poster id : 828 dna barcoding and phylogenetic analysis of ......dna barcoding and phylogenetic...
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DNAbarcodingandphylogenetic analysisofSearsia
MakhadoTshililo,MichellevanderBankandSaneleN.S.ShibaTheAfricanCentreforDNABarcoding(ACDB),DepartmentofBotany&PlantBiotechnology,UniversityofJohannesburg,SouthAfrica
Anacardiaceae also known as the “cashew family” consists of 82genera represented by around 800 species. The largest genus in thisfamily is Rhus L. (senso lato) for which issues around its delimitationhas caused a lot of controversy among taxonomists. Searsia, a newlyaccepted genus within Anacardiaceae, was originally part of the Rhuscomplex but was recently removed along with five other genera. Thegenus Searsia has approximately 120 species and is mainlycharacterised by trifoliate leaves (Yang et al., 2016). It is widelydistributed across Africa with 111 species occurring in southern Africaand 3 species occurring in Asia. The African species all belong to thesection Gerontogeae. Recent phylogenetic studies have definedSearsia as a monophyletic group but these findings need to beconfirmed by conducting a broader sampling of the genus (Yang et al.,2016).
The aim of this study is to investigate the phylogenetic relationshipsamong the seven genera within the Rhus complex as well as resolvethe systematics within Searsia.
MATERIAL&METHODS
DNA extraction, amplification and sequencingDNA was extracted using the 10X CTAB method of Doyle & Doyle(1987). Standard protocols were followed for PCR and sequencingusing the core barcoding regions (rbcLa & matK).
Data analysisA phylogenetic tree was generated using Maximum Parsimony (MP) inPAUP* (Swofford, 2002).
INTRODUCTION
RESULTS&DISCUSSION
The results obtained in this study support the monophyly of African representatives ofSearsia and illustrates the inability of core barcoding regions to provide speciesdelimitation within this genus. Further investigation is required with additionalmarkers (ITS, ETS and trnL-F) to provide better delimitation and resolution within thegenus.
Figure 2 (Clade A) shows high support (99%) for the monophyly ofsouthern African Searsia species which supports the findings of Yanget al. (2016).
Clade B shows high support (100%) for the monophyly of the genusSchinus, having two highly supported sister clades. These findings backGuala (2016) who differentiated S. terebinthifolius from S. areira and S.molle by the presence of pubescent leaflets.
Figure 2 (Clade C) shows weak support (53%) for the monophyly ofLannea. There is also moderate support (80%) for the monophyly ofClade D, with the African species Protorhus nestled within it. Thissupports findings by Von Teichman and Van Wyk (1996) who groupedOzoroa and Protorhus due to morphological similarities.
Results also show no support for the monophyly for the Rhus speciesincluded in this study (Figure 2). In 2007, Yi et al. cited themorphological heterogenousity of Rhus sensu Barkley (1937).
CONCLUSION
Doyle, J.J. & Doyle, J.L. 1987. A rapid isolation procedure for small amounts of leaf tissue. Phytochemical Bulletin, 19: 11-15.Guala, M.S., Lapissonde, M.O., Elder, H.V., van Baren, C.M., Bandoni, A.L. & Dellacassa, E. 2006. Essential Oils in FoodPreservation, Flavor and Safety.Swofford, D.L. 2002. PAUP* - Phylogenetic Analysis Using Parsimony (*and Other Methods). Version 4. SinauerAssociates, Sunderland, MA.Yang, Y., Meng, Y., Wen, J., Sun, H. & Nie, Z. 2016. Phylogentic analyses of Searsia (Anacardiaceae) from eastern Asia andits biogeographic disjunction of its African relatives. South African Journal of Botany, 106: 129-136.Yi, T., Miller, A. J. & Wen, J. 2007 . Phylogeny of Rhus (Anacardiaceae) based on sequences of nuclear NIA-i3 intron andchloroplast trnCtrnD. Systematic Botany 32: 379-391.Von Teichman, I. & Van Wyk, A. E. 1996. Taxonomic significance of pericarp and seed structure in Heeria argentea(Thunb.) Meisn. (Anacardiaceae), including reference to pachychalazy and recalcitrance. Botanical Journal of the LinneanSociety, 122: 335-352.
REFERENCES
CotinusMill.
Cotinuscoggygria
ToxicodendronMill.
Toxicodendronradicans
Lannea Engl.
Lannea schweinfurthii
Schinus L.
Schinus terebinthifolia
Rhus L.(S.str.)
RhuschinensisSearsia dentata
Searsia F.A.Barkley
SamplingA total of 205 taxa were included in the study. These includerepresentatives of the genera: Searsia, Rhus, Lannea, Schinus, Cotinusand Toxicodendron (Figure 2).
FIGURE 2: Bootstrap consensus tree generated using PAUP of the combined plastid datasethighlighting Clades A – D, representatives of genus Rhus as well as some representatives of othergenera included in the study.
FIGURE1: DistributionmapoffieldcollectedSearsiasamples.
ACKNOWLEDGEMENTS
PosterID:828
Background. The family Anacardiaceae includes approximately 800 species in 82 genera. Rhus L. is the largest and most widely distributed genus in Anacardiaceae. However, controversy hassurrounded the delimitation of the genus. Searsia F.A. Barkley was originally included into the Rhus complex but recent molecular studies have separated it along with six other genera from the Rhuscomplex and recognised them as separate genera. The genus Searsia, which is the focus of the current study, is represented by approximately 120 species and is widely distributed in Africa with onlythree species currently known from Asia. Here we include an extensive sampling and molecular analyses of Searsia species across its distribution range in Africa to access the relationships within thegenus. Results. In total 205 taxa, representing 35 species of Searsia, were sequenced for the core barcoding regions (matK and rbcLa) along with additional markers, i.e. ITS, ETS, trnL-F and ndhF.Significance. This study contributes a first large sampling of Searsia. An infrageneric classification for the genus will be presented.
ABSTRACT
Searsia chirendensis OM1987
Searsia ciliata Hahn2981
Searsia pyroides OM713 1Searsia tumulicola OM3907Searsia tumulicola var meeseana OM3818
Searsia zeyheri OM2256Searsia leptodictya RBN205Searsia leptodictya RL1645
Searsia leptodictya RL1655
Searsia tenuinervis Hahn2999
Searsia nebulosa Abbott9106
Searsia pyroides var integrifolia OM2477
Searsia discolor OM3911
Searsia fastigiata Abbott9135
Searsia lancea OM1942
Searsia lancea PG0001
Searsia pendulina OM1984
Searsia pondoensis Burrows10242
Searsia pyroides OM1236
Searsia transvaalensis OM0943
Searsia transvaalensis RL1427
Searsia tumulicola var tumulicola OM2028
Searsia tumulicola var tum
ulicola OM3813
Searsia wilmsii OM
3910
Searsia gueinzii OM
0248
Searsia gueinzii OM
0265
Searsia gueinzii OM
0336
Searsia gueinzii OM
3371
Searsia natalensis OM
2655
Searsia pentheri OM
0942
Searsia pentheri OM
0945
Searsia longispina AM0243
Searsia pallens OM
1976
Searsia undulata O
M2940
Searsia lucida M
WC
05809
Searsia crenata O
M1986
Searsia crenata O
M3209
Sea
rsia
neb
ulos
a O
M22
85
Sea
rsia
aco
cksi
i Abb
ott9
154
Sea
rsia
chi
rinde
nsis
OM
2284
Sear
sia
angu
stifo
lia O
M28
47
Sear
sia
diss
ecta
JL
802
4
Sear
sia
inci
sa v
ar in
cisa
OM
3059
Sear
sia
luci
da A
bbot
t923
0
Sear
sia
tom
ento
sa S
M 7
31 3
Sorin
deia
jugl
andi
folia
PM
4827
Sorin
deia
jugl
andi
folia
PM
5106
Sorin
deia
jugl
andi
folia
PM
4949
Loxo
styli
s al
ata
OM
1827
Ozoro
a ob
ovat
a su
bsp
ellipt
ica O
M24
25
Ozoro
a ob
ovat
a su
bsp
ellipt
ica O
M25
11
Ozoro
a ob
ovat
a su
bsp
ellipt
ica O
M2733
Protor
hus l
ongif
olia M
WC28
088
Protorh
us lo
ngifo
lia O
M1764
Ozoroa
barab
etoren
s Burr
ows0
8069
Ozoroa
engle
rii OM11
54
Ozoroa englerii
OM1169
Ozoroa englerii RL1277
Ozoroa insignis s
ubsp reticu
lata OM3580
Ozoroa paniculosa OM1948
Ozoroa sphaerocarpa OM0940
Ozoroa sphaerocarpa OM1106
Heeria argentea PG0016
Rhus thouarsii AY594484Schinus areira AY594488Schinus molle DMP145Schinus molle DMP200Schinus molle DMP224Schinus molle DMP231
Schinus molle JG052Schinus terebinthifolius BS0098
Schinus terebinthifolius DMP237
Schinus terebinthifolius DMP256
Schinus terebinthifolius OM1982
Schinus terebinthifolius PPRI 0027
Mosquitoxylum jamaicens AY594490
Rhus aromatica AY594486
Rhus aromatica AY594494
Rhus copallina AY594485
Rhus hirta AP184
Rhus hypoleuca HQ427342
Astronium graveolens AY594492
Schinopsis brasiliensis AY594477
Thyrsodium puberulum FJ514664
Thyrsodium puberulum FJ514723
Toxicodendron radicans AP422
Toxicodendron rydbergii BAB 0354
Amphipterygium adstring AY594458
Comocladia glabra HM446677
Cotinus coggygria HE966907
Lithrea molleoides AY594470
Loxopterygium huasango AY594471
Pachycormus discolor AY594493
Toxicodendron sylvestre HQ415319
Toxicodendron vernix AY594495
Trichoscypha klainei Tricklai PM5051
Trichoscypha preussii Tricpreu PM5027
Trichoscypha patens Tricpate PM4897
Trichoscypha patens Tricpate PM5076
Trichoscypha acuminata M
PADN
0001
Trichoscypha acuminata M
PADN
0004
Trichoscypha acuminata M
PADN
0277
Trichoscypha arborea MPAD
N0003
Anacardium excelsum
GQ
981932Anacardium
occidentale AY594459A
nacardium occidentale G
H0152
Anacardium
occidentale OM
3645G
luta wallichii A
Y594468
Man
gife
ra in
dica
AY
5944
72
Fegi
man
ra a
frica
na A
Y59
4489
Sem
ecar
pus
aust
ralie
nsi A
Y594
479
Dob
inea
vul
garis
AY
5944
66Pe
gia
nitid
a AY
5944
73Sp
ondi
as m
ombi
n AY
5944
80La
nnea
ant
isco
rbut
ica
OM
2704
Lann
ea d
isco
lor O
M33
22La
nnea
edu
lis O
M19
71La
nnea
edu
lis O
M19
91
Lann
ea s
chwe
infu
rthii O
M34
13
Lann
ea s
chwe
infu
thii O
M13
55
Lann
ea s
chwe
infu
thii O
M24
46
Lann
ea sc
hwein
futh
ii OM
2505
Lann
ea sh
impe
ri OM
2521
Draco
ntom
elon
dao
AY59
4467
Pleiog
ynium
timor
iense
AY59
4474
Pleiog
ynium
timori
ense
AY72
4340
Harpep
hyllu
m caffru
m BS00
58
Harpep
hyllu
m caffru
m DMP20
4
Harpephyllum ca
ffrum O
M1555
Tapirira bethanniana AY594481
Tapirira obtusa AY594482
Tapirira obtusa JQ626383
Choerospondias axillari AY594463
Choerospondias axillari HQ427341
Antrocaryon amazonicum AY594460
Cyrtocarpa procera AY594464
Pseudospondias microcarpa PM5455
Sclerocarya birrea AY594478
Sclerocarya birrea subsp caffra OM0278
Sclerocarya birrea subsp caffra OM0498
Sclerocarya birrea subsp caffra RL1117
Commiphora africana OM0334
Commiphora glandulosa RBN160
Commiphora schimperi OM1361100
71
9589
50
65
88 99
59
77
50
93 62
88
7256
64
62
95
9963
83
8085
90
100
98
85
73
5999
82
75
97
97
9676 89
5175
97
81
53
85
54
76
89
65
CladeC
CladeA
CladeD
CladeB
KEY
Rhus Representatives