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Jatindra Nath Mohanty, PhD scholar
Developmental
control
Signal
transduction
1. Flowering induction
2. Specification of inflorescence
3. Specification of flower meristem
4. Est. of floral organ identity
5. Regulation of fruit , seed &
embryo development
1. Senescence
2. Stress response
3. Growth & polarity of
cell.
4. Differentiation.
etc
This family of transcription factors is defined by the presence
of a conserved domain, the MADS box, in the N-terminal
region, involved in DNA binding and dimerization with other
MADS-box proteins.
MIKC-
M-MADS domain
I - weakly conserved intervening domain
K – conserved Keratin like domain(transcriptional activation )
C – highly variable C-terminal domain
MADS box
genes
1. Class A gene – AP1
2. Class B genes – AP3 & PI
3. Class C gene – AG
4. Class D genes -
STK/AGL11 & SHP
5. Class E genes - SEP1,
SEP2, SEP3, SEP4.
6. Regulatory genes –
SOC1and SVP are involved
in the regulation of
flowering transition
+ve – SOC1 & AGL 24
-ve – FLC & SVP, AGL 12,
AGL 15, AGL 17
 MIKC* structure is very similar to MIKCC genes but the
K-domain is poorly conserved in its last part and
gene structure show an exon duplication in its 5’
region.
 Very small size in all plant species two genes in the
eudicot (Eschscholzia) , the basal angiosperm
(Aristolochia) and six genes in Arabidopsis thaliana
 Phylogenetic analyses of MIKC* genes from a broad
variety of vascular plants confirm the existence of
two clades S( AGL66 and AGL104) and P (AGL30,
AGL65, AGL94)
 MIKC* genes seem to retain a conserved and essential
role in gametophyte development during the
evolution of land plants
 11-229 genes are there in flowering plants.
 61 members are found in Arabidopsis
 Expression of type I Arabidopsis genes in central cell,
antipodal cell and chalazal endosperm of the embryo
sac, indicate that they play an important role in
female gametophyte and early seed development.
 Mα-type proteins preferentially form heterodimers
with Mβ or Mγ-type proteins whereas interactions
within the same subclass are rare.
Mα(25) Mβ(20) Mγ(16)
 The complete MIKCC type genes set with the
discovery of 10 new genes, mainly belonging
to the SVP, AGL17, BSister (BS) and AGL6
subfamilies, as well as the characterization of
the complete set of MIKC* and type I MADS-
box genes of grapevine.
 This work also represents a direct application
of the published grapevine nomenclature
recommendations on the annotation of a
complex gene family
Phylogenetic analysis ontology
 Sequence comparison analysis shows 169 genome region
homology with at least one of the gene.
 42 genes without having MADS domain
 37 genes/region having truncated with stop codon
 15 regions are non functional due to in complete sequence in
the data assembly or by natural genetic variation within
grapevines
 Finally there is 90 MADS-box genes with functional structure are
identified in grapevines.
37 new genes with compare to previous study
30 new genes compared to the CRIBI V1annotation
19 new genes with respect to reference sequence
14 new genes never been detected before are there.
 By the recommendation of Super-Nomenclature
Committee for Grape Gene Annotation(sNCGGa)
here is the Phylogenetic tree of MADS box
proteins

42 grapevines genes corresponds to MIKCC
6 grapevines genes corresponds to MIKC *
23 grapevines genes corresponds to Mα(type 1)
19 grapevines genes corresponds to Mγ(type 1)
MIKC *(Mδ) – MADSD
Mα(type 1) – MADS1A
Mγ(type 1) – MADS1G
MIKC *(Mδ) –MADSD-1,2 ,3
clade
MADS1A- 1, 2, 3 clade of
genes
MADS1G- 1, 2, 3 clade of
genes
 MADS-box genes were located on 17 of the
19 grapevine chromosomes
 There are still three genes located in the
unknown chromosome
 The ten new MIKCC genes compared to 37 are
definitely positioned in the chromosome.
 Here different members of several
subfamilies are located in chromosomal
regions that might represent paralogous
segments resulting from ancestral
polyploidization events
The MIKC* MADS-box genes
Mα Type 1 MADS-box genes:
M α2 Type 1 MADS-box genes:
 Six gene in A.thaliana matches here
MADSD1a & MADS1c only K domain
 MADS3a homolog to AGL 66, 67,104 in MADS region only
 3 clades are here(Mα1,α2,α3)
In Mα1, 7 genes in grapevines (MADS1A1a,1b,1c,1d,1e, 1f,1g etc)
These results suggest that duplication of
some members of this family is relatively old predating species
divergence.
In Mα2, 9 genes in grapevines like MADSA2F,2G, 2H etc are here.
These sequences homology with soybean, Medicago, poplar and
papaya.
Mγ Type 1 MADS-box genes:
Mγ Type= Mγ1 Type+ Mγ2Type+ Mγ3 Type
Mγ1Type : 8 genes are located in grapevines
chr 17,15,5 with no clear homology to genes
from other species.
Mγ2Type: 7 genes are located in grapevines.
Out of that 5 genes located in chr 5 and 2 genes
are in chr 2.
Mγ3 Type: only one gene is found in
grapevines which has homology with
watermelon.
10 new MIKC gene they got
 TM8b not coming fit in Type-II MIKC only it contain a
MADS domain.
 SVPSI, SVPS4, SVPS5 contain only one exon follwed by 30
AA.
 SVPS2, SVPS3 having large exon site
 The orthology analyses indicated that SVPS sequences
were not present in the monocot species.
 Interestingly, the new grapevine genes found in the
subfamilies BS (VviBS3), AGL17 (VviAGL17c,VviAGL17d),
AGL6 (VviAGL6b) and TM8 (VviTM8b), did not showed
orthology with other species.
1 belonging to BS sub family
2 belonging to AGL17 sub family
1 belonging to AGL6 sub family
1 Vvi TM 8b belonging to TM8a
Five new genes related to SVP in
chromosome 3 & 15
 They have performed an exhaustive analysis of
MADS-box genes on the 12x grapevine genome
based on the isolation of the complete set of genes
identified in PN40024.
 This supposes the identification of 90 functional
genes what adds 37 new genes to previous studies.
 A new type of MADS-box genes not yet
characterized in other plant systems are found that is
VviSVPS
 Their genomic analysis of MADS-box genes in
grapevine allowed the discovery of genes belonging
to the BS, AGL17, AGL6 and TM8 subfamilies that had
no homologs in other plant species.
Thank you

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Structural and functional annotation of mads box gene in grape vines

  • 2. Developmental control Signal transduction 1. Flowering induction 2. Specification of inflorescence 3. Specification of flower meristem 4. Est. of floral organ identity 5. Regulation of fruit , seed & embryo development 1. Senescence 2. Stress response 3. Growth & polarity of cell. 4. Differentiation. etc
  • 3. This family of transcription factors is defined by the presence of a conserved domain, the MADS box, in the N-terminal region, involved in DNA binding and dimerization with other MADS-box proteins. MIKC- M-MADS domain I - weakly conserved intervening domain K – conserved Keratin like domain(transcriptional activation ) C – highly variable C-terminal domain
  • 5. 1. Class A gene – AP1 2. Class B genes – AP3 & PI 3. Class C gene – AG 4. Class D genes - STK/AGL11 & SHP 5. Class E genes - SEP1, SEP2, SEP3, SEP4. 6. Regulatory genes – SOC1and SVP are involved in the regulation of flowering transition +ve – SOC1 & AGL 24 -ve – FLC & SVP, AGL 12, AGL 15, AGL 17
  • 6.  MIKC* structure is very similar to MIKCC genes but the K-domain is poorly conserved in its last part and gene structure show an exon duplication in its 5’ region.  Very small size in all plant species two genes in the eudicot (Eschscholzia) , the basal angiosperm (Aristolochia) and six genes in Arabidopsis thaliana  Phylogenetic analyses of MIKC* genes from a broad variety of vascular plants confirm the existence of two clades S( AGL66 and AGL104) and P (AGL30, AGL65, AGL94)  MIKC* genes seem to retain a conserved and essential role in gametophyte development during the evolution of land plants
  • 7.  11-229 genes are there in flowering plants.  61 members are found in Arabidopsis  Expression of type I Arabidopsis genes in central cell, antipodal cell and chalazal endosperm of the embryo sac, indicate that they play an important role in female gametophyte and early seed development.  Mα-type proteins preferentially form heterodimers with Mβ or Mγ-type proteins whereas interactions within the same subclass are rare. Mα(25) Mβ(20) Mγ(16)
  • 8.  The complete MIKCC type genes set with the discovery of 10 new genes, mainly belonging to the SVP, AGL17, BSister (BS) and AGL6 subfamilies, as well as the characterization of the complete set of MIKC* and type I MADS- box genes of grapevine.  This work also represents a direct application of the published grapevine nomenclature recommendations on the annotation of a complex gene family Phylogenetic analysis ontology
  • 9.  Sequence comparison analysis shows 169 genome region homology with at least one of the gene.  42 genes without having MADS domain  37 genes/region having truncated with stop codon  15 regions are non functional due to in complete sequence in the data assembly or by natural genetic variation within grapevines  Finally there is 90 MADS-box genes with functional structure are identified in grapevines. 37 new genes with compare to previous study 30 new genes compared to the CRIBI V1annotation 19 new genes with respect to reference sequence 14 new genes never been detected before are there.
  • 10.
  • 11.  By the recommendation of Super-Nomenclature Committee for Grape Gene Annotation(sNCGGa) here is the Phylogenetic tree of MADS box proteins  42 grapevines genes corresponds to MIKCC 6 grapevines genes corresponds to MIKC * 23 grapevines genes corresponds to Mα(type 1) 19 grapevines genes corresponds to Mγ(type 1) MIKC *(Mδ) – MADSD Mα(type 1) – MADS1A Mγ(type 1) – MADS1G MIKC *(Mδ) –MADSD-1,2 ,3 clade MADS1A- 1, 2, 3 clade of genes MADS1G- 1, 2, 3 clade of genes
  • 12.  MADS-box genes were located on 17 of the 19 grapevine chromosomes  There are still three genes located in the unknown chromosome  The ten new MIKCC genes compared to 37 are definitely positioned in the chromosome.  Here different members of several subfamilies are located in chromosomal regions that might represent paralogous segments resulting from ancestral polyploidization events
  • 13.
  • 14. The MIKC* MADS-box genes Mα Type 1 MADS-box genes: M α2 Type 1 MADS-box genes:  Six gene in A.thaliana matches here MADSD1a & MADS1c only K domain  MADS3a homolog to AGL 66, 67,104 in MADS region only  3 clades are here(Mα1,α2,α3) In Mα1, 7 genes in grapevines (MADS1A1a,1b,1c,1d,1e, 1f,1g etc) These results suggest that duplication of some members of this family is relatively old predating species divergence. In Mα2, 9 genes in grapevines like MADSA2F,2G, 2H etc are here. These sequences homology with soybean, Medicago, poplar and papaya.
  • 15. Mγ Type 1 MADS-box genes: Mγ Type= Mγ1 Type+ Mγ2Type+ Mγ3 Type Mγ1Type : 8 genes are located in grapevines chr 17,15,5 with no clear homology to genes from other species. Mγ2Type: 7 genes are located in grapevines. Out of that 5 genes located in chr 5 and 2 genes are in chr 2. Mγ3 Type: only one gene is found in grapevines which has homology with watermelon.
  • 16.
  • 17.
  • 18.
  • 19. 10 new MIKC gene they got  TM8b not coming fit in Type-II MIKC only it contain a MADS domain.  SVPSI, SVPS4, SVPS5 contain only one exon follwed by 30 AA.  SVPS2, SVPS3 having large exon site  The orthology analyses indicated that SVPS sequences were not present in the monocot species.  Interestingly, the new grapevine genes found in the subfamilies BS (VviBS3), AGL17 (VviAGL17c,VviAGL17d), AGL6 (VviAGL6b) and TM8 (VviTM8b), did not showed orthology with other species. 1 belonging to BS sub family 2 belonging to AGL17 sub family 1 belonging to AGL6 sub family 1 Vvi TM 8b belonging to TM8a Five new genes related to SVP in chromosome 3 & 15
  • 20.  They have performed an exhaustive analysis of MADS-box genes on the 12x grapevine genome based on the isolation of the complete set of genes identified in PN40024.  This supposes the identification of 90 functional genes what adds 37 new genes to previous studies.  A new type of MADS-box genes not yet characterized in other plant systems are found that is VviSVPS  Their genomic analysis of MADS-box genes in grapevine allowed the discovery of genes belonging to the BS, AGL17, AGL6 and TM8 subfamilies that had no homologs in other plant species.