“Head to tail” Tool Analysis through

ClustalW Alignment Algorithm and

Construction of Distance Method

Neighbor-Joining Trees Based

on Genus Fusarium Genomic DistancesJuan D. Henao, Melissa Rincón, Juan J.

FilgueiraNueva Granada Military University

(UMNG).Bogotá. Colombia.

Juanhenao.sanchez@gmail.com

Gibberella clad e

Clade V

Clade III

Clade VII

Clade VI

Biological information

Fusarium oxysporum f.sp. dianthi

Morphological and genetic information. Deleting all gaps to create haplotipe. More Mathematical support but less biological support.

Mol. Biol. Evol. 4(4):406-425. 1987.

Mathematical process

Neighbor-Joining (Saitou & Nei)

Fusarium graminearum (FG)

Fusarium avenaceum (FA)

Fusarium foetens (FF)

Fusarium culmorum (FC)

Fusarium equiseti (FE)Fusarium verticillioides (FV)

Fusarium proliferatum (FP)

Fusarium subglutinans (FSG)

Fusarium solani (FS)

Fusarium sporotrichioides (FSP)

Fusarium oxysporum (FO)

Species of genus Fusarium

SECTION Fusarium SPECIE

Discolor culmorumgraminearum

Elegans oxysporumfoetens

Gibossum equiseti

Liseolaproliferatumsubglutinansverticillioides

Martiella solaniRoseum avenaceum

Sporotrichiella sporotrichioides

Eleven Fusarium species Seven morphological sections.

Morphological sections

Six genes.

Ten sequences.

GENE PRIMER SEQUENCE ANNEALING

β-tubulin

Bt1a (Glass y Donaldson, 1995) 5'TTCCCCCGTCTCCACTTCTTCATG3'

58.6Bt1b (Glass y Donaldson, 1995) 5'GACGAGATGGTTCATGTTGAACTC3'Bt2a (Glass y

Donaldson, 1995) 5'GGTAACCAAATCGGTGCTGCTTTC3'60.6Bt2b (Glass y

Donaldson, 1995) 5'ACCCTCAGTGTAGTGACCCTTGGC3'

Cytochrome oxidase

AHyFuF (Barcode, Forward from F.

oxysporum AY874423)5'CTTAGTGGGCCAGGAGTTCAATA3'

57.2AHyFuR (Barcode, Forward from F.

oxysporum AY874423)5'ACCTCAGGGTGTCCGAAGAAT3'

Translation Elongation Factor

EF1H (O’Donell et al., 1998) 5'ATGGGTAAGGAAGACAAGAC3'

57EF2T (O’Donell et al., 1998) 5'GGAAGTACCAGTGATCATTGTT3'

Histone 3H31a (Glass y

Donaldson, 1995) 5'ACTAAGCAGACCGCCCGCAGG3'61.3H31b (Glass y

Donaldson, 1995) 5'GCGGGCGAGCTGGATGTCCTT3'

Histone 4H41a (Glass y

Donaldson, 1995) 5'GCTATCCGCCGTCTCGCT3'57.8H41b (Glass y

Donaldson, 1995) 5'GGTACGGCCCTGGCGCTT3'

Internal Transcribed

Spacer

ITS FuR (White et al., 1990) 5' GCGACGATTACCAGTAACGA3'

61ITS FuF (White et al., 1990) 5'CAACTCCCAAACCCCTGTGA3'

ITS1 (White et al., 1990) 5'TCCGTAGGTGAACCTGCGG3'

64ITS2 (White et al., 1990) 5'GCTGCGTTCTTCATCGATGC3'

ITS1 (White et al., 1990) 5'TCCGTAGGTGAACCTGCGG3'

61ITS4 (White et al., 1990) 5'TCCTCCGCTTATTGATATGC3'

ITS4 (White et al., 1990) 5' TCCTCCGCTTATTGATATGC3'

56.5ITS5 (White et al., 1990) 5'GGAAGTAAAAGTCGTAACAAGG3'

Primers

Frequency of get any topology. You use only the tree-type topology determined to phylogeny studies.

Likelihood Mapping Analysis (TREE-PUZZLE)

Options used in Neighbor-Joining process.

Neighbor-Joining (MEGA5)

Bt1a/Bt1b Bt2a/Bt2b Ahy-FuF/AHyFuR EF-1H/EF-2T H3-1a/H3-1b H4-1a/H4-1b ITS FuR/ITS FuF ITS-1/ITS-2ITS-1/ITS-4 ITS-4/ITS-5Bt2a/Bt2bBt1a/Bt1b Ahy-FuF/AHyFuR EF-1H/EF-2T H3-1a/H3-1b H4-1a/H4-1b ITS FuR/ITS FuF ITS-1/ITS-2ITS-1/ITS-4 ITS-4/ITS-5

a)

b)

A single change in the order of concatenates.

Mathematical support.

Mathematical support of the gaps

a)

b)

The molecular inferences are very sensitive with a single change of the order.

Molecular Inferences

Bt1a/Bt1b Bt2a/Bt2b

Bt2a/Bt2bBt1a/Bt1b

Discolor

Glibossum

Sporotrichiella

Liseola

Elegans

Liseola

Roseum

Martiella

a)

Sections and bootstrap

Liseola

Liseola

Elegans

Elegans

Roseum

Discolor

Sporotrichiella

Martiella

Glibossum

b)

Sections and bootstrap

Gaps are very important because contain information about evolutionary transformations of the sequences.

Gaps have the necesary mathematical power for establish calculus in phylogenetic process.

The gaps don’t have the necessary mathematical power for establish sturdiness (stability and confidence) in phylogenetic trees.

Something to say

UMNG Molecular Phytopathology Group

Juan J FilgueiraMelissa Rincón Guillermo BuragliaAngelica Zambrano Michel FigueredoKatherin VelasquezFelipe CristanchoAngela Niño

Sponsor:

Vicerectoría de Investigaciones UMNG