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Plant Physiol, October 1999, Vol. 121, pp. 479-488
Cloning, Expression, and Molecular Characterization of a Small
Pea Gene Family Regulated by Low Levels of Ultraviolet B Radiation and
Other Stresses1
Mikael
Brosché and
Åke
Strid*
Biokemi och Biofysik, Göteborgs Universitet, P.O. Box 462, S-40530 Göteborg, Sweden
A pea (Pisum sativum)
DNA fragment (termed MB3) was isolated by differential
display of cDNAs obtained from total leaf RNA of ultraviolet B (UV-B)
radiation-treated plants. Longer cDNAs were cloned by rapid
amplification of cDNA ends in the 3' to 5' direction. Three different,
but very similar, cDNAs were cloned, sadA, sadB, and
sadC, the major difference between them being a 36-bp
deletion in the coding region of sadB. Southern blotting confirmed the occurrence of at least three genes in the pea genome. Database comparisons of the SAD protein sequences revealed high identity (46%) and similarity (77%) with a putative tomato
(Lycopersicon esculentum) short-chain alcohol
dehydrogenase. Very low levels of UV-B radiation (the biologically
effective radiation normalized to 300 nm = 0.08 W
m 2) was shown to up-regulate expression, a dose
considerably lower than that needed to induce expression of the
well-known UV-B defensive chalcone synthase and phenylalanine ammonia
lyase genes. RNase protection assay revealed that primarily
sadA and sadC mRNA accumulation was
enhanced by UV-B. In addition to UV-B irradiation, ozone fumigation, wounding, aluminum stress, and salt stress induced increased transcript levels of the sad genes in pea.
1
This work was supported by grants to Å.S. from
the Swedish Natural Science Research Council, the Crafoord Foundation,
the Carl Trygger Foundation, and the Swedish Research Council for Engineering Sciences and to M.B. from the Lawski Foundation,
Hierta-Retzius fond för vetenskaplig forskning, and from Kungliga
och Hvitfeldtska stipendiestiftelsen.
*
Corresponding author; e-mail ake.strid{at}bcbp.gu.se; fax
46-31-7733910.
© 1999 American Society of Plant Physiologists
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