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RNAi-mediated silencing of spermidine synthase gene results in reduced reproductive potential in tobacco.

ABSTRACT: Spermidine belongs to a class of polycationic compounds known as polyamines. Polyamines are known to be involved in a wide range of biological processes but the exact role and contribution of different polyamines to these processes are still not clear. In the present study, we have tried to understand the contribution of triamine spermidine to the growth and development of tobacco by downregulating spermidine synthase gene (SPDS) using RNA interference. Down-regulatioin of SPDS gene resulted in decreased spermidine levels and a slight increase in the levels of its precursor, the diamine putrescine and the molecule downstream of Spd, the tetraamine spermine. While the vegetative growth of the transgenics remained largely unaffected, SPDS down-regulation resulted in smaller size of flowers, decreased pollen viability and seed setting, and a reduced and delayed seed germination. When subjected to abiotic stress, the transgenics showed an increased tolerance to salinity and drought conditions owing to a steady intracellular pool of putrescine and spermine. The results not only highlight the importance of spermidine in determining reproductive potential in plants but have also help delineate its function from that of putrescine and spermine.

SUBMITTER: Choubey A

PROVIDER: S-EPMC6214437 | biostudies-literature | 2018 Nov

REPOSITORIES: biostudies-literature

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RNAi-mediated silencing of spermidine synthase gene results in reduced reproductive potential in tobacco.

Choubey Ami A Rajam M V MV

Physiology and molecular biology of plants : an international journal of functional plant biology 20180630 6

Spermidine belongs to a class of polycationic compounds known as polyamines. Polyamines are known to be involved in a wide range of biological processes but the exact role and contribution of different polyamines to these processes are still not clear. In the present study, we have tried to understand the contribution of triamine spermidine to the growth and development of tobacco by downregulating spermidine synthase gene (<i>SPDS</i>) using RNA interference. Down-regulatioin of <i>SPDS</i> gen ...[more]

PMID: 30425424

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Project description:BACKGROUND: Gene silencing by RNA interference (RNAi) is a powerful tool for functional genomics. Although RNAi was first described in Caenorhabditis elegans, several nematode species are unable to mount an RNAi response when exposed to exogenous double stranded RNA (dsRNA). These include the satellite model organisms Pristionchus pacificus and Oscheius tipulae. Available data also suggest that the RNAi pathway targeting exogenous dsRNA may not be fully functional in some animal parasitic nematodes. The genus Panagrolaimus contains bacterial feeding nematodes which occupy a diversity of niches ranging from polar, temperate and semi-arid soils to terrestrial mosses. Thus many Panagrolaimus species are adapted to tolerate freezing and desiccation and are excellent systems to study the molecular basis of environmental stress tolerance. We investigated whether Panagrolaimus is susceptible to RNAi to determine whether this nematode could be used in large scale RNAi studies in functional genomics. RESULTS: We studied two species: Panagrolaimus sp. PS1159 and Panagrolaimus superbus. Both nematode species displayed embryonic lethal RNAi phenotypes following ingestion of Escherichia coli expressing dsRNA for the C. elegans embryonic lethal genes Ce-lmn-1 and Ce-ran-4. Embryonic lethal RNAi phenotypes were also obtained in both species upon ingestion of dsRNA for the Panagrolaimus genes ef1b and rps-2. Single nematode RT-PCR showed that a significant reduction in mRNA transcript levels occurred for the target ef1b and rps-2 genes in RNAi treated Panagrolaimus sp. 1159 nematodes. Visible RNAi phenotypes were also observed when P. superbus was exposed to dsRNA for structural genes encoding contractile proteins. All RNAi phenotypes were highly penetrant, particularly in P. superbus. CONCLUSION: This demonstration that Panagrolaimus is amenable to RNAi by feeding will allow the development of high throughput methods of RNAi screening for P. superbus. This greatly enhances the utility of this nematode as a model system for the study of the molecular biology of anhydrobiosis and cryobiosis and as a possible satellite model nematode for comparative and functional genomics. Our data also identify another nematode infraorder which is amenable to RNAi and provide additional information on the diversity of RNAi phenotypes in nematodes.

Dataset Information

RNAi-mediated silencing of spermidine synthase gene results in reduced reproductive potential in tobacco.

Publications

RNAi-mediated silencing of spermidine synthase gene results in reduced reproductive potential in tobacco.

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