A systematic revision of the genus Manekia (Piperaceae)

Heather Schubert

Outline• Introduction of

Piperaceae• Taxonomic history of

Piperaceae• Introduction of Manekia• Materials and methods• Results• Discussion

(Bornstein)

Piper obliquum

Piperaceae (Agardh)

• Pantropical family• Consists of 5 genera• 2,400-3,600 species (Callejas 2001; Bornstein 2005; Wanke et

al. 2007; Smith et al. 2008)

• Largest genera Peperomia and Piper

Piperaceae

• Aromatic• Perennial

– Shrubs, trees or vines (pipers)

– Herbs (peperomias) • Leaves

– Alternate, opposite, whorled or basal

– Sessile or petiolate

P. arboretum

(Bornstein)

Piperaceae

• Inflorescence– Spicate or racemose– Terminal or leaf-opposed

P. arboreum (Bornstein)

(Bornstein)

Piperaceae

• Flower– Small, perfect

(Neotropical) or imperfect (Old World tropics), lack a perianth

– Subtended by a glabrous to pubescent bract

– Unilocular ovary with single, basal ovule

• Fruit– drupe

P. abalienatum

(Bornstein)

P. schiedeanum P. pseudofuligineum

(Bornstein)(Bornstein)

P. arboreum

(Bornstein)

P. arboreum

(Bornstein)

Piper nigrum

• Black pepper• Spice/seasoning

(Bornstein)

Piper betle

• “Betle leaf” chewed with the areca nut

• Used as herbal medicine and for narcotic properties

(Bornstein)

The Problem in Piperaceae

• Circumscription– Large number of described species– Based on characters of questionable taxonomic

value, or fragmented material lacking flowers– Based on occurrence in a distinct political

distributional unit

Circumscription in Piper

• Several generic segregates established (Kunth 1839; Miquel 1843-33)

• Reduced to synonymy or recognized at sectional level (C. de Candolle 1869)

• Additional genera created (Trelease and Yuncker 1950; Yuncker 1958)

• Phylogenetic analysis confirmed Manekia generic status (Jaramillo et al. 2004)

Circumscription: Manekia vs Sarcorhachis

• Sarcorhachis and Manekia recognized (Trelease 1927)– Placed in tribe Pothomorpheae

• Single species of Manekia, M. urbanii– Haiti

• Six species and two varieties of Sarcorhachis (Steyermark 1971)– Costa Rica to Colombia, Ecuador, Peru, Venezuela,

Brasil and the Lesser Antilles

• Sarcorhachis and Manekia– Shared a multitude of features– Taxonomic synonyms (Bornstein 1996)

• Principle III of the International Code of Botanical Nomenclature– Earlier name deserves priority– Bornstein (1996) proposed conserving the name

Sarcorhachis– Arias et al. (2006) provided most new name

combinations

Manekia• Scandent habit• Alternate, palmately veined

leaves• Petiole vaginate• Spikes usually axillary,

occasionally terminal (solitary or paired)

• Stamens 4• Stigmas 3-5• Fruits laterally compressed

and immersed in fleshy rachis• Rachis pubescent

(http://images.search.yahoo.com/central and southamerica)

M. naranjoanaM. venezuelana

M. obtusaM. sydowii

M. incurva

M. urbanii

• Systematic revision of Manekia– Morphological study

• Examination of type specimens and other herbarium material

– Molecular investigation• Examined three DNA regions

– Internal transcribed spacer (ITS)– psbj-petA intergenic intron– rpl32 spacer

• Phylogenetic analysis pending

ITS = a piece of non-functional RNA situated between structural ribosomal RNAs (rRNA)

http://www.google.com/images

Diekmann, K. et al. DNA Res 2009 0:dsp008v1-8; doi:10.1093/dnares/dsp008

Circular structure of the chloroplast genome of Lolium perenne

Herbarium Materials

• A, BM, C, ILL, F, G, GH, K, MO, NY, P, RB, US and WIS

• Destructive sampling– MO, NY, US

• Collected after 1980• Wide distribution

Morphological Characteristics

Vegetative• Leaf

– Shape– Base– Apex– Venation– Pubescence– Texture/Color

• Stipule• Prophyll • Stem

Reproductive• Inflorescence/Infructescence

– Position– Rachis – Bract– Stamen position– Anther– Carpel– Stigma– Fruit

Morphological Measurements

• Internode length• Prophyll length• Leaf blade length and width• Petiole length• Petiole vagination distance• Inflorescence/

Infructescence length• Peduncle length• Anther length• Fruit length

Manekia naranjoanaPetiolevagination

Methods: Extraction and amplification

• Quiagen Dneasy extraction kit• PCR/Amplification

– ITS– psbj-petA – rpl32

• Amplifications run in MJ Research programmable thermal cyclers PTC-100-60 and PTC-100-96

Methods: PCR and cloning

• Visually verified PCR products on 0.7% agarose gels

• Gel purification using Millipore Montage DNA Gel Extraction kit

• Cloned using a Promega PGEM T-easy cloning kit

Methods: Sequencing

• Li-Cor LongreadIR 4200 automated sequencer• Verified with BLAST searches• Edited using Geneious Pro 4.7.6• Aligned using CLUSTAL 2.0.12• Submitted to Genbank upon publication

Statistical and Phylogenetic Analyses

• Canonical Discriminant Analysis• Maximum Likelihood Phylogenetic Analysis

with Bootstrap values

P. friedrichsthalii

(Bornstein)

Canonical Discriminant Analysis

Canonical Axis Eigenvalue

CumulativePercent

Likelihood Ratio

Approximate F Value Num DF Den DF p

1 4.7047 43.82% 0.00518292 5.47 175 506.14 <0.0001

2 2.5564 67.63% 0.02956688 4.27 136 408.69 <0.0001

3 1.6936 83.40% 0.10515160 3.50 99 309.27 <0.0001

4 1.1061 93.70% 0.28323568 2.86 64 208 <0.0001

5 0.6763 100.00% 0.59653614 2.29 31 105 0.0010

Petiole vagination stops prior to blade

Bract partly ciliate

Petiole vagination extends onto blade

Bract ciliate

5 main veins

7 main veins

5 main veins 7 main veins

Leaf base cordate

7 main veins

Leaf base obtuse, deltoid, oblique

5 or 9 main veins

naranjoana (Costa Rica) sydowii (Peru)

sydowii (Peru) sydowii (Peru)

sydowii (Colombia) sydowii (Ecuador)

naranjoana (Panama) venezuelana (Venezuela)

naranjoana (Costa Rica)venezuelana (Venezuela)

sydowii (Peru) sydowii (Peru)

venezuelana (Venezuela) incurva (Dominica)

incurva (Dominica) obtusa (Brasil)obtusa (Brasil) naranjoana (Panama)

sydowii (Peru) sydowii (Colombia)

naranjoana (Costa Rica) obtusa (Brasil)

obtusa (Brasil) naranjoana (Costa Rica)

0.5 substitutions/site

75

100

86

52

92

67

100

69

rpl32 spacer550 nucleotides

Discussion

• 4 distinct taxa– M. incurva– M. obtusa– M. urbanii– M. naranjoana

• Basionym: Piper naranjoanum C. DC., Linnaea 37: 363. 1872.• Basionym: Sarcorhachis sydowii Trel., Repert. Spec. Nov.

Regni Veg. 48: 16. 1940.• Basionym: Sarcorhachis venezuelana Steyermark, Pittiera 3:

33. 1971.

Discussion

• Better resolution – Use ITS, psbj-petA and rpl32 individually and

combined– M. venezuelana - morphological analysis– M. urbanii – genetic analysis

• Future Studies– Establish phylogenetic and biogeographic

relationships between Manekia and Zippelia

AcknowledgementsI thank Southeast Missouri State University for allowing me to

conduct this research. I acknowledge Dr. Allan J. Bornstein and Dr. James F. Smith for obtaining specimens. I extend thanks to Boise State University (BSU), especially Dr. James F. Smith, for allowing me to use the necessary equipment and teaching me the techniques needed for this study. Also, I thank Dr. Michael S. Taylor for allowing me to use the necessary equipment in his lab.

I thank Dr. Bornstein, Dr. Smith and Dr. Taylor for comments and discussions; Dr. John S. Scheibe for assistance with statistical analyses; my committee members, Dr. Bornstein, Dr. Taylor and Dr. Michael Aide, for assistance and encouragement throughout this study; and Jay Zimmers from BSU for helping with molecular work.

This study was funded by the College of Graduate Studies, the College of Science and Mathematics, the Department of Biology, Dr. Allan J. Bornstein, Dr. Michael S. Taylor, and Dr. James F. Smith (BSU).

ReferencesArias, T., R. Callejas, and A. Bornstein. 2006. New combinations in Manekia, an earlier name for Sarcorhachis (Piperaceae). Novon 16: 205-208.Bornstein, A. 1989. Taxonomic studies in the Piperaceae—I. The pedicellate pipers of Mexico and Central America (Piper subg. Arctottonia). Journal of the

Arnold Arboretum 70: 1-55. . 1991. The Piperaceae in the southeastern United States. Journal of the Arnold Arboretum, Supplementary Series 1: 349-366. . 1996. Proposal to conserve the name Sarcorhachis against Manekia (Piperaceae). Taxon 45: 323-324.

. 2005. Piperaceae. Pp. 453-457 in A Tropical Garden Flora, eds. G. Staples & D.R. Herbst. Honolulu: Bishop Museum Press.Callejas, R. 2001. Piperaceae. Pp. 1928-1984 in Flora de Nicaragua, Tomo III, eds. W. D. Stevens, C. Ulloa, A. Pool, and O. M. Montiel. St. Louis: Missouri

Botanical Garden Press.Candolle, C. de. 1869. Piperaceae. DC. Prodr. Syst. Nat. Veg. 16(1): 235-471.Committee for Spermatophyta. 1998. (1223) Conserve Sarcorhachis Trel. Against Manekia Trel. (Piperaceae). Taxon 47: 870.Holmgren, P. K., N. H. Holmgren and L. C. Barnett (editors). 1990. Index Herbariorum Part I: The Herbaria of the World, 8th edition. New York Botanical

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Questions?

P. bredemeyeri

(Bornstein)

HERBARIUM CODE

INSTITUTION LOCATION

A Arnold Arboretum, Harvard University U.S.A. Massachusetts. CAMBRIDGE.

BM The Natural History Museum U.K. England. LONDON.

C University of Copenhagen Denmark. COPENHAGEN.

ILL University of Illinois U.S.A. Illinois. URBANA.

F Field Museum of Natural History U.S.A. Illinois. CHICAGO.

G Conservatoire et Jardin botaniques de la Ville de Genève

Switzerland. GENÈVE.

GH Harvard University U.S.A. Massachusetts. CAMBRIDGE.

K Royal Botanic Gardens U.K. England. KEW.

MO Missouri Botanical Garden U.S.A. Missouri. SAINT LOUIS.

NY New York Botanical Garden U.S.A. New York. BRONX.

P Muséum National d'Histoire Naturelle France. PARIS.

RB Jardim Botânico do Rio de Janeiro Brazil. Rio de Janeiro. RIO DE JANEIRO.

US Smithsonian Institution U.S.A. District of Columbia. WASHINGTON.

WIS University of Wisconsin U.S.A. Wisconsin. MADISON.

Herbarium codes for herbaria loaned from for this study.