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STUDY OF MEDICAGO TRUNCATULA GENE EXPRESSION IN TIME AND SPACE

STUDY OF MEDICAGO TRUNCATULA GENE EXPRESSION IN TIME AND SPACE. Iryna Sanders, Fares Z. Najar, Bruce A. Roe Department of Chemistry and Biochemistry, Stephenson Research and Technology Center, University of Oklahoma, 101 David L. Boren Blvd., Norman, Oklahoma, 73019. Abstract.

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STUDY OF MEDICAGO TRUNCATULA GENE EXPRESSION IN TIME AND SPACE

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  1. STUDY OF MEDICAGOTRUNCATULA GENE EXPRESSION IN TIME AND SPACE Iryna Sanders, Fares Z. Najar, Bruce A. Roe Department of Chemistry and Biochemistry, Stephenson Research and Technology Center, University of Oklahoma, 101 David L. Boren Blvd., Norman, Oklahoma, 73019

  2. Abstract • Medicago truncatula flavonoid genes exist in families that occur as single, duplicated or clustered genes. The flavonoid genes within a family share a high level of homology, both at the sequence and exon structure levels, implying homologous functions of duplicated genes. However even when flavonoid genes occur in clusters, all members are not necessarily expressed at the same time and place as a response to the same environmental condition. In majority of instances however, the phylogenetically related genes of a particular family have similar time and spatial expression profiles. Furthermore, not all members of a cluster or other genes from the same family are overexpressed as a result of stress. Since is known that flavonoid genes often are expressed as a result of pathogenic attack or extreme environmental conditions (starvation, irradiation, etc.), EST-based expression profiling reveals that PAL, 4CL, CHS, IFR are expressed in leaves, roots, seedlings, nodules, reproductive tissues; FS-II is expressed in roots, leaves, seeds, reproductive tissues; FLS is expressed in leaves, roots, nodules, seeds, shoots; F3H is expressed in roots, nodules, seeds; UFGT is expressed in leaves; CHR is expressed in seedlings, roots; and DFR is expressed in roots, nodules, leaves. • These observations are consistent with the hypothesis that duplicated flavonoid genes have highly specialized function, and this specialization is necessary to give an adequate response to a variety of different environmental stresses.

  3. Location of flavonoid genes on M.truncatula chromosomes in cM Source: http://www.medicago.org

  4. Comparison of two clusters of chalcone synthase from different chromosomes AC146575 Chr. V AC137823 Chr. VII The results from Artemis Comparison Tool (ACT) show high level of exon duplications between the two clusters from chromosomes 5 and 7. This conservation suggests that both of these clusters evolved to encode enzymes with the same function.

  5. Analysis of expression profile of Chalcone synthase genes based on Blast against EST database Members of the same cluster may have different expression profile. (Identity >98%)

  6. Correlation of expression profile with phylogeny of chalcone synthase                 Colors represent different chalcone synthase duplicated genes

  7. 1,3 4,7 3-deoxyanthocyanidins 9 9 9 2 2 2 PAL C4H 4CL + malonyl-CoA p-coumaric acid 4-coumaroyl-CoA phenylalanine cinnamic acid 2 1,23 1,23 1,2 1,2 1 1 1 1 1 CHS/CHR CHS 2,5,7 2,5,7 1,2 1,2 1 1 CHI trihydrochalcone tetrahydroxychalcone Flavan-4-ols 7,4’-dihydroxyflavanone IFS CHI DFR DFR IOMT IFS formononetin daidzein naringenin eridiodictyol F3’H FS I,II I2’H I2’H 2 4,14 FS I,II 2,4 5,6 2,4 5,6 1,2 1 flavones 1 3 3 2’-hydroxydaidzein 2 2 2,6,7 FLS IFR IFR 2,5 6 2,5 6 1 1 1 Flavonol glycosides 2 2 1 1 1,2 1,2 RT,OMT,UFGT F3H flavonols vestitone demethylvestitone 7 VR VR F3’H F3’5’H 2,3-dihydrokaempferol 2,3-dihydroquercetin 2,3-dihydromyricetin DMI demethyl-DMI DFR DFR DFR DMID leucocyanidin leocopelargonidin leucodelphinidin isoflavonoids ? LDOX LCR 3-OH-anthocyanidins Flavan-3-ols OMT o Proanthocyanidins (condensed tannins) UFGT anthocyanines 7 o RT Expression profile of flavonoid biosynthesis enzymes. identity cutoff >98%

  8. DFR-dihydroflavonol 4-reductase CHS-chalcone synthase CHR-chalcone reductase CHI-chalcone isomerase PAL-phenylalanine ammonia lyase C4H-cinnamic acid 4-hydroxylase 4CL-coumarate: coenzymeA ligase IFS-isoflavone synthase IOMT-isoflavonoid O-methyltransferase F3’-H-flavonoid 3’-hydroxylase I2’-H-isoflavone 2’-hydroxylase IFR-isoflavone reductase VR-vestitone reductase F3H-flavanone 3-hydroxylase F3’5’H-flavonoid 3’,5’-hydroxylase FLS-flavonol synthase FS I, II - flavone synthases LDOX-leucoanthocyanidin dioxygenase LCR-leucoanthocyanidin reductase OMT-O-methyltransferase RT-rhamnosyl transferase UFGT-flavonoid glucosyl transferase

  9. Conclusions • Flavonoid biosynthesis genes are scattered all over the Medicago genome. • Duplicated flavonoid genes seem to share highly conserved exon structure and organization but might have different expression profile. • Duplicated flavonoid genes within a cluster might have different expression profiles. • Most flovonoid genes are expressed in Roots. • Phylogenetically related genes (often not from the same cluster) have similar expression profile

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