Spermine facilitates recovery from drought but does not confer drought tolerance in transgenic rice plants expressing Datura stramonium S-adenosylmethionine decarboxylase

Polyamines are known to play important roles in plant stress tolerance but it has been difficult to determine precise functions for each type of polyamine and their interrelationships. To dissect the roles of putrescine from the higher polyamines spermidine and spermine, we generated transgenic rice plants constitutively expressing a heterologous S-adenosylmethionine decarboxylase (SAMDC) gene from Datura stramonium so that spermidine and spermine levels could be investigated while maintaining a constant putrescine pool. Whereas transgenic plants expressing arginine decarboxylase (ADC) produced higher levels of putrescine, spermidine and spermine, and were protected from drought stress, transgenic plants expressing SAMDC produced normal levels of putrescine and showed drought symptoms typical of wild type plants under stress, but the transgenic plants showed a much more robust recovery on return to normal conditions (90% full recovery compared to 25% partial recovery for wild type plants). At the molecular level, both wild type and transgenic plants showed transient reductions in the levels of endogenous ADC1 and SAMDC mRNA, but only wild type plants showed a spike in putrescine levels under stress. In transgenic plants, there was no spike in putrescine but a smooth increase in spermine levels at the expense of spermidine. These results confirm and extend the threshold model for polyamine activity in drought stress, and attribute individual roles to putrescine, spermidine and spermine

Development of a novel gene transfer system for Cajanus cajan and expression of a monocot arginine decarboxylase cDNA in transformed cell lines

We describe the development of a novel transformation system for pigeonpea (Cajanus cajan (L.) Millsp.) cell lines using particle bombardment. The aim of our study was not only to develop a transformation system for this important but recalcitrant species but to do so using a gene of interest and to study the behaviour of this transgene in a legume background. The gene we chose for our studies was the heterologous oat arginine decarboxylase cDNA. Transgenic lines engineered with this cDNA had a small, but significant increase in arginine decarboxylase activity (ADC; EC 4.1.1.17). We detected an increase in putrescine levels in all the lines we analysed irrespective of whether major or minor changes in ADC activity were observed. Spermidine and spermine levels were also increased in some of the lines. We confirmed that small changes in the adc message (mRNA) had a significant effect on enzyme activity and more importantly on polyamine levels. This is the first report describing modulation of a gene involved in a metabolic pathway in transgenic pigeonpea tissues

Molecular characterization of the Arginine decarboxylase gene family in rice

Arginine decarboxylase (ADC) is a key enzyme in plants that converts arginine into putrescine, an important mediator of abiotic stress tolerance. Adc genes have been isolated from a number of dicotyledonous plants but the oat and rice Adc genes are the only representatives of monocotyledonous species described thus far. Rice has a small family of Adc genes, and OsAdc1 expression has been shown to fluctuate under drought and chilling stress. We identified and characterized a second rice Adc gene (OsAdc2) which encodes a 629-amino-acid protein with a predicted molecular mass of 67 kDa. An unusual feature of the OsAdc2 gene is the presence of an intron and a short upstream open reading frame in the 50-UTR. Sequence comparisons showed that OsAdc2 is more closely related to the oat Adc gene than to OsAdc1 or to its dicot homologs, and mRNA analysis showed that the two rice genes are also differently regulated. Whereas OsAdc1 is expressed in leaf, root and stem, OsAdc2 expression is restricted to stem tissue. Protein expression was investigated with specific antibodies against ADC1 and ADC2, corroborating the mRNA data. We discuss the expression profiles of OsAdc1 and OsAdc2 and potential functions for the two corresponding proteins

Transgenic wheat plants expressing an oat arginine decarboxylase cDNA exhibit increases in polyamine content in vegetative tissue and seeds

We generated and characterized a trans- genic wheat ( Triticum aestivum L.) population expressing constitutively an oat arginine decarboxylase ( ADC ) cDNA. Transgenic wheat plants expressing the transgene had a significant increase in ADC activity with no concomitant changes in ODC activity. We measured 2-fold increases in putrescine, spermidine and spermine content in leaves of transgenic plants com- pared to wild-type. These plants also exhibited significant increases in the three polyamines in seeds (7-fold putrescine, 2.5-fold spermidine and 1.8-fold spermine). This significant polyamine accumulation was heritable in subsequent generations. In order to characterize molecularly this population we cloned a partial TaADC cDNA. This sequence was used as a probe to ascertain the presence of two distinct ADC transcripts in wheat. Further DNA analyses confirmed the presence of at least two distinct ADC genes in the wheat genome. Our results demonstrate that polyamine levels can be altered in transgenic wheat plants resulting in phenotypically normal and fertile plants. This transgenic wheat population will serve as useful germ- plasm to embark on further in depth studies to ascertain the role of polyamines in abiotic stress responses