Abstract
An asymmetric (top vs. bottom halves of pulvini) induction of invertase mRNA by gravistimulation was analyzed in oat shoot pulvini. Total RNA and poly(A)+ RNA, isolated from oat pulvini, and two oligonucleotide primers, corresponding to two conserved amino acid sequences (NDPNG and WECPD) found in invertase from other species, were used for the polymerase chain reaction (PCR). A partial length cDNA (550 bp) was obtained and characterized. A 62% nucleotide sequence homology and 58% deduced amino acid sequence homology, as compared to β-fructosidase of carrot cell wall, was found. Northern blot analysis showed that there was an obviously transient induction of invertase mRNA by gravistimulation in the oat pulvinus system. The mRNA was rapidly induced to a maximum level at 1 h after gravistimulation treatment and gradually decreased afterwards. The mRNA level in the bottom half of the oat pulvinus was significantly higher than that in the top half of the pulvinus tissue. The kinetic induction of invertase mRNA was consistent with the transient accumulation of invertase activity during the graviresponse of the pulvinus. This indicates that the expression of the invertase gene(s) could be regulated by gravistimulation at the transcriptional level. Southern blot analysis showed that there were two to three genomic DNA fragments which hybridized with the partial-length invertase cDNA.
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References
Brock TG, Kaufman PB: Growth regulators: an account of hormones and growth regulation. In: Bidwell PGS (ed) Growth and Development Plant Physiology, vol. 10, pp. 277–326. Academic Press, New York (1991).
Gilbeaut DM, Karuppiah N, Chang S-R, Brock TG, Vadlamudi B, Kim D, Ghosheh NS, Rayle DL, Carpita NC, Kaufman PB: Cell wall and enzyme changes during the graviresponse of leaf-sheath pulvinus of oat (Avena sativa). Plant Physiol 94: 411–416 (1990).
Innis MA, Myambo KB, Gelfand DH, Brow MAD: DNA sequencing with Thermus aquaticus DNA polymerase and direct sequencing of polymerase chain reaction-amplified DNA. Proc Natl Acad Sci USA 85: 9436–9440 (1988).
Karuppiah N, Vadlamudi B, Kaufman PB: Purification and characterization of soluble (cytosolic) and bound (cell wall) isoforms of invertase in barley (Hordeum vulgare) elongating stem tissue. Plant Physiol 91: 993–998 (1989).
Kim WT, Silverstone A, Yip WK, Dong JG, Yang SF: Induction of 1-amino-cyclopropane-1-carboxylate synthase mRNA by auxin in mung bean hypocotyls and cultured apple shoots. Plant Physiol 98: 465–471 (1992).
Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press. Cold Spring Harbor, NY (1990).
Sturm A, Chrispeels MJ: cDNA cloning of carrot extracellular β-fructosidase and its expression in response to wounding and bacterial infection. Plant Cell 2: 1107–1119 (1990).
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Wu, LL., Song, I., Karuppiah, N. et al. Kinetic induction of oat shoot pulvinus invertase mRNA by gravistimulation and partial cDNA cloning by the polymerase chain reaction. Plant Mol Biol 21, 1175–1179 (1993). https://doi.org/10.1007/BF00023613
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DOI: https://doi.org/10.1007/BF00023613