Reduction in the endogenous arginine decarboxylase transcript levels in rice leads to depletion of the putrescine and spermidine pools with no concomitant changes in the expression of downstream genes in the polyamine biosynthetic pathway

Trung-Nghia, Pham; Bassié, Ludovic; Safwat, Gehan; Thu-Hang, P.; Lepri, O.; Rocha, Paulo Alexandre Costa; Christou, Paul; Capell, Teresa

doi:10.1007/s00425-003-1079-3

Cited by 25 publications

(20 citation statements)

References 30 publications

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“…Labeling of the probe, hybridization, washes, and chemiluminescent detection were carried out as described in ref. 10. Membranes were then incubated with CSPD Chemiluminescent Substrate (Roche) and exposed to x-ray film (Fuji) for 30 min at 37°C.…”

Section: Methodsmentioning

confidence: 99%

“…Hybridization with the Datura adc probe was carried out as described above. Probes for the rice adc and samdc genes were generated by PCR as described (10), and reprobing was performed as described in ref. 22.…”

Section: Methodsmentioning

confidence: 99%

“…Plant polyamine content has been modulated by the overexpression͞down-regulation of arginine decarboxylase (adc), ornithine decarboxylase (odc), and S-adenosylmethionine decarboxylase (samdc) (6)(7)(8)(9)(10). Overexpression of heterologous adc or odc cDNAs in plants generally results in the production of high levels of putrescine (11)(12)(13).…”

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confidence: 99%

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Modulation of the polyamine biosynthetic pathway in transgenic rice confers tolerance to drought stress

Capell

Bassié

Christou

2004

Proc. Natl. Acad. Sci. U.S.A.

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We have generated transgenic rice plants expressing the Datura stramonium adc gene and investigated their response to drought stress. We monitored the steady-state mRNA levels of genes involved in polyamine biosynthesis (Datura adc, rice adc, and rice samdc) and polyamine levels. Wild-type plants responded to the onset of drought stress by increasing endogenous putrescine levels, but this was insufficient to trigger the conversion of putrescine into spermidine and spermine (the agents that are believed to protect plants under stress). In contrast, transgenic plants expressing Datura adc produced much higher levels of putrescine under stress, promoting spermidine and spermine synthesis and ultimately protecting the plants from drought. We demonstrate clearly that the manipulation of polyamine biosynthesis in plants can produce drought-tolerant germplasm, and we propose a model consistent with the role of polyamines in the protection of plants against abiotic stress.A biotic stresses such as drought represent some of the most significant constraints to agricultural productivity. Transgenic approaches can be used in combination with conventional breeding strategies to create crops with enhanced drought tolerance, and one way in which this can be achieved is through the manipulation of polyamine metabolism. Polyamines are small, ubiquitous, nitrogenous compounds that have been implicated in a variety of stress responses in plants (1). The link between polyamines and abiotic stress was first documented through putrescine accumulation in response to suboptimal potassium levels in barley (2). Since then, a connection has been suggested between increased putrescine levels and abiotic stress (3). Similar phenomena have been described in animals, e.g., during ischemic and postischemic responses in neurons (4). The physiological role of putrescine in abiotic stress responses is a matter of controversy. It has been very difficult to establish a direct cause-and-effect relationship between increased putrescine levels in plants and abiotic stress. Elevated putrescine might be the cause of stress-induced injury or, alternatively, a protective response resulting from stress (5).The genetic manipulation of polyamine metabolism has become a valuable tool for studying their physiological roles in plants (6). Plant polyamine content has been modulated by the overexpression͞down-regulation of arginine decarboxylase (adc), ornithine decarboxylase (odc), and S-adenosylmethionine decarboxylase (samdc) (6-10). Overexpression of heterologous adc or odc cDNAs in plants generally results in the production of high levels of putrescine (11-13). In most cases, this is accompanied by a relatively small increase in spermidine and spermine concentrations (7,14). Such findings suggest that the levels of spermidine and spermine are under strict homeostatic regulation (15). Therefore, the study of plants transformed with genes involved in polyamine biosynthesis may shed light on the importance of polyamines, their role in the acquisition of stress ...

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Modulation of the polyamine biosynthetic pathway in transgenic rice confers tolerance to drought stress

Capell

Bassié

Christou

2004

Proc. Natl. Acad. Sci. U.S.A.

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536

293

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“…In subsequent experiments we used an antisense transgene to reduce OsADC mRNA levels and hence ADC activity, but OsODC, OsSAMDC and OsSPDS mRNA levels remained unchanged. We concluded that altering polyamine levels had no effect on the steady-state mRNA levels of endogenous pathway genes, demonstrating that the endogenous and transgenic pathways were uncoupled Trung-Nghia et al 2003). These data were used to derive a uniform model in which the putrescine pool must exceed a certain threshold to induce the synthesis of spermidine and spermine under stress.…”

Section: Introductionmentioning

confidence: 98%

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

et al. 2009

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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.

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“…All the genes in the metabolic pathway were shown to function coordinately to produce b-carotene in the endosperm, revealed by the yellow color of the polished rice seed. Capell and colleagues have also used particle bombardment to engineer a complex metabolic pathway, and have created a diverse range of rice germplasm with various genes in the polyamine biosynthetic pathway either over-expressed or suppressed through antisense mechanisms (Capell et al 1998Bassie et al 2000a, b;Noury et al 2000;Lepri et al 2001;Thu-Hang et al 2002;Trung-Nghia et al 2003). Most recently, they have used the data from these numerous lines to synthesize a uniform model of polyamine regulation in plants, based on the simultaneous measurement of endogenous and heterologous gene expression, enzyme activities and polyamine levels (Capell et al 2004).…”

Section: Particle Bombardment Facilitates Simultaneous Multiple Gene mentioning

confidence: 99%

Particle bombardment and the genetic enhancement of crops: myths and realities

et al. 2005

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DNA transfer by particle bombardment makes use of physical processes to achieve the transformation of crop plants. There is no dependence on bacteria, so the limitations inherent in organisms such as Agrobacterium tumefaciens do not apply. The absence of biological constraints, at least until DNA has entered the plant cell, means that particle bombardment is a versatile and effective transformation method, not limited by cell type, species or genotype. There are no intrinsic vector requirements so transgenes of any size and arrangement can be introduced, and multiple gene cotransformation is straightforward. The perceived disadvantages of particle bombardment compared to Agrobacterium-mediated transformation, i.e. the tendency to generate large transgene arrays containing rearranged and broken transgene copies, are not borne out by the recent detailed structural analysis of transgene loci produced by each of the methods. There is also little evidence for major differences in the levels of transgene instability and silencing when these transformation methods are compared in agriculturally important cereals and legumes, and other non-model systems. Indeed, a major advantage of particle bombardment is that the delivered DNA can be manipulated to influence the quality and structure of the resultant transgene loci. This has been demonstrated in recently reported strategies that favor the recovery of transgenic plants containing intact, singlecopy integration events, and demonstrating high-level transgene expression. At the current time, particle bombardment is the most efficient way to achieve plastid transformation in plants and is the only method so far used to achieve mitochondrial transformation. In this review, we discuss recent data highlighting the positive impact of particle bombardment on the genetic transformation of plants, focusing on the fate of exogenous DNA, its organization and its expression in the plant cell. We also discuss some of the most important applications of this technology including the deployment of transgenic plants under field conditions.

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Reduction in the endogenous arginine decarboxylase transcript levels in rice leads to depletion of the putrescine and spermidine pools with no concomitant changes in the expression of downstream genes in the polyamine biosynthetic pathway

Cited by 25 publications

References 30 publications

Modulation of the polyamine biosynthetic pathway in transgenic rice confers tolerance to drought stress

Modulation of the polyamine biosynthetic pathway in transgenic rice confers tolerance to drought stress

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

Particle bombardment and the genetic enhancement of crops: myths and realities

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