Herbarium introduction - Natural Resources Institute

Introduction to
Herbaria
Professor Philip Stevenson
RRoyal l BBotanic t i GGardens, d KKew
Natural Resources Institute, University of Greenwich

The Herbarium
• A collection of dried plants
• A store of reference material
• A means of identification
• An arbiter of correct names
• A comprehensive data-bank
“The essential working tool for systematics”

Role of the Herbarium
• Identification
• Taxonomic research
• Data repatriation (including databasing)
• IUCN Conservation ratings
• Voucher specimens
– Ecological
– DNA sequencing
– Phytochemical
y

Fieldwork and the Herbarium
• Fi Field ld Id Identifications tifi ti diffi difficult lt
– Tropical habitats very diverse
– Family or genus level
– Floras often not available (tropics ( p especially) p y)
– Floras bulky
• EEcological l i l fi fieldwork: ld k
– Plots, often sterile vouchers, shorter-term
• Taxonomic fieldwork:
– General collecting, collecting fertile material material, long long-term
term

Herbaria around the world
• General or international herbaria
• National and regional
– E.g., Forest Research Institute Malaysia (FRIM)
FForest t Herbarium, H b i SSarawak k (SAR)
Forest Research Centre, Sabah (SAN)
• University herbaria
• Local botanists = local contacts
• Index Herbariorum

HHerbaria b i ffrom around d th the world
ld

Field to specimen

Specimen
Determinavit slip
Loose parts
Label
Barcode
Herbarium stamp

The Kew Herbarium
• Approx Approx. 5 million specimens
• Comprehensive collections from all regions
Wing C - 1857
Wing A - 1903

The Kew herbarium contd contd.
• Specimen arrangement
– Phylogenetic y g according g to Bentham and
Hooker (1880)
– Re-arrangement Re arrangement with the new wing
• Mabberley’s Plant Book (new edition)
• St Staff ff ‘arrangement’
‘ t’
– Regional teams
– Systematic teams

Regional Teams
• South-East Asia and Pacific
• Drylands Africa
• Africa Wet tropics
• South America
• Temperate
• Name all material except systematic families
• Sorts Sorts, identification identification, field guides guides, and research

Systematic Teams
• MMalpighiales l i hi l (E (Euphorbs) h b )
•Myrtaceae y (Eucaplypts ( pyp etc.) )
• Labiatae/Lamiaceae (Mints)
• Rubiaceae (Coffee)
• Leguminosae (Peas and beans)
• Monocots
• Monographic and phylogenetic research

Data repatriation of
4000 specimens
ffrom Mexico M i and d
Central America
Digitisation progress
Nearly 50,000
legume specimens
on-line
32 legume
“species pages”

Tephrosia p vogelii g
Plants reportedly used (%) in
Malawi for pest control (n=167)
Some farmers report no activity from Tephrosia

Callosobruchus maculatus

eggs
ber of e
n numb
Mean
Oviposition by fecund bruchid beetles
on cowpea seeds treated with
powdered plant @1% w/v after 48 h.

Rotenoids from Tephrosia vogelii leaves
O
Deguelin R=H
Tephrosin R = OH
O
O
O
H
R
O
OMe
OMe
O R
OMe O
O
O
H
Rotenone R=H
12α-hydroxyrotenone R = OH
O
O
O
O
OH
OMe
OMe OH O
Sarcolobine Toxicarol
O
O
OMe
O
OMe
OMe
OMe

Is T. . candida effective?
Tephrosia
vogelii.
Controls bruchids
Tephrosia
candida
Promoted for soil
improvement (N2 fixing
& green mulch)
and (assumed) pest
control properties.
(growing at an International Agroforestry Centre)

1
Oh – its actually TT.
. vogelii
Tephrosia
vogelii.
Controls bruchids
Tephrosia
candida
Promoted for soil
improvement (N2 fixing
& green mulch)
2
and (assumed) pest
control properties.
Plastid Ltrn region, ITS nuclear DNA sequences &
morphology indicate both to be T. vogelii

Numbber
of inseects
(%)
100
90
80
70
60
40
30
Effect on C. maculatus of cowpea
ttreated t d with ith acetone t extracts t t of f
T. vogelii chemotypes after 48 h.
50 Dead
20
10
0
2% 0.20% 0.02% 2% 0.20% 0.02% AcO
Chemotype 1 Chemotype 2 Control
Sick insects are alive but paralysed
Sick
Healthy

1
2
LC LC-MS MS chromatograms of
T. vogelii chemotypes 1 & 2
Tvogelii herb specimen 34mg/ml
3 x Fig 150 mm 1a Phenomenex HPLC Luna C18(2), chromatogram 95:0:5(0 min) 0:95:5(20 min) of 0:95:5(25 T. vogelii min), water : MeOH chemotype : ACN +1% formic acid 1
PhilSAPP
22-Oct-2009
PhilSAPP
Tvogelii herb specimen 1 3: Diode Array
19.40
Range: 4.337e+2
AU
3.5e+2
3 3.0e+2 0e+2
2.5e+2
2.0e+2
1.5e+2
1.0e+2
5.0e+1
12.03
12.62
15.12
deguelin
tephrosin
tephrosi
sarcolobine 18.75
sarcolobin n
e
deguelin
rotenone
rotenone
11.50 12.00 12.50 13.00 13.50 14.00 14.50 15.00 15.50 16.00 16.50 17.00 17.50 18.00 18.50 19.00 19.50 20.00 20.50 21.00 21.50 22.00
Tcandida BI19309 3: Diode Array
20.68
Range: g 6.261e+1
AU
Fig 5.0e+1 1b HPLC chromatogram of T. vogelii
chemotype 2 Obovatin
Obovatin
5-methylether
O
O
5-methylether
4.0e+1
3.0e+1 12 12.00 00 12.58
2.0e+1
1.0e+1
0.0
13.40
OMe
O
15.12
20.35
toxicarol
Z-tephrostachin Z-
4 4 & & 5
tephrostachin
1 & 2 3
19.93 5
6
Time
11.50 12.00 12.50 13.00 13.50 14.00 14.50 15.00 15.50 16.00 16.50 17.00 17.50 18.00 18.50 19.00 19.50 20.00 20.50 21.00 21.50 22.00
Compound IDs based on 700MHz NMR and Orbitrap HR-EI MS
20.37

Flavanones and flavones from
T. vogelii chemotype 2 (inactive)
O O
OMe
O
OMe
O
2
O
O
H
R
O
HO
OMe
Obovatin 5-methylether 3
OMe
Deguelin R=H
Tephrosin R=OH R = OH
Z-tephrostachin
O
OMe
O
O
OH
O
OMe
*two two of 6 new
flavonoid aglycones
Yukovanol 55-methylether methylether * TTephrovogelone* h l *
Stevenson et al., Phytochemistry (submitted)
O
O
O O
O

Mildbraediodendron excelsum
Herbarium sheet:
Specimen collected by
Johannes Mildbraed in 1928
Living specimen:
Grown from seed collected
iin 1996 1996, Mt Mt. KKupe, CCameroon

AU
AU
LC-UV Analysis of Mildbraediodendron excelsum
Living specimen
Mildbraed 10643
2 4 6 8 10 12 14 t (min)
ANALYTES: 50% aq. MeOH
LC chromatogram (total scan PDA)
extracts of leaflet material

MILDBRAEDIN: a flavonol tetraglycoside from
Mildbraediodendron excelsum
HO
OH
O
O OH 7
α-RhaI
O CH
1
3
3 O
OH
3 O
OH O O
1
1 2
O
O
OH
β-Gal
OHOH CH3
CH HO
3 6 HO
CH HO α-RhaII
2
α-RhaIII O O
HO 1
HO
Main phenolic component of:
(1) Herbarium leaf fragment (1928)
(2) Li Living i specimen i
Veitch et al. (2005) Tetrahedron Lett. 46, 8595

Abundance
Relative
100
50
Flavonol pentaglycosides of Cordyla haraka
LC m/z = 1033 UV
m/z = 1047
265
348
200 300 400
λ (nm)
0
0 5 10 15 20 25 30 35
t (min)
Relative Abunndance
100
Ion current chromatogram 40
Cordyla haraka
D. Du Puy
90
80
70
60
50
30
20
10
0
MS
-146 146
287
-162 162
433
-146 146
595
-146 146
741
-146 146
887
1033
m/z
200 400 600 800 1000 m/z
Electrospray mass spectrum

CH 3
HO
HO
Distribution of flavonol pentaglycosides in Cordyla s.l.
HO
O
α-RhaIV
1
O
7
α-RhaIII
OH
O
O
CH 3
HO
HO
Amburana
Mildbraediodendron
Cordyla
M. excelsum
Cordyla CC. africana f i
C. densiflora
Aldina
C. haraka
C. madagascariensis
C. pinnata
C. richardii
OH
C. somalensis
3
β-Gal
HO
O
O
O
O
OH
1
1 2
3
O
OHOH
6
O
CH 2
R = OH, Cordylasin B
R = H, Cordylasin A
1
CH 3
α-RhaI
OH
O
1
O α-RhaII α RhaII
HO
HO
OH
CH3
Dupuya haraka
Dupuya madagascariensis
CONCLUSIONS:
C. haraka allied with
C. pinnata & C. richardii
NNo support t ffor transfer t f off
C. haraka to Dupuya
Veitch, Kite & Lewis (2008) Phytochemistry 69, 2329