of sulfur assimilation in higher plants: a sulfate transporter induced in sulfate-starved roots plays a central role in Arabidopsis thaliana. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[11]  D. Clarkson,et al.  Partitioning and redistribution of sulphur during S-stress in Macroptilium atropurpureum cv. Siratro , 1995 .

[12]  G. Church,et al.  Genomic sequencing. , 1993, Methods in molecular biology.

[13]  M. Hawkesford Transporter gene families in plants: the sulphate transporter gene family: redundancy or specialization? , 2003 .

[14]  M. Hawkesford,et al.  Distribution of sulfur within oilseed rape leaves in response to sulfur deficiency during vegetative growth , 1998, Plant physiology.

[15]  F. W. Smith,et al.  Plant members of a family of sulfate transporters reveal functional subtypes. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[16]  A. Hoekema,et al.  A small-scale procedure for the rapid isolation of plant RNAs. , 1989, Nucleic acids research.