PHYLOGENETIC SYSTEMATIC OF

GENUS LACTOBACILLUS AND

SALMONELLA
Group : 1
Entourage : I

Member :
Herlina Wahyuningrum (B1A017036)
Alfiani Rahmawati (B1A017038)
Wiwi Meilani (B1A017101)
Bunga Giftha Nurhamsry (B1B017004)
Alif Arfa Maulida Firdauzi (B1B017008)
 INTRODUCTION
• The phylogenetic system is a system that emerged after the birth of the theory of
evolution. This system requires that classification not only results in a summary of the
world of living things, but can also reflect a description of the order in which living
creatures are based on their phylogenetic history, and the close relations of kinship
between one taxon and another (Emerson et al., 2008). 16S rRNA sequences are
increasingly being used. The ribosomal RNA coding gene (rRNA) is the most sustainable
(preserved) gene. The portion of rDNA sequences of each species related to genetics.
Thus each culture that has a certain kinship can be aligned so that it is easier to determine
the differences in the sequences that are characteristic of these cultures (Rinanda, 2011).
• The genus Lactobacillus comprises a phenotypically heterogeneous group of
facultatively anaerobic, catalase-negative, non-spore-forming, Gram-positive, rod-shaped
organisms which produce lactic acid as a major end-product of metabolism (Falsen et al.,
1999).
 INTRODUCTION
PROBLEM FORMULATION
1. How is the kinship between bacterial strains based on 16S rRNA.
2. What is the level of similarity and dissimilarity between bacterial strains based on 16S
rRNA.

OBJECTIVES
3. To know the evolutionary kinship between strains based on 16S rRNA with phylogenetic
systematics.
4. To know the level of similarity and dissimilarity between bacterial strains based on 16S
rRNA.
 MATERIALS AND METHODS
TOOLS
Tools used in this agenda were laptops with ClustalX, MEGA 7, Phydit, and
Microsoft Excel program.

MATERIALS
10 Lactobacillus strains that obtained from NCBI as ingroup and 1 strain of
Salmonella from NCBI as outgroup.
 METHODS
Data of 16S rRNA strains with a
base length of 1000-1500 bp out
of 10 strains of microorganisms
(ingroup) and 1 strain (out group)
were taken from NCBI. saved on
notepad.
The Phydit application was opened for
the matrix similarity construction and
the 16S rRNA nucleotide differences
of 10 strains of microorganisms
(ingroup) and 1 strain of
microorganisms (out group) using data
stored in the form of GDE. the table
that appears appears in Excel.
ClustalX application is opened for the
preparation of 16S rRNA
microorganism sequences. The data
obtained is saved in the form "aln".
Data that has been processed in
Clustalx and saved in "aln" format is
opened in the MEGA application to
construct phylogenic trees. data in
the "aln" format are renamed and
saved in MEGA format. The
resulting phylogeny tree is copied
into Word
 RESULT AND DISCUSSION
Table 1. Similarity Table from Lactobacillus and Salmonella
SIMILARITY TABLE
YELLOW: The highest
LC5 LC9 LC6 LC10 LC1 LC3 LC7 LC4 LC8 LC2 SM1 percentage of similarity.
LC5 --- GREEN: The lowest percentage
LC9 94,37 --- of similarity.
LC6 89,08 91,47 ---
LC10 89,58 91,06 90,07 ---
LC1 85,29 87,36 88,42 87,53 ---
LC3 88,54 89,32 88,1 87,45 85,58 ---
LC7 88,74 89,39 88,36 87,52 85,97 99,42 ---
LC4 86,49 87,6 87 87,47 85,57 91,58 91,41 ---
LC8 86,99 88,12 87,63 86,94 84,58 92,24 91,99 91,82 ---
LC2 78,94 79,75 78,78 80,62 79,59 79,89 80,07 79,03 78,58 ---
SM1 75,29 76,27 76,41 77,29 77,02 76,37 76,49 76,88 74,93 74,39 ---
 RESULT AND DISCUSSION
Table 2. Dissimilarity Table from Lactobacillus and Salmonella
DISSIMILARITY TABLE
LC5 LC9 LC6 LC10 LC1 LC3 LC7 LC4 LC8 LC2 SM1
LC5 --- YELLOW: The lowest
LC9 84/1491 --- nucleotide differences.
163/149 127/148
LC6 --- GREEN: The highest
3 9
155/148 133/148 148/149 nucleotide differences.
LC10 ---
8 7 0
213/144 183/144 168/145 181/145 These are the amount of
LC1 ---
8 8 1 2 different base from both
172/150 160/149 179/150 192/153 210/145
LC3
1 8 4 0 6
--- strain.
168/149 158/148 174/149 190/152 203/144
LC7 9/1564 ---
2 9 5 3 7
201/148 184/148 194/149 191/152 210/145 129/153 131/152
LC4 ---
8 4 2 4 5 2 5
193/148 176/148 184/148 194/148 224/145 116/149 119/148 122/149
LC8 ---
4 2 7 6 3 5 6 2
303/143 291/143 307/144 280/144 290/142 292/145 288/144 304/145 310/144
LC2 ---
9 7 7 5 1 2 5 0 7
362/146 347/146 347/147 340/149 330/143 363/153 359/152 348/150 368/146 369/144
SM1 ---
5 2 1 7 6 6 7 5 8 1
 RESULT AND DISCUSSION
Figure 1. Dendrogram of Phylogenetic Systematics of Lactobacillus and Salmonella Strain
100
Lactobacillus
LC5 sanfransisco
0.03

52
0.02 Lactobacillus
LC9 fructivorans
0.01
0.00
53 Lactobacillus
LC10 ruminus
0.00 0.04
Lactobacillus
LC6 sake
0.04
96
0.02
Lactobacillus
LC3 gasseri
100 0.00
0.03 Lactobacillus johnsonii
LC7
100 0.00
99 0.03 Lactobacillus
LC4 lactis
0.02 63 0.04
0.00
Lactobacillus
LC8 acetotolerans
0.03
0.02
Lactobacillus
LC1 thermophilus
0.06

Lactobacillus
LC2 vitulinus
0.14
Salmonella bongori
SM1
0.16
 RESULT AND DISCUSSION
Based on the result, it showed that Lactobacillus gasseri and Lactobacillus johnsonii
have the highest similarity up to 99,42% similarity among the others, this is in
accordance with Falsen et al. (1999), which states that treeing analysis confirmed this
placement and showed bacteria that phylogenetically most closely are related to
Lactobacillus gasseri and Lactobacillus johnsonii (approx. 96% sequence similarity).
Bootstrap resampling (value 100%) showd that the association between Lactobacillus
gasseri and Lactobacillus johnsonii was statistically significant.
 CONCLUSION
Based on the result and discussion above can be concluded that the closest
evolutionary kinship of Lactobacillus is between Lactobacillus gasseri and
Lactobacillus johnsonii. While the furthest evolutionary kinship is between
Lactobacillus sanfransisco and Lactobacillus vitulinus.
 REFERENCES
Emerson., David., agulto, L., Liu, H. & Liu, L., 2008. Identifying and Characterizing
Bacteria in a an Era of Genomics and Proteomics. Bioscience, 58(10), pp. 925-936.
Rinanda, T., 2011. Analisis Sekuensing 16s rRNA Di Bidang Mikrobiologi. Jurnal
Kedokteran Syiah Kuala, 11(3), pp. 1-10.
Falsen, E., Pascual, C., Sjödén, B., Ohlén, M. & Collins, M. D., 1999. Phenotypic and
phylogenetic characterization of a novel Lactobacillus species from human sources:
description of Lactobacillus iners sp. nov. International Journal of Systematic and
Evolutionary Microbiology, 49(1), pp. 217-221.
THANK YOU