Ficha Catalográfica Online

Home , Blysmus compressus, Bolboschoenus caldwellii, Bolboschoenus maritimus, Bulbostylis hispidula, Carex hostiana, Carex monostachya

UNIVERSIDADE ESTADUAL DE CAMPINAS

INSTITUTO DE BIOLOGIA – IB

SUZANA MARIA DOS SANTOS COSTA

SYSTEMATIC STUDIES IN CRYPTANGIEAE (CYPERACEAE)

ESTUDOS FILOGENÉTICOS E SISTEMÁTICOS EM CRYPTANGIEAE

CAMPINAS, SÃO PAULO

2018

SUZANA MARIA DOS SANTOS COSTA

SYSTEMATIC STUDIES IN CRYPTANGIEAE (CYPERACEAE)

ESTUDOS FILOGENÉTICOS E SISTEMÁTICOS EM CRYPTANGIEAE

Thesis presented to the Institute of Biology of the University of Campinas in partial fulfillment of the requirements for the degree of PhD in Plant Biology

Tese apresentada ao Instituto de Biologia da Universidade Estadual de Campinas como parte dos requisitos exigidos para a obtenção do Título de Doutora em Biologia Vegetal

ESTE ARQUIVO DIGITAL CORRESPONDE À VERSÃO FINAL DA TESE DEFENDIDA PELA ALUNA Suzana Maria dos Santos Costa E ORIENTADA PELA Profa. Maria do Carmo Estanislau do Amaral (UNICAMP) E CO- ORIENTADA pelo Prof. William Wayt Thomas (NYBG).

Orientadora: Maria do Carmo Estanislau do Amaral

Co-Orientador: William Wayt Thomas

CAMPINAS, SÃO PAULO

2018

Agência(s) de fomento e nº(s) de processo(s): CNPq, 142322/2015-6; CAPES

Ficha catalográfica Universidade Estadual de Campinas Biblioteca do Instituto de Biologia Mara Janaina de Oliveira - CRB 8/6972

Costa, Suzana Maria dos Santos, 1987- C823s CosSystematic studies in Cryptangieae (Cyperaceae) / Suzana Maria dos Santos Costa. – Campinas, SP : [s.n.], 2018.

CosOrientador: Maria do Carmo Estanislau do Amaral. CosCoorientador: William Wayt Thomas. CosTese (doutorado) – Universidade Estadual de Campinas, Instituto de Biologia.

Cos1. Savanas. 2. Campinarana. 3. Campos rupestres. 4. Filogenia - Aspectos moleculares. 5. Cyperaceae. I. Amaral, Maria do Carmo Estanislau do, 1958-. II. Thomas, William Wayt, 1951-. III. Universidade Estadual de Campinas. Instituto de Biologia. IV. Título.

Informações para Biblioteca Digital

Título em outro idioma: Estudos filogenéticos e sistemáticos em Cryptangieae (Cyperaceae) Palavras-chave em inglês: Savannas Campinaranas Rupestrian fields Phylogeny - Molecular aspects Cyperaceae Área de concentração: Biologia Vegetal Titulação: Doutora em Biologia Vegetal Banca examinadora: Maria do Carmo Estanislau do Amaral [Orientador] Julie Henriette Antoinette Dutilh André dos Santos Bragança Gil Aparecida Donisete de Faria Marcelo Reginato Data de defesa: 18-04-2018 Programa de Pós-Graduação: Biologia Vegetal

Campinas, 18 de abril de 2018.

COMISSÃO EXAMINADORA

Maria do Carmo Estanislau do Amaral (Orientadora)

Julie Henriette Antoinette Dutilh

André dos Santos Bragança Gil

Aparecida Donisete de Faria

Marcelo Reginato

Os membros da Comissão Examinadora acima assinaram a Ata de defesa, que se encontra no processo de vida acadêmica do aluno.

AGRADECIMENTOS

Antes de quaisquer pessoas, agradeço aos meus pais, Ozana e Lucas, por todo o apoio e investimento que deram a minha Educação: sou grata pelo exemplo que são para a formação do meu caráter e pela oportunidade de me dedicar exclusivamente a minha formação acadêmica. Às minhas irmãs, Rosana e Luciana: eternas companheiras neste aprendizado sobre os diferentes modos de ser mulher, negra e independente neste mundo. Ao Emerson: companheiro de não-tão-longa data que parece me acompanhar já há muito tempo e que aguentou com respeito e empatia meus conflitos internos nestes passos finais da tese. Aos meus familiares, antepassados e contemporâneos, obrigada!

Agradeço grandemente a Profa. Maria do Carmo por ter aceitado me orientar em mais esta etapa de pós-graduação (depois de dois anos e meio de mestrado, ainda me aguentou por mais quatro anos! =P). Ao Prof. Wayt Thomas, pelo voto de confiança quando o convidei para coorientação nesta pesquisa. Aos dois agradeço pela excelência de orientação, que me permitiu entender quais modelos quero seguir quando tiver alunos sob minha tutela. Por me mostrarem que é possível uma pós-graduação onde o fator humano não é esquecido ou subjulgado, ainda que haja prazos e recursos restritivos.

Agradeço à Rosemeri Morokawa, pela preciosa colaboração nesta pesquisa: espero que venham muito mais trabalhos pela frente! À Nathália Streher e Marina Wolowski, pela parceria com os trabalhos de polinização (não incluí na tese, mas vamos finalizar?! =P).

Agradeço ao PPG – Biologia Vegetal, professores e funcionários. Aos professores, pelos preciosos conhecimentos transmitidos nas aulas. Aos funcionários que dentro da sobrecarga de trabalho certamente fazem o possível para atender as necessidades discentes (especialmente o João Galvão, Rafael Pessoa e Maria Roseli!). Agradeço a todos os colegas e funcionários da Unicamp e demais instituições pelo coleguismo ao longo dessa jornada. Agradeço a todas as equipes de herbários e unidades de conservação por onde passei, por facilitarem tanto minhas expedições e visitas. Agradeço pela oportunidade de aprender e conhecer tanto sobre o Brasil e sobre as pessoas que nele habitam (êta! Povo bonito e guerreiro!).

Aos amigos que carrego desde a adolescência (Driele), graduação (Thiago Ranniery, Zé Junior, Dani “sis”) e mestrado (Kátia, Martinha, Gabi, Anna, Luciana, Shimizu, Nah, Talitex, Larissa, Marcelinho, Rosemeri, Milena, Marcela, Edson Junior, Murilo e tantos

outros). As minhas primeiras amizades em Campinas, especialmente Tiago “Padre”, Gabi Atique, Anna Abrahão e Luciana Franci. E aos que conheci e/ou me aproximei no doutorado (Ana Claúdia, Kyria, Elimar, Edimar, Amanda, João Nazário, Raquel, Pavarotti, João do Carmo, Andreza (malvadeza), Batata, Juliana, Nicoll, Rebeca, Maíra, Guilherme, Kaire, Pedro Joel e tantos outros que agora fogem da memória!). [Desculpa se não coloquei seu nome! Se você acha é meu amigo, seu nome deveria estar aqui! =P].

Agradeço a Alba Luz Arbeláez e aos colegas que me receberam durante minha viagem ao The Missouri Botanical Garden. Agradeço ao prof. Thomas e Ina Ina Vandebroek pela recepção calorosa durante o doutorado-sanduíche em Nova Iorque, no The New York Botnaical Garden. Agradeço a Ina por dividir comigo sua morada e o convívio com o Koko.

Finalmente, agradeço a todas as medidas de inclusão social que permitem que mulheres, afrodescendentes, pobres, pessoas com limitações físicas e cognitivas ou que simplesmente estão distantes dos grandes centros urbanos tenham acesso a uma educação gratuita e de qualidade. Mesmo que eu não tenha sido diretamente favorecida por essas medidas, elas são impreteríveis em uma sociedade que pretenda ser justa.

Muito Obrigada!

Caçador de Mim Milton Nascimento Sergio Magrão e Luiz Carlos Sá (compositores)

Por tanto amor, por tanta emoção A vida me fez assim Doce ou atroz, manso ou feroz Eu, caçador de mim Preso a canções Entregue a paixões que nunca Tiveram fim Vou me encontrar longe do meu lugar Eu, caçador de mim Nada a temer Senão o correr da luta Nada a fazer Senão esquecer o medo Abrir o peito à força Numa procura Fugir às armadilhas da meta escura Longe se vai sonhando demais Mas onde se chega assim Vou descobrir o que me faz sentir Eu, caçador de mim

RESUMO

A pesquisa desenvolvida nesta tese de doutorado tem como objetivo esclarecer relações filogenéticas envolvendo a tribo Cryptangieae (Cyperaceae), além de contribuir para o reconhecimento de seus representantes. O trabalho encontra-se organizado em duas etapas. Na primeira parte (CAPÍTULO 1 e 2) são realizados estudos filogenéticos moleculares e morfológicos buscando reafirmar Cryptangieae como uma tribo e esclarecer o limite de seus gêneros. A segunda etapa traz o material necessário para o reconhecimento de novas espécies (CAPÍTULO 3), especialmente ocorrentes no Escudo das Guianas, e um tratamento atualizado de Cephalocarpus (CAPÍTULO 4). Ao final, impressões sobre os resultados obtidos e possíveis projetos futuros com a tribo são brevemente apresentados.

Palavras-chave: White-sand savanas, campinaranas, campos rupestres, Neotrópicos

ABSTRACT

I developed this PhD project aiming to clarify filogenetic relationships involving the sedge tribe Cryptangieae (Cyperaceae), and also to contribute to the recognition of its representatives. We organized the research in two steps. In the first part (CHAPTER 1 and 2), we perform molecular phylogenies and morphology studies to assert Cryptangieae as a tribe ans clarify its genera boundaries. Secondly we provide the essential material to the recognition of new species (CHAPTER 3), especially occuring in the Guiana Shield, and an updated treatment to the Cephalocarpus (CHAPTER 4). Finally, I discuss briefly about the results and possible future projects with the tribe.

Keywords: White-sand savanas, campinaranas, campos rupestres, Neotropics

Sumário INTRODUÇÃO ...... 13 Figura 1 – Cryptangieae morphological aspects (images): A-C) Inflorescences: A. spike-like inflorescence at Exochogyne amazonica, B. head-like inflorescence at Cephalocarpus confertus, C. panicle-like inflorescence at Lagenocarpus bracteosus. D-F) Inflorescence insertion: D. lateral and head-like inflorescence, E. lateral and panicle-like inflorescence, F. terminal and panicle-like inflorescence. G-H) Paracladia branching: G. with branches of many orders at Lagenocarpus sabanensis and, E. with just the 1st order at Lagenocarpus cf. glomerulatus...... 15 Tabela 1 – Circunscrição dos representantes de Cryptangieae de acordo com as revisões taxonômicas de Koyama (2003, 2004) e Vitta (2005) e sua apresentação em listagem para o Brasil (Alves et al. 2009) ...... 16 CHAPTER 1 - What about Cryptangieae (Cyperaceae)—a molecular hypothesis about its tribal status and circumscription ...... 17 Introduction ...... 18 Material and methods ...... 19 Results ...... 20 Discussion ...... 21 TABLE 2. List of taxa sampled with vouchers and Genbank accession numbers. Species name following Govaerts et al. (2017) and Elliott & Muasya (2017) [Acronyms according to Thiers (2016 – continuously updated)] ...... 24 TABLE 3. Molecular markers, primers and protocols applied during PCR experiments ...... 36 FIGURE 2. Phylogenetic hypothesis based on rbcL + trnL-F + ITS sequences to Cyperaceae. [Consensus tree of PP=0.5 branches; black – Thurniaceae + Juncaceae ; purple – Mapanioideae; pink– Trilepideae; red – Schoeneae; green – Sclerieae + Bisboeckelereae; orange – Cryptangieae; dark blue–Rhynchosporeae; light blue – other Cyperaceae tribes; full circle – PP = 1; empty circle – PP ≥ 0.95; full star– BS ≥ 90%; empty star – 75%≤ BS<90% . We describe PP and BS values to the mainly discussed branches] ...... 37 FIGURE 3. Summary of phylogenetic reconstruction of Cyperaceae based on rbcL, trnL-F and ITS matrices. [Consensus tree of PP=0.5 branches; black – Thurniaceae + Juncaceae; purple – Mapanioideae; pink– Trilepideae; red– Schoeneae; green – Sclerieae + Bisboeckelereae; orange – Cryptangieae; dark blue –Rhynchosporeae; light blue – other Cyperaceae tribes; full circle – PP = 1; empty circle – PP ≥ 0.95. We include PP values to the mainly discussed branches] ...... 38 CHAPTER 2 – “The more the merrier”: a molecular and morphological approach to the Cryptangieae (Cyperaceae) genera circumscription ...... 39 Introduction ...... 40 Material and Methods ...... 41 Results and Discussion ...... 43 Taxonomic results, Identification Key.– ...... 46 Taxonomic results, Genera description, accepted species and new combinations.– ...... 47 Ancestral Character State reconstruction.– ...... 54

General Conclusions ...... 58 Table 4 – Cryptangieae species and sequences at the Maximum Parsimony and Bayesian Inference analysis (type-species of previously described genera in bold; type-species of Koyama’s (2004) sections with an asterisk*) ...... 59 Table 5 – Molecular markers, primers and protocols applied during PCR experiments ...... 62 Table 6 – Diagnostic characters and character status selected to the Ancestral Character State analysis ...... 63 Figure 4 – Phylogenetic hypothesis on Cryptangieae genera based on a combined matrix with two chloroplast (rbcL and trnL-F) and three nuclear ribosomal (ITS, ETS and 5S-NTS) regions. [full circle – PP≥0.95; one star - BS=50%-<75%; two stars - BS≥75%. Group/genus color: black – outgroup; yellow – Cryptangium; pink – Didymiandrum; brownish-green – Exochogyne; purple – Krenakia; blue and red– Cephalocarpus (including Everardia); green - Lagenocarpus] ...... 64 Figure 5 – Mapping of all diagnostic characters applied the Ancestral Character State analyses and Ancestral Character States recovered to Cryptangieae (see selected diagnostic characters at Table 3; character n: distribution area, white – C&S America, green – campos rupestres, blue – Guiana Shield and Amazon, black – S America and Africa)...... 65 Figure 6 – Cryptangieae general morphology. A. Cephalocarpus confertus (habit), B-C. Cephalocarpus montanus habit (B) and caudex in cross section (C); D-E. Krenakia minarum habit (D) and detail of leaves reduced to sheaths (E); F-G. Exochogyne amazonica with mature male (F) and female (G) spikelets; H. Cryptangium verticillatum male spikelets; I. Krenakia sp. spikelets. J. Lagenocarpus rigidus inflorescence (female spikelets apical and in rigid peduncles and male spikelets basal and in flexible peduncles); K. Cephalocarpus confertus inflorescence detail (male and female spikelets at the same cluster); I. Cephalocarpus montanus uniflowered spikelets with immature achenes...... 66 CHAPTER 3 - Ten Cryptangieae (Cyperaceae) new species, the diversity buried at herbaria collections ...... 67 Introduction ...... 68 Material and Methods ...... 68 Description of the new species ...... 69 Conclusions ...... 83 Figure 7:– Male and female spikelets and achene of: A. Cephalocarpus chimantensis (voucher: J.A. Steyermark 128931), B. Cephalocarpus illustrata (voucher: O. Huber 10949), C. Cephalocarpus “lowlandia” (voucher: B. Maguire 42341), D. Cephalocarpus martinhae (voucher: J.A. Steyermark 129574), E. Cephalocarpus neblinensis (voucher: B. Maguire 42107), F. Cephalocarpus “chimanta 2” (voucher: J.A. Steyermark 75480) [scale (black bar)= 1 mm] (S.M.Costa original drawings)...... 84 Figure 8:–: A. Lagenocarpus setaefolius (voucher: W.W. Thomas 2656a & G.K. Rogers, NY! – holotype) ...... 85 Figure 9:– Lagenocarpus “aff glomerulatus” (photos; voucher: F. Guanchez 3401): A. male and female spikelets and achene, B. hypogynous scale (detail); Lagenocarpus rubriglumis (photos): C. male and female spikelets and achene (voucher: C.C. Berg P19505), D. hypogynous scale (detail; Zaruchi 2021) [scale (black bar)= 1 mm]...... 86

CHAPTER 4 – An updated taxonomic treatment to Cephalocarpus Nees (Cryptangieae, Cyperaceae) ...... 88 Introduction ...... 89 Material and Methods ...... 89 Results ...... 90 Taxonomic Treatment ...... 90 Final Comments ...... 132 CONCLUSÕES FINAIS ...... 139 APPENDIX 1 – Ancestral Character State analysis to each diagnostic morphological character ...... 148 APPENDIX 2 – Phylogenetic hypothesis on Cryptangieae genera based single-marker matrix. [PP values above and BS values below and between brackets. Group/genus color: black – outgroup; light green – Cryptangium; pink – Didymiandrum; brownish-green – Exochogyne; purple – Krenakia; red – Everardia; blue – Cephalocarpus; yellow - Lagenocarpus] ...... 161 APPENDIX 3 – Illustrations of Cephalocarpus from the treatments of Gilly (1942, 1943), Koyama & Maguire (1965) and Koyama (2003) ...... 166 ANEXO 1- Declaração de bioética e/ou biossegurança ...... 181 ANEXO 2- Declaração referente aos direitos autorais ...... 182

INTRODUÇÃO Cyperaceae Juss. é uma família cosmopolita que abarca mais de 5500 espécies de plantas herbáceas (Govaerts et al. 2017) e cujos representantes habitam diferentes fitofisionomias, áreas abertas a florestais (Goetghebeur 1998), desde ambientes áridos a aquáticos e palustres. Numa classificação infra-familiar baseada em caracteres morfológicos são aceitas quatro subfamílias e 14 tribos (Goetghebeur 1998). Esta classificação foi revisada após a elaboração de hipóteses filogenéticas com base em dados moleculares (Simpson et al. 2007, Muasya et al. 2009, Hinchliff & Roalson 2013). As hipóteses recentes foram realizadas com diferentes quantidades de marcadores moleculares, espécimes e metodologias estatísticas e corroboram para a aceitação de apenas duas subfamílias em Cyperaceae: Mapanioideae e Cyperoideae; além de novas propostas e indicações de pendências para a organização das tribos. Embora a amostragem da família seja suficiente para o entendimento de relações mais amplas em Cyperaceae, algumas tribos permanecem insuficientemente amostradas (Simpson et al. 2007, Muasya et al. 2009, Hinchliff & Roalson 2013). Ora por sua ampla diversidade, como no caso de Schoeneae, ora por tratar-se de uma tribo de localização restrita e/ou pouco estudada do ponto de vista taxonômico, como no caso de Cryptangieae. Cryptangieae está distribuída em áreas neotropicais extra-andinas, especialmente no Escudo das Guianas e campos rupestres, no Escudo Brasileiro (Goetghebeur 1998, Koyama 2003, 2004, Vitta 2005). Seus representantes são descritos em Goetghebeur (1998) como possuindo espiguetas unissexuadas com glumas espiraladas e perianto composto por três escamas hipóginas fimbriadas e de inserção opostas aos lados de aquênios trígonos. Nessa classificação, a tribo possui quatro gêneros (Cephalocarpus Nees, Didymiandrum Gilly, Everardia Ridley e Lagenocarpus Nees) e 50 espécies. Mais tarde, com os estudos moleculares, sugere-se que Cryptangieae engloba outro gênero (Exochogyne C.B. Clarke) (Muasya et al. 2009), o que lhe confere características adicionais: espiguetas com glumas (sub)dísticas, aquênios biconvexos e sem escamas hipóginas. Outros caracteres morfológicos importantes para diagnosticar os membros da tribo são relacionados a inflorescência, paracládios e aquênios (Figura 1). Existem inconsistências sobre quais gêneros e espécies devem ser aceitos em Cryptangieae (Koyama 2003, 2004, Vitta 2005, Alves et al. 2009; Tabela 1), mas os trabalhos taxonômicos recentes baseados em morfologia realizados por Koyama (2003, 2004) reconhecem três gêneros (Cephalocarpus, Everardia e Lagenocarpus incluindo

Didymiandrum e Exochogyne) e 47 espécies. Outro estudo morfológico e anatômico importante realizado com a tribo foi apresentado por Vitta (2005). Esse trabalho propõe uma nova classificação para os representantes de Lagenocarpus sensu Koyama (2004), mas seus resultados não foram publicados em periódicos científicos e permanecem disponíveis apenas no formato de tese. Cryptangieae possui incongruências no reconhecimento taxonômico de seus gêneros e espécies e uma distribuição restrita em áreas neotropicais, como o Escudo da Guiana e campos rupestres do sudeste brasileiro. Concomitantemente, os estudos moleculares disponíveis não incluem espécies de Cephalocarpus e nem de Everardia, o que mantem Cryptangieae como uma incógnita para o entendimento sistemático e evolutivo da família. O status de tribo de Cryptangieae foi contestado em Hinchliff & Roalson (2013), que sugere sua inclusão na tribo Schoeneae, seu monofiletismo e relação com demais tribos de Cyperaceae, dúbios devido à baixa amostragem de sua diversidade, e a dificuldade de reconhecimento taxonômico, devido à incongruência entre propostas recentes, permanecem como barreira ao entendimento das tribos mais basais de Cyperoideae. A obscuridade sobre os limites e relações entre seus gêneros e espécies impedem estudos incluindo evolução de caracteres macro e micro morfológicos, biologia reprodutiva, entre outros, sob a abordagem da biologia molecular. Devido a isso, esta tese foi desenvolvida com o objetivo geral de esclarecer relações filogenéticas de Cryptangieae com as demais tribos de Cyperaceae e entre seus gêneros. No primeiro capítulo, publicado no periódico Phytotaxa Journal em abril/2018, é testado e defendido o status monofilético e tribal de Cryptangieae a partir de uma maior amostragem da diversidade de seus gêneros e tribos, através do uso de ferramentas moleculares filogenéticas. O segundo capítulo é elaborado para reavaliar a circunscrição e relações de gêneros em Cryptangieae, com base em dados moleculares e morfológicos. Neste capítulo são fornecidas informações morfológicas e taxonômicas atualizadas, incluindo chaves para reconhecimento morfológico dos gêneros como aqui propostos. Os dois capítulos seguintes são tratamentos taxonômicos para dez espécies novas (sete Cephalocarpus e três Lagenocarpus) descobertas durante a tese (Capítulo 3) e para Cephalocarpus Nees (sensu Costa et al., Capítulo 2) (Capítulo 4).

Figura 1 – Cryptangieae morphological aspects (images): A-C) Inflorescences: A. spike-like inflorescence at Exochogyne amazonica, B. head-like inflorescence at Cephalocarpus confertus, C. panicle-like inflorescence at Lagenocarpus bracteosus. D-F) Inflorescence insertion: D. lateral and head-like inflorescence, E. lateral and panicle-like inflorescence, F. terminal and panicle-like inflorescence. G-H) Paracladia branching: G. with branches of many orders at Lagenocarpus sabanensis and, E. with just the 1st order at Lagenocarpus cf. glomerulatus. A B C

D E F

G H

Tabela 1 – Circunscrição dos representantes de Cryptangieae de acordo com as revisões taxonômicas de Koyama (2003, 2004) e Vitta (2005) e sua apresentação em listagem para o Brasil (Alves et al. 2009)

CHAPTER 1 - What about Cryptangieae (Cyperaceae)—a molecular hypothesis about its tribal status and circumscription AUTHORS: Suzana M. Costa, Rosemeri Morokawa, Volker Bittrich, William W. Thomas & Maria do Carmo E. Amaral [published at Phytotaxa 347 (2): 127–149]

Abstract

We endorse the tribal status of Cryptangieae and discuss its most probable sister-group based on phylogenetic analyses using rbcL, trnL-trnL-trnF and ITS sequences. Cryptangieae were poorly sampled in previous molecular phylogenies of Cyperaceae: only three of about 50 species were sampled and all of them are currently included in the genus Lagenocarpus. We gathered more data about the tribe to verify its monophyletic status and to clarify the relationships among the tribes of subfamily Cyperoideae. Here we present a phylogeny with 18 species of Cryptangieae including all three currently recognized genera. Cryptangieae appears as monophyletic with great support in our Maximum Parsimony and Bayesian Inference analyses based on single and combined matrices. We emphasize the need for morphological, molecular and biogeographical data to achieve a broadly supported interpretation of the early divergent taxa of subfamily Cyperoideae. Keywords: Cephalocarpus, Everardia, Lagenocarpus, Neotropics, sedges

Resumo

É defendido o status de tribo para Cryptangieae e discutido o seu grupo-irmão mais provável baseado em análises filogenéticas usando sequências de rbcL, trnL-trnL-trnF e ITS. Cryptangieae foi pouco amostrada nas filogenias moleculares anteriores de Cyperaceae: apenas três de cerca de 50 espécies foram amostradas e todas elas atualmente inclusas em Lagenocarpus. Foram reunidos mais dados sobre a tribo para verificar seus status monofilético e para esclarecer as relações entre as tribos da subfamília Cyperoideae. Aqui, é apresentada uma filogenia com 18 espécies de Cryptangieae incluindo os três gêneros atualmente reconhecidos. Cryptangieae aparece como monofilética e com bom suporte em nossas análises de Máxima Parcimônia e Inferência Bayesiana baseadas em matrizes de região única e combinada. Enfatiza-se a necessidade de dados morfológicos, moleculares e biogeográficos para permitir uma interpretação mais abrangente sobre os táxons que divergiram primeiro na subfamília Cyperoideae.

Introduction

Cyperaceae are cosmopolitan and the third largest family among the monocots, with about 93 genera and 5616 species (Govaerts et al. 2017). Goetghebeur (1998) reported four subfamilies (Mapanioideae, Sclerioideae, Caricoideae and Cyperoideae) and 14 tribes for Cyperaceae based on non-molecular characters. Cryptangieae was included in Sclerioideae (Goetghebeur 1998) and historically grouped with Trilepideae, based on morphological characters such as unisexual spikelets, achene morphology, and fimbriate hypogynous scales (Bruhl 1995, Koyama & Maguire 1965, Goetghebeur 1998).

Currently, Cryptangieae comprises about 47 species and three genera (Cephalocarpus Nees, Everardia Ridley and Lagenocarpus Nees—sensu Koyama 2003, 2004) occurring from Central America and Cuba south to subtropicalregions of South America (Koyama 2003, 2004, Govaerts et al. 2017). The tepuis of the Guiana Shield and the campos rupestres of the Brazilian Shield in southeastern Brazil are particularly species rich and include numerous endemic species (Koyama 2003, 2004). The tribe comprises species with unisexual spikelets, spirally arranged glumes and three fimbriate hypogynous scales (Goetghebeur 1998). Most of the species are monoecious and present the male spikelets at the base and the female ones at the apex of the inflorescence (Goetghebeur 1998, Koyama 2003, 2004, Vitta 2005).

Phylogenetic hypotheses developed using different molecular markers and broader sampling clarified relationships at different levels within Cyperaceae (Simpson et al. 2007, Muasya et al. 2009, Hinchliff & Roalson 2013) and led to the recognition of only two subfamilies: 1. Mapanioideae, comprising the same tribes as in the classical circumscription, and 2. Cyperoideae, comprising all other tribes (including Cryptangieae). The results highlighted the need to reappraise the circumscription and relationship among the Cyperoideae tribes (Muasya et al. 2009). Also, the close relationship between Cryptangieae and Trilepideae was rejected (Hinchliff & Roalson 2013, Spalink et al. 2016), although sometimes with only weak support (Simpson et al. 2007, Muasya et al. 2009).

Other doubts arose after the molecular studies, including: 1. the relationship among Schoeneae, Bisboeckelereae, Sclerieae and Cryptangieae; 2. the circumscription of Schoeneae and, 3. the tribal status of Cryptangieae. Muasya et al. (2009) presented an imprecise relationship among these four tribes, while Hinchliff & Roalson (2013) found Bisboeckelereae + Sclerieae as a separate clade and suggested the inclusion of Cryptangieae

in Schoeneae. In both studies, Cryptangieae were poorly sampled, both in numbers of species sampled and the morphological diversity within the tribe. The sampled species (Lagenocarpus albo-niger (A. St.-Hil.) C.B.Clarke, Exochogyne amazonica C.B.Clarke = Lagenocarpus amazonicus (C.B.Clarke) H.Pfeiffer, and Didymiandrum stellatum (Boeckeler) Gilly = Lagenocarpus stellatus (Boeckeler) O.Kuntze) essentially represent Lagenocarpus sensu Koyama (2003, 2004), with no specimens of Cephalocarpus or Everardia sampled. Other molecular phylogenies involving infrafamiliar relationships show neither a different placement nor a better sampling of Cryptangieae (Jung & Choi 2013).

Here we present a molecular phylogenetic study based on a greater sampling diversity within Cryptangieae aiming to test its monophyly and tribal status. We also add information about relationships among early divergent Cyperoideae tribes.

Material and methods

To investigate if the tribe Cryptangieae (sensu Koyama 2003, 2004) is monophyletic, we selected the rbcL, trnL-trnL-trnF (trnL-F) and ITS regions as these are widely available for Cyperaceae in online databases, useful in answering the questions raised, and relatively easy to amplify (Muasya et al. 2009, Hinchliff & Roalson 2013). We obtained sequences of one Thurniaceae species and four Juncaceae species from GenBank to use as outgroups. We included almost all Cyperaceae genera, based mainly on the list found in Muasya et al. (2009); we preferentially included species with both rbcL, trnL-F and ITS sequences available. The Cryptangieae samples represent the three currently recognized genera of the tribe (sensu Koyama 2003, 2004) and added sequences from GenBank as well as our own new sequences. (see Table 2)

We extracted total DNA from vouchers and silica-gel dried specimens using DNA extraction kits for plants (NucleoSpin® Plant II, Macherey-Nagel, DNeasy® Plant Mini kit, Qiagen). The amplification experiments followed modified protocols from Muasya et al. (2002) and Roalson & Friar (2004) (Table 3). The new sequences of Cryptangieae were deposited in the GenBank database (Table 2).

We aligned the sequences using MAFFT (Katoh et al. 2002) at default settings and made some manual adjustments with Mesquite 3.04 (Maddison & Maddison 2007). We

performed Maximum Parsimony (MP) and Bayesian Inference (BI) analyses using single and combined matrices.

We elaborated the MP hypothesis using TNT 1.5 (Goloboff et al. 2008, 2016): 1.. A heuristic search with 1000 RASR holding 10 trees at each step, TBR branching swapping with MULT REES and steepest descent in effect saving max. 50 trees/ replicate; 2. Another heuristic search including retained trees, and finally; 3. A Bootstrap Search (BS) with 300 (single matrices) or 900 (combined matrix) replicates, 100 RASR with three trees held per step, TBR branch swapping with steepest descent and MULT REES in effect, saving 10 trees at each RASR. We considered those clades having a Bootstrap Support values (BS) ≥ 75% as supported.

We elaborated the BI hypothesis using JModeltest online (Guindon & Gascuel 2003, Darriba et al. 2012) to select the best evolutionary model (by AICc criterion) and MrBayes 3.2.2 (Huelsenbeck & Ronquist 2001) on the CIPRES platform (www.phylo.org) to perform the MCMC analysis, with the following settings: nchains = 4; nruns = 2; ngen = 107 (to single matrices) / 5 × 107 (combined matrix); burnin = 0.25; samplefreq = 1,000. We considered those clades as supported when the Posterior Probability (PP) ≥ 0.95.

Results

Our analyses include 44 new sequences of 18 Cryptangieae species among the three markers (rbcL, trnL-F and ITS), together with 381 sequences/ 68 genera/ 186 species of Cyperaceae from GenBank (Table 1). The aligned matrix of each molecular marker presents the following lengths: 1424 base pairs (bp) for the rbcL matrix; 2279 bp for the trnL-F matrix and 1013 bp for the ITS matrix.

The MP analyses based on single-marker matrices showed Cryptangieae with BS = 97 - 100% and the combined matrix shows Cryptangieae with BS = 93%. T he best evolutionary models were: 1. rbcL – GTR + I + G; 2. trnL-F – TVM + I + G; and, 3. ITS – GTR + I + G. The clades recovered in our different analyses (single/combined matrix; MP/BI analyses) are similar and mainly match with the ones recovered in Muasya et al. (2009), Simpson et al. (2007) and Hinchliff & Roalson (2013) (Fig. 2).

The subfamily Mapanioideae is supported in all analyses (BS values = 86–97%; PP values = 1) while Cyperoideae is reconstructed in most of them (Fig. 2). Trilepideae, here

represented only by rbcL sequences, is the earliest divergent clade of subfamily Cyperoideae (BS values = 100% in the rbcL-only analysis and 67% in the combined analysis; PP values = 1; Fig. 2, 3a). Bisboeckelereae and Sclerieae are sister-clades (BS values = 71–92%; PP values = 1) except in the trnL-F analysis, where Bisboeckelereae is paraphyletic without the inclusion of Sclerieae, a result also recovered by Hinchliff & Roalson (2013). The circumscription of Schoeneae is unclear and some taxa are separate from the species richer clade, including Cladium, Gymnoschoenus, and Carpha + Trianoptiles; additionally the tribe frequently is not well supported in the analyses (Fig. 2, Fig. 3a–c).

Cryptangieae is strongly supported as monophyletic in all analyses: MP (BS values = 97–100% in single-marker analyses and BS = 93% in combined analysis) and BI (PP values = 1), either with single-markers or a combined matrix.

Rhynchosporeae and the remaining Cyperoideae tribes are gathered together in almost all phylogenetic hypotheses (BS values = 56–91%, but absent in the rbcL reconstruction; PP values = 0.95–1). A closer look at the Cryptangieae combined-matrix BI analysis shows Lagenocarpus (sensu Koyama 2003, 2004) as paraphyletic without Cephalocarpus and Everardia. A study including more species and markers is in progress (Costa et al., unpublished-CHAPTER 2).

As our phylogenetic hypotheses show similar clades and support values, we focus our discussion on the BI reconstruction of the combined matrix, with additional comments on MP support values (Fig. 2). [The single and combined phylogenetic hypotheses with complete terminals are available as “Supplementary Material”]

Discussion Cryptangieae sensu Muasya et al. (2009) is monophyletic using Maximum Parsimony (BS values = 97–100 % in single-marker analyses and BS = 93% in combined analysis) or BI (PP values = 1) methods, but it still is uncertain as to what is its sister-clade (Fig. 1).

Although relationships among some early divergent clades of Cyperoideae are weakly supported, the hypothesis of a close relationship between Cryptangieae and Trilepideae is rejected as in previous molecular works (Simpson et al. 2007, Muasya et al. 2009, Hinchliff & Roalson 2013). According to our data, the closest Cryptangieae

relationships may be among the following tribes: Bisboeckelereae, Schoeneae, or Sclerieae. This confirms the results of Simpson et al. (2007) and Muasya et al. (2009), but is less conclusive than the analysis of Hinchliff & Roalson (2013), which points to an indirect relationship among Cryptangieae and a Bisboeckelereae + Sclerieae clade. They also suggest the inclusion of Cryptangieae in Schoeneae, based on the results of their supermatrix analysis.

Here, we present the most complete sampling of Cryptangieae, an adequate sampling of other genera of Cyperaceae, and at least two independent sources of molecular data (two chloroplast and one nuclear ribosomal region). We support the maintenance of Cryptangieae as a tribe because it is consistently monophyletic in all analyses, with few or many taxa, different numbers of markers, or different outgroups (Simpson et al. 2007, Muasya et al. 2009, Hinchliff & Roalson 2013, Spalink et al. 2016, and our own analyses). Also, species of Cryptangieae are morphologically coherent, grouping taxa with unisexual spikelets, spirally arranged glumes, and with fimbriate hypogynous scales opposite the achene sides (Goetghebeur 1998).

The question of Schoeneae circumscription and the need for more data emerged before, as in Goetghebeur’s (1998) morphology-based work, and persist in recent papers, where Schoeneae are frequently polyphyletic (Simpson et al. 2007, Muasya et al. 2009, Hinchliff & Roalson 2013); this is reflected in our own analysis. The most recent phylogeny of Schoeneae is from Viljoen et al. (2013) but also, due to lack of data, they decided to delay publication of their conclusions on a Schoeneae rearrangement. Some additional research on the Schoeneae tribe is being carried out (Isabel Larridon, pers.comm.), but we do not have access to the results or discussion yet.

An overall approach studying these five tribes (Schoeneae, Cryptangieae, Sclerieae, Bisboeckelereae, and Trilepideae) using molecular, morphological and biogeographical information, may give us a better understanding of their circumscription and relationships. This will be a large task, however, as these genera are spread all over the globe and present cases of endemism, each with its own particularities.

This is the first phylogenetic molecular study to confirm Cephalocarpus and Everardia within the Cryptangieae. But even if our dataset is currently the largest, it is insufficient to discuss the relationships within Cryptangieae. Also, our analyses highlight that the generic circumscriptions need particular attention. The delimitation of Lagenocarpus has

changed over time based on morphological data, with repeated inclusion and exclusion of taxa, and the genus, sensu Koyama (2003, 2004), is paraphyletic in our phylogenies. A work in progress, focusing on Cryptangieae with a broader sampling and including morphological and molecular data, may lead us to a clearer circumscription of the genera and of the tribe (Costa et al. unpublished data-CHAPTER 2).

TABLE 2. List of taxa sampled with vouchers and Genbank accession numbers. Species name following Govaerts et al. (2017) and Elliott & Muasya (2017) [Acronyms according to Thiers (2016 – continuously updated)] (Family, Subfamily) Tribe and Species Voucher and/or Reference GenBank Reference rbcL trnL-F ITS (Thurniaceae) Prionium serratum (L. f.) Drège F.Getliffe Norris, s.n. U49223.1 AY344155 - (Nat.Bot.Garden, Cape Town, SA)/ .1 Muasya s.n. (K)/ - (Juncaceae) Juncus effusus L. Simpson et al. (2003)/ Chase et al. L12681 AY344156 AY727792 (1993)/ China:SABE 805 .1 Juncus gerardii Loisel. Simpson et al. (2003)/ Drabbkova et AY216613 AY344157 - al. (unpublished)/ - Luzula multiflora (Ehrh.) Lej. Bremer (2002)/ Simpson et al. (2003)/ AJ419945 AY344158 - - Luzula sylvatica (Huds.) Gaudin A - Simpson et al. (2003)/ B - AY216637 AY344159 FJ213851. Drabbkova et al. (unpublished) A A 1B (Cyperaceae, Mapanioideae C.B. Clarke) Tribe Chrysitricheae Lestib. ex Fenzl Capitularina involucrata (J. V. Suringar) Kern Simpson et al. (2003) AY344168 - - Chorizandra cymbaria R. Br. Bremer (2002) AJ419940 - - Chorizandra enodis Nees Bremer (2002) AJ419939 - - Chorizandra sphaerocephala R. Br. Simpson et al. (2003) - AY344170 - Chrysitrix capensis L. Bremer (2002)/ S. Africa: Muasya AJ419938 AY344171 - 1242 (K)/- Exocarya sclerioides (F. Muell.) Benth. Costion (unpubl.)/Simpson et al. KF496341. AY344167 - (2003)/- 1 .1 Lepironia articulata (Retz.) Domin. Malaysia: Simpson 1236 (K) Y12957 AY344169 - Tribe Hypolytreae Presl ex Fenzl

Diplasia karatifolia Rich. Simpson et al. (2003) - AY344166 - Hypolytrum bullatum C.B. Clarke Brazil: Thomas et al. 10318 (NY) Y12956 - - Hypolytrum nemorum (Vahl) Spreng. Malaysia: Simpson 1379 (K) Y12958 AJ295816 AY242046 .1 Hypolytrum testui Cherm. Simpson et al. (2003) - AY344163 - Mapania cuspidata (Miq.) Uittien Brunei: Marsh 4 (K) Y12955 AJ295817 - Mapania lorea Uitt. Simpson et al. (2003) AY344161 - - Mapania meditensis D.A. Simpson Brunei: Simpson et al. 2515 (K) Y12954 AY344160 - Mapania tenuiscapa C.B. Clarke Simpson et al. (2003) AY344162 - Scirpodendron bogneri S.S. Hooper Malaysia: Simpson 2650 (K) Y12946 AY344164 - Scirpodendron ghaeri (Gaertn.) Merrill Simpson et al. (2003) AY344165 - (Cyperaceae, Cyperoideae Suess.) Tribe Abildgaardieae Lye Actinoschoenus composita K. L. Clarke 213 (NE) - AY506702 AY506755 .1 .1 Actinoschoenus repens Raynal Zambia: Robinson 3643 (K) EF178537 - - Arthrostylis aphylla R. Br. Australia: Wilson 8249 (NSW)/ K.L. AY725939 - AY506757 Clarke 183 (NE) .1 Bulbostylis juncoides (Vahl) Kük. ex Herter E.H. Roalson 1350 (RSA) - AF285063. AF190615. 1 1 Bulbostylis barbata (Rottb.) C.B. Clarke A - Christin, P.A.(unpubl.)/ B - K.L. AM99979 AY506709 AY506764 Clarke 113 (NE) 0.1A .1B .1B Bulbostylis atrosanguinea (Boeckeler) C.B. Clarke Kenya: Muasya 1037 (EA, K) Y12992 - - Bulbostylis hispidula (Vahl) R.W. Haines Kenya: Muasya 1025 (EA, K) Y12944 - - Crosslandia setifolia W. Fitzg. A - Australia: Wilson 10147 (K)/ B - EF178538 EF178592 AY506768 K. L. Clarke 246 (NE) A A .1B Fimbristylis complanata (Retz.) Link Kenya: Muasya 1029 (EA, K) Y13009 - AY242051 .1 Fimbristylis dichotoma (L.) Vahl A - Kenya: Muasya 1006 (EA, K)/ B – Y13008A AJ295755 JX644882. Jung & Choi (2013) A 1B

Fimbristylis ovata (Burm. f.) J.Kern A - Kenya: Muasya et al. 684 (EA, K)/ Y12985A AJ295754 AB250638 B – Japan: Tsusaka 19707 (OKAY) A .1B Nemum spadiceum (Lam.) Desv. ex Ham. West Africa: Baldwin 9766 (K) Y12945 - - Trachystylis stradbrokensis (Domin) Kük. Australia: Wilson 8175 (K) EF178539 EF178591 - Tribe Bisboeckelereae Pax ex L.T. Eiten Becquerelia cymosa Brongn. A - Brazil: Thomas et al. 10284 (NY)/ Y12948A KF553496. LN886915 B – Suriname: Van Andel T.R. & al. 1A .1B 4473 (GENT) Calyptrocarya sp K 11301 (Viljoen et al. 2013) - - KF553442. 1 Calyptrocarya glomerulata (Brongn.) Urb. BioBot 05962 (iBOL data)/ Guyana: JQ591219. - LN886843 De Wilde Bart 7 (TCD), 1 .1 Calyptrocarya bicolor (H. Pfeiff.) T. Koyama RBGKEW DNA 10389 EF178540 - Diplacrum africanum C.B. Clarke Tanzania: Vollensen 3967 (K) AY725942 - - Diplacrum caricinum R. Br. Jung & Choi (2013) JX644670. JX644810. JX644865. 1 1 1 Cariceae Kunth ex Dumort. Carex cephalophora Muhl. ex Willd. Kress et al. (2005)/ USA: Olmstead DQ006089 AF164942. AY757408 s.n. (WTU)/ USA: Waterway 2001.092 1 .1 (MTMG) Carex deweyana Schwein. AP333 (OAC 90174)/ USA: Yen 94 HQ589994 AF164941. AY757412 (WTU)/ Canada: Waterway 98.005 .1 1 .1 (MTMG) Carex echinochloe Kunze A - Kenya: Muasya 1051 (K)/ B - Y12997A AF191818 EU288585 Berit et al., BG-Af 161, Kenya (ZH) A .1B Carex hostiana DC. Chase et al. (1993) L12672 - - Carex monostachya A. Rich. Kenya: Muasya 1052 (K)/ - / Berit et Y12998 - EU288605 al., BG-Af 069 (H) .1 Carex nemoralis (K.L.Wilson) K.L.Wilson Australia: Wilson et al. 9533 (K) AY725956 - - Carex simpliciuscula Wahlenb. Plunkett et al. (1995)/Yen et al. (2000) U49232 AF164948 - Carex sylvatica Huds. De Vere et al. (2012)/ Simpson et al. JN893542. AY344175 AY278306

(2003)/Germany: W Mainer 2015 1 .1 .1 Tribe Cryptangieae Benth. Cephalocarpus rigidus Gilly ex Gleason & Killip Brazil: Costa 1113, 1130 (UEC) MF564249 MF564233 - /MF56425 /MF56423 0 4 Everardia diffusa T. Koyama & Maguire B. Maguire 27825 (NY) MF564260 MF564236 MF564221 Everardia disticha T. Koyama & Maguire O. Huber 10060 (NY) MF564251 MF564237 Everardia flexifolia (Gilly) T. Koyama & Maguire B. Maguire 29528 (F) - MF564238 - Everardia maguireana T. Koyama Brazil: Costa 1133, 1121 (UEC) MF564253 MF564239 MF564219 /MF56425 /MF56422 2 0 Everardia montana Ridl. Maguire, B. et al. 53624 (K) FN870820. 1 Lagenocarpus albo-niger (A. St.-Hil.) C.B. Clarke Brazil: Thomas 11111 (NY); Costa AY725949 - - 1240 (UEC) / MF564256 Lagenocarpus amazonicus C.B. Clarke Colombia: Arbeláez 929 (MO). Brazil: EF178545/ MF564240 MF564232 Aparecida da Silva 1986 (GENT - MF564265 / GenBank); Costa 1031 (UEC) MF564241 Lagenocarpus celiae T.Koyama & Maguire Brazil: Costa 1044 (UEC) MF564254 MF564242 MF564225 Lagenocarpus comatus (Boeckeler) H. Pfeiff. Brazil: Valente 612 (NY) MF564262 MF564243 MF564222 Lagenocarpus glomerulatus Gilly Brazil: Costa 1047 (UEC) MF564229 MF564244 MF564257 Lagenocarpus guianensis Nees Brazil: Costa 1071 (UEC) MF564258 - MF564228 Lagenocarpus humilis (Nees) Kuntze Zappi 2633 (NY) MF564261 MF564245 MF564224 Lagenocarpus minarum (Nees) Kuntze Brazil: Costa 1274 (UEC) - MF564246 - Lagenocarpus rigidus (Kunth) Nees Brazil: Costa 1045 (UEC) MF564259 MF564247 MF564226 Lagenocarpus rigidus subsp. tenuifolius (Boeckeler) Brazil: Costa 1269 (UEC) MF564255 MF564248 MF564227 T.Koyama & Maguire Lagenocarpus stellatus (Boeckeler) Kuntze Venezuela: Liesner 23562 (GENT) EF178544 - - Lagenocarpus verticillatus (Spreng.) T. Koyama & Brazil: Costa 1076, 1167 (UEC) MF564264 - MF564230 Maguire /

MF564231 Tribe Cypereae Dumort. Androtrichum giganteum (Kunth) H. Pfeiff. Argentina: Tressens et al. 4292 (K) EF178546 - - Androtrichum trigynum (Spreng.) H. Pfeiff. Argentina: Goetghebeur 4764 (GENT) EF178547 - - Cyperus alternifolius subsp. flabelliformis Kük. A - Madagascar: Kew Acc. 6136603/ Y12967A AJ295758 JX644851. B – Jung & Choi (2013) A 1B Cyperus ascocapensis Bauters Kenya: Muasya 1009 (EA, K) Y13003 AJ295757 AB685858 .1 Cyperus blepharoleptos Steud. Zambia: Richards 13318 (K) Y13006 AY040602 JX566741. 1 Cyperus brevifolius (Rottb.) Hassk. Australia: Coveny et al. 17459 (K) AF449515 AF449564/ - AF449576 Cyperus compressus L. A - Thailand: Muasya 1375 (K)/B - AF449506 AF449555 KF193575. LSU:Reid and Carter 7761 A A 1B Cyperus distichus (Merxm. & Czech) Bauters Namibia: Muller et al. 4245 (K) EF178561 - - Cyperus erinaceus (Ridl.) Kük. Tanzania: Faden et al. 96/338 (K) AJ404699 AJ295764 - Cyperus flavescens L. A - Kenya: Muasya 1022 (EA, K)/ B - Y13005A AJ295763 KF150598. LSU:Reid 7576 A 1B Cyperus hyalinus Vahl A - Kenya: Mwachala 296 (EA)/ B - AY725953 - KF150602. LSU:Carter 18737 A 1B Cyperus isolepis (Nees) Bauters Thailand: Muasya 1217 (K) AF449516 AF449565/ AB685866 AF449577 .1 Cyperus kyllingiella Larridon Zimbabwe: Muasya et al. 1118 (K) AY040592 AJ295807 - Cyperus leptocarpus (F.Muell.) Bauters Australia: Wilson et al. 3383 (K)/ K: Y12991 JX644797. JX644895. Muasya 1217/ Jung & Choi (2013) 1 1 Cyperus mundii (Nees) Kunth Thailand: Muasya 1464 (K) AF449517 AF449566/ - AF449578 Cyperus nuerensis Boeckeler Tanzania: Muasya 940 (EA, K) Y13004 AY040603 - Cyperus papyrus L. Chad: Hepper 4213 (K) Y12966 AJ295759 AY242048 .1 Cyperus pedunculatus (R.Br.) J.Kern B - Tanzania: Faden et al. 96/48 (K)/ A AY040593 AY040604 KX306854

- LSU:Reid 8666 A A .1B Cyperus persquarrosus T.Koyama Kenya: Muasya 972 (EA, K) Y12990 AJ295762 - Cyperus proteus (Welw.) Bauters Congo: Fay 2700 (K) Y13002 - - Cyperus sanguinolentus Vahl A - Australia: Coveny et al. 17461 (K)/ AF449567 AF449579 AB261671 B - OKAY:Komagine & Masyo 17655 A A .1B Ficinia gracilis Schrad. A - S. Africa: Muasya 2355/ B - EF178589 EF178595 KM46232 Muasya 2742 A A 8.1B Ficinia paradoxa (Schrad.) Nees S. Africa: Verboom 534 (BOL) DQ058354 DQ058317 KF553447. 1 Ficinia spiralis (A.Rich.) Muasya & de Lange A - New Zealand: Ford 44/94 (NU)/ B AJ404701 AJ295753 DQ385562 - AK 283483 A A .1B Ficinia tristachya (Rottb.) Nees S. Africa: Muasya 2337 (BOL) EF200588 EF178593 AB685861 .1 Hellmuthia membranacea (Thunb.) R. W. Haines & S. Africa: Weerderman et al. 269 (K)/ Y13000 AJ295815 GU012384 Lye Muasya 1145 (K)/ Muasya 3081 .1 (BOL) Scirpoides burkei (C. B. Clarke) Goetgh., Muasya & A - S. Africa: Hargreaves 3361 (K)/ B Y13001A AJ295810 KM46229 D. A. Simpson - Muasya 3762 A 4.1B Scirpoides holoschoenus (L.) Soják S. Africa: Acocks s.n. (K) Y12994 AJ295811 KC677967 .1 Scirpoides holoschoenus subsp. thunbergii (Schrad.) A - S. Africa: Muasya 1205 (K)/ B - AJ404727 AJ295812 KM46229 Soják Muasya 4569 A A 6.1B Tribe Dulicheae Rchb. ex J. Schultze-Motel Blysmus compressus (L.) Panz. Afghanistan: Dobson 221 (K) AJ404700 AJ295766 - Dulichium arundinaceum (L.) Britton A - USA: Goetghebeur 9914 (GENT)/ AY725943 AF285067 DQ998949 B - Waterway, M.J. (unpubl.) A A .1B Tribe Eleocharideae Goetgh. Eleocharis atropurpurea (Retz.) J.Presl & C.Presl Kenya: Muasya et al. 752 (EA, K)/ - / Y13012 - AB180694 Japan – Hatooka et al. 14426 (OKAY) .1 Eleocharis gracilis R.Br. Australia: Wilson et al. 9462 (K)/ - EF178563 - DQ385565 /AK 284498 .1

Eleocharis marginulata Hochst. ex Steud. Kenya: Muasya 1039 (EA, K) Y13011 AJ295768 AY242056 .1 Eleocharis quinqueflora (Hartmann) O.Schwarz USA: Mastrogiuseppe 7461 (WS)/ G. U49229 GQ244896 AF190606. K. Helmkamp s.n. (RSA) .1 1 Eleocharis uniflora O.Seberg Chile: Grau 1433 (GENT) EF178562 - - Tribe Fuireneae Reichenb. ex Fenzl Actinoscirpus grossus (L. f.) Goetgh. & D.A. A - Malaysia: Simpson 2660 (K)/ B - Y12953A AJ295765 AB261672 Simpson Japan: Katsuyama et al 19915 A .1B (OKAY) Bolboschoenus caldwellii (V.J. Cook) Soják A - Australia: Wilson et al. 9530 (K)/ EF178564 AY506698 AY506752 B - K. Ghamkhar A1 (NSW) A .1B .1B Bolboschoenus maritimus (L.) Palla A - Botswana: Smith 2452 (K)/ B - Y12996A JF313191. JF313179. Korea:C.Kim s.n. 1B 1B Bolboschoenus nobilis (Ridl.) Goetgh. & D.A. S. Africa: Leistner 144 (K) Y12995 - - Simpson Fuirena abnormalis C. B. Clarke Tanzania: Faden et al. 96/118 (K) EF178565 - - Fuirena breviseta (Coville) Coville A - Neubig 23-2003 (FLAS)/ B - USA: KX397802 KC678035 KC677981 Shiels 37 (CMC) .1A .1B .1B Fuirena ciliaris (L.) Roxb. Tanzania: Muasya 951 (EA, K)/ - Y12971 - JX644891. /Jung & Choi (2013) 1 Fuirena coerulescens Steud. S. Africa: Muasya 2322 (K) EF178566 - - Fuirena hirsuta (P.J. Bergius) P.L. Forbes A - S. Africa: Muasya 2324 (K)/ B - EF178567 KM46253 KM46226 Muasya 5637 A 0.1B 2.1B Fuirena umbellata Rottb. A - BioBot05964 (iBOL)/ B - Jung & JQ591247. JX644792. JX644892. Choi (2013) 1A 1B 1B Fuirena welwitschii Ridl. Kenya: Muasya 1024 (EA, K) Y12993 EF178605 - Schoenoplectiella articulata (L.) Lye Tanzania: Muasya 947 (EA, K) Y12987 - JX566742. 1 Schoenoplectiella juncea (Willd.) Lye Kenya: Muasya et al. 775 (K) Y12952 - - Schoenoplectiella mucronata (L.) J.Jung & H.K.Choi Thailand: Muasya et al. 1290 (K)/ EF178572 JF313200. KC677979 Japan: J.Jung 908034 /USA: Rothrock 1 .1

4253 (CMC) Schoenoplectiella senegalensis (Steud.) Lye Kenya: Muasya et al. 2440 (EA) EF178568 EF178606 - Schoenoplectus confusus (N. E. Br.) Lye Kenya: Muasya et al. 2438 (EA) EF178569 - - Schoenoplectus corymbosus (Roth ex Roem. & Kenya: Muasya 1004 (EA) EF178570 EF178607 JX566743. Schult.) J. Raynal 1 Schoenoplectus lacustris (L.) Palla A - Britain: Muasya 1043 (K)/ B - U. Y12943A AJ295809 KC677958 K. no collector information (K) A .1B Schoenoplectus litoralis (Schrad.) Palla A - Hong Kong: Shaw 883 (K)/ B - EF178571 AY506697 AY506753 K.L. Clarke 26 (NE) A .1B .1B Tribe Rhynchosporeae Nees Pleurostachys sp. Brazil: Kallunki et al. 513 (NY) Y12989 - - Rhynchospora alba (L.) Vahl A - Simpson et al. (2003)/ AY344174 KX659156 KX166668 Buddenhagen et al (2017)/NMW3143 .1 .1 Rhynchospora rugosa subsp. brownii (Roem. & S. Africa: Verboom 616 (BOL) DQ058353 DQ058316 KF553455. Schult.) T.Koyama 1 Rhynchospora fascicularis (Michx.) Vahl Plunkett et al. (1995) U49233 - - Rhynchospora nervosa (Vahl.) Boeckeler Brazil: Kallunki et al. 512 (NY)/ Y12977 KX659166 AY242050 Buddenhagen et al (2017)/ Kallunki et .1 .1 al. 512 (NY) Tribo Schoeneae Dumort. Capeobolus brevicaulis (C. B. Clarke) J. Browning S. Africa: Verboom 646 (BOL) DQ058343 DQ058303 KF553443. 1 Carpha alpina R. Br. Wardle et al. (2001); Zhang et al. AF307909 AY230012 DQ385557 (2004)/ AK289109 .1 Carpha glomerata (Thunb.) Nees. S. Africa: Muasya 1176 (K)/ - / AY725941 - KF553444. Muasya 5863 (BOL) / 1 AM99979 7.1 Caustis dioica R. Br. Australia: Chase 2225 (K) Y12976 KF553498. - 1

Cladium mariscus (L.) Pohl Locality unknown: MJC 292 (K) DQ058338 DQ058298 - Cladium mariscus subsp. jamaicense (Crantz) Kük. Brazil: Thomas et al. 10403 (NY) Y12988 - - Costularia arundinacea (Sol. ex Vahl) Kük. Zhang et al. (2004) - AY230036 - Costularia fragilis (Däniker) Kük. New Calendonia: McKee NSW41617 EU828589 - - (K) Costularia nervosa J.Raynal Zhang et al. (2004) - AY230032 - Costularia pubescens J.Raynal Zhang et al. (2004) - AY230037 - Cyathochaeta diandra (R. Br.) Nees Zhang et al. (2004) - AY230042 - Cyathocoma bachmannii (Kük.) C. Archer S. Africa: Browning 835 (GENT) EF200590 EF178604 - Cyathocoma hexandra (Nees) J. Browning S. Africa: Verboom 648, BOL) DQ058344 DQ058304 - Evandra aristata R. Br. A - Australia: Wilson et al. 8974 AY725944 KF553508. KX963553 (NSW)/B - PERTH:Barrett 5356 A 1B .1B Gahnia baniensis Benl. Malaysia: Simpson 2737 (K) DQ058342 DQ058302 - Gahnia deusta (R. Br.) Benth. Australia: Alcock 11198 (WS) U49231 - - Gymnoschoenus sphaerocephalus (R. Br.) Hook. f. Australia: Wilson et al. 9463 (K)/ AY725945 AY230033 - Zhang et al. (2004) Lepidosperma tortuosum F. Muell. Australia: Coveny et al. 17470 (K)/ AY725950 - KF553451. PERTH:Bruhl 2357 1 Machaerina mariscoides (Gaudich.) Kern Tahiti: Sachet 2636 (GENT) EF178575 - - Machaerina juncea (R. Br.) T.Koyama Verboom 1229 (BOL) KF553610. 1 Machaerina rubiginosa (Biehler) T.Koyama Australia: Wilson et al. 9471 (K)/ AY725940 KF553517. AB261679 PERTH:Bruhl 1859/ Hoshino et al. 1 .1 19254 (OKAY) Machaerina sp. New Guinea: Johns 9195 (K) DQ058340 DQ058300 - Mesomelaena pseudostygia (Kük.) K. L. Wilson Australia: Chase 2226 (K) Y12959 DQ058301 - Mesomelaena tetragona (R. Br.) Benth. Australia: Chase 2227 (K) Y12949 KF553518. - 1 Morelotia gahniiformis Gaudich. Hawaii: Herbst 1167 (GENT)/ EF178576 KF553519. KF553452. PERTH:Morden s.n./ PERTH:Morden 1 1 2117

Neesenbeckia punctoria (Vahl) Levyns A - S. Africa: Muasya 1214 (K)/ B - AY725952 DQ058306 KF553453. PERTH:Bruhl 1731 A A 1B Oreobolus kuekenthalii Steenis ex Kük. Malaysia: Simpson 2659 (K) Y12972 EF178536 AY242047 .1 Oreobolus obtusangulus Gaudich. Wardle et al. (2001)/ - /Argentina, AF307926 - DQ450472 Moore 2817 (K) .1 Oreobolus oligocephalus W. M. Curtis Zhang et al. (2004) - DQ456963 DQ450473 .1 .1 Oreobolus pectinatus Hook. f. Wardle et al. (2001)/ New Zealand: AF307927 DQ456965 DQ385589 Menville & Connor 6528/ AK289108 .1 .1 Ptilothrix deusta (R. Br.) K. L. Wilson Coveny R.G. 17471 (K)/ Zhang et al. FN870912. AY230041 KF553454. (2004)/ PERTH:Bruhl 2055 1 .1 1 Schoenus compactus (Levyns) T.L.Elliott & Muasya S. Africa: Verboom 614 (BOL) DQ058351 DQ058313 - Schoenus compar L. S. Africa: Verboom 549 (BOL) DQ058350 DQ058312 - Schoenus crassus (Levyns) T.L.Elliott & Muasya S. Africa: Verboom 507 (BOL) DQ058352 DQ058314 - Schoenus nigricans L. Saudi A - Arabia: Edmondson 3382 (K)/ B - Y12983A AJ295814 KF553460. Haase et al. s.n. (K) A 1B Schoenus quadrangularis Boeckeler S. Africa: Verboom 636 (BOL) DQ058349 DQ058311 - Tetraria crinifolia (Nees) C. B. Clarke S. Africa: Verboom 638 (BOL) DQ058348 DQ058309 - Tetraria microstachys (Vahl) Pfeiffer S. Africa: Verboom 640 (BOL) DQ058347 DQ058307 - Trianoptiles capensis (Steud.) Harv. S. Africa: Muasya 3160 (BOL) - KF553532. KF553463. 1 1 Trianoptiles solitaria (C. B. Clarke) Levyns Zhang et al. (2004) - AY230028 - Tricostularia pauciflora (R. Br.) Benth. Australia: Coveny et al. 17484 (K)/ AY725954 AY230038 - Zhang et al. (2004) Tribe Scirpeae Kunth ex Dumort. Amphiscirpus nevadensis (S. Watson) Oteng-Yeboa Argentina: Charpin et al. 20572 DQ317926 DQ317925 AF190618. (GENT)/ G. Helmkamp s.n. (RSA) .1 1 Eriophorum angustifolium Honckney M.Kuzmina, K.Johnson JN965529. AY344177 DQ998950 (BIO)/Simpson et al. (2003)/ 1 .1 .1 Waterway (unpubl.)

Eriophorum vaginatum L. Poland: Beyer et al. 2 (K) Y12951 AJ295769 JX566737. 1 Eriophorum viridicarinatum (Engelm.) Fernald A - USA: Boufford 23053 (WS)/ B - U49230A KR902947 KR902925 MTMG:M.J. Waterway 2001/ .1B .1B Oreobolopsis clementis (M. E. Jones) Dhooge & Dhooge (2005) AJ811011 - - Goetgh. Oreobolopsis inversa Dhooge & Goetgh. Ecuador: Laegaard 21492 (GENT) AJ811009 DQ317923 - Oreobolopsis tepalifera T. Koyama & Guagl. Dhooge et al. (2003) AJ575932 AJ576035 - Phylloscirpus acaulis (Phil.) Goetgh. & D. A. Dhooge et al. (2003) AJ575926 AJ576029 - Simpson Phylloscirpus boliviensis (Barros) Dhooge & Goetgh. Ecuador: Laegaard 102805 (GENT) AJ566081 AJ566082 - Phylloscirpus deserticola (Phil.) Dhooge & Goetgh. Ecuador: Laegaard et al. 21478 AJ704785 AJ704786 - (GENT) Scirpus flaccidifolius (Fernald) Schuyler Nacsi 9774 (DOV) EF178582. - - 1 Scirpus polystachyus F. Muell. A - Australia: Pullen 4091 (K)/ B - Y12974A AJ295813 AH012953 Wilson s.n. (MWC 5927), (K) A .2B Trichophorum alpinum (L.) Pers. A - Canada: Waterway 2002.95 AJ810999 DQ317924 AY757432 (GENT)/ B - Canada:Waterway A A .1B 2001.110 (MTMG) Trichophorum cespitosum (L.) Hartm. British Isles: Nelmes 954 (K)/ Y12969 DQ999004 AB180721 Waterway (unpubl.)/ Japan: Hosoi .1 .1 16191 (OKAY) Trichophorum clintonii (A.Gray) S.G.Sm. Canada: Baldwin 4856 (K) Y12982 - - Trichophorum planifolium (Spreng.) Palla Dhooge (unpub.) AJ811001. - - 1 Trichophorum rigidum (Steud.) Goetgh., Muasya & Argentina: Renvoize et al. 5021 (K) AJ297509 AJ295808 - D. A. Simpson subsp. rigidum Zameioscirpus atacamensis (Phil.) Dhooge & Bolivia: Ruthsatz & Budde 10328 AJ575929 AJ576032 - Goetgh. (Trier) Zameioscirpus gaimardioides (E. Desv.) Dhooge & Argentina: Ruthsatz 9212 (GENT); AJ575938 AJ576031 - Goetgh. Dhooge et al. (2003)

Zameioscirpus muticus Dhooge & Goetgh. Dhooge et al. (2003) AJ575927 AJ576030 - Tribe Sclerieae Kunth ex Fenzl Scleria distans Poir. Kenya: Muasya 1023 (EA, K) Y12968 DQ058299 KF553462. 1 Scleria foliosa Hochst. ex A.Rich. Tanzania: Muasya 939 (EA, K)/ Y12986 - LN886855 Malaisse F. & Goetghebeur P. 418 .1 (GENT) Scleria rugosa R.Br. Jung & Choi (2013) JX644705. JX644811. JX644899. 1 1 1 Scleria terrestris (L.) Fassett Malaysia: Simpson 2658 (K)/ Y12947 - LN886822 Goetghebeur P. 12129 (GENT) .1 Tribe Trilepideae Goetgh. Coleochloa abyssinica (Hochst. ex A.Rich.) Gilly Ethiopia: Vollesen 80/2 (K) Y12975 - - Microdracoides squamosa Hua Bonn Acc. 150 AY725951 - - Trilepis lhotzkiana Nees Bonn Acc. s.n. AY725955 - - Uncertain tribe aff. Cariceae Khaosokia caricoides D.A.Simpson, Chayam. & Thailand: Simpson et al. 1886 (K) AY725948 EF178535 - J.Parn.

TABLE 3. Molecular markers, primers and protocols applied during PCR experiments Region Primers (and its combination) Protocols (temperature; time) trnL- c –CGA AAT CGG TAG ACG CTA Premelt – 95°C; 2’ trnL - CG Denature – 95°C; 30” trnF d – GGG GAT AGA GGG ACT TGA Anneal – 55°C; 1’ AC Extension – 72°C; 1’ e – GGT TCA AGT CCC TCT ATC CC Final extension – 72°C; 7’ f – ATT GAA CTG GTG ACA CGA G Number of cycles – 35

Additives: (none)

[c-f or c-d/e-f] rbcL 1F – ATG TCA CCA CAA ACA GAA Premelt – 95°C; 2’ ACT AAA GC Denature – 95°C; 30” 636F – TCC TTT TAG TAA AAG ATT Anneal – 55-60°C; 1’ GGG CCG AG Extension – 72°C; 1’ 674R – GAT TTC GCC TGT TTC GGC Final extension – 72°C; 7’ TTG TGC TTT ATA AA Number of cycles – 35 1460R – TCC TTT TAG TAA AAG ATT GGG CCG AG

[1F-674R and 636F-1460R] Additives: (none) ITS 5i – AGG TGA CCT GCG GAA GGA Premelt – 95°C; 2’ TCA TT Denature – 95°C; 30”/95°C; 4i – GGG TAG TTC CCG CCT GAC 30”/95°C; 30” CTG G Anneal – 56°C; 30”/53°C; 30”/50°C; 30” Extension – 72°C; 1’15”/72°C; 1’15”/72°C; 1’15” Final extension – 72°C; 7’ Number of cycles – 4/4/35 (“touchdown”) [5i-4i] Additives: DMSO 5% and betaine 2M Observations: we performed “nested” PCR experiments on some samples due to the low concentration of DNA after the first PCR experiment, mainly in PCR products from herbarium specimens.

FIGURE 2. Phylogenetic hypothesis based on rbcL + trnL-F + ITS sequences to Cyperaceae. [Consensus tree of PP=0.5 branches; black – Thurniaceae + Juncaceae ; purple – Mapanioideae; pink– Trilepideae; red – Schoeneae; green – Sclerieae + Bisboeckelereae; orange – Cryptangieae; dark blue–Rhynchosporeae; light blue – other Cyperaceae tribes; full circle – PP = 1; empty circle – PP ≥ 0.95; full star– BS ≥ 90%; empty star – 75%≤ BS<90% . We describe PP and BS values to the mainly discussed branches]

FIGURE 3. Summary of phylogenetic reconstruction of Cyperaceae based on rbcL, trnL-F and ITS matrices. [Consensus tree of PP=0.5 branches; black – Thurniaceae + Juncaceae; purple – Mapanioideae; pink– Trilepideae; red– Schoeneae; green – Sclerieae + Bisboeckelereae; orange – Cryptangieae; dark blue –Rhynchosporeae; light blue – other Cyperaceae tribes; full circle – PP = 1; empty circle – PP ≥ 0.95. We include PP values to the mainly discussed branches]

CHAPTER 2 – “The more the merrier”: a molecular and morphological approach to the Cryptangieae (Cyperaceae) genera circumscription

Additional co-authors: Rosemeri Morokawa, Volker Bittrich, Muthama Muasya, Fabio A. Vitta & George J. Shepherd

(Intended submission to the Journal Taxon)

(Short title: What about Cryptangieae II)

Abstract: Cryptangieae was asserted as a monophyletic tribe in a recent molecular hypothesis that also raised questions about the circumscription of its genera. We performed additional sampling and provide new analyses using single and combined matrices from two chloroplast (rbcL and trnL-F) and three nuclear ribosomal (ITS, ETS and 5S-NTS) regions with Maximum Parsimony and Bayesian Inference to clarify the genera limits and relationships, including about 74% of Cryptangieae species. We also performed ancestral character state analyses, with the diagnostic morphological characters traditionally applied in Cryptangieae taxonomy, using parsimony reconstruction methods. The molecular analyses show Lagenocarpus in the current circumscription as polyphyletic, and Cephalocarpus as paraphyletic without the inclusion of Everardia. We propose an updated circumscription following the phylogeny and discuss the phylogenetic significance of the morphological characters in Cryptangieae taxonomy. Additionally, we provide an identification key, a brief description of genera under the new proposal and a list of accepted species. Keywords “Acrocarpus”; Cryptangium; Dydimiandrum; Exochogyne; Neotropics

Resumo: Cryptangieae foi reafirmada como uma tribo monofilética emu ma hipótese molecular recente que concomitantemente levantou questionamentos quando a circunscrição de seus gêneros. A amostragem foi ampliada e novas analises são fornecidas utilizando matrizes de marcador-único e concatenada para duas regiões de cloroplasto (rbcL e trnL-F) e três regiões nuclear ribossomais (ITS, ETS e 5S-NTS) utilizando Máxima Parcimônia e Inferência Bayesiana para esclarecer os limites e relações entre os gêneros, incluindo cerca de 74% das espécies de Cryptangieae. Também foram realizadas analyses de estado ancestral de carater, com os caracteres morfológicos tradicionalmente aplicados na taxonomia de Cryptangieae, utilizando métodos de reconstrução parcimoniosa. As analyses moleculares demonstram Lagenocarpus na circunscrição atual como polifilético, e Cephalocarpus como parafilético sem a inclusão de Everardia. É proposta uma nova circunscrição seguindo a hipótese filogenética e discutido o significado taxonômico dos caracteres morfológicos na taxonomia de Cryptangieae. Adicionalmente, são fornecidas chaves de identificação, breves descrições dos gêneros como aqui propostos e uma lista de espécies aceitas.

Introduction Cryptangieae is a Neotropical tribe mainly distinguished from other Cyperaceae tribes by the unisexual flowers and spikelets, the spirally arranged glumes, the common presence of three fimbriate hypogynous scales at the base of its trigonous, rarely biconvex, achenes (Koyama & Maguire 1965; Goetghebeur 1998; Koyama 2003, 2004; Muasya et al. 2009; Costa et al. – CHAPTER 1); and the presence of conspicuous reddish-pink stigmas in almost all species/populations. Its representatives occur from Cuba to southern Brazil and the tribe is especially diverse on the Guiana Shield and at the campos rupestres, in southeastern Brazil (Goetghebeur 1998; Koyama 2003, 2004; Vitta 2005 – PhD thesis).

Cryptangieae taxa have been described and rearranged since the “Synopsis Generum Cyperacearum” (Nees von Esenbeck 1834), which included the description of Lagenocarpus (with the only species then known: L. guianensis Nees). The genus was first classified in the tribe Sclerieae and then, Cryptangieae was described in Bentham (1886) with Lagenocarpus as typical genus. The main characters used to circumscribe its genera are (Gilly 1941a, 1941b, 1942, 1951; Koyama & Maguire 1965; Koyama 2003, 2004; Vitta 2005 – PhD thesis): breeding system, caudex development (a more or less lignified vertical stem covered by sheaths remains and adventitious roots), inflorescence general morphology, and various achene characters (shape in cross section, hypogynous scales development, ornamantation, achene beak shape). These diagnostic characters were enough to distinguish the genera, but we found some inconsistencies in a closer look and some species have been frequently moved from one genus to another, depending on the taxonomist morphological interpretation and the weight given to each character.

The morphology-based works of Koyama (2003, 2004) about Cryptangieae representatives are the broadest and the most comprehensive due to the number of species and specimens included and his generic circumscription is still in use. The author recognizes three genera in Cryptangieae: Cephalocarpus Nees (four species), Everardia Ridley (11 spp.) and Lagenocarpus Nees (26 spp.). Koyama (2004) also proposed an infrageneric classification for the largest genus, Lagenocarpus, with the following sections: Lagenocarpus section Lagenocarpus, with L. guianensis Nees as the type species; L. sect. Imberbitae H. Pfeiffer, with L. albo-niger (A. St.-Hil.) C.B.Clarke as type-species; L. sect. Corymbosae H. Pfeiffer1, type-species: L. verticillatus (Sprengel) T. Koyama & Maguire; L. sect. Exochogyne (C.B.Clarke) T.Koyama2, with L. amazonicus (C.B.Clarke) H. Pfeiffer as type-species; L. sect. Acrocarpus (Nees) T.Koyama, type-species: L. polyphyllus (Nees)

O.Kuntze3; L. sect. Junciformes T. Koyama3, type species: L. junciformis (Kunth) O.Kuntze; L. sect. Didymiandrum (Gilly) T.Koyama4, with L. stellatus (Boeckeler) O.Kuntze as type-species. These sections are primarily based on distribution patterns, plant indument, inflorescence structure and achene (and beak) characters and sometimes are directly equivalent to previously accepted genera (1 – Cryptangium Schrader ex Nees, 2 – Exochogyne C.B.Clarke, 3 – Acrocarpus Nees, nom.illeg., 4 – Didymiandrum Gilly) (Koyama 2004).

Another important work worth is Fabio Vitta’s PhD thesis of 2005, with his own morphological studies and a new circumscription to Lagenocarpus and Cryptangium. He analyzed more specimens than Koyama (2004), but his works remains unpublished. Afterwards, one species of Everardia and another of Cephalocarpus were described (Strong 2016).

Until recently Cryptangieae were neglected in molecular phylogenies of Cyperaceae and remained a gap in the knowledge of infrafamilial relationships and tribe circumscriptions (Simpson et al. 2007; Muasya et al. 2009; Hinchliff & Roalson 2013). The phylogenetic hypothesis of Costa et al. (2018 - CHAPTER 1) reconstructed the tribe as monophyletic, but a broader sampling, including more species and more DNA regions, is necessary to clarify genera limits. Also, to avoid misinterpretations that may yield circumscriptions of unnatural taxa, the morphological data should be analyzed concomitantly with molecular phylogenies, as we aim at in this manuscript.

Here, we propose a re-circumscription of the genera of tribe Cryptangieae based on a molecular phylogeny and discuss the use of diagnostic morphological characters in Cryptangieae taxonomy. We also provide taxonomical material to their recognition as well as a list of accepted species.

Material and Methods Taxonomic sampling.– We made expeditions to localities with great or significant diversity of Cryptangieae species and collected specimens as vouchers, as well as silica-dried and alcohol samples (specimens deposited at herbarium UEC – state University of Campinas, Brazil). We analyzed Cryptangieae specimens and type collections at herbaria in Brazil (ASE, MIRR, MG, IAN, INPA, SPF, UFG, UFRR) and in the U.S.A. (F, MO, NY and US), and images of additional specimens at JStor Global Plants (plants.jstor.org/), and Species

Link (http://splink.cria.org.br/).

Molecular sampling.– We chose two chloroplast (rbcL and trnL-F) and three nuclear ribosomal (ITS, ETS and 5S-NTS) regions as they are relatively easy to amplify and with enough potential variation at the investigated taxonomical level. The outgroup includes sequences of Trilepis (Trilepideae), Diplacrum (Bisboeckelereae) and Scleria (Sclerieae), one terminal each; the Trilepis sp. terminal is a chimera of two different species (T. lhotzkiana at rbcL-matrix and T. ciliatifolia at ETS-matrix) to avoid an abnormal topology due to the data gaps. The ingroup includes 35 species of Cryptangieae, which covers about 75 % of its known specific diversity. (Table 4)

Molecular procedures.– We extracted total DNA from silica-dried and herbarium material using NucleoSpin® Plant II – Macherey-Nagel and DNeasy® Plant Mini kit – Qiagen for plants. We amplified the targeted regions trough modified PCR protocols based on Muasya et al. (2002), Roalson & Friar (2004) and Cox et al. (1992) (Table 5). We purified the PCR products using a mix of Exonuclease I (10u/µL) and Shrimp Alkaline Phosphatase (rSAP - 1u/µL) (1:2) and sequenced the samples at the Labotatório Multiusuário de Genotipagem e Sequenciamento (CBMEG/Unicamp) using a ABI 3500xL da Life Technologies sequencer and deposited the sequences at GenBank.

Phylogenetic analyses.– We reconstructed the phylogenetic hypotheses using the Maximum Parsimony (MP) and the Bayesian Inference (BI) approaches for single-marker and combined matrices. We aligned the sequences using MAFFT (Katoh et al. 2002) and made some manual adjustments with Mesquite 3.04 (Maddison & Maddison 2007). We elaborated the MP hypothesis using TNT 1.5 (Goloboff et al. 2008, 2016): 1.. A heuristic search with 1000 RASR holding 10 trees at each step, TBR branching swapping with MULT REES and steepest descent in effect saving max. 50 trees/ replicate; 2. Another heuristic search including retained trees, and finally; 3. A Bootstrap Search (BS) with 200 (single matrices) or 700 (combined matrix) replicates, 100 RASR with three trees held per step, TBR branch swapping with steepest descent and MULT REES in effect, saving 10 trees at each RASR. We considered those clades having a Bootstrap Support values (BS) ≥ 75% as supported.

perform the MCMC analysis, with the following settings: nchains = 4; nruns = 2; ngen = 107 (to single matrices) / 108(combined matrix); burnin = 0.25; samplefreq = 1,000. We considered those clades as supported when the Posterior Probability (PP) ≥ 0.95. Ancestral Character Reconstruction.– We selected 13 diagnostic characters used for Cryptangieae taxa in previous works and our own studies (Table 6, a–m). We recovered the maximum credibility tree of the BI combined hypothesis using the TreeAnnotator on XSEDE (1.8.4) available at the CIPRES platform (burnin = 0.25). We constructed the character matrix (Appendix 1) based on the analyses of herbaria specimens and performed the Ancestral Character Reconstruction using Mesquite 3.04 (Maddison & Maddison 2007), with parsimony reconstruction methods. We present a phylogeny with summarized character status mapping and the individual reconstructions are presented as supplementary material. The character “n” represents the distributional area of each species plotted to facilitate the discussion, but wasn’t included in the analysis.

Results and Discussion Phylogenetic findings.– We performed the analyses with 137 sequences of 37 species of Cryptangieae from all three currently accepted genera, and 8 sequences as outgroups (Table 4). The aligned matrix of each molecular marker presents the following lengths: 1541 bp for the rbcL matrix, 1017 bp for the trnL-F matrix, 651 bp for the ITS matrix; 668 bp for the ETS matrix, and 571 bp for the 5S-NTS matrix. Each marker shows the following values of constant characters (cc), uninformative characters (uninfc) and informative characters (infc): 1. rbcL – cc = 1324, uninfc = 119, infc = 96 (~0.06 %); 2. trnL-F – cc = 676, uninfc = 255, infc = 86 (~0.08%); 3. ITS – cc = 394, uninfc = 142, infc = 115 (~0.17%); 4. ETS – cc = 250, uninfc = 244,infc = 174 (~0.26%), and; 5. 5S-NTS – cc =327, uninfc = 91, infc = 153 (0.27%).

In the Maximum Parsimony analyses, Cryptangieae was reconstructed in all single-marker matrices analyses with Bootstrap support (BS) = 99--100% and in the combined matrix with BS = 77% (Figures 4, Appendix 2). At the rbcL-only analysis just two groups of Cryptangieae had BS ≥ 50%, first, Lagenocarpus sect. Exochogyne (BS = 56%) and second including L. sect. Acrocarpus and L. sect. Junciformes species (BS = 77%). The trnL- F-only analysis recovered Everardia + Cephalocarpus with BS = 100% and Lagenocarpus sect. Lagenocarpus + L. sect. Imberbitae with BS = 84%. The ITS-only analysis recovered the

L. sect. Acrocarpus + L. sect. Junciformes clade (BS = 100%), the Cephalocarpus + Everardia group (BS = 71%) and a Lagenocarpus s.s. clade (BS = 93%). The ETS-only analysis recovered the following BS values to the clades: BS = 99% for the L. sect. Junciformes; BS = 98% for the Cephalocarpus + Everardia clade; and BS = 100% for the L.sect. Lagenocarpus + L. sect. Imberbitae clade. The 5S-NTS-based analysis supports the following clades: Cephalocarpus + Everardia (BS = 90%) and L. sect. Acrocarpus + L. sect. Junciformes (except L. triqueter (Boeckeler) O.Kuntze) (BS = 94%).We can recognize the following groups with the combined matrix MP analysis (Figure 5): 1. Lagenocarpus sect. Exochogyne (BS = 55%); 2. L. sect. Acrocarpus + L. sect.Junciformes (BS = 90%); 3. Cephalocarpus + Everardia (BS = 69%); L. sect. Lagenocarpus + L. sect. Imberbita (BS=98%). Lagenocarpus sect. Corymbosae and L. sect. Didymiandrum species do not cluster with any other group.

The best evolution models according to AICc criterion to each marker were: rbcL – HKY+I+G; trnL-F – TIM2+G; ITS – TIM3+G; ETS – GTR+G, and; 5S-NTS – GTR+G. Cryptangieae is supported with PP = 1 at all single-marker and combined matrices analyses, except for the trnL-F analysis (Figure 4, Appendix 2).

The rbcL dataset analysis recovered the following clades with Posterior Probability (PP) = 0.99--1: Lagenocarpus sect. Exochogyne, L. sect. Acrocarpus + L. sect. Junciformes, and Everardia. Species of Cephalocarpus and L.sect. Lagenocarpus + L. sect. Imberbitae form a group with PP = 0.94 and L. sect. Lagenocarpus + L. sect. Imberbitae shows PP = 0.83. Lagenocarpus sect. Corymbosae and L. sect. Didymiandrum again do not cluster with any other group. In the analysis based on the trnL-F matrix, the only reconstructed clade was Cephalocarpus+Everardia.

The ITS-only analysis recovered Lagenocarpus sect. Acrocarpus + L. sect. Junciformes and and L. sect. Lagenocarpus + L. sect. Imberbitae (both with PP = 1). Cephalocarpus + Everardia with PP = 0.96 and L. sect. Corymbosae does not cluster with any other group. The ETS BI analysis recovered the Lagenocarpus sect. Junciformes clade and Cephalocarpus + Everardia (both with PP = 1) and it shows a group of L.sect. Lagenocarpus + L. sect. Imberbitae with PP = 0.73. Lagenocarpus sect. Corymbosae and L. sect. Exochogyne do not cluster with any other group. The 5S-NTS BI analysis shows the lowest PP values for the above-mentioned groups: L. sect. Acrocarpus + L. sect. Junciformes (PP = 0.89), Cephalocarpus + Everardia (PP = 0.76), a species rich group with L. sect.

Lagenocarpus + L. sect. Imberbitae species (PP = 0.93), and a clade with three species of L. sect. Lagenocarpus + L. sect. Imberbitae (PP = 0.99). Lagenocarpus sect. Corymbosae, L. sect. Exochogyne, L. sect. Didymiandrum and L. celiae T. Koyama & Maguire and L. sect. Imberbitae do not cluster with any other group.

Finally, the combined matrix BI analysis recovered the following clades with PP = 1 (Figure 1): Lagenocarpus sect. Acrocarpus + L. sect. Junciformes, Everardia + Cephalocarpus and L. sect. Lagenocarpus + L. sect. Imberbitae; and a group with L. sect. Exochogyne with PP = 0.94. Lagenocarpus sect. Corymbosae, L. sect. Didymiandrum do not group with any other clade. We mainly focus our discussion at the combined matrix BI analysis with plotted BS values at important reconstructed clades (Figure 4).

The Maximum Parsimony analyses show clades with weaker support than the Bayesian Inference analyses, a statistical frequent and expected outcome. The ITS and ETS single-marker analyses were the clearest to show the relationships among taxa; the remaining markers were less helpful at the aimed taxonomic level, due to the lower (rbcL and trnL-F) or higher (5S-NTS) variability at each region in Cryptangieae taxa. The combined analyses are more reliable as they gather at least three independent sources of characters, and also show supported clades that can be supported as morphologically recognizable genera.

In our analyses, we can recognize six major groups in the Cryptangieae even though relationships are sometimes unclear: A. Lagenocarpus sect. Corimbosae (= Cryptangium Schrader ex Nees); B. L. sect. Didymiandrum (= Didymiandrum Gilly); C. L. sect. Exochogyne (= Exochogyne C.B.Clarke); D. Everardia + Cephalocarpus; E. L. sect. Acrocarpus + L. sect. Junciformes (= Acrocarpus Nees, illegitimate name), and; F. L. sect. Lagenocarpus + L. sect. Imberbitae. The first three lineages contain only one accepted species each which do not group among them or with any other clades in the analyses. Cephalocarpus is frequently inserted in (or grouped with) Everardia, except for the BI rbcL-only analysis. Lagenocarpus sect. Acrocarpus + L. sect. Junciformes is supported in the analyses with enough resolution using our dataset; this also happens mostly with L. sect. Lagenocarpus + L. sect. Imberbitae (Figure 4, Appendix 2).

Koyama (2003, 2004) morphological interpretation of Cryptangieae genera is not completely recognizable in our analyses. Especially about Lagenocarpus, Koyama (2004) does not describe a monophyletic genus and his subgeneric sections are questionable according to our data. Vitta (2005) proposed the restatement of Cryptangium, apart of

Lagenocarpus, but could not recover other blur relationships in the tribe, as the case of Lagenocarpus glomerulatus.

Therefore, we advocate classifying the tribe into six monophyletic and morphologically recognizable genera (Figures 4, 6): 1. Cephalocarpus Nees (including Everardia Ridley), 2. Cryptangium Schrader ex Nees, 3. Didymiandrum Gilly, 4. Exochogyne C.B.Clarke, 5. Krenakia S.M.Costa, and 6. Lagenocarpus Nees. Below we provide an identification key to the genera, a brief morphological description. We also provide a list of accepted species combining the molecular data and our own interpretation of morphology of the unsampled species. There is no detail on synonym; we will provide this information in future taxonomic treatments to the genera (Cephalocarpus, Costa et al. unpublished - Chapter 3 and 4).

Taxonomic results, Identification Key.– 1. Spike-like inflorescence; glumes 2-ranked; achene biconvex; hypogynous scales absent …...…………….…………………………….……….…..……….… Exochogyne C.B.Clarke 1’. Panicle-like or head-like inflorescences; achene trigonous; glumes spirally or sub- distichally arranged; hypogynous scales inconspicuous or conspicuous …………………. 2

2. Caudex elongate; inflorescence lateral; hypogynous scales with fimbriate margins …………………………………………………………………….…….. Cephalocarpus Nees 2’. Caudex absent or inconspicuous; inflorescence terminal; hypogynous scales glabrous, with sparse ciliate to ciliate margins or pubescent at margin and dorsal surface …………………. 3

3. Male spikelets with (1)4--5 stamens; achene sessile, obscurely trigonous (trisulcate or inconspicuously ridged) …...……………………..……………...………. Lagenocarpus Nees 3’. Male spikelets with 2(3) stamens; achene stipitate, trigonous with sharp ridges ………. 4

4. Dioecious herbs; involucral bracts (sterile or fertile) solitary or forming pseudo-whorls with elliptic to oblong-elliptic blades …….…………………..……………... Didymiandrum Gilly 4’. Monoecious herbs; involucral bracts (sterile or fertile) never forming pseudo-whorls ...... 5

5. Leaf with normally developed blade, rarely reduced to sheath; inflorescence with conspicuously whorled paracladia; female spikelets with 1(2) flowers and 3 glumes; achene with three conspicuous cavities at the base …………….……. Cryptangium Schrad. ex Nees 5’. Leaf always reduced to sheath; inflorescence branched or nearly unbranched but not with whorled paracladia; female spikelets with 1--5 flowers and 3--7 glumes; achene cavities absent or inconspicuous ………………………………………….…… Krenakia S.M.Costa

Taxonomic results, Genera description, accepted species and new combinations.– Cephalocarpus Nees, Flora brasiliensis (Martius) 2(1): 162, pl.18. 1842. Type species: Cephalocarpus dracaenula Nees Everardia Ridley, Timehri 5: 210. 1886. Lagenocarpus subgen Cephalocarpus (Nees) H. Pfeiff., Repertorium Specierum Novarum Regni Vegetabilis 18: 91. 1922. Monoecious, rarely dioecious herbs, with elongate caudex: male and female spikelets separate or mixed in the paracladia of monoecious species; female spikelets most frequently at a terminal position. Leaf blades developed; linear to lanceolate. Inflorescence lateral, solitary or many at a prophyll, head or panicle-like. Involucral bracts leaf-like, with linear- lanceolate to lanceolate blades. Paracladia superficially monomorphic. Male spikelets 1--9 flowered; stamens 2--6 (7--9). Female spikelets 1-flowered, 4--7(9) glumes; stigmas 3 (rarely 2 or 4-fid). Glumes spirally or sub-distichally arranged. Achenes sessile or stipitate, obovoid, ellipsoid, oblong or ovoid, trigonous, generally trisulcate, glabrous or minutely to densely ciliate especially at the apex (including the beak), main body continuous to the beak or with a constriction zone between the beak and the main body; hypogynous scales well- developed, long or shortly fimbriate (rarely sparsely ciliate); beak long or short conical, glabrous, minute ciliate to densely ciliate, persistent or caducous.

Cephalocarpus occurs on the Guiana Shield and some border areas in northern South America, in Venezuela, Guyana, Colombia, Brazil, Peru, Ecuador and Suriname. Its species are more frequent and diverse at higher elevations on nutrient poor sandy or rocky soils. Total number of species: 18.

According to our phylogenetic analysis, Cephalocarpus (sensu Koyama 2003) should include Everardia (sensu Koyama 2003) to become monophyletic. Cephalocarpus was morphologically supported mainly by the head-like inflorescence and

the constricted region between the beak and achene main portion (Koyama & Maguire 1965; Koyama 2003). A careful look at the inflorescence reveals that it is just an extremely contracted version of the Everardia (sensu Koyama 2003) inflorescence. Six recently discovered species (Costa et al., unpublished data) reinforce our results: the new species present mixed inflorescence and achene character states of Everardia (sensu Koyama 2003) and Cephalocarpus (sensu Koyama 2003).

1. Cephalocarpus angustus (N.E. Brown) S.M. Costa, comb. nov. Basionym: Everardia angusta N.E. Brown, Trans. Linn. Soc. of London, 2nd series: Botany 6: 73. 1901. 2. Cephalocarpus confertus Gilly, Bull. Torrey Bot. Club 69 (4): 293-294, fig. 1a-c.1942. 3. Cephalocarpus debilis (T. Koyama & Maguire) S.M. Costa, comb. nov. Basionym: Everardia debilis T. Koyama & Maguire, Memoirs of the New York Botanical Garden 12: 32, f. 7, A–B. 1965. 4. Cephalocarpus diffusus (T. Koyama & Maguire) S.M. Costa, comb. nov. Basionym: Everardia diffusa T. Koyama & Maguire, Mem. New York Bot. Gard. 12: 21, f. 4 D– E. 1965. 5. Cephalocarpus distichous (T. Koyama & Maguire) S.M. Costa, comb.nov. Basionym: Everardia disticha T. Koyama & Maguire, Mem. New York Bot. Gard. 12: 25, f. 5 D – F. 1965. 6. Cephalocarpus dracaenula Nees, Flora brasiliensis (Martius) 2 (1): 162–163, plate 18. 1842. 7. Cephalocarpus duidae (Gilly) S.M. Costa, comb. nov. Basionym: Everardia duidae Gilly, Bull. Torrey Bot. Club 68: 30, f. 3 g–g’. 1941. 8. Cephalocarpus erecto-laxus (T. Koyama & Maguire) S.M. Costa, comb. nov. Basionym: Everardia erecto-laxa T. Koyama & Maguire, Mem. New York Bot. Gard. 12: 27, f. 5, H–J.1965. 9. Cephalocarpus flexifolium (Gilly) S.M. Costa, comb. nov. Basionym: Didymiandrum flexifolium Gilly, Bull. Torrey Bot. Club 68: 331. 1941. 10. Cephalocarpus longibracteatus Gilly, Brittonia 3: 153, f. 2d–j. 1939. 11. Cephalocarpus longifolius (Gilly) S.M. Costa, comb. nov. Basionym: Everardia longifolia Gilly, Brittonia 3(2): 153, f. 2g–h. 1939. 12. Cephalocarpus maguireanus (T. Koyama) S.M. Costa, comb. nov.

Basionym: Everardia maguireana T. Koyama, Mem. New York Bot. Gard. 18: 23. 1969. 13. Cephalocarpus montanus (Ridley) S.M. Costa, comb. nov. Basionym: Everardia montana Ridley, Timehri 5:210. 1886. 14. Cephalocarpus obovoideus T. Koyama, Mem. New York Bot. Gard. 18 (2):22-23, f.4 h-o. 1969. 15. Cephalocarpus ptariensis (Gilly) S.M. Costa, comb. nov. Basionym: Everardia ptariensis Gilly, Fieldiana Bot. 28(1): 55.1961. 16. Cephalocarpus recurviglumis (T. Koyama & Maguire) S.M. Costa, comb. nov. Basionym: Everardia recurvigluma T. Koyama & Maguire, Mem. New York Bot. Gard. 12: 23, f. 4, F–G. 1965. 17. Cephalocarpus rigidus Gilly ex Gleason & Killip, Brittonia 3: 152, f.2 a-c. 1939. 18. Cephalocarpus vareschii (Maguire) S.M. Costa, comb. nov. Basionym: Everardia vareschii Maguire, Acta Biologica Venezuelica 2(6): 43. 1957.

Cryptangium Schrad. ex Nees, Flora brasiliensis (Martius) 2(1): 163. 1842. Type species: Cryptangium verticillatum (Sprengel) Vitta & S.M.Costa Lagenocarpus section Corymbosae H. Pfeiffer, Repert. Sp. Nov. 18: 87 .1922.; emend. T. Koyama & Maguire, Mem. N.Y. Bot. Gard. 12 (3): 48. 1965. Monoecious herbs, with short and/or creeping rhizome: male and female spikelets mixed in the paracladia (female generally distal). Leaf blades reduced to sheaths or well-developed; long lanceolate to linear when developed. Inflorescence terminal, solitary, panicle-like. Involucral bracts leaf-like, with linear-lanceolate to lanceolate blades. Paracladia monomorphic. Male spikelets 6--7 flowered; stamens 2, rarely 3. Female spikelets 1(2)- flowered, 3 glumes; stigmas

3. Glumes spirally or sub-distichally arranged. Achenes stipitate, obovoid, trigonous with noticeable ridges, glabrous and with conspicuous cavities at the base; hypogynous scales reduced and glabrous; beak short conical and glabrous, persistent.

The genus occurs in non-Andean South America regions, from of Guyana and Brazilian Shields southwards to southern areas of Brazil (probably São Paulo state), from western Brazilian coast to eastern Colombian Vaupés, on sandy soils. Total number of species: 1 (one).

1. Cryptangium verticillatum (Sprengel) Vitta [comb. nov. 2015] Basionym: Fuirena verticillata Sprengel, Novi Provent. 47. 1818.

Didymiandrum Gilly, Bulletin of the Torrey Botanical Club 68 (5): 330-332. 1941. Type species: Didymiandrum stellatum (Boeckeler) Gilly Lagenocarpus sect. Didymiandrum (Gilly) T.Koyama, Makinoa 4:70–71. 2004. Dioecious herbs, with short creeping rhizome. Leaf blades reduced to sheaths. Inflorescence terminal, solitary, panicle-like. Involucral bracts leaf-like, with elliptic to oblong-elliptic blades. Paracladia superficially monomorphic. Male spikelets 2--7 flowered; stamens 2. Female spikelets 1 flowered, 6--7 glumes; stigmas 3. Glumes spirally or sub-distichally arranged. Achenes stipitate, trigonous with sharp ridges, glabrous; hypogynous scales reduced and ciliate; beak short conical and glabrous, persistent.

The genus occurs on the Guiana shield, in Colombia, Venezuela and Guiana and Brazil in borders of rain forest areas. Total number of species: 1 (one).

1. Didymiandrum stellatum (Boeckeler) Gilly, Bull. Torr. Bot. Club 68: 331. 1941.

Exochogyne C.B.Clarke, Verh. Bot. Vereins Berlin Brandenburg 47: 101. 1905. Type species: Exochogyne amazonica C.B. Clarke Lagenocarpus section Exochogyne (C.B. Clarke) T. Koyama, Makinoa, new series 4: 47. 2004.

Monoecious herbs, with short creeping rhizome: male and female spikelets mixed in the paracladia. Leaf blades well-developed, linear-lanceolate. Inflorescence terminal, solitary, congested or laxly spike-like. Involucral bracts leaf-like, with linear-lanceolate to lanceolate blades, expanded at base and enclosing the paracladia. Paracladia monomorphic. Male spikelets multiflowered; stamens 2 (rarely 4 or 6). Female spikelets 1 flowered, 4 glumes; stigmas 2. Glumes distichally arranged. Achenes abruptly or gradually attenuate at base, ovoid, ellipsoid or obovoid, biconvex with distinguishable ridges, glabrous; hypogynous scales absent; beak conical, persistent.

This genus occurs in northern South America, on the eastern Guyana Shield and in the Amazon basin, mainly in open vegetation, but is also found in forested white-sand savannas, on nutrient poor sandy soils and among rocks. Total number of species: 2.

Exochogyne lacks both the trigonous achene and the hypogynous scales typical of Cryptangieae and presents a spike-like inflorescence uncharacteristic for the tribe. We can highlight as typical morphological characters of the tribe that also occur in the genus: the unisexual spikelets and the reddish-pink stigmas; and also the distribution more centered in the Amazon biome on poor nutrient soils. The genus was considered as incertae sedis by Goetghebeur (1998) but its species are included in Lagenocarpus in Koyama’s morphology based works (2004) and included in Cryptangieae in Muasya et al. (2009), based on molecular data.

1. Exochogyne amazonica C.B. Clarke, Verh. Bot. Vereins Berlin Brandenburg 47: 101. 1906. 2. Exochogyne steyermarkii Gilly, Fieldiana, Bot. 28(1): 53, f. 6M. 1951.

Krenakia S.M.Costa, nom. nov. Type: Krenakia polyphylla (Nees) S.M.Costa Acrocarpus Nees, Flora brasiliensis (Martius) 2(1): 157. 1842, nom. illegit. Lagenocarpus section Acrocarpus T. Koyama, Mem. New York Bot. Gard. 50 . 1965. Based on Acrocarpus Nees, syn.nov.

Replacement name for Acrocarpus Nees, Flora brasiliensis (Martius) 2(1): 157 (1842), nom illegit., non Wight ex Arn. (1838)

Etymology.– The replacement name is proposed in honor of the indigenous ethnicity that lives nearby the Serra do Cipó National Park (an important protected area of campos rupestres in the Espinhaço Range, Minas Gerais, Brazil), in the “Terra Indígena Fazenda Guarani”, the Krenák people. The obstacles faced by the Krenák, as by many other indigenous ethnicity descendants are troublesome and include culture devaluation, low access to education and health services, land disputes, among others.

Monoecious herbs, with short creeping rhizome: male and female spikelets in separate or mixed paracladia; female spikelets mostly at the apical portion and male ones basal. Leaf blades reduced to sheaths. Inflorescence terminal, panicle-like. Involucral bracts leaf-like, with linear- lanceolate to lanceolate blades. Paracladia monomorphic. Male spikelets (3)5-- 11(22) flowered; stamens 2. Female spikelets 1--5 flowered, 3--7 glumes; stigmas 3. Glumes spirally or sub-distichally arranged. Achenes stipitate, elliptic, obovoid or obtrullate, conspicuously trigonous with sharp ridges, sometimes winged and with

ornamented surface and inconspicuous cavities at the base; hypogynous scales reduced, glabrous; beak short conical and glabrous, persistent.

The genus occurs in Brazil and is especially species rich in the campos rupestres (SE Brazil), with two cases of disjunction, one in the Venezuelan highlands and one in the Caribbean (Cuba). Its species are found on mountain top habitats, mainly in open or riparian vegetation on nutrient poor soils. Total number of species: 10.

The genus comprises species of Acrocarpus Nees (Flora brasiliensis (Martius) 2(1): 157, 1842 – illegitimate name). As the name is illegitimate due to the existence of a homonymous legume genus described five years earlier: Acrocarpus Wight ex Arn., Magazine of Zoology and Botany 2: 547 (1838), we propose a new name for it: Krenakia S.M.Costa.

1. Krenakia claussenii (C.B. Clarke) S.M. Costa, comb. nov. Basionym: Cryptangium claussenii C.B. Clarke, Bull. Misc. Inform, Addit. Ser. 8: 65. 1908. 2. Krenakia comata (Boeckeler) S.M. Costa, comb. nov. Basionym: Cryptangium comatum Boeckeler, Flora 65: 351. 1882. 3. Krenakia cubensis (Kukenthal) S.M. Costa, comb. nov. Basionym: Lagenocarpus cubensis Kükenthal, Repert. Sp. Nov.Regni Veg. 23: 219. 1926. 4. Krenakia humile (Nees) S.M. Costa, comb. nov. Basionym: Acrocarpus humilis Nees, Flora Brasiliensis (Martius) 2(1): 161. 1842. 5. Krenakia junciforme (Kunth) S.M. Costa, comb. nov. Basionym: Scleria junciforme Kunth, Enum. Pl. 2: 56.1837. 6. Krenakia minarum (Nees) S.M. Costa, comb. nov. Basionym: Acrocarpus minarum Nees, Flora Brasiliensis (Martius) 2(1): 159. 1842. 7. Krenakia polyphylla (Nees) S.M. Costa, comb. nov. Basionym: Acrocarpus polyphyllus Nees, Flora Brasiliensis (Martius) 2(1): 160 (1842); lectotype designated by T. Koyama & B. Maguire (1965). 8. Krenakia subaphylla (T. Koyama) S.M. Costa, comb. nov. Basionym: Lagenocarpus subaphyllus T. Koyama, Brittonia 24(3): 279. 1972. 9. Krenakia triquetra (Boeckeler) S.M. Costa, comb. nov. Basionym: Cryptangium triquetrum Boeckeler, Linnaea 38: 413. 1874. 10. Krenakia venezuelensis (Davidse) S.M. Costa, comb. nov

Basionym: Lagenocarpus venezuelensis Davidse, Novon 3: 244. 1993, replacement name for amazonicus Davidse, Novon 2: 319. 1992, non L. amazonicus (C.B. Clarke) H. Pfeiff., Repert. Spec. Nov. Regni Veg. 21: 35. 1925.

Lagenocarpus Nees, Linnaea 9: 304. 1834. Type species: Lagenocarpus guianensis Nees Anogyna Nees, Hooker Journ. Bot. 2: 395. 1840. Adamantogeton Schrad. ex Nees, Flora Brasiliensis (Martius) 2(1): 165. 1842, nom. inval. (pro syn.). Orobium Schrad. ex Nees, Flora Brasiliensis (Martius) 2(1): 165. 1842, nom. inval. (pro syn.). Phaenopyrum Schrad. ex Nees, Flora Brasiliensis (Martius) 2(1): 166. 1842, nom. inval. (pro syn.). Microlepis Schrad. ex Nees, Flora Brasiliensis (Martius) 2(1): 167. 1842, nom. inval. (pro syn.). Lerisca Schltdl. Bot. Zeit. 3: 476. 1845. Monoecious, rarely dioecious herbs, with short or long creeping rhizome, rarely with caudex: male and female spikelets mostly at separate paracladia; female spikelets mostly in apical paracladia. Leaf blades developed, linear-lanceolate. Inflorescence terminal, solitary, panicle- like. Involucral bracts leaf-like, with linear-lanceolate to lanceolate blades. Paracladia mostly dimorphic. Male spikelets (3)4--7flowered; anthers\ 1, rarely 2 or 4--5. Female spikelets 1 flowered, (4)5--6(7) glumes; stigmas 3 (rarely 4--5). Glumes spirally or sub-distichally arranged. Achenes sessile, ovoid or less frequently ellipsoid, obscurely trigonous with inconspicuous ridges or trisulcate, glabrous or rarely pubescent, surface generally irregularly rugose at maturity or after dry, main body continuous to the beak; hypogynous scales reduced or well-developed (in L. guianensis subsp. hypocrateus), mostly with glabrous or with sparsely ciliate margins, rarely densely pubescent at surface and margin; beak short conical or pyramidal, glabrous (rarely pubescent), persistent.

Lagenocarpus occurs from Caribbean to southern South America (Rio Grande do Sul state, Brazil), in different non-Andean biomes but mainly in soils with low nutrient availability, like sandy, rocky or waterlogged soils. Total number of species: 15.

1. Lagenocarpus albo-niger (A. St.-Hil.) C.B. Clarke, Bull. Misc. Inform., Addit. Ser.8: 64.1908.

2. Lagenocarpus bracteosus C.B. Clarke, Repert. Spec. Nov. Regni Veg. 2: 145. 1906. 3. Lagenocarpus celiae T. Koyama & Maguire, Mem. New York Bot. Gard. 12(3): 46. 1965. 4. Lagenocarpus eriopodus T. Koyama & Maguire, Mem. New York Bot. Gard. 12(3): 47. 1965. 5. Lagenocarpus glomerulatus Gilly, Fieldiana Bot. 28(1): 57. 1951. 6. Lagenocarpus griseus (Boeckeler) H. Pfeiff. Ber. Deutsch. Bot. Ges. 39: 131. 1921. 7. Lagenocarpus guianensis Nees, Linnaea 9: 304. 1834. 8. Lagenocarpus inversus C.B. Clarke, Kew Bull. Addit. Ser. 8: 64. 1908. 9. Lagenocarpus lanatus (T. Koyama & Maguire) T. Koyama, Makinoa New Series 4:39- 40. 2003. 10. Lagenocarpus pendulus T. Koyama, Mem. New York Bot. Gard. 12: 44. 1965. 11. Lagenocarpus rigidus (Kunth) Nees, Flora Brasiliensis (Martius) 2(1): 167. 1842. 12. Lagenocarpus sabanensis Gilly, Fieldiana Bot. 28(1): 57. 1951. 13. Lagenocarpus tenuifolius (Boeckeler) C. B. Clarke, Kew Bull. Addit. Ser. 8: 64. 1908. 14. Lagenocarpus tristis (A. St.-Hil.) Vitta [comb. nov., 2015] Basionym: Scleria tristis A. St.-Hil. Voy. Distr. Diam. 1: 370. 1833. 15. Lagenocarpus velutinus Nees, Flora Brasiliensis (Martius) 2(1): 166. 1842.

Ancestral Character State reconstruction.– The maximum credibility tree recovered the following clades (Figure 5): Cephalocarpus, Cryptangium, Exochogyne, Didymiandrum, Krenakia, and Lagenocarpus. Exochogyne appears as the first divergent genus, followed by Krenakia and then a clade with Cryptangium and Didymiandrum as sister group to Cephalocarpus and Lagenocarpus.

We recovered the following ancestral states for the selected diagnostic characters in Cryptangieae clade: monoecy* (character a); without caudex* (character b); terminal* (character c) and panicle-like* (character d) inflorescence, with monomorphic paracladia* (character e); and the female flowers with a reddish-pink stigma (character m). The hypogynous scales were absent (character f), and once present it is uncertain if they were conspicuous or inconspicuous (character g) but probably glabrous* or short ciliate (character h). The ancestral achene’s transversal shape in cross section is uncertain (character i), but it probably had a surface projections and/or wings* (character j), and a beak (character k) continuous with the main achene body (character l). If we could identify this hypothetical ancestral, it would fit better to Krenakia representatives (observe the

characters with and asterisk -*- in the identification key and Krenakia description). (Figure 5, Table 3)

For the first selected character, “a. monoecy vs. dioecy”, only three species were stated as dioecious, and these are scattered among clades. The ancestral state of this character to Cryptangieae is “monoecious” (Figures 5, Appendix 1). The dioecism is considered a “dead end” in the evolution of breeding system in plants, especially when no reduced nonfunctional structure, like staminodes or pistiloids, is observed in the flower (Ainsworth 2000); so it is expected that the ancestral state for this character is “monoecy” in the tribe.

Gilly (1941) united Didymiandrum stellatum and Cephalocarpus flexifolius together in Dydimiandrum based mostly on the dioecism despite other morphological differences. Koyama & Maguire (1965) described another dioecious species later (Lagenocarpus celiae) and Koyama (2004) included Didymiandrum under his Lagenocarpus concept, while recognizing Everardia flexifolia (=Cephalocarpus flexifolius) (Koyama 2003). After our molecular phylogenetic analysis, each dioecious species belongs to different clades and belongs to a different genus in our proposal: Dydimiandrum stellatum, Cephalocarpus flexifolius and Lagenocarpus celiae; all three species only occur in the Amazon region (see character “n. distribution region”).

Trilepis spp., Cephalocarpus spp. and two Lagenocarpus spp. develop a caudex (a narrow perennial stem, more or less lignified, covered by old leaf sheaths and frequently with adventitious roots (Goetghebeur 1998) (character b). The presence/absence of this structure was frequently used to distinguish Lagenocarpus (caudex absent) and Everardia (caudex present) species, and led Vitta (2005) to propose the transfer of two species from Lagenocarpus to Everardia (=Cephalocarpus). In the present proposal the development of a caudex is common to all Cephalocarpus spp.but also occurs in two species of Lagenocarpus; these two species are from the Amazon region and are early divergent within the clade of Lagenocarpus. Another interesting thing to notice is the occurrence of a caudex in Trilepideae, a tribe not-so-closely related to Cryptangieae but which occurs in an area similar to the Guiana Shield, the South-Africa Greater Capes (Hopper 2009).

The character state “lateral inflorescence” is a novelty common to all Cephalocarpus spp. (in the circumscripition here proposed) (character c). The panicle-like

inflorescence morphology (character d) is common to Cryptangieae, except to some species of Cephalocarpus, Exochogyne and maybe Krenakia comata, depending on the interpretation of the inflorescence by the researcher. Individually, these inflorescence characters (c and d) are useful to distinguish some Cephalocarpus species, but not the other genera. Together, a “terminal” and “spike-like” inflorescence distinguish E. amazonica, as maybe K. comata.

The different morphology between male and female paracladia (character e) is a novelty of Lagenocarpus spp. from the campos rupestres, except L. inversus, and two species with broader distribution (L. guianensis and L. rigidus). All other Cryptangieae genera and species have monomorphic paracladia, and the male and female spikelets are often mixed even if the female spikelets are mostly in terminal position (in the paracladia or in the whole inflorescence). Lagenocarpus inversus is an interesting species with inflorescences shorter than the leaves and basal female spikelets, while the other species of the group show inflorescences conspicuously higher than the leaves and terminal female spikelets in the inflorescence. The pattern within the Lagenocarpus group from campos rupestres are in agreement with a hypothesis about anemophilous species, the most probable system Lagenocarpus spp., that the capture of conspecific pollen may be optimized through male/female dimorphism (Friedman 2011). Anyway, while not all Lagenocarpus have dimorphic paracladia, all species with dimorphic paracladia are in Lagenocarpus.

The achene characters are the most often used in Cyperaceae taxonomy, and mature achene characters are essential for the recognition of many species. Here, we reconstructed seven achene characters used in Cryptangieae taxonomy (characters f – k). The absence/presence of hypogynous scales (character f) is one of the characters that joint most representatives of Cryptangieae, except for Exochogyne. The conspicuous scales (character g) are common to Cephalocarpus while the inconspicuous scales are typical of other clades (excluding Exochgyne – scales absent). Most Lagenocarpus species have inconspicuous scales, except for some species, from Amazonian areas, and the subspecies L. guianensis Lindley & Nees subsp. hypochrateus T.Koyama. The hypogynous scales ornamentation is a good character to segregate the genera (character h). The “fimbriate margin” is probably a novelty of Cephalocarpus spp., but some Lagenocarpus species (essentially from Amazon) present pubescent hypogynous scales, not only on the margins but on the dorsal surface too.

The achene shape in transversal section (character “i”) is important to distinguish Exochogyne species from others, and also helpful to distinguish Krenakia and Didymiandrum from Cryptangium, Cephalocarpus and Lagenocarpus species. The ornamentation of the achene (character “j” – (0) smooth and glabrous (1) smooth with trichomes (2) with projection and/or wings) is constant within Didymiandrum, Cryptangium, Exochogyne and Krenakia; but the achenes of Cephalocarpus spp. and Lagenocarpus spp. may be smooth/glabrous or with trichomes. The presence of trichomes seems to be a novelty that distinguishes the Cephalocarpus + Lagenocarpus clade. The presence of an achene beak is acharacter common to all Cryptangieae representatives, but only some Cephalocarpus species show a constriction zone between the beak and the main body of the achene (character l)). Almost all Cryptangieae representatives (except for some populations) present a reddish-pink stigma, and it is the character state recovered as a synapomorphy of the tribe.

All that said, the morphological recognition of the genera after the addition of the informations from the molecular phylogeny, cannot rely on a single character, but depends on the combination of two or more characters. When those characters were used isolated or with different weights by various specialists (Gilly 1941a, 1941b, 1942, 1951; Koyama & Maguire 1965; Koyama 2003, 2004; Vitta 2005 - PhD thesis) they led to an artificial circumscription of the genera. Different combinations of the above-mentioned characters are the better way to identify Cryptangieae specimens and a traditional dichotomous key developed to the tribe should present more than a character at each step, as done in our own identification key.

We did not perform an ancestral area analysis, but it is worth to mention that Cephalocarpus, Exochogyne and Didymiandrum mostly occur in the Amazon region, while Krenakia is mostly restricted to the campos rupestres. Cryptangium is spread in C & S America and Lagenocarpus shows widespread species, a group of species from Amazon areas only and another group restricted to the campos rupestres.

Two intriguing cases are Krenakia venezuelensis, that occurs on the Guiana Shield, and K. cubense, that occurs only in Cuba, and may be investigated in the future. These are two cases of disjunction in the distribution of Krenakia. Both also present unique morphological character states within the genus: K. cubense is probably dioecious and K. venezuelensis has densely lanose leaf sheaths and connate stamens. Unfortunately, there is

still no DNA sequence available of these species to support their position in the genus without doubt.

General Conclusions With the development of molecular tools and the respective analyses, it is increasingly and inevitably common to apply these data for a hypothesis about the classification of various plant groups. For some of those groups, there is a correspondence between molecular and morphological data, but incongruences between them may occur.

Cryptangieae genera were traditionally distinguished by a more or less short list of morphological characters (Gilly 1941a, 1941b, 1942, 1951; Koyama & Maguire 1965; Koyama 2003, 2004; Vitta 2005 – PhD thesis), but as suggested by the present molecular analyses not all character states are exclusive of each Cryptangieae genera in the phylogeny. Luckily, even if there is no single character able to distinguish the genera, different sets of vegetative and reproductive character may be useful for this task; even in depauperized specimens (for example, a specimen with terminal inflorescence remains plus caudex development probably belongs to the Amazon group of Lagenocarpus).

There still are many things to do. Future studies in early-divergent tribes of subfamily Cyperoideae (Cyperaceae) should include this molecular + morphological approach and representatives of tribe Schoeneae. Also a study to investigate the correlation among characters related to pollination process in representatives of Cryptangieae could enrich the discussion about anemophilous pollination and characteristics of the inflorescence.

Table 4 – Cryptangieae species and sequences at the Maximum Parsimony and Bayesian Inference analysis (type-species of previously described genera in bold; type-species of Koyama’s (2004) sections with an asterisk*) Taxa Specimen/Accession number Molecular marker rbcL trnL-F ITS ETS 5S- NTS (Trilepideae) Trilepis lhotzkiana AY725955.1 (GenBank) X Trilepis sp. S.M. Costa 1125 X

(Bisboeckelereae) Diplacrum sp. S.M. Costa 1083 X X X

(Sclerieae) Scleria amazonica Camelbeke, M.T. Strong & S.M. Costa 1054 X X X Goetghebeur

(Cryptangieae – spp. sensu Koyama 2003, 2004) Cephalocarpus confertus Gilly S.M. Costa 1113 X X X X X Cephalocarpus rigidus Gilly ex Gleason & Killip Maguire & Maguire 35389 (?) X

Everardia debilis T. Koyama & Maguire Maguire 31735 (F) X Everardia diffusa T. Koyama & Maguire B. Maguire 27825 (NY) X X X Everardia disticha T. Koyama & Maguire O. Huber 10060 (NY) X X Everardia flexifolia (Gilly) T. Koyama & Maguire R.Liesner 24939 (???) X Everardia montana Ridley Vareschi et Foldats ??? (NY), X X X FN870820-1 (GenBank) Everardia recurvigluma T. Koyama & Maguire J.A. Steyermark 608 (F) X X X Everardia vareschii Maguire O. Huver 9540 (NY) X Everardia sp1 S.M. Costa 1121, S.M. Costa 1133 X X X X X Everardia sp2 Rodriguez 2890 (UEC)

(Lagenocarpus sect. Lagenocarpus) Lagenocarpus guianensis Nees* S.M. Costa 1085 X X X X Lagenocarpus inversus C.B.Clarke S.M. Costa 1235 X X X X Lagenocarpus pendulus T.Koyama J.M. CArdiel 1011 (NY) X Lagenocarpus rigidus (Kunth) Nees S.M. Costa 1011, S.M. Costa 1263, X X X X X S.M. Costa 1014 (rixte) Lagenocarpus sabanensis Gilly S.M. Costa 1077 X X X X X Lagenocarpus tenuifolius (Boeckeler) Kuntze S.M. Costa 1009 X X X X X

(Lagenocarpus sect. Imberbitae H.Pfeiffer) Lagenocarpus adamantinus Nees S.M. Costa 1251, A. V. Scatigna ??? X X X X X Lagenocarpus albo-niger (A. St.-Hil.) C. B. S.M. Costa 1240, GB X X X X X Clarke* Lagenocarpus bracteosus C.B.Clarke S.M. Costa 1238 X X X X Lagenocarpus celiae T. Koyama & Maguire S.M. Costa 1044 X X X X X Lagenocarpus eriopodus T. Koyama & Maguire S.M. Costa 1046 X X X X X Lagenocarpus aff glomerulatus Gilly S.M. Costa 1047, S.M. Costa 1049 X X X X X Lagenocarpus griseus (Boeckeler) H.Pffeif. J.A. Oliveira 664128 X X X X X Lagenocarpus velutinus Nees Leitão Filho 9762 (UEC) X

(Lagenocarpus sect..Corymbosae H.Pfeiffer) Lagenocarpus verticillatus (Sprengel) T. Koyama S.M. Costa 1076, S.M. Costa 1051, X X X X X & Maguire* S.M. Costa 1034

(Lagenocarpus sect.. Exochogyne (C.B.Clarke) T.Koyama) Lagenocarpus amazonicus C.B.Clarke* S.M. Costa 1032, Aparecida da Silva X X X X 1986 (GENT)

(Lagenocarpus sect.. Acrocarpus (Nees) T.Koyama)

Lagenocarpus comatus (Boeckeler) H. Pfeiff. Valente 612 X X Lagenocarpus humilis (Nees) Kuntze Zappi 2633 (NY) X X X Lagenocarpus polyphyllum (Nees) Kuntze* W.R. Anderson 11678 X

(Lagenocarpus sect. Junciformes T.Koyama) Lagenocarpus junciformis (Kunth) Kuntze* S.M. Costa 1230 X X X Lagenocarpus minarum (Nees) Kuntze S.M. Costa 1080; S.M. Costa 1271 X X X X X Lagenocarpus triqueter (Boeckeler) Kuntze Meirelles 312 (UEC) X X X Lagenocarpus subaphyllus T.Koyama G. Sheperhd 75826 (UEC) X

(Lagenocarpus sect. Didymiandrum (Gilly)T.Koyama) Lagenocarpus stellatus (Boeckeler) Kuntze* T. Koyama 7476, GB X X

Table 5 – Molecular markers, primers and protocols applied during PCR experiments Region Primers (and its combination) Protocols (temperature; time) trnL- F c –CGA AAT CGG TAG ACG CTA CG Premelt – 95°C; 2’ d – GGG GAT AGA GGG ACT TGA AC Denature – 95°C; 30” e – GGT TCA AGT CCC TCT ATC CC Anneal – 55°C; 1’ f – ATT GAA CTG GTG ACA CGA G Extension – 72°C; 1’ Final extension – 72°C; 7’ Number of cycles – 35

[c-f or c-d/e-f] Additives: (none) rbcL 1F – ATG TCA CCA CAA ACA GAA Premelt – 95°C; 2’ ACT AAA GC Denature – 95°C; 30” 636F – TCC TTT TAG TAA AAG ATT Anneal – 55-60°C; 1’ GGG CCG AG Extension – 72°C; 1’ 674R – GAT TTC GCC TGT TTC GGC Final extension – 72°C; 7’ TTG TGC TTT ATA AA Number of cycles – 35 1460R – TCC TTT TAG TAA AAG ATT GGG CCG AG

[1F-674R and 636F-1460R] Additives: (none) ITS 5i – AGG TGA CCT GCG GAA GGA TCA Premelt – 95°C; 2’ TT Denature – 95°C; 30”/95°C; 30”/95°C; 4i – GGG TAG TTC CCG CCT GAC CTG 30” G Anneal – 56°C; 30”/53°C; 30”/50°C; 30” Extension – 72°C; 1’15”/72°C; 1’15”/72°C; 1’15” Final extension – 72°C; 7’ Number of cycles – 4/4/35 (“touchdown”)

[5i-4i] Additives: DMSO 5% and betaine 2M ETS f – CTG TGG CGT CGC ATG CGT TG Premelt – 95°C; 2’ 18Sr – AGA CAA GCA TAT GAC TAC Denature – 95°C; 30” TGG CA Anneal – 62°C; 1’ Extension – 72°C; 1’30” Number of cycles – 30

Additives: DMSO 10% [f-18Sr] 5S-NTS PI – TGG GAA GTC CTY GTG TTG Premelt – 95°C; 3’ PII – KTM GYG CTG GTA TGA TCG CA Denature – 95°C; 30” Anneal – 53°C; 40” Extension – 72°C; 45” Number of cycles – 40

[F-R] Additives: DMSO 5% and betaine 2M Observation: we performed “nested” PCR experiment on some samples due to the low DNA concentration after the first standard PCR experiment, mostly at samples from herbarium specimens

Table 6 – Diagnostic characters and character status selected to the Ancestral Character State analysis Diagnostic Character Possible States of Character

(a) Sexual system (0) monoecious; (1) dioecious (b) Caudex development (0) absent; (1) present (c) Inflorescence position (0) terminal; (1) lateral (d) Inflorescence general morphology (0) head-like; (1) panicle-like; (2) spike-like (e) Paracladia sexual morphology (0) monomorphic; (1) dimorphic (f) Hypogynous scales (0) absent; (1) present (g) Hypogynous scales development (0) conspicuous; (1) inconspicuous (h) Hypogynous scales ornamentation (0) glabrous or short ciliate margins; (1) short or long fimbriate margin; (2) pubescent margin and dorsal surface (i)Achene shape in cross section (0) circular; (1) obscurely trigonous; (2) triangular with sharp ridges; (3) biconvex (j) Achene ornamentation (0) smooth and glabrous; (1) smooth with trichomes; (2) surface with projections and/or winged (k) Achene beak presence (0) absent; (1) present (l) Achene constriction area (0) absent; (1) present (m) Stigma color (0) white; (1) reddish-pink

Figure 4 – Phylogenetic hypothesis on Cryptangieae genera based on a combined matrix with two chloroplast (rbcL and trnL-F) and three nuclear ribosomal (ITS, ETS and 5S-NTS) regions. [full circle – PP≥0.95; one star - BS=50%-<75%; two stars - BS≥75%. Group/genus color: black – outgroup; yellow – Cryptangium; pink – Didymiandrum; brownish-green – Exochogyne; purple – Krenakia; blue and red– Cephalocarpus (including Everardia); green - Lagenocarpus]

Figure 5 – Mapping of all diagnostic characters applied the Ancestral Character State analyses and Ancestral Character States recovered to Cryptangieae (see selected diagnostic characters at Table 3; character n: distribution area, white – C&S America, green – campos rupestres, blue – Guiana Shield and Amazon, black – S America and Africa).

Figure 6 – Cryptangieae general morphology. A. Cephalocarpus confertus (habit), B-C. Cephalocarpus montanus habit (B) and caudex in cross section (C); D-E. Krenakia minarum habit (D) and detail of leaves reduced to sheaths (E); F-G. Exochogyne amazonica with mature male (F) and female (G) spikelets; H. Cryptangium verticillatum male spikelets; I. Krenakia sp. spikelets. J. Lagenocarpus rigidus inflorescence (female spikelets apical and in rigid peduncles and male spikelets basal and in flexible peduncles); K. Cephalocarpus confertus inflorescence detail (male and female spikelets at the same cluster); I. Cephalocarpus montanus uniflowered spikelets with immature achenes.

CHAPTER 3 - Ten Cryptangieae (Cyperaceae) new species, the diversity buried at herbaria collections Additional author: Volker Bittrich

(Intended submission at Phytotaxa Journal)

Abstract Many of the new angiosperm species to be discovered by botanical taxonomists may already be deposited in the collections of great herbaria. In this paper we describe ten new species of sedge tribe Cryptangieae (seven Cephalocapus spp. and three Lagenocarpus spp.) based on specimens collected before the 1990’s. These species are from the Venezuelan Guiana Shield mainly, but also adjacent areas in Brazil and Colombia. We also provide illustrations of diagnostic characters, distribution maps and additional comments to the species. Keywords:— Cephalocarpus, Lagenocarpus, white-sand savannas, campinaranas

Resumo Muitas das novas espécies de angiospermas a serem descobertas por taxonomistas botânicos podem já estar depositadas em coleções de herbários importantes. Neste manuscrito, são descritas des novas espécies da tribo Cryptangieae (sete Cephalocapus spp. e três Lagenocarpus spp.) baseados em espécies coletados antes da década de 1990. Estas espécies são principalmente de áreas venezuelanas do Escudo das Guiandas, mas também ocorrem em áreas adjacentes do Brasil e Colômbia. Tambpem são fornecidas ilustrações de caracteres diagnósticos, mapas de distribuição e comentários adicionais para as espécies.

Introduction A recent study asserted that most descriptions of new plant species come from older collections rather than recent ones, due to reasons that include diminished investment in taxonomy and the difficult access to specialized bibliography (Bebber et al. 2010). As pointed by the authors, many new species stay stocked at great herbaria collection till broad and profound monographic studies, including the analyses of the numerous specimens and taxa at each focused group. Additionally, some important collecting effort in high diversity areas somehow aimed to increase the collections available at herbaria even if there were no prompt intention to taxonomically study them, as the case of expeditions to the Venezuelan Guiana Shield areas.

Cryptangieae is endemic to the Neotropics, from Cuba to southern Brazil, and is especially rich at the highlands of Guiana Shield and the campos rupestres, in southeastern Brazil (Koyama 2003, 2004, Costa et al. CHAPTER1, CHAPTER2). Recent studies aimed to clarify the status of Cryptangieae and circumscription of its genera (Costa et al. CHAPTER1, CHAPTER2) and monographic treatments of each genus will soon be published (Costa et al. CHAPTER4). During these studies, the number of accepted species for the tribe was estimated to 47 species distributed in six genera (Cephalocarpus Nees, Cryptangium Schrad. ex Nees, Didymiandrum Gilly, Exochogyne C.B.Clarke, Krenakia S.M.Costa and Lagenocarpus Nees). None the less, during the recent herbaria studies between 2014 and 2017, we could find some unidentified or missindentified specimens that we understand as new species. These new species are from northern South America and their distribution are related to the Guiana Shield environments, mainly in Venezuela.

Here we describe seven new species of Cephalocarpus and three new species of Lagencarpus increasing the known diversity of Cryptangieae to 57 species. We also provide comments on taxonomy, morphology and distribution and illustration of the diagnostic characters of each species.

Material and Methods We visited herbaria at Brazil (ASE, MG, INPA, SPF, UEC) and U.S.A. (NY, F, MO, US) and analyzed more than 1400 specimens of Cryptangieae, including several type collections. The acronyms cited follow Thiers et al. (continuously updated). We examined the specimens

using binocular dissecting microscope available at the visited institutions. We obtained the multifocal photographs of details of some species using a multifocal stereoscope Leica DFC295 (Leica Biosystems, Wetzlar, Germany) and the software LAS® (Leica Biosystems, Wetzlar, Germany). We used the softwares BRAHMS (available at: http://herbaria.plants.ox.ac.uk/bol/brahms/) and ArcGis 10.3 (ESRI 2015) to build a database including geographical coordinates and to elaborate distributional maps.

Description of the new species 1. Cephalocarpus chimantensis S.M. Costa, sp. nov. (Figures 7A, 10A).

Similar to Cephalocarpus longibracteatus and C. rigidus, regarding the arrangement of leaves and inflorescence, but differs from both species by the developed peduncle of the spikelets, achene (up to 0.6 mm long) with truncate apex and short conical beak without narrow neck between main body and beak.

Type specimen:— VENEZUELA. Bolívar: Piar, Macizo del Chimantá, Sección oriental del Chimantá-tepui, cabeceras del afluente derecho superior del río Tirica (“Caño del Grillo”), 2450 m, 5°18’N 62°03’W, 7-9 February 1983, J.A.Steyermark 128931 & O. Huber, V. Carreño E (NY! – holotype; VEN? - isotype)

Monoecious herb: sexes intermingled at paracladia, monomorphic. Caudex ~1.3 mm diam. (4 –5 mm diam. including sheaths remains and adventitious roots). Leaf: sheath pubescent, greenish or brown; contraligule indistinct or a straight line with shiny and smooth surface below; blade 3–4.5 × 0.13–0.15 cm, linear-lanceolate, margins and midrib glabrous or sparsely pubescent, concolorous, greenish to reddish-brown after drying. Inflorescence panicle-like, with 2 paracladia; culm 3 –4.5 × 0.05 –0.07 cm, compressed, glabrous or with rare trichomes. Paracladia (lowermost) up to 1 cm long, 1 order; peduncle up to 0.6 cm long, compressed, glabrous to sparsely ciliate. Involucral bract (lowermost) leaf-like: sheath up to 3 mm long, pubescent, reddish-brown; blade 10–13 mm long, “M”-shaped, pubescent, reddish-brown to brownish-green. Male spikelet 3.5–4.5 × 0.7–0.9 mm, 5–6 glumes, 1–2 flowers; glume (outermost) –2.5 mm long, mucronate (mucro up to ~0.1 mm long); stamens 2, connective projection not observed. Female spikelet ~4 × ~1 mm, 4–5 glumes; glume (outermost) –2.5 mm long, mucronate (mucro up to ~0.1 mm long); stigmas 3. Achene 1.3– 1.5 × 0.4–0.6 mm, obtriangular, smooth, short ciliolate at apex, reddish-brown, with truncate

apex; hypogynous scale ~0.3 mm long, elliptic to orbicular, trichomes <0.01 mm long (sparse); beak ~0.3 mm long, short conical, glabrous.

Etymology:— The name of the species refers to the type-locality, the Chimantá Massif (Bolívar, Venezuela).

Distribution:— This species is known only from its type-locality, in the Chimantá Massif (Bolívar, Venezuela), at altitudes above 2450 m high.

Taxonomic notes:— Cephalocarpus chimantensis is morphologically similar to C. longibracteatus and C. rigidus, but differ from both by the presence of a peduncle, up to 0.6 mm long, below the spikelets (sessile at the other two species), and the achene with a short conical beak and lacking the narrow neck between the beak and the main body.

Examined Material (additional):— VENEZUELA. Bolívar: Piar, Macizo del Chimantá, Seccion oriental del Chimantá-tepui, cabeceras del afluente derecho superior del rio Tirica (“Caño del Grillo”), 2450 m, 5°18’N 62°03’W, 7–9 February 1983, O. Huber 7183 & J.A. Steyermark, (NY!); Chimantá Massif, east-south portion of summit of Apacará-tepui, 2450– 2500 m, 21–22 June 1953, J.A. Steyermark 75933 (F! – 2 sheets).

2. Cephalocarpus illustrata S.M. Costa, W.W. Thomas, sp.nov. (Figures 7B, 10B)

This remarkable species presents an open panicle-like inflorescence (as typical for Everardia sensu Koyama 2003) and an achene with a constriction zone between the main body and the beak (as Cephalocarpus spp. sensu Koyama 2003). The combination of these two characters is unique in Cephalocarpus.

Type specimen:— VENEZUELA. Bolivar: Cedeño, Serranía Guanay, sector nororiental,Altiplanicie poco disectada, inclinada hacia el S y SW, em las cabeceras más orientales del Río Parguaza, 1700 m, 5°55’N 66°23 W, 20–28 October 1985, O. Huber 10949 (NY!)

Monoecious herb: sexes mostly separate (female spikelets at the apex), monomorphic. Caudex ~9 mm diam. including sheaths remains and adventitious roots. Leaf: sheath glabrous, reddish-brown; contraligule indistinct; blade 5–10 × 0.25–0.38 cm, linear-lanceolate, margins and midrib (abaxial) sparsely pubescent, reddish-brown. Inflorescence panicle-like, 3–5

paracladia; culm 6–10 × ~0.1 cm, compressed, margin minute ciliate. Paracladia (lowermost) up to 2.5 cm long, 2–3 orders; peduncle up to 4 cm long, compressed, glabrous. Involucral bract (lowermost) leaf-like: sheath –7 mm long, surface ciliate and mouth pubescent, light reddish-brown to reddish-brown; blade –10.5 × –1,6 mm, “V”-shaped, margin and midrib (ab) ciliate. Male spikelet –~7 × ~0.9 mm, 5 glumes, 1–2 flowers, 1–2 clusters; glume (outermost) –~5 mm long, mucronate (mucro –~3 mm long); stamen 2, connective projection oval with minute straight trichomes. Female spikelet –~6.5 × ~1 mm, 4–5 glumes; glume (outermost) – ~4 mm long, mucronate (–~1.8 mm long); stigma 3. Achene 4–5 × 0.6–0.8 mm, oblong- ovoid, smooth, ciliate at the apex, with a constriction zone between main body and beak; hypogynous scales 1–1.2 mm, orbicular, trichomes 0.75–1 mm long; beak ~1–1.4 mm long, conical, pubescent.

Etymology:— This species was the first new species discovered presenting mixed morphological characters of Cephalocarpus Nees (sensu Koyama 2003) and Everardia Ridley (sensu Koyama 2003) and started to clarify the morphological link between these two widely accepted genera after the rearrangement based on molecular studies (Costa et al. CHAPTER 1, CHAPTER 2). So the species illustrates the connection between Cephalocarpus and Everardia.

Distribution:— This species registers are from the Chimantá-tepui and the Serranía Guanay (Bolívar, Venezuela)

Taxonomic notes:— This species is the only that presents an open panicle-like inflorescence and an achene with a constriction zone between the main body and beak. Mr. Koyama examined its specimens and believed it was an immature Everardia plant, probably due to its narrow spikelet. But the collections had mature achenes completely enclosed by the glumes.

3. Cephalocarpus “lowlandia” S.M. Costa, sp.nov. (Figures 7C, 10C)

Similar to Cephalocarpus angustus by the densely pubescent, shiny and light green leaves. Cephalocarpus lowlandia differs by its visibly smaller female spikelets (5.8–6.9 mm long in C. angustus) and dark-brown to blackish glumes (instead of paleaceous in C. angustus).

Type specimen:— VENEZUELA. Amazonas: Rio Negro, Neblina Massif, 500–700 m, 0°50’N 66°06’W, 15 July 1984, G. Davidse 27378 & J.S. Miller (NY! – holotype; MO! - isotype)

Monoecious herb: sexes mostly at separate paracladia (male on basal and female on apical ones), monomorphic . Caudex ~1–2 mm (2–7 mm including fibrous sheaths remains and adventitious roots). Leaf: sheath glabrous, dark-brown; contraligule indistinct; blade (8) 20– 25 × 0.2–0.3 cm, linear-lanceolate, margin sparsely pubescent, generally concolorous, light green to green. Inflorescence panicle-like, (5) 8–12 paracladia; culm (20) 45–60 × (0.06) 0.09–0.15 cm, compressed, glabrous. Paracladia (lowermost) up to 12 cm long, 3–4 orders; peduncle –8.5 cm long, compressed, glabrous. Involucral bract (lowermost) leaf-like or reduced to sheath: sheath 1–2 cm long, glabrous, dark-brown; blade 1–3 × 0.07 cm, “V”- shaped, glabrous, dark-brown. Male spikelet 3–5.5 × 0.8–1.1 mm, 7–9 glumes, 4–5 flowers, 1–2 each cluster; glume (outermost) ~1.5–3.7 mm long, glabrous, mucronate (mucro ~0.3–1.5 mm long), dark-brown; stamens 4, connective projection short-conical with short trichomes. Female spikelet (1.8) 2–3.5 × (0.45) 0.6–0.8 mm, 4–5 (7) glumes; glume (outermost) ~1.9–3 mm long, glabrous, mucronate (mucro ~0.3–0.5 mm long), dark-brown; stigma 3. Achene 3– 5.5 × 0.8–1.7 mm, oblong-elliptic, smooth, ciliate to pubescent apex, main body continuous to the beak; hypogynous scales 1–1.4 mm long, elliptic, trichome 0.7–1 mm long; beak 1–2.5 mm long, narrow-conical, ciliate or pubescent especially at base.

Distribution: — its registers are from Cerro de la Neblina (Amazonas, Venezuela

Taxonomic notes:— Cephalocarpus lowlandia morphologically resembles C. angustus, but it is quite different due to the narrower and less pubescent leaves, the smaller female spikelets, and the achene with pubescent beak. It is possible to notice a gradient increase of pubescence and lowermost male glume size and decrease of the leaf length at higher altitudes at this species.

Examined Material (additional):— VENEZUELA. Amazonas: Rio Yatua, 11 December 1957, B. Maguire 42341 & J.J.Wurdack & C.K. Maguire (NY!); Cerro de la Neblina, slope N of Rio Mawarinuma, 500–700 m, 0°50’N 66°05’W, 25 April 1984, B.A. Stein 1671 & A. Gentry (NY!, MO); Cerro de la Neblina camp 2, Neblina Massif, 2100 m, 0°49’40”N 65°59’W, 15 April 1984, B.A. Stein 1572 & A. Gentry (NY!, MO); Rio Negro, Cerro de La

Neblina above Puerto Chimo Camp on Rio Mawarinuma, 150–500 m, 00°50’N 66°07’W, 14 February 1984, R. Liesner 15902 (NY!, MO).

4. Cephalocarpus martinhae S.M. Costa, sp.nov. (Figures 7D, 10D)

Similar to Cephalocarpus rigidus due to its head-like inflorescence, but clearly distinguished by its narrower leaf without main midrib and achene with round apex and linear beak without a narrow neck between main body and beak.

Type specimen:— VENEZUELA. Amazonas: Atabapo, Cerro Marahuaca, cumbre, sección suroriental, vecinidades dek zanjón, 2685 m, 3°37’N 65°21’W, 15 January 1981, B. Maguire 65631 & J.A. Steyermark, C. Brewer-Carías, C.K. Maguire, V.C. Espinosa (NY02480927! – holotype, VEN?).

Monoecious herb: sexes intermingled at paracladia, monomorphic. Caudex 4.5–6 mm diam. including sheaths remains and adventitious roots. Leaf: sheath greenish, margin glabrous or sparse-pubescent; contraligule indistinct; blade 3–5 cm × 0.08–0.13 cm, linear, no main midrib, margin and midrib glabrous or sparse pubescent, concolorous, greenish. Inflorescence reduced panicle-like, single at each prophyll, 2 paracladia; culm 1.3–1.5 × 0.05-0.07 cm, compressed, glabrous. Paracladia (lowermost) 0.5–1.5 cm long, 1–2 orders; peduncle up to 1 cm, compressed, glabrous. Involucral bract (lowermost) leaf-like: sheath 0.3 cm, mouth pubescent, yellowish/reddish-brown; blade 1–1.2 × 0.09 cm, “M”-shaped, glabrous to sparse long ciliate, yellowish/reddish-brown. Male spikelet 3.5–4 × 0.7–0.8 mm, 1–2 flowers, 5–6 glumes; glume (outermost) –2,5 mm long, margin ciliate, mucronate (ciliate mucro up to ~0.6 mm long); stamen, 2, connective projection short ciliate. Female spikelet ~4 × ~1mm, 4–5 glumes; glume (outermost) –2,5 mm long, margin ciliate, mucronate (ciliate mucro up to ~0.6 mm long); stigmas 3. Achene 2–4 × ~1 mm, obovoid, smooth, apex ciliate, light brown to brown, apex rounded; hypogynous scale –1.2 mm long, orbicular or obtriangular, trichome –1 mm long; beak –0.8 mm long, linear, glabrous.

Etymology: — This species is named in honor of Marta Regina Silva Pereira (a.k.a. Martinha), an enthusiastic bryologist, an exciting biology teacher (at various campus of the Universidade do Estado do Amazonas – UEA Brazil) and captivating person. She is clearly in love with her Calymperaceae green friends and is a real friend to all her students and

colleagues. Despite all the obstacles in her life, especially in the last decade, she enjoys the good oportunities of life and always has a smile and a big hug for us.

Distribution:— Only registered from its type-locality and probably restricted to Cerro Marahuaca (Amazonas, Venezuela) at altitudes above 2400m.

Taxonomic notes:— This species is easily distinguishable by its leaves (without conspicuous midrib) and rounded achene apex with a linear beak, without the constriction zone between beak and the main body.

Examined Material (additional):— VENEZUELA. Amazonas: Cerro Marahuaca (cumbre), 3-4 February 1982, J.A. Steyermark 126085 & M. Guariglia, N. Holmgren, J. Luteyn, S. Mori (NY!, US!).

5. Cephalocarpus neblinensis S.M. Costa, sp.nov. (Figures 7E, 10E)

Similar to Cephalocarpus rigidus due to its head-like inflorescence, but it is a dioecious species with longer male and female spikelets (C. rigidus – 3.5–4 mm long; C. neblinensis – 6–7 mm long).

Type specimen:— VENEZUELA. Amazonas: Cerro de la Neblina, Rio Yatua, more often in open places about Cumbre Camp, 1800 m, 15 November 1957, B. Maguire 42107 & J.J. Wurdack, C.K. Maguire (F! –holotype; NY!, US! – isotypes)

Dioecious herb. Male plants: Caudex 4–8 (10) mm diam. including sheaths remains and adventitious roots. Leaf: sheath pubescent, same color as leaf blades; contraligule indistinct; blade (7)10–20 × 0.09–0.2 cm, linear-lanceolate, margins and midrib (ab) sparsely ciliate, concolorous, reddish-brown to dark-brown. Inflorescence head-like or reduced panicle-like, single at each prophyll, 0–2 paracladia; culm 3–11 × ~0.1 cm, compressed, margins glabrous, pubescent or pilose. Paracladia up 1.5 cm long, 1 order; peduncle –0.8 mm long, compressed, glabrous to sparsely scabrous. Involucral bract (outermost) leaf-like: sheaths –-0.3 cm long, mainly pubescent (sometimes glabrous or pilose); blade 1-2 × ~0.2 cm, margin and surface (ab) glabrous, pubescent or pilose. Male spikelet 6.5–7 × ~1 mm, 1 flower, 5 glumes; glume (outermost) 4–4.5 mm long, glabrous or ciliolate, mucronate (mucro 0.1–1.5 mm long, glabrous or sparse ciliolate); stamen 2, connective projection short ciliate. Female plants:

Caudex ~3.8–5 mm diam. including sheaths remains and adventitious roots. Leaf: sheath pubescent, reddish-brown; contraligule indistinct; blade 7–10 × 0.1–0.12 cm, linear- lanceolate, margins and midrib (ab) sparse ciliate, concolorous, reddish-brown. Inflorescence panicle-like, 1–3 paracladia; culm –6 × ~0.1 cm, compressed, glabrous. Paracladia (lowermost) 1–2 cm long, 1–2(3) orders; peduncle –1 × 0.05 mm, compressed, margin glabrous. Involucral bract (lowermost) leaf-like: sheath up to 0.5 cm long, mouth pubescent, reddish-brown; blade –1 × ~0.15 cm, “V”-shaped, margin sparse ciliate, reddish-brown. Female spikelet ~6 × ~0.8 mm, 6 glumes, solitary; glume (outermost) 3.2–4.5 mm long, margin glabrous or ciliate, mucronate (mucro 0.8–1.7 mm long, glabrous or sparse ciliate); stigmas 3. Achene ~3.5 × ~0.9 mm, obovoid, smooth, middle and apex sparse ciliate, light yellow to reddish-brown, with a constriction zone between main body and beak; hypogynous scale up to 0.85 mm long, elliptic to oboval, ciliate, trichome –0.7 mm long (transduced); beak ~1.3 mm, conical with rounded apex, ciliate.

Etymology:— The name is a reference to the type and only known occurrence locality.

Distribution:— All collections of Cephalocarpus neblinensis are from Cerro de la Neblina, at the Amazonas (Venezuela) – Amazonas (Brazil) border. All specimens are from the Venezuelan side of Cerro de la Neblina, but this species may occur at the Brazilian side too.

Taxonomic notes:— This is another dioecious species of Cephalocarpus. The male and female plants superficially show similar morphologies, but a careful look reveals differences in the inflorescences: male inflorescences are head-like or highly reduced panicle-like and have with up to 40 spikelets; female inflorescences have 1–3 distinguishable levels of paracladia and bear relatively lax and fewer spikelets (up to 15).

Just few collections have both sexes but as the female/male individuals of these collections show nearly the same vegetative morphology, I believe that the morphological variation observed in the male plants may be similar in the female ones, e.g., regarding leaf blade length, width and ornamentation. The collections are from many different expeditions to the Cerro de la Neblina, between 1953 and 1985. Koyama (2003) had access to three collections of this species (with an asterisk - *) but did not realize that they belong to a different species and included them in Cephalocarpus rigidus Gilly ex Gleason & Killip.

Cephalocarpus neblinensisis easily distinguishable from C. rigidus due to its notably longer spikelets (up to ~6 mm long in the female spikelets and up to ~7 mm long in male spikelets) and, of course, the dioecy.

Examined Material (additional):— VENEZUELA. Amazonas: Cerro de la Neblina: Río Yatua, 30 December 1953, B. Maguire 37021 & J.J. Wurdack, G.S. Bunting (NY!)*; Rio Yatua, 15 November 1957, B. Maguire 42107 & J.J. Wurdack, C.K. Maguire (NY! – holotype; US!, F! - isotype)*; Rio Yatua, 21 December 1957, B. Maguire 42455 & J.J.Wurdack, C.K. Maguire (NY!,US!’)*; Rio Negro, 13 February 1985, B.M. Boom 5808 & A.L.Weitzman (NY!) Rio Negro, Camp III, Neblina and Massif, 16 February 1984, R. Liesner 16083 (NY!, MO!- masc and fem); Rio Negro,15 March 1984, Liesner, R. 16651 (NY!, MO!) Rio Negro, 7 February 1984, J.L. Luteyn 9429 & J.A. Steyermark (F!); Rio Negro, 26 February 1985, M. Nee 31107[-A] (NY!, MO!); Río Negro, río Mawarinumma, Rio Baria, 7 February 1984, J.L. Luteyn 9429 & J.A. Steyermark (MO!, US!); Cerro de la Neblina, 7-8 February 1984, J.A.Steyermark 129827 & J.L. Luteyn (MO!)

6. Cephalocarpus “chimanta 2” S.M. Costa, sp.nov. (Figures 7F, 10G)

Similar to Cephalocarpus confertus, but it has bigger leaves (leaf blades ~15 × ~0.5 cm, versus (3)6–10 × 0.2–0.25 cm in C. confertus), the achene with a triangular beak without a constriction zone between the main body and the beak (the achene is conical to short cylindrical and with the constriction zone in C. confertus)

Type specimen:— VENEZUELA. Bolivar: Chimantá Massif, among southwest-facing sandstone bluffs of Chimantá-tepuí (Torono-tepui) (near Southern corner), 1700 m, 19–20 May 1953, J.A. Steyermark 75480 (NY! – holotype, F! - isotype)

Monoecious herb: sexes intermingled. Caudex 5–12 mm diam. including sheaths remains and adventitious roots. Leaf: sheath glabrous, reddish-brown after dry; contraligule indistinct; blade ~15 × ~0.5 cm, linear-lanceolate, “V”-shaped and flattened, sparsely minute ciliate or glabrous except for the blade base margins sparsely minute ciliate, concolorous, reddish- brown. Inflorescence head-like (appearing sessile), single at each prophyll; culm <1 cm, compressed, margins ciliate. Involucral bract (lowermost) leaf-like: sheath –1.3 mm long, glabrous, reddish-brown after dry; blade –6 × –1.1 mm, “V” in cross section, margin sparsely

minute ciliate, reddish-brown after dry. Male spikelet ~6 × ~1mm, 5–6 glumes, 1 flower; glume (outermost) ~3.2 mm long, glabrous, mucronate (mucro ~1.2 mm long); stamens 3, connective projection short conical and glabrous. Female spikelet ~4.5 × ~1.2 mm, 5–6 glumes, 1 flower; glume (outermost) ~3 mm long, glabrous, mucronate (mucro 1 mm long); stigmas not observed. Achene ~3.2 × ~1.1 mm, ellipsoid-hexagonal, smooth, ciliate at the apex, main body continuous to the beak; hypogynous scales ~3 mm, lanceolate, trichomes up to 1.7 mm long; beak ~0.85 mm long, triangular, pubescent.

Distribution:— Known only to the type-locality.

Taxonomic notes:— This intriguing species resembles a plus-sized Cephalocarpus confertus in its vegetative morphology, but additionally to its clearly bigger size, the achene characters are notably different, lacking the constriction zone between the body and the beak, with lanceolate hypogynous scales and a triangular beak. It is only registered for the type-locality and by a single collection.

7. Cephalocarpus “neblina dioica 2” S.M.Costa, sp.nov. (Figures 10F)

Similar to Cephalocarpus confertus but it is a dioecious species and has broader leaf size ( 0.35-0.48 cm width, while 0.2–0.25 cm in C. confertus). Also, female inflorescences are laterally and terminally inserted at the plants

Type specimen:— VENEZUELA. Amazonas: Depto Río Negro, Cerro de la Neblina, 1880 m, 0°52–053’N 66°05’W, 7–8 February 1984, J.A. Steyermark 129812 & J.L. Luteyn (MO! – holotype, female plant).

Dioecious herb. Caudex up to 3 cm including sheaths remains and adventitious roots. Leaf congested at the caudex apex; sheath glabrous with sparse cilia at mouth; contraligule indistinct; blade 6–9 × 0.35–0.48 cm, linear-lanceolate, “V”-shaped and flattened, margins and midrib (ab) ciliate and midrib (ad) minute ciliate, concolorous or discolorous, reddish- brown after drying. Male inflorescence head-like, 1–3 at each prophyll; culms –5 × ~0.2 cm, compressed, margins minutely ciliate or glabrous. Male involucral bract (lowermost) leaf- like: sheath <1 mm long, glabrous; blade –9 × 0.9 mm, “V”-shaped, margin sparsely minute ciliate. Male spikelet ~6 × ~1mm, 4–5 glumes, 1 flower; glume (outermost) ~5mm long, glabrous, mucronate (mucro ~1 mm long); stamen (not available). Female inflorescence head-

like, lateral and terminal, solitary when lateral and in groups of up to 10 when terminal; culms <1 cm, compressed, margins ciliate. Female involucral bract (lowermost) absent. Female spikelet ~3.5 × ~0.9 mm, 5–6 glumes; glume (outermost) ~3.5 mm long, glabrous, mucronate (mucro ~3.5 mm long); stigmas not observed. Achene immature, ciliate at apex, with a narrow neck between the main body and the beak; hypogynous scales shortly ciliate; beak clavate, ciliate.

Distribution:— Only registered from the type-locality, Cerro de la Neblina, at the Amazonas (Venezuela) – Amazonas (Brazil) borders, about 1800–1900 m alt. All specimens are from the Venezuelan side of Cerro de la Neblina, but this species may also occur at the Brazilian side.

Taxonomic notes:— This species resembles a magnified Cephalocarpus confertus at its vegetative characters, but besides the bigger caudex and leaf size, it is a dioecious species. Another interesting feature is the lateral and terminal inflorescences placement at the female plant. All achenes available are immature, but already show the clavate beak typical of Cephalocarpus (sensu Koyama 2003).

Examined Material (additional):— VENEZUELA. Amazonas: Depto Río Negro, Cerro de la Neblina, 1820–1880 m, 0°52’N 66°05’W, 1 February 1985, B.M. Boom 5507 & A.L. Weitzman, C. Brewer-Carías, (NY! – male plant)

8. Lagenocarpus setaefolius S.M.Costa (Figures 8, 10H)

Superficially similar to Lagenocarpus rigidus and L. guianensis, but with distinguishably reduced and superficially monomorphic paracladia that bear spikelets of both sexes, unlike the other two species.

Type specimen:— VENEZUELA. Amazonas: (savannas near) Santa Cruz, a small village Rio Atabapo 1 km N of the confluence of the Rio Temeni and the Rio Atacavi, 17–18 November 1979, W.W. Thomas 2656a & G.K. Rogers (NY! – holotype; MICH (not seen)- isotype)

Monoecious herb. Shoot structures not observed. Leaf: sheaths brown; contra-ligule not observed; blades (unable to measure) × ~1 cm, linear-lanceolate, margins and surface (ab)

rarely long-ciliate, margins minutely scabrous. Inflorescence 7 paracladia, sexes intermingled at clusters (apparently male ones only at basal paracladia), cluster composed of 2–3 female and 2–3 male spikelets, 5 sterile bracts similar to fertile ones; culm 125 × 0.5 cm, compressed at base and becoming trigonous toward the apex, glabrous. Paracladia (lowermost) up to 5.5 cm long, 1–3 order at the basal paracladia, monomorphic; peduncle up to 2.5 cm long, compressed, sparsely ciliate. Involucral bract (lowermost) 7 × 0.4 cm, leaf-like, “V”-shaped, sparsely scabrous, erect and rigid; sheath 2 mm long to the mouth (2.5 mm including “lingueta”), sparsely ciliate at the side opposite to the blade, mouth glabrous, light brownish- green. Male spikelet ~3.5 × ~1 mm, 5–7? glumes, 1–2 flowers; stamens 2, connective projection minute, conical, with tiny trichomes. Female spikelet ~3 × ~0.5 mm, 5glumes, 1 flower; stigma 3. Glume (outermost) ~2.8 mm long (male) and ~2.4 mm long (female), glabrous, thin membranaceous, only female mucronate; mucro <0.1mm long. Achene (immature) 4.4 × 1.6 mm, ovoid, smooth; hypogynous scales 0.6 mm long, oboval, rare trichomes <0.1 mm long; beak ~2 mm long, conical, glabrous.

Etymology:— The name refers to the rigid, sharp and arrow shaped bracts (and probably also the leaves) of this species

Distribution:— Known only from the type-locality. There was no coordinate information at the specimen, the coordinates inserted at the map was estimated using the locality information available in the label.

Taxonomic notes:— I (S.M.C.) only examined a depleted specimen deposited at NYBG herbarium, donated by the Herbarium of Michigan University but due to such peculiar inflorescence character states this is a new species with no doubt. Lagenocarpus setaefolius is possibly related to L. guianensis, L. sabanensis and the “L. rigidus” complex, but differs from all these by its short paracladia and bisexual clusters.

9. Lagenocarpus rubriglumis S.M. Costa, sp. nov (Figures 9C-D, 10I)

Similar to Lagenocarpus pendulus, but L. rubriglumis differs by the less branched lowermost paracladia and the obovoid achene with irregularly rugose surface and pubescent beak and side ridges.

Type-specimen:— BRAZIL. Roraima: Caracaraí, PARNA Viruá, “Estrada Perdida”, 1°16’46”N/60°59’20”W 192 m, 21 September 2010, S.M. Costa 793 & T.D.M. Barbosa, K.G. Cangani (UEC! – holotype, INPA! – isotype)

Monoecious herbs. Leaf basal; sheaths white-ciliate, light reddish-brown after drying; contraligule densely white ciliate/pubescent, pilose at mouth region; blades 50–60 × 0.5–1 cm, linear-lanceolate, margins minute scabrous, reddish-brownish after dry. Inflorescence 5– 10 paracladia, sexes at different paracladia (female apical, male basal); culms –120 × 0.3– 0.45 cm, compressed at base, trigonous at the apex, glabrous. Paracladia (lowermost) up to – 15 cm long, 3 orders; peduncle up to 5.5 cm long, compressed, ciliate and densely ciliate at base. Involucral bract (lowermost) –13 × ~0.45 cm, leaf-like, “V”-shaped, green, margin sparsely scabrous; sheath 2-3 mm long, ciliate, yellowish-brown. Male spikelet –2.2 × –0.55 mm,5–6 glumes, 2–3 flowers; anthers 4, connective projection short with few trichomes at the apex. Female spikelet –5.25 × –1.6 mm, 7 glumes, 1 flower, solitary or rarely 2; stigmas 3. Glume (outermost) –2.8 mm long (male) and ~3 mm long (female), margin ciliate, mucronate?; mucro –0.5 mm (male) and 0.9 mm (female) long. Achene 3.2 × 1.2 mm, balloon-like, irregularly rugose (after dry?!), ciliate at beak and lateral ridges; hypogynous scales 1 mm long, orbicular, trichomes 0.6 mm long; beak 1.2 mm long, conical, ciliate

Etymology:— The name refers to the ocher coloration of the glumes at spikelet maturity, especially female ones. This character state remains conspicuous in the dried herbaria specimen

Distribution:— This species occurs in Brazilian and Venezuelan white-sand savannas, a vegetational formation biogeographically related to the Guiana Shield highlands. The white- sand savannas areas, and its nutrient poor soils, are scattered in the Northern margin of the Negro River associated with the erosion of the Guiana Shield.

Taxonomic notes:— This species can be distinguished from similar ones (L. pendulus, L. glomerulatus) by the branching of the lowermost paracladia: mostly 3 orders, against (5) 6–8 in L. pendulus and 1(2–3) in L. glomerulatus; and achene shape: obovoid achene with irregularly rugose surface and pubescent beak and lateral ridges, compared with orbicular achenes with smooth surface and pubescent beak and glabrous ridges in L. pendulus and elliptic to orbicular achene with irregularly rugose surface and pubescent beak and glabrous ridges in L. glomerulatus. The mature diaspores of L. rubriglumis are composed by the

achene and subtending glumes and a fragment of its peduncle and presents a daisy-like aspect, due to the increased length and divergent glume and a vivid reddish-brown color of the glumes.

Examined Material (additional):— BRAZIL: Amazonas: Manaus, Campina das Pedras, BR 174 km 115, Corrêa, M.P.F. 76 & Miranda, F.E.; Miranda, M.C.C.,28 June 1984 (INPA!); Manicoré, Proj RADAM/BRASIL SB 20 XD, Mota, C.D.A. s/n ,21 August 1976 (INPA!); Nova Prainha, Proj RADAM/Brasil, SB 20 ZD, pto 87, Mota, C.A.D. & Monteiro, O.P. ,23 July 1976 (INPA!); Presidente Figueiredo, Igarapé das Lajes, Miranda, F.E. 890 & Miranda, M.C.C.; Correa, M.P.F. ,21 June 1984 (INPA!); Presidente Figueiredo, Rebio Uatumã, Zartman, C.E. 5974 & Carvalho-Sobrinho, J.G. de; Silva, K.M.; Assunção, P.A. ,11 August 2006 (INPA!); Roraima: Cantá, Costa, S.M. 1075 (UEC!); Caracaraí, PARNA Viruá, Barbosa, T.D.M. 1275 & Costa, S.M. (UEC, INPA!); Caracaraí, along Manaus-Caracaraí rd, km 130, Berg, C.C. P19505 & Lleras, E.; Holley, C.; Monteiro, O.P. ,13 November 1973 (INPA , NY, MO!); Caracaraí, PARNA Viruá, Carvalho, F.A. 943 , (INPA!); Caracaraí, Manaus-Caracaraí road km 130, Conant, D.S. 1159 & Tryon, Rolla, Tryon, A.; Windish, P. ,27 September 1974 (INPA!); Caracaraí, PARNA do Viruá, Grade do PPBio, Costa, F.R.C. 1691 ,15 January 2006 (INPA!); Caracaraí, PARNA Viruá, Costa, F.R.C. 790 (INPA!); Caracaraí, PARNA Viruá, Estrada Perdida, Costa, S.M. 1047 (UEC!); Caracaraí, PARNA Viruá, Estrada Perdida, Costa, S.M. 1049 (UEC!); Caracaraí, PARNA Viruá, Costa, S.M. 790 &Barbosa, T.D.M.; Cangani, K.G. (UEC, INPA!); Caracaraí, PARNA Viruá, Costa, S.M. 793 &Barbosa, T.D.M.; Cangani, K.G. (UEC, INPA!); Caracaraí, PARNA Viruá, Holanda, A.S.S. 524 (INPA!); Caracaraí, Estrada Manaus-Caracaraí km 125, Rodrigues, W. 10471 &Coelho, L.; Guedes, J. ,3 November 1983 (INPA!); Caracaraí, Estrada Manaus-Caracaraí Km 130, Igarapé das Lajes, Rodrigues, W. 9292 & Loureiro, A.; Coelho, D. ,25 May 1974 (INPA!); Mucajaí, 6km após Mucajaí, rumo a Colônia do Apiau, Miranda, I.S. 177 & Eiten, G. ,19 September 1993 (INPA!); São Luiz, BR 174 km 350 (Manaus-Caracaraí), Teixeira, L.O.A. 1650 ,15 September 1985 (INPA!); VENEZUELA: Amazonas: , Rio Guainia, Maguire, B. 41761 & Wurdack, J.J.; Keith, W.M. , (NY!); Rio Guainia, Maguire, B. 41761 & Wurdack, J.J.; Keith, W.M., 7 October 1957 (NY, F!); Atures, caño "Cabeza de Manteco", Guanchez, F. 3503 & Melgueiro, E. (NY!); Atures, caño "Cabeza de Manteco", Guanchez, F. 3503 & Melgueiro, E., 12 November 1984 (NY!); San Carlos de Rio Negro, Clark, H.L. 6782 (NY!); San Carlos de Rio Negro, Rio Negro, Brazo Casiquiare, Clark, H.L. 6782, 11 September 1978 (INPA, NY, F!).

10. Lagenocarpus “glomerulatus-like” S.M. Costa, sp. nov. (Figures 9A-B, 10J)

Similar to Lagenocarpus glomerulatus, but differs by the larger culms, regularly branched and longer paracladia, and obovoid and completely glabrous achenes with smooth surface.

Type specimen:— VENEZUELA. Amazonas: Atures, sobre afloramiento granitico en raudal “Pereza”, en el Río Autana, 100–115 m, 4°48’N 67°18’W, 9 November 1984, F. Guanchez 3401 & E. Melgueiro (NY! – holotype)

Monoecious herbs. Stem structute rhizome?caudex?. Leaves: sheaths ciliate towards the apex, apparently glaucous; contra-ligule not distinguishable?; blades –120? × –1.8 cm, linear- lanceolate, margin and midrib (ab) sparsely scabrous, pale brownish-green after dry. Inflorescence terminal, open, panicle-like, –13? paracladia, sexes intermingled in paracladia (female generally at apex of paracladia and/or inflorescences), one sterile bracts similar to fertile ones; culms –150 × 0.6 cm, trigonous, glabrous. Paracladia up to 14 cm long, 3 order at the basal paracladia, monomorphic (spikelets dimorphic); peduncle up to 10 cm long, compressed, ciliate, the ones that ends in spikelets mostly glabrous with a densely white ciliate area where it usually breaks. Involucral bract (lowermost) –80 × –1.5 cm, leaf-like, “V”-shaped, pale brownish green, margin and midrib (ab) sparsely scabrous; sheath -6 mmm long, mostly glabrous to minute ciliate especially at mouth, light greenish-brown. Male spikelet ~3 × ~0.6 mm, glumes 3–5, 2–3 flowers; stamens 1–2, connective projection conical with trichomes. Female spikelet –~2.8 × ~1.5 mm, 5 glumes, 1 flower, 1–2; stigmas 3. Glume (outermost) –2.35 mm long (male) and –2.2 mm long (female), margin ciliate, female one mucronate; mucro 0.5 mm long (female only). Achene 3.3–3.7 × 1.7–1.8 mm, obovoid, smooth, glabrous; hypogynous scales ~0.5–1 mm long, irregular or orbicular, trichomes ~0.3; beak –1.9 mm long, round, glabrous, larger than the achene body, with a rigid, abruptly narrow, conical style residue at apex.

Distribution:— known only from the type-locality.

Taxonomic notes:— This species probably belongs to the Amazonian Lagenocarpus clade, it resembles L. glomerulatus in gross morphology and its achene looks like the one of L. celiae. Even if it is known only by the type-specimen, it has sufficient distinguishable characters to permit its recognition as a new species.

Conclusions Except for Lagenocarpus rubriglumis, that has been recently collected, the above described species are based on collections from the 1950’s to the 1980’s. At these years, there were intense collecting efforts focused on the Guiana Shield tepuis. Some of the specimens had annotations highlighting the unique states of their characters and even draft descriptions and sketches, mostly probably made by the collectors and herbarium researchers (for example, J.A. Steyermark, G. Davidse). Although most of those characters were enough to recognize the new species, the lack of specialists in the tribe Cryptangieae along with the scattered bibliography about Cryptangieae taxa may have delayed their proper description.

Figure 7:– Male and female spikelets and achene of: A. Cephalocarpus chimantensis (voucher: J.A. Steyermark 128931), B. Cephalocarpus illustrata (voucher: O. Huber 10949), C. Cephalocarpus “lowlandia” (voucher: B. Maguire 42341), D. Cephalocarpus martinhae (voucher: J.A. Steyermark 129574), E. Cephalocarpus neblinensis (voucher: B. Maguire 42107), F. Cephalocarpus “chimanta 2” (voucher: J.A. Steyermark 75480) [scale (black bar)= 1 mm] (S.M.Costa original drawings).

Figure 8:–: A. Lagenocarpus setaefolius (voucher: W.W. Thomas 2656a & G.K. Rogers, NY! – holotype)

Figure 9:– Lagenocarpus “aff glomerulatus” (photos; voucher: F. Guanchez 3401): A. male and female spikelets and achene, B. hypogynous scale (detail); Lagenocarpus rubriglumis (photos): C. male and female spikelets and achene (voucher: C.C. Berg P19505), D. hypogynous scale (detail; Zaruchi 2021) [scale (black bar)= 1 mm]. A

B C

Figure 10:– Distribution maps of: A. Cephalocarpus chimantensis, B. Cephalocarpus illustrata, C. Cephalocarpus lowlandia; D. Cephalocarpus martinhae, E. Cephalocarpus neblinensis, F. Cephalocarpus “neblina dioica 2”; G. C. Cephalocarpus “chimanta 2”; H. Lagenocarpus setaefolius, I. Lagenocarpus rubriglumis; J. Lagenocarpus aff glomerulatus.

A B

C D

E F

G H

I J

CHAPTER 4 – An updated taxonomic treatment to Cephalocarpus Nees (Cryptangieae, Cyperaceae) Additional authors: Volker Bittrich

(Intended submission at ????)

ABSTRACT – We provide an updated taxonomic treatment of Cephalocarpus, Cyperaceae. This sedge genera is mostly restricted to highlands at Guiana Shield and gather 25 species of perennial herbs with caudex development, lateral inflorescences, unisexual flowers and spikelets and conspicuous hypogynous scales generally with long trichomes.We provide descriptions and synonyms of the genus and its species, identification key, comments on taxonomy, morphological and geographical, a complete list of examined material and illustrations. Keywords – Cryptangieae, Everardia, white-sand savanas

RESUMO – Esse manuscrito traz o tratamento taxonômico atualizado de Cephalocarpus, Cyperaceae. Esse gênero é especialmente restrito às terras altas do Escudo das Guianas e reune 25 espécies de ervas perenes com desenvolvimento de cáudex, inflorescências laterais, flores e espiguetas unissexuadas e escamas hipóginas conspícuas geralmente com longos tricomas. São fornecidas descrições e sinonímia para o gênero e suas espécies, chaves de identificação, comentários taxonômicos, sobre morfologia e distribuição geográfica, uma lista de material examinado e ilustrações.

Introduction The sedge genus Cephalocarpus Nees is mostly restricted to Venezuelan-Guyanese highlands, with some scattered occurences in Brazil, Colombia, Ecuador, Peru and Surinam (Gilly 1941a, 1941b, 1942, 1951; Koyama & Maguire 1965; Koyama 2003, Costa et al. CHAPTER 3, Figure 1A). As other representatives of the tribe Cryptangieae, its species present unisexual spikelets and achenes with hypogynous scales, but can be distinguished from the other genera by the lateral inflorescence and generally long fimbriate hypogynous scales (Gilly 1941a, 1941b, 1942, 1951; Koyama & Maguire 1965; Koyama 2003, Costa et al. CHAPTER 3).

Recently, Costa et. al. (CHAPTER 1 – 3) investigated and reorganized the Cryptangieae diversity using molecular and morphological data. Cephalocarpus Nees (Flora brasiliensis 2(1): 162, pl.18. 1842) circumscription now includes the species formerly accepted in Everardia Ridley (Timehri 5: 210. 1886) (CHAPTER 2) and other seven new species that were described to the genus (CHAPTER 3). After that, Cephalocarpus became the most species rich genus of Cryptangieae (25 Cephalocarpus spp. in a total of 57 Cryptangieae spp.).

Here, we provide an updated taxonomic treatment to Cephalocarpus Nees, including the necessary information and key to a clear recognition of the species (descriptions to genus and species, identification keys to species, synonym, taxonomical, morphological and geographical comments, examined material and illustrations and distribution maps).

Material and Methods We analyzed about 500 herbaria specimens at Brazil (MG, INPA, UEC) and U.S.A. (NY, F, MO, US) and additional type collections in the JStor Global Plants Platform (http://plants.jstor.org/, last access: December 2017). The type-specimens analyzed directly have the herbaria acronym followed by an exclamation point (“!”) and the type-specimens analyzed via online platforms have the herbaria acronym followed by a hash sign (“#”).We accessed the original description of Cephalocarpus and its species and specialized bibliography to Cryptangieae at physical libraries (MOBOT, NYBG and UNICAMP) and online platforms. The descriptions and identification key include the morphological variation of examined specimens and we include additional information under comments, when necessary. Diagnostic characters and states of characters exclusive of a single species are

mentioned only at the identification key and/or “taxonomic notes” to avoid to extensive descriptions. The illustrations are based on examined material and we also refer to the illustrations available in previous treatments (see Appendix 4A). We based the geographical information in the literature and in our own material list, as also the maps. We used the software BRAHMS 7.7 (University of Oxford 2015) and ArcGIS 10.5 (ESRI 2015) to help with the data management of examined material, coordinates and elaboration of distribution maps.

Results

Taxonomic Treatment Cephalocarpus Nees, Flora brasiliensis 2(1): 162, pl.18. 1842. Everardia Ridley, Timehri 5: 210. 1886. Lagenocarpus subgen Cephalocarpus (Nees) H. Pfeiff., Repertorium Specierum Novarum Regni Vegetabilis 18: 91. 1922.

[based on Gilly (1942), Koyama (2003) and Costa et al. (CHAPTER 2)] Monoecious or dioecious terrestrial and perennial herbs, growing on rocks or epiphytic; at monoecious plants, the sexes can be intermingled or separate at paracladia or cluster level. Caudex erect or decumbent, sometimes branched, covered by adventitious roots and residues of leaf-sheaths usually disintegrating into parallel fibers. Leaf blades linear to lanceolate, 0-1- costate, generally “V”-shaped in cross section (sometimes “M”-shaped at apex), completely glabrous to lanose, concolorous or discolorous, green or reddish-brown after drying; loosely to densely grouped at apices of caudices. Inflorescence lateral (also terminal in Cephalocarpus “neblina dioica 2” female plants), lax to highly congested, single or up to 8 at each prophyll, panicle-like or head-like. Involucral bract (lowermost) leaf-like, subulate; sheaths of the same color of blades or darker-colored. Male spikelets with lower glumes empty and upper glumes fertile, glumes imbricated; stamens 2-6 (7-9), exserted, the connective prolonged between the thecae into a glabrous to pubescent conical projection. Female spikelets 1-flowered, solitary (rarely 2-3/cluster), 4-7 (-9) glumes, irregularly imbricated; style trifid (abnormally 2 or 4-branched). Achene broad or narrow obovoid, ellipsoid, oblong or ovoid, trigonous to obscurely trigonous in transverse section, generally trisulcate, main body continuous to the beak of the achene or with a constriction zone between

the main body and the beak of achene, rarely the apex of main body truncate or rounded, generally pubescent at the apex; hypogynous scales 3, short or long fimbriate; beak linear, triangular, conical or cylindrical, glabrous to pubescent.

Type species: Cephalocarpus dracaenula Nees

Its species can be distinguished from other Cryptangieae representatives by the lateral inflorescence and the achenes with conspicuous hypogynous scales with short to long fimbriate margins. The genus is mostly restricted to Venezuela, Guyana, Colombia and northern Brazilian Amazon, but also occurs in Ecuador, Peru and Suriname localities. Its species are more diverse at higher altitudes and generally occur on nutrient poor sandy or rocky soils. Total species: 25 species. (MAPS 1A-B) Fig. 2B-C (Koyama & Maguire 1965)

Identification key to species

1. Dioecious species ...... 2 1’. Monoecious species ...... 4

2. Inflorescence always lax and panicle-like ...... 10. Cephalocarpus flexifolium 2’. Inflorescence head-like (highly congested panicle-like) ...... 3

3. Leaf up to 0.2 mm width; female inflorescence lateral; female spikelets ~6 mm long ...... 18. Cephalocarpus neblinensis 3’. Leaf 0.35–0.4 mm width; female inflorescences lateral and terminal in the same plant; female spikelets ~3.5 mm long ...... 25. Cephalocarpus “neblina dioica 2”

4. Inflorescence 3-8 at each prophyll ...... 5 4’. Inflorescence single at each prophyll ...... 7

5. Inflorescence lax panicle-like for the most part, with accessoral 1--2 head-like inflorescences; achene beak continuous to the main body of achene. 4. Cephalocarpus debilis 5’. Inflorescence only head-like; achene with a constriction zone between the main body and the beak……………………………………………………………..………………………….6

6. Loosely aggregated leaves at the caudex apex; female spikelet glume with glabrous or sparsely ciliate carena; stamen connective projection ciliate .… 7. Cephalocarpus dracaenula 6’. Highly congested leaves at the caudex apex; female spikelet glume with pubescent to densely pubescent carena; stamen connective projection glabrous 3. Cephalocarpus confertus

7. Inflorescence head-like (or reduced panicle-like) ...... 8 7’. Inflorescence lax panicle-like ...... 12

8. Achene with a constriction zone between the main body and the beak; beak conical ...... 9 8’. Achene main body continuous to the beak or with rounded apex; beak pyramidal or linear ………………………………………………………………………………………………...11

9. Female spikelets generally in clusters of three; achene broadly obovoid…………………….………………………………..... 19. Cephalocarpus obovoideus 9’. Female spikelets generally single; achene oblong to broadly ellipsoid …………………..10

10. Plants (almost) completely glabrous; male spikelet 3.5–3.7 mm long; achene broadly ellipsoid with long-conical beak (~1 mm long, 1/3 of the achene) .22. Cephalocarpus rigidus 10’. Plants pubescent to densely pubescent; male spikelet 4.5–6 mm long; achene oblong with short-conical beak (~0.7 mm long, 2/5 of the achene) ….. 12. Cephalocarpus longibracteatus

11. Leaves with no conspicuous midrib; spikelets peduncles up to 1 mm long; achene oblong; beak linear ...... 16. Cephalocarpus martinhae 11’. Leaves with conspicuous midrib; spikelets sessile; achene ellipsoid-hexagonal; beak pyramidal/triangular ...... 24. Cephalocarpus “chimanta 2”

12. Inflorescences with culms up to 5 cm long; achene with truncate apex and short linear beak ………………………………………………...……….. 2. Cephalocarpus chimantensis 12’. Inflorescences with culms more than 5 cm long; achene with a constriction zone below or continuous to a apical conical beak ...... 13

13. Achene with a constriction zone bellow the conical beak ..... 11. Cephalocarpus illustrata 13’. Achene continuous into a short or long conical beak ...... 14

14. Glumes of female spikelets recurved at maturity ...... 21. Cephalocarpus recurviglumis 14’. Glumes of female spikelets not recurved at maturity ………………….……………… 15

15. Achene completely glabrous ……………………………………………………………. 16 15’. Achene ciliolate, ciliate or pubescent, at least the apex ………………….……………. 17

16. Leaf adaxial surface glabrous and shiny; with a bromeliad aspect; involucral bract blade up to 2 cm long ………………………………….………...…… 20. Cephalocarpus ptariensis 16’. Leaf adaxial surface ciliate to pubescent; without a bromeliad aspect; involucral bract blade >2–3 cm long …………………………...…………....… 9. Cephalocarpus erecto-laxus

17. Female spikelets ≥6 mm long ...... 18 17’. Female spikelets <6mm long ...... 20

18. Female spikelets 7.5–10 mm long; glumes dark-brown ...... 13. Cephalocarpus longifolius 18’. Female spikelets ~6–7 mm long; glumes paleaceous or reddish-brown ...... 19

19. Leaves conspicuously pubescent; glumes paleaceous ...... 1. Cephalocarpus angustus 19’. Leaves always glabrous; glumes reddish-brown ...... 5. Cephalocarpus diffusus

20. Leaves with rugose surface ...... 21 20’. Leaves with smooth surface …………………...... 22

21. Female spikelets 1.5–2.5 mm long; achene ellipsoid ………...... 8. Cephalocarpus duidae 21’. Female spikelets 3–6 mm long; achene broad-ellipsoid, obovoid, ovoid ……………………………………………..…………...……… 17. Cephalocarpus montanus

22. Achene shorter than subtending glumes at maturity ...... 15. Cephalocarpus maguireanus 22’. Achene longer than subtending glumes at maturity …….…………….…...... 22

23. Leaves conspicuously distichally arranged ...... 6. Cephalocarpus distichous 23’. Leaves tristichously arranged, rosette-like ...... 24

24. Hypogynous scales >2 mm long (including the trichomes) …………...... 25 24’. Hypogynous scales less than 1.5 mm long (including the trichomes) …………………. 26

25. Leaf adaxial surface pubescent; hypogynous scales ~2.5 mm long (including the trichomes) ...... 23. Cephalocarpus vareschii 25’. Leaf adaxial surface glabrous and shiny; hypogynous scales 0.5-0.7 mm long (including the trichomes) ………………………………….……...………. 20. Cephalocarpus ptariensis

26. Leaves up to 0.4 cm wide ...... 14. Cephalocarpus lowlandia 26’. Leaves more than 0.5 cm wide……………………………. 17. Cephalocarpus montanus

Species descriptions

1. Cephalocarpus angustus (N.E. Brown) S.M. Costa, (CHAPTER 2)

Basionym: Everardia angusta N.E. Brown, Transactions of the Linnean Society of London, 2nd series: Botany 6: 73. 1901. Everardia gracilis Gilly, Bull. Torrey Club 68: 26, f.2, b-b1. 1941. Type specimen: British Guyana, Mt. Roraima, G.H.H. Tate 426 (NY!, MSC#-achenes only)

Everardia steyermarkii Gilly, Fieldiana Bot. 28 (1): 55. 1951. Type specimen: Venezuela, Ptari-tepui, J.A. Steyermark 59781 (F!, holotype; NY!, US!, VEN#, MSC# - spikelets only, ISC#) [Cryptangium stamineum N.E. Brown ex C.B. Clarke, Kew Bull. Add. Ser. 8: 135. 1908. (nomen nudum)]

Type specimen: BRITISH GUYANA: summit of Mt. Roraima, 8600ft., McConnell & Quelch 676, (Autumn)1898 (#K000881766 – lectotype; #K000881765 - isolectotype)

Fig. 2 a–b’ (Gilly 1941); Fig. 5 A-C (Koyama & Maguire 1965); Fig. 6 (Koyama 2003) (Appendix 3);

MAP 1C

Monoecious herb: sexes at separate paracladia (male basal/female apical), paracladia monomorphic. Caudex ~4 mm diam. ((7)10–20(25) mm diam. including fibrous sheaths remains and adventitious roots), mainly resembling a rhizome. Leaf: sheath glabrous or sparsely ciliate, brownish to dark-brown; contraligule indistinct; blade (10)18–30(40) ×(0.15)0.25–0.4 cm, linear to linear-lanceolate, margins, midrib and abaxial surface sparsely to densely pubescent (especially at young leaves), generally discolorous, light green (adaxial) and yellowish-green (abaxial). Inflorescence single at each prophyll, panicle-like, 4-7(9) paracladia; culms (25) 30–50 (80) ×(0.1) 0.13–0.2 (0.25) cm, compressed, glabrous to densely pubescent. Paracladia (lowermost) -10(20) cm long, 1-3 orders; peduncle 3.5–7.5(19.5) cm long, compressed, glabrous to pubescent. Involucral bract (lowermost): leaf-like; sheath(1.3) 1.5–2.5(3) cm long, glabrous to densely pubescent, dark-brown; blade (1)1.5–2.5(3.5) ×0.1– 0.15(0.2) cm, no midrib or “M”-shaped, green, glabrous to pubescent. Male spikelet (6)7–8 × 1.5–1.9 mm, 6-7 glumes, 2-4 flowers, 1-2 each clusters; glume (outermost) ~5.3 mm long, glabrous to pubescent, mucronate (mucro 1.6 mm long); stamen 4-5, connective projection short-linear with long and short trichomes. Female spikelet ~6–7 × (1) 1.4–1.7 mm, 4-5(7) glumes; glume (outermost) ~6.2 mm long (female), glabrous to pubescent, mucronate (mucro 1.6 mm long); stigma 3 (4). Achene ~6 × 1.4 mm, oblong-elliptic, smooth, paleaceous, main body continuous to the beak; hypogynous scale ~1.6 mm long (only trichomes visible); beak ~3 mm long, narrow-conical, pubescent.

Distribution: This species occurs at Venezuela and Guayana southern regions and may occur at the borders with Brazil.

Taxonomic notes: This species is recognizable by its nearly completely pubescent leaves (except adaxial surface), generally green after drying, conspicuously dark-brown to blackish involucral bracts sheaths, female spikelet 5–7 mm long, paleaceous glumes and achenes ~6 mm long. The morphologically closest species are Cephalocarpus longifolius and C. lowlandia.

We can distinguish Cephalocarpus angustus from C. longifolius and C. lowlandia according to the leaf characters: C. longifolius has margins and midrib (abaxial) sparsely scabrous to scabrous and ciliate surface, generally brownish-green after drying and C. lowlandia has only the leaf margins sparsely pubescent. Cephalocarpus longifolius and C. lowlandia present dark-brown to blackish glumes.

The spikelets lengths are also different: C. longifolius shows female spikelets 8–10 mm long and male spikelets 8-11(13) mm long, and C. lowlandia shows female spikelets 2– 3.5 mm long and male spikelets 3–5.5 mm long. The achenes are 7–9.5 mm long in Cephalocarpus longifolius specimens and 3–5.5 mm long in C. lowlandia.

Examined material: GUYANA: Cuyuni-Mazaruni region: Mt Maringma summit, Clarke, H.D. 11722, 15 June 2004 (NY!, US!) , Mt Maringma, summit, Clarke, H.D. et al. 11758, 17 June 2004 (US!); Mt. Roraima, Persaud, R. 159, 2 November 1973 (NY!); Mt Roraima, Warrington, J.F. et al. 129, 1978 (NY!). VENEZUELA: Amazonas: Atabapo, Plateau Cerro Marahuaca above Salto Los Monos, Liesner, R.L. 17963, 25 February 1985 (NY!); Atabapo, Cerro Marahuaca, cumbre, Maguire, B. et al. 65565, 15 January 1981 (NY!); Atabapo, Cima del Cerro Marahuaca sur, Rodriguez, H. 2673, 13 October 1988 (NY!); Atabapo, Cerro Marahuaca, cumbre, Steyermark, J.A. et al. 124437, 16 February 1981 (NY!, US!); Atabapo, Cerro Marahuaca, Steyermark, J.A. et al. 126034,2 February 1982 (NY!); Atabapo, Cerro Marahuaca, Steyermark, J.A. et al. 126325, 9 February 1982 (NY!); Atabapo, Cerro Marahuaca, Steyermark, J.A. 129073 & Delascio, F., 26 March 1983 (NY!); Atabapo, Cerro Marahuaca, Steyermark, J.A. 129251 & Delascio, F., 30 March 1983 (NY!) Atabapo, Cerro Marahuaca, Steyermark, J.A. 129304 & Delascio, F., 30 March 1983 (F!); Atabapo, Cerro Marahuaca, cumbre, Steyermark, J.A. 129518, 10 October 1983 (NY!); Atabapo, Cerro Marahuaca, Steyermark, J.A. 130777-A & Hoslt, B., 26 February 1985 (US!); Atabapo, Cerro

Marahuaca, Steyermark, J.A. 130830 & Holst, B., 26 February 1985 (NY!); Atabapo, Cerro Marahuaca, Steyermark, J.A. et al. 216034, 2 February 1982 (NY!); Bolivar: Ilu-tepui, Liesner, R. 23340, 15 April 1988 (US!); PN Canaima, Roraima Tepuy, Michelangeli, F.A. 004, 10 October 1992 (INPA!); Ptari-tepui, Moore Jr, H.E. et al. 9816,14 August 1970 (US!); Cerro Roraima, headwaters of Rio Arabapó, Steyermark, J.A. et al. 112542, 26 August 1976 (NY, US!); Cerro Roraima, Steyermark, J.A. et al. 112625, 26 August 1976 (NY!); Cumbre del Ptari-tepui, Steyermark, J.A. et al. 115720, 23 February 1978 (NY!); Mission Sta Teresita de Kavanayén, Steyermark, J.A. et al. 115725, 23 February 1978 (NY!); Cumbre de Aprada- tepui, Steyermark, J.A. et al. 115899, 25 February 1978 (NY!); cumbre de Aprada-tepui, Steyermark, J.A. et al. 115923, 25 February 1978 (NY!); Mt Roraima, Summit Camp, Steyermark, J.A. 58829 ,27 September 1944 (F!); Ptari-tepui, Steyermark, J.A. 59781 ,2 November 1941 (F!, US!)Gran Sabana, Macizo del Chimantá, Huber, O. 12038, 3 March 1987 (US!); Gran Sabana, Sierra de Lema, Kral, R. et al. 81883, 22 January 1993 (US!); Gran Sabana, Ilu-tepui, Maguire, B. 33399 ,13 March 1952 (F!); Gran Sabana, Cumbre del Macizo de Chimantá, Steyermark, J.A. et al. 115859, 24 February 1978 (NY!); Piar, Murisipan-tepui summit, Holst, B. et al. 2911, 26 May 1986 (NY!); Piar, Cumbre occidental del Angasima- ("Adanta")-tepui, Huber, O. 11691, 9 August 1986 (NY!); Piar, Macizo del Chimantá, Steyermark, J.A. et al. 129001, 7 February 1983 (NY!, US!); Roscio, (ahora Sifontes), Huber O. 10059 , 23 January 1985 (NY!); Roscio, Kukená-(Matauí)-tepui, Huber, O. 10534 & Alarcon, C.,15 June 1985 (NY!); Roscio, Cumbre del Yuruaní-tepui, Huber, O. 9087, 29 February 1984 (NY!).

2. Cephalocarpus chimantensis S.M. Costa, Costa et al. (CHAPTER 3)

Type specimen:— VENEZUELA. Bolívar: Piar, Macizo del Chimantá, Sección oriental del Chimantá-tepui, cabeceras del afluente derecho superior del río Tirica (“Caño del Grillo”), 2450 m, 5°18’N 62°03’W, J.A.Steyermark 128931 & O. Huber, V. Carreño E, 7-9 February 1983 (NY! – holotype; VEN? - isotype)

Fig. 7A, 10A (CHAPTER 3)

(vide Costa et al. CHAPTER 3)

Distribution: This species is known only from its type-locality, on the Chimantá Massif (Bolívar, Venezuela), at altitudes above 2450 m.

Taxonomic notes: Cephalocarpus chimantensis is morphologically similar to C. longibracteatus and C. rigidus, but differ from both by the disposition of spikelets in peduncle (sessile at the other two species), and achene with truncate apex abruptly turning into a short conical beak.

Examined material: VENEZUELA, Bolívar: Piar, Macizo del Chimantá, Sección oriental del Chimantá-tepui, cabeceras del afluente derecho superior del río Tirica (“Caño del Grillo”), 5°18’N/62°03’W 2450 m alt., J.A.Steyermark 128931 & O. Huber, V. Carreño E., 7-9 February 1983 (NY! - holotype); Piar, Macizo del Chimantá, Seccion oriental del Chimantá- tepui, cabeceras del afluente derecho superior del rio Tirica (“Caño del Grillo”), 5°18’N/62°03’W 2450 m alt., O. Huber 7183 & J.A. Steyermark, 7-9 February 1983 (NY!); Chimantá Massif, east-south portion of summit of Apacará-tepui, 2450 – 2500 m alt., J.A. Steyermark 75933, 21-22 June 1953 (F! – 2 sheets).

3. Cephalocarpus confertus Gilly, Bulletin of the Torrey Botanical Club 69 (4): 293-294, fig. 1a-c (1942)

Type specimen: VENEZUELA: Amazonas: Mount Duida, dry ridge tops, Savanna Hills, alt. 4400 ft., G.H.H. Tate 800, August 1928 (NY! - holotype).

Fig. 1 a–c (Gilly 1942); Fig. 7 K–L (Koyama & Maguire 1965); Fig. 2C 1-6 (Koyama 2003); MAP 1D

Monoecious herb: sexes intermingled at paracladia, coarsely monomorphic. Caudex 3 mm diam. (6–8(12) mm diam. including sheaths remains and adventitious roots). Leaf densely congested: sheath glabrous, ferruginous (same color of blades); contraligule indistinct or shiny under a straight line; blade (3)6–10 × 0.2–0.25 cm, linear-lanceolate, margins and midrib (abaxial and adaxial surface) pubescent, discolorous, generally grayish (adaxial) and reddish-brown (abaxial) after drying. Inflorescence 3-5 at each prophyll, head-like; culm 0.5– 3 × –0.1 cm, compressed, margins pubescent. Paracladia (lowermost) highly reduced; peduncle absent to highly reduced. Involucral bract (lowermost) leaf-like; sheath up to 2 mm long, long ciliate, reddish-brown; blade 5–6 × –1 mm, “V”-shaped, pubescent, reddish-brown.

Male spikelet 3.5–6 × ~0.8-1.2 mm, 5–7 glumes, 1–3 flowers; glume up to 5 mm long, ciliate at apex, mucronate (mucro up to 4 mm long); stamen 2, connective projection short conical and glabrous. Female spikelet 3–6 × ~1 mm, 4–5 glumes; glume up to 5 mm long, costae sparsely pubescent to pubescent, mucronate (mucro up to 0.5 mm long, sparsely pubescent to pubescent); stigma 3. Achene 2–3 × ~1 mm, ellipsoid, smooth, glabrous or glabrescent, olive- brown, with a constriction zone between main body and beak; hypogynous scale ~0.25 mm long, triangular, trichomes ~0.55 mm long; beak ~1 mm long, conical to short cylindrical, glabrous or glabrescent.

Distribution: Cephalocarpus confertus mostly occurs in the Venezuelan tepuis, with confirmed occurences at Serra do Aracá (Brazil) and Mount Roraima (including Guyanese side).

Taxonomic notes: This species is morphologically related to Cephalocarpus dracaenula, but can be distinguished by the pubescent external glumes of male/female spikelets (glabrous in C. dracaenula). A detailed discussion is available at the C. dracaenula taxonomic notes.

Examined material: BRAZIL: Amazonas: Barcelos, Serra do Aracá, subida pelo Igarapé Pretinho, Costa, S.M. et al. 1113, 22 ago 2014 (UEC!); Barcelos, Serra do Aracá, arredores do acampamento, Costa, S.M. et al. 1124, 22 ago 2014 (UEC!); Barcelos, Serra do Aracá, Cachoeira El Dorado, Costa, S.M. et al. 1130, 23 ago 2014 (UEC!); Barcelos, Serra Aracá, Prance, G.T. et al. 2911, 16 February 1984 (F, INPA, MG, NY!); Barcelos, Serra do Aracá, Rosa, N.A. 2375 & Lira, S.B., 4 February 1978 (INPA, MG, MO, NY, UEC!); Barcelos, Serra do Aracá, face norte, topo da encosta, Vicentini, A. 1839 & Mesquita, R. de C., 26 August 2001 (INPA!); Barcelos, Serra do Aracá, do Amaral, I.L. 1534, 13 February 1984 (INPA!); Roraima: Serra Parima, Prance, G.T. et al. 21566, 30 July 1974 (MG, NY!); GUYANA: Cuyuni-Mazaruni region: Mt Roraima, Tate, G.H.H. 285, 7 November 1927 (NY!); VENEZUELA: Amazonas: Cerro Duida, Savanna Hills, Steyermark, J.A. 58238 ,2 September 1944 (F, NY!) , Summit of Mount Duida, Tate, G.H.H. 800, August 1928 (NY!); Atabapo, cabeceras del Río Yudi, frente de trabajo no. 17, Delgado, L. 1716, March 1992 (NY!); Rio Negro, Cerro la Neblina, Liesner, R. 15870 & Brewer, C., 11 February 1984 (US!); Bolívar: , Gran Sabana, Ilu-tepui, Maguire, B. 33394 ,14 March 1952 (NY!); Ptari- tepui, Moore Jr., H.E. et al. 9733, 14 August 1970 (NY, US!); Gran Sabana, Steyermark, J.A. et al. 104126, 21 December 1970 (MO, NY!); Ptari-tepui, Steyermark, J.A. 59625, 1

November 1944 (F, NY!); Gran Sabana, Huber, O. 11959, 19 February 1987 (NY!); Roscio, Uei-tepui (Cerro del Sol), Huber, O. 10021, 22 January 1985 (NY, US!).

4. Cephalocarpus debilis (T. Koyama & Maguire) S.M. Costa, (CHAPTER 2)

Basionym:Everardia debilis T.Koyama & Maguire, Memoirs of the New York Botanical Garden 12: 32, f. 7, A–B. 1965.

Fig. 7 A–B (Koyama & Maguire 1965); Fig. 11 M–R (Koyama 2003); MAPS 1E

Type specimen: VENEZUELA, Amazonas: Rio Manapiare, Cerro Yutaje, from Northwest Ridge at 1400 m alt., B. Maguire 35156 & C.K. Maguire, 11 February 1953 (NY! – holotype; F!, VEN#)

Monoecious herb: sexes intermingled at paracladia, monomorphic. Caudex –2 mm diam. (–12 mm diam. including fibrous sheaths remains and adventitious roots). Leaf: sheaths glabrous, light reddish-brown; contraligule indistinct or lighter in color below a rounded line; blades 15–40 × 0.16– 0.5 cm, linear to linear-lanceolate, margins, midrib (abaxial) and surface (adaxial) minute scabrous to ciliate, concolorous, green or reddish-brown after drying. Inflorescence 2-3 at each prophyll, panicle-like (with one or two head-like inflorescences), 3- 5 paracladia; culm 15–30 × 0.04–0.1 cm, compressed, glabrous, ciliate or pubescent;. Paracladia (lowermost) up to 8 cm long, 3–4 order; peduncle up to 6 cm long, compressed, glabrous. Involucral bract (lowermost) leaf-like: sheath -10 mm long, glabrous, brown; blade 1–1.5 × –0.06 cm, no midrib, glabrous, brown. Male spikelet 4–4.8(5.7) × 0.9–1.2 mm, 8–11 glumes, (4)6–8 flowers, solitary; glume ˜2.5 mm long, glabrous, mucronate; mucro -0.8 mm long; stamen 2–4, connective projection conical-ovoid with few trichomes. Female spikelet 3.8–5.5 × 0.8–1.2 mm, 5–7 glumes; glume ~2.5 mm long, glabrous, mucronate (mucro -0.8 mm long); stigma 3. Achene 3–4.5 × ˜1 mm, ellipsoid to oblong, smooth, pubescent at the apex, dark-brown to blackish with paleaceous side lines, main body continuous to the beak; hypogynous scales -2.5 mm long, oval to triangular (sometimes as long as the main achene body), trichomes up to 1.7 long; beak ~0.7 mm long, conical, pubescent to pilose.

Distribution: It occurs in areas near Serranía Yutaje, in Venezuela.

100

Taxonomic notes: This species is morphologically related to C. flexifolium (dioecious species) and they were both in Everardia section Pseudo-everardia (Gilly) T. Koyama & Maguire (Koyama 2003), but it is a monoecious species. The achene morphology is similar between these two species, but the achene in C. debilis is ellipsoid to oblong, while it is more obovate at C. flexifolium.

Cephalocarpus debilis also presents a different inflorescence aspect, with head-like fascicles and a panicle-like rising from the same prophyll (a single panicle-like inflorescence at each prophyll in C. flexifolium). Though this state of character doesn’t get much attention at its first description (Koyama & Maguire 1965) or at the Everardia previous treatment (Koyama 2003), it illustrates some similarity with inflorescences of Cephalocarpus (sensu Koyama 2003) and it is quite interesting under the new circumscription of the genus.

Examined material: VENEZUELA: Amazonas: Caño Guaviarito, Rio Manapiare, Rio Ventuari, Maguire, B. et al. 31735, 2 February 1951 (F!, US!); Serrania Yutaje, Rio Manapiare, Maguire, B. 35156 & Maguire, C.K., 11 February 1953 (F!, US!).

5. Cephalocarpus diffusus (T. Koyama & Maguire) S.M. Costa, (CHAPTER 2)

Basionym: Everardia diffusa T.Koyama & Maguire. Mem. New York Bot. Gard. 12: 21, f. 4 D – E. 1965.

Type specimen: VENEZUELA, Amazonas: Cerro Sipapo (Paráque), from West Peak Caño at 1200 m alt., B. Maguire 27825 & L. Politi, 20 December 1948 (NY00051120! – lectotype; NY03091114!, US!, F!, US!, VEN#, S# – isolectotypes).

Fig 4 D-E (Koyama & Maguire 1965); Fig. 3 (Koyama 2003); MAPS 2A

Monoecious herb: sexes at separate paracladia (male basal and female apical, rarely mixed at the middle), monomorphic. Caudex ~6 mm diam. (18 – 25 mm diam. including fibrous sheaths remains and adventitious roots). Leaf: sheaths glabrous, light brown; contraligule indistinct; blades 30–60 × 0.5–1(1.2) cm, long-lanceolate, glabrous, concolorous or slightly discolorous, yellowish-green to light-green after drying. Inflorescence single at each prophyll, panicle-like, -8 paracladia (one sterile bract at the base); culms 45–62 × 1.4–2 cm, compressed, glabrous. Paracladia (lowermost) up to 15 cm long, 1–3(4) order; peduncle up to

101

6 cm long, terete or compressed, glabrous. Involucral bract (lowermost) leaf-like; sheath 1.7– 2 cm long, glabrous, reddish-brown; blade 2–4x 1.2–1.6 cm, no conspicuous midrib, green, glabrous. Male spikelet 6–8 × 1.2–1.4 mm, 6–7 glumes, 5–7 flowers, single or 2/cluster; glume (outermost) -5.4 mm long, glabrous, mucronate (mucro –2.6 mm long); stamen 3-6, connective projection short-conical with short trichomes. Female spikelet 6–7 × ~1 mm, 5 glumes; glume (outermost) –5.7 mm long, glabrous, mucronate (mucro –2.6 mm long); stigma 3. Achene ~6 × ~1 mm, oblong-elliptic, smooth, rarely with sparse trichomes at the apex of the main body, main body continuous to the beak; hypogynous scales 3 mm long, oval to elliptic, trichomes up to 2.6 mm long; beak ~3.2 mm long, narrow-conical, glabrous.

Distribution: Its registers are scattered in Venezuelan tepuis.

Taxonomic notes: Cephalocarpus diffusus is easily distinguishable by its slender inflorescence and spikelets, with major peduncle carrying a single spikelet (either male or female). It is also distinguished by the spikelets and achene length (up to 7 mm), wich are similar as in C. angustus and C. longifolius, but the plants are completely glabrous, maintain the greenish aspect after drying (brownish in C. longifolius) and have reddish-brown glumes (paleaceous in C. angustus and dark-brown in C. longifolius).

Examined Material: VENEZUELA: Caroní, Cerro Uaipán, Cardona, F. 2035, 26 November 1946 (US!); Amazonas: Cerro Sipapo (Paráque), Maguire, B. 27825 & Politi, L., 20 December 1948 (F!); Atabapo, Cerro de Marahuaca, Salto Los Monos, Liesner, R.L. 17995, 26 February 1985 (F!); Bolivar: Sierra de Lema, Steyermark, J.A. 89638, 29 August 1961 (US!).

6. Cephalocarpus distichous (T.Koyama & Maguire) S.M. Costa, (CHAPTER 2)

Basionym: Everardia disticha T.Koyama & Maguire. Mem. New York Bot. Gard. 12: 25, f. 5 D – F. 1965.

Type specimen: VENEZUELA, Bolivar: Chimantá Massif, Central Section, east branch of headwaters of Río Tirica 2150-2200 m alt., J.A. Steyermark 808 & J.J. Wurdack, 12 February 1955 (NY00051121! – lectotype; F!, NY00051122!,US!,VEN# – isolectotypes)

Fig.5 D-F (Koyama & Maguire 1965); Fig. 7(Koyama 2003); MAPS 2B

102

Monoecious herb: sexes at separate paracladia (male basal, female apical), monomorphic. Caudex (4)5–10 mm diam. (20–25 mm diam. including fibrous sheaths remains and adventitious roots). Leaf distichally arranged: sheaths glabrous, brown to dark-brown; contraligule indistinct or a darker colored straight line; blades 13–25(30) × 1–2 cm, linear- lanceolate, margin and midrib (abaxial) ciliate to pubescent, concolorous, green after drying. Inflorescence single at each prophyll, panicle-like, 3-8 paracladia; culm 25–40 × (0.15)0.25– 0.4 cm, compressed, glabrous. Paracladia (lowermost) up to 10 cm long, 3(5) orders; peduncle up to 5.5 cm long, compressed, glabrous. Involucral bract (lowermost) leaf-like: sheath 1-2.5 mm long, glabrous to sparsely ciliate, brown; blade 1.5–2 × 0.2–0.25 cm, “V”-shaped, margin and midrib (abaxial) glabrous to sparsely ciliate, green to brown. Male spikelet 5–6 × 1.5–1.7 mm, 6–7 glumes, 3–4 flowers, 1-3 at each cluster; glume (outermost) 4.1–4.5 mm long, surface sparsely ciliate, mucronate (mucro 1.3–1.8 mm long, sometimes sparsely ciliate); stamen 3-4, connective projection linear-compressed with trichomes (sometimes star-like). Female spikelet 4.5-5 × ~1 mm, 5 glumes; glume (outermost) 4.1–4.5 mm long, surface sparsely ciliate, mucronate (mucro 1.3–1.8 mm long, sometimes sparsely ciliate); stigma 3. Achene 4.5–6.5 × 0.8–1 mm, oblong-ellipsoid, smooth with pubescent apex, main body continuous to the beak; hypogynous scales 2.4 mm long, obtriangular, trichomes ~2.4 mm long; beak 1.5–3.2 mm long, linear to narrowly conical, pubescent.

Distribution: This species occurs at highlands in Venezuela-Guyana frontier vicinity, and may occur in Brazil, as there are specimens from Mt. Roraima in the (Venezuela-Brazil-Guyana border).

Taxonomic notes: According to Koyama (2003), young plants of this species may be misidentified as C. montanus, but differs from it by: more densely and regularly distichous leaf arrangement, and larger spikelets and cymules (according to the author: 5 mm in C. distichous vs. 3.5 mm in C. montanus).

After I analyzed an isotype of Cephalocarpus montanus, deposited at the US herbarium and which I think Mr Koyama didn’t examine, I am inclined to accept C. distichous as another subspecies of the “C. montanus” complex. Not only the leaf disposition is not a strong character to maintain C. distichous as a separate species, but the length measurements of the spikelets between of the two species are overlapping (the spikelets of C. montanus isotype are up to 6 – female – or 7 – male – millimeters long). Though, unhappily,

103

this isotype had no achene available and thus it is not possible to investigate this case more deeply. (also see a discussion under 17. Cephalocarpus montanus)

Examined Material: GUYANA: Cuyuni-Mazaruni region: Pakaraima Mtns, Hoffman, B. 3219 & Henkel, T., 3 November 1992 (US!); Marazuni-Potaro region: Roraima, Summit, Liesner, R. 23290, 14 April 1988 (US!) VENEZUELA: Bolivar: Cerro Apacará, cumbre, Río Caroní, Cardona, F. 1578, 8 July 1946 (US!); Ilu-tepui, Liesner, R. 23331, 15 April 1988 (US!); Auyan-tepui, Steyermark, J.A. 93972, 15 May 1964 (US!); Gran Sabana, Cumbre del Caraurín-tepui, Huber, O. 12024 & Picon, G.,21 February 1987 (US!); Piar, Murisipan-tepui, Steyermark, J.A. et al. 132136, 26 May 1986 (US!); Bolívar: Chimantá massif, Central Section, Steyermark, J.A. 808 & Wurdack, J.J., 12 February 1955 (F!).

7. Cephalocarpus dracaenula Nees, Flora brasiliensis 2 (1): 162-163, plate 18 (1842). Cryptangium dracaenula (Nees) Boeckeler, Linnaea 38: 415. 1874. Lagenocarpus dracaenula (Nees) H.Pfeiff., Repert. Spec. Nov. Regni Veg. 18: 92. 1922. Type specimen: Mons Cupati at fluvium Japura, In sumo Monte Cupati, Martius 3218, 16 February 1820 (M0008753# -lectotype; M0264603#, M0264604#, M00087654#, BR672799, K000584600, GH0027596; NY! – photos of M0008753# M00087654#)

Fig. 1 d–e (Gilly 1942); Fig. 7 G–H (Koyama & Maguire 1965); Fig. 1(Koyama 2003); MAPS 2C

Monoecious herb: male and female spikelets separated at the same paracladia or cluster, monomorphic. Caudex 3–4.5 (7) mm diam. including sheaths remains and adventitious roots. Leaf loosely congested: sheath glabrous, glabrescent or pubescent, reddish-brown; contraligule indistinct; blade 2–12 × (1.5)2–3.5(4) mm, linear-lanceolate, margins and midrib glabrous, glabrescent, or pubescent (mainly at the base) to glabrous, greyish to dark- ferruginous (adaxial) and ferruginous to light ferruginous (abaxial) after drying. Inflorescence 3–8 at each prophyll, head-like or highly reduced panicle-like, 0–2 paracladia; culm 1–5 cm long, compressed, glabrescent or minute pubescent. Paracladia (lowermost) highly reduced or 1–2 orders; peduncle absent or 2–5 (6.5) mm long. Involucral bract (outermost) leaf-like; sheath up to 0.2 mm long, glabrous, reddish-brown after drying; blade 5–8(12) × ~0.1–0.2 mm, “V” – shaped or no main midrib , margins sparsely long ciliate, reddish-brown after

104

drying. Male spikelet 3–5.5 × 1–1.5 mm, 1–4 flowers, 5–7 glumes; glume margins glabrous or sparsely ciliolate, carena glabrous, mucronate (mucro 0.2–0.4(0.9) mm long); stamen 2, connective projection short conical with few trichomes. Female spikelet 3–5(6) × ~1 mm, 5– 7 glumes; glume margins glabrous or sparsely ciliolate, costae glabrous, mucronate (mucro 0.2–0.4(0.9) mm long); stigma 3, dark red-vinaceous. Achene 2–2.5 × ~1 mm, obovoid to oblong, smooth, glabrous or glabrescent, paleaceous or brown, with a constriction zone between main body and beak; hypogynous squamellae ~0.1–0.2 mm long, orbicular or obtriangular, short ciliolate or ciliate, trichomes ~0.1–0.5 mm; beak ~0.5 mm, narrowly conical, glabrous or sparsely pubescent.

Distribution: This species occurs in Colombia and here we present evidence for its occurrence in Venezuela and Brazil.

Taxonomic notes: This species is morphologically close to Cephalocarpus confertus, from which it can be distinguished by: its loosely congested leaves at caudex apex (highly congested in C. confertus); its glabrous/glabrescent external spikelet glumes (margins and main nervure pubescent to densely pubescent in C. confertus); and the ciliate connective projection (glabrous in C. confertus). Additionally, C. dracaenula may present some short ramifications of the head-like inflorescences, while none were observed in C. confertus.

Cephalocarpus dracaenula shows a gradient of: leaves length and trichomes density and at the size of hypogynous scales and its trichomes. Colombian specimens have longer and glabrous leaves and shorter hypogynous scales, while Brazilian and Venezuelan specimens are glabrescent to sparsely pubescent and have bigger hypogynous scales. But there is no accentuated gap between the two groups and these differences may be explained by the geographical distribution.

It is worth to mention that previously to this manuscript, only Colombian specimens were analyzed in specialists’ works (Gilly 1942; Koyama & Maguire 1965; Koyama 2003): Gilly (1942) only examined one specimen, an isotype at herbarium GH; Koyama & Maguire (1965) and Koyama (2003) examined fewer collections at the US herbarium, but all from Colombia.

Examined material: BRAZIL: Amazonas: Barcelos, Serra do Aracá, Amaral, I.L. do 1534, 13 February 1984 (MG!, NY!); Barcelos, Serra do Aracá, Prance, G.T. 29591 & Guedes, J. ,16 July 1985 (INPA!, MG!, MIRR!, NY!); Roraima: Auaris, Serra Parima, Prance, G.T. et al.

105

9797,10 February 1969 (F!, INPA!, MG!, NY!); COLOMBIA: Mons Cupati at fluvium Japura, Martius 3218, 16 February 1820 (NY!); Rio Kananari, Cerro Isibukuri, Schultes, R.E. 14735 & Cabrera, I., 4 December 1951 (F!, US!); Rio Caqueta, La Pedrera and vicinity, Schultes, R.E. 17677 & Cabrera, I., 2 October 1952 (F!, US!); Rio Caquetá, vicinity of La Pedrera, Schultes, R.E. 5858, April 1944 (F!, US!); Amazonas: Rio Caquetá, Cerro de La Pedrera, Schultes, R.E. 16305 & Cabrera, I., 2 May 1952 (US!); La Pedrera, Río Caquetá, Cerro Yupati, Galeano, G. et al. 1989, 8 March 1990 (NY!); La Pedrera, Rio Caquetá, Cerro de La Pedrera, Schultes, R.E. 16305 & Cabrera, I., 2 May 1952 (NY!); Vaupes, Rio Kananari y Cerro Isibukuri, Schultes, R.E. 14474 & Cabrera, I., 28 September 1951 (US!); Guainia: Maimachi, Serranía de Naquen, Cortés, R. et al. 124, 24 July 1992 (NY!); Vaupés: Río Kananarí y Cerro Isibukuri, García-Barriga, H. 13799, 29 November 1951 (NY!, US!); Rio Piraparaná, García-Barriga, H. 14332, 28 August 1952 (US!); Rio Guainía, Rio Naquieni, Schultes, R.E. 10066 & Lopez, F., June 1948 (US!); La Pedrera, camino al Cerro Yupati, Cortés, R. 488, 15 July 1993 (NY!); VENEZUELA: Amazonas: Sierra de Unturán, Henderson, A. et al. 957, 4 February 1989 (NY!); Rio Negro, Neblina Massif, Canyon Grande, Davidse, G. 27357 & Miller, J.S., 9 July 1984 (F!, MG!, MO!, NY!); Rio Negro, Neblina Massif, Davidse, G. 27406 & Miller, J.S.,15 June 1984 (MO!, NY!, US!); Rio Negro, Cerro Aracamuni (slope), Liesner, R. 22333 & Carnevali, G., 22 October 1987 (MO!, NY!); Rio Negro, Cerro de la Neblina, Stein, B.A. 1672 & Gentry, A., 25 April 1984 (NY!); Bolívar: Chimantá Massif, Steyermark, J.A. 75480 ,19 May 1953 (F!).

8. Cephalocarpus duidae (Gilly) S.M. Costa, (CHAPTER 2) Basionym: Everardia duidae Gilly, Bull. Torrey Club 68: 30, f. 3 g-g’. 1941 [Everardia neblinae Metcalfe, Advances in Bot. Res. Ed. Preston 1: 124,132 & 133. 1963. Nomen nudum ] Type specimen: VENEZUELA, Amazonas: summit of Mt. Duida, alt. 7100 ft., G.H.H. Tate 638, August 1928 to April 1929 (NY!- holotype, US! isotype)

Fig. 3 g–g’ (Gilly 1941); MAPS 2D Monoecious herb: sexes at intermingled at paracladia (mostly female at apex and male at base) , monomorphic. Caudex 5–12 mm diam. including fibrous sheaths remains and adventitious roots. Leaf: sheath minute ciliate, brownish; contraligule shiny under a straight line; blade 10–30 × 0.3–0.9 cm, linear-lanceolate, margins and midrib (abaxial) ciliate,

106

surface (abaxial and adaxial) slightly to markedly rugose, reddish-brown. Inflorescence single at each prophyll, panicle-like, 5–8 paracladia; culms 14–45 × 0.08–0.2 cm, compressed, minute ciliate. Paracladia (lowermost) up to 10(14) cm long, 3–4(6) orders; peduncle up to 5.5 cm long, compressed, minute ciliate. Involucral bract (lowermost) leaf-like: sheath 8–20 mm long, minute ciliate, dark brown; blade 0.5–1.5 × 0.1–0.15 cm, “V”-shaped, minute ciliate, reddish-brown to brownish-green. Male spikelet 3–4 × 0.8–1.3 mm, 6-8 glumes, 3-5 flowers, solitary; glume (outermost) 1.5–1.9 mm long, ), glabrous or margins sparsely ciliate, mucronate (mucro 1 mm); stamen 5, connective projection short-conical with trichomes. Female spikelet 1.5–2.5 × 0.5–1 mm, 5 glumes; glume (outermost) 1.5–1.6 mm long, glabrous or margins sparsely ciliate, mucronate ( mucro 0.3–0.45 mm long); stigma 3. Achene 2.5–3 × 0.6–0.85 mm, ellipsoid, smooth, ciliate, main body continuous to the beak; hypogynous scales 0.65 – 1.5 mm long, orbicular to flattened orbicular, trichomes 0.5–1.3 mm long; beak 0.8-1.2 mm long, conical, ciliate.

Distribution: This species occurs in eastern Venezuelan tepuis.

Taxonomic notes: This species was once synonymized under Cephalocarpus montanus, but the coherence and constancy of the vegetative and reproductive characters are good arguments to maintain its species status; even if the morphological similarity between the two species is evident. It also presents a broad geographical distribution.

Cephalocarpus duidae is distinguishable by the leaf ornamentation (margins and midrib ciliate and surface slightly to markedly rugose), spikelet length (male: 3–4 mm long, and female: 1.5–2.5 mm long). The achene character status is also constant, with length between 2.5–3 mm long, a ciliate surface at the apex and a dark-brown to blackish general color with lighter yellowish side-lines. (also see a discussion under 17. Cephalocarpus montanus)

Examined Material: VENEZUELA: Amazonas: Cerro Moriche, Rio Ventuari, Maguire, B. et al. 30954, 15 January 1951 (US!); Bolívar: , Cerro Pauo, Alto Caura, Cardona, F. 2979, January 1963 (US!); Uaipan-tepui, Koyama, T. 7529 & Agostini, G., 6 March 1967 (US!); Cerro Pitón, Cordillera Epicara, Maguire, B. et al. 53624, 5 September 1962 (US!); Cerro Pitón, Cordillera Epicara, Maguire, B. et al. 53714, 9 November 1962 (US!); Cerro el Venado, Prance, G.T. 28437 & Huber, O., 31 August 1983 (US!); Sierra Ichún, Steyermark, J.A. 90226, 27 December 1961 (F, US!); Sierra Ichún, Steyermark, J.A. 90231 ,27 December

107

1961 (US!); Auyan-tepui, Steyermark, J.A. 94195, 18 May 1964 (US!); Cerro Guaiquinima, rio Carapo, Steyermark, J.A. et al. 117240, 23 May 1978 (F!); Raul Leoni, Fernandez, A. 4717, 1 September 1988 (US!)

9. Cephalocarpus erecto-laxus (T.Koyama & Maguire) S.M. Costa, (CHAPTER 2)

Basionym: Everardia erecto-laxa T.Koyama & Maguire, Memoirs of the New York Botanical Garden 12: 27, f. 5, H–J. 1965.

Type specimen: VENEZUELA, Bolívar: Ilu-tepui, Gran Sabana, Mesa Ridge, 1950 m alt., B. Maguire 33399, 13 March 1952 (NY00051124! – lectotype; NY00051125!, VEN# - isolectotypes)

Fig. 5 H- J (Koyama & Maguire 1965); Fig. 8 (Koyama 2003); MAPS 3A

Monoecious herb: sexes intermingled, monomorphic. Caudex ~5 mm diam. (10–20 mm diam. including fibrous sheaths remains and adventitious roots). Leaf: sheath glabrous, brown to dark-brown; contraligule darker under a rounded line; blade 12–30 × 0.3–0.7 cm, linear- lanceolate, margin, surface (ad) and midrib (abaxial) ciliate to pubescent, concolorous or discolorous, light-green to green [or reddish-brown]. Inflorescence single at each prophyll, panicle-like, 5-7 paracladia (0-2 sterile bracts at the base); culm 15–60 × 0.15–0.18 cm, compressed, glabrous to ciliate. Paracladia (lowermost) up to 9.5 cm long, 1-4 orders; peduncle up to 5 cm long, compressed, glabrous to ciliate. Involucral bract (lowermost) leaf- like: sheath 1.5–2 cm long, ciliate, dark-brown to blackish; blade 2–3 × 0.1–0.15 cm, “V”- shaped, compressed, green, glabrous to ciliate. Male spikelet ~4[6] × ~0.8[1.64] mm, 5–8 glumes, 4–5 flowers, 1–2 per cluster; glume (outermost) up to 4 mm long, glabrous, mucronate (mucro up to 1.5 mm long); stamen (4)6, connective projection compressed and truncate with translucent short trichomes. Female spikelet 3.5–3.7 × 0.8–1 mm, 5–7 glumes; glume (outermost) up to 4 mm long, glabrous, mucronate (mucro up to 1.5 mm long); stigma 3. Achene 4–5.7 × 1.3–1.7 mm, elliptic to oblong-elliptic, smooth, glabrous, main body continuous to the beak; hypogynous scales ~1 mm long, orbicular, trichome ~0.9 mm long; beak ~1.7 mm long, conical to narrow conical, glabrous.

Distribution: The only record of this species is from Ilu-tepui, a table mountain 40 km NE from Mt. Roraima.

108

Taxonomic notes: This is another species closely related to the “Cephalocarpus montanus” complex. Here, as in previous cases, there is an overlap of states at vegetative and reproductive characters.

Koyama & Maguire (1965) and Koyama (2003) highlight as distinctive characters the glabrous achene (sparse ciliate to ciliate at the apex in Cephalocarpus montanus), its more likely ovate shape (that tends to be obovate in Cephalocarpus montanus) and the whitish squamelae (reportedly yellowish in Cephalocarpus montanus). In my opinion, these characters alone or combined aren’t enough to justify the recognition of C. erecto-laxus as a separate species and, in addition, the paratypes have bigger spikelets and achenes. But, due to the lack of specimens, I choose to maintain it species status until more detailed analyses are possible.

Examined Material: VENEZUELA: Gran Sabana, Ilu-tepui, Maguire, B. 33398, 13 March 1952 (US!)

10. Cephalocarpus flexifolium (Gilly) S.M. Costa, (CHAPTER 2) Basionym: Didymiandrum flexifolium Gilly. Bull. Torrey Club 68: 331. 1941. Pseudoeverardia flexifolia (Gilly) Gilly. Fieldiana Bot 28(1):59. 1951. Everardia flexifolia (Gilly) T.Koyama & Maguire, Memoirs of the New York Botanical Garden 12(3): 34, f. 7F. 1965. [Everardia brevis. Nomem nudum?]

Type specimen: VENEZUELA, Amazonas: Cerro Duida, Río Cunucunuma, Caçno Culebra, 1000-1100 m alt., G.H.H. Tate 542, 18 November 1950 (NY! - holotype, US!, VEN# - isotypes)

Fig. 7 F (Koyama & Maguire 1965); Fig. 11A–L (Koyama 2003); MAPS 3B Dioecious herb: male and female plants with similar vegetative morphology and coarsely similar reproductive morphology. Caudex 2–4 mm diam. (5–10 mm diam. including fibrous sheaths remains and adventitious roots). Leaf: sheath glabrescent to ciliate, light reddish- brown; contraligule rounded, shiny, glabrous or densely pubescent; blades (10)20–30 × 0.16– 0.2 cm, linear-lanceolate, margins, midrib and surface sparsely ciliate to pubescent, concolorous, light reddish-brown. Male inflorescence single at each prophyll, panicle-like,

109

open or congested, 1–3 paracladia; culm 7.5–15 × 0.05–0.1 cm, compressed, ciliate to pubescent. Paracladia (lowermost - male) up to 4 cm long, (1)3 orders; peduncle up to 2 cm long, compressed, ciliate to pubescent. Involucral bract (lowermost - male) leaf-like: sheath – ~3.5 mm long, ciliate to pubescent, reddish-brown; blade –0.1 × ~0.06 cm, no midrib, ciliate to pubescent, reddish-brown. Male spikelet (3.2)4.2–6 × 0.8–1.5(1.7) mm, 7–10(12) glumes, 3–5(9) flowers; glume (outermost) 2 mm long, glabrous, mucronate (mucro ~1 mm long); stamen 2(3), connective projection oval with few trichomes at the apex. Female inflorescence single at each prophyll, panicle-like, 2 – 5 paracladia; culm 6–210 × 0.05–0.07 cm, compressed, ciliate. Paracladia (lowermost -female) up to 3 cm long, 3–4 order; peduncle up to 1 cm long, compressed, ciliate. Involucral bract (lowermost - female) leaf-like: sheath up to ~4 mm long, ciliate, same color of leaf and culms; blade –1.7 × 0.04 cm, “V-shaped”, ciliate. Female spikelet 3–3.5 × ~0.8 mm, 6–9 glumes; glume (outermost) 2 mm long, glabrous, mucronate (mucro ~1 mm long); stigma 3. Achene ~2.5 × 1 mm, obovoid, smooth, apex ciliate, main body continuous to the beak; hypogynous scales ~1 mm long, oval, trichomes ~0.7 mm long; beak ~0.3 mm long, short conical, ciliate.

Distribution: All registers are from the Marahuaka and Duida tepuis, in central-southern Venezuela.

Taxonomic notes: This is one of the three dioecious Cephalocarpus species and can be distinguished by the others (C. neblinensis and C. “neblina dioica 2”) using the following characters: inflorescences panicle-like (head-like in C. “neblina dioica 2” and head-like or highly reduced panicle-like in C. neblinensis); male and female inflorescences lateral (lateral and terminal female inflorescences in C. “neblina dioica 2”); male spikelets 4–6 mm long (~6 mm long in C. “neblina dioica 2” and 6.5–7 mm long in C. neblinensis); female spikelets 3– 3.5 mm long (~3.5 in C. “neblina dioica 2” and ~6mm long in C. neblinensis); achene up to 2 mm long (4–4.5 mm long in C. neblinensis) and without a constriction zone between the main body and beak (present in both remaining species).

Three male specimens from Cerro Duida and Cerro Marahuaca do not totally fit the C. flexifolium morphology, but as I could not find their female companion it is difficult to defend it as a different and new species. These specimens do not presents distinguishable contraligule and have consistently smaller spikelets (all around 3 mm long). This could be a local/environmental variation, but the plants were collected each one in a different place and where typical C. flexifolium plants are also present (exc. Cerro Marahuaca).

110

Examined Material: VENEZUELA: Amazonas: Cerro Duida, Culebra Peak, Maguire, B. 29085 & Maguire Jr., B., 23 April 1949 (US!); Cerro Duida, Culebra Peak, Maguire, B. 29127 & Maguire Jr., B., 23 April 1949 (F, US!); Cerro Duida, Rio Cunucunuma, Maguire, B. et al. 29513, 18 November 1950 (US!); Maguire, B. et al. 29528, 18 November 1950 (F!, US!); Cerro Duida, Brocchina Hills, Steyermark, J.A. 58194, 1 September 1944 (F!); Atabapo, Cerro Duida, cumbre, Steyermark, J.A. et al. 126427, 10 February 1982 (US!); Depto Atabapo, Falda del extremo Norte del Cerro Duida, Steyermark, J.A. et al. 126109, 6 February 1982 (MG, US!); Mt Duida, summit, Tate, G.H.H. 542, 20 December 1929 (US!).

11. Cephalocarpus illustrata S.M.Costa, M.C.E. Amaral & W.W. Thomas, Costa et al. (CHAPTER 3)

Figs. 7B, 10B (CHAPTER 3)

(vide Costa et al. CHAPTER 3)

Distribution: Known only to the Chimantá-tepui and the Serranía Guanay (Bolívar, Venezuela).

Taxonomic notes:- This species is the only that presents an open panicle-like inflorescence and an achene with a constriction zone between the main body and beak. Mr. Koyama examined its specimens and believed it was an Everardia immature specimen, probably due to its narrow spikelet. But the collections had developed and mature achenes completely enclosed by the glumes.

12. Cephalocarpus longibracteatus Gilly, Brittonia 3: 153, f. 2d–j. 1939. Cephalocarpus lineariifolius Gilly. Bull. Torrey Bot. Club 69 (4): 295, f. 1f. 1942 Type specimen: Venezuela, Amazonas, summit of Mt. Duidae at Central Camp, G.H.H. Tate 1035 (NY! holotype; US!-isotype)

111

Cephalocarpus maguirei Gilly, Bull. Torrey Club 76: 292. 1949. Type specimen: Guyana, Kaieteur savanna, B. Maguire 23453 & D.B. Fanshawe (NY00011594! lectotype; NY00011595!, F!, US! - isolectotypes). Cephalocarpus steyermarkii Gilly. Fieldiana Bot 28 (1): 54. 1951.Type specimen: Venezuela, Bolívar, Ptari-tepui, J.A. Steyermark 59640 (F! - holotype, NY!, GH#, VEN#) Cephalocarpus rigidus subsp. mucronatus Gilly. Bull. Torrey Bot. Club 69(4): 296, f. 1 j–l. 1942. Type specimen: Venezuela, Amazonas, Mount Duida, G.H.H. Tate 721 (NY! – holotype; US!)

Type specimen: VENEZUELA, Bolívar: Mount Auyan-tepui, alt. 2200 m, G.H.H. Tate 1342, December 1937 (NY!)

Fig. 1 f, j–m (Gilly 1942); Fig. 7 M–N (Koyama & Maguire 1965); Fig 2B 1-6 (Koyama 2003); MAPS 3C Monoecious herb: male and female spikelets intermingled, monomorphic. Caudex up to 20 mm diam. including sheaths remains and adventitious roots. Leaf: sheath mouth ciliate to densely pilose, generally dark-green after drying; contraligule indistinct or pubescent, rounded; blade 3–15 × 0.17–0.28 cm, linear to linear-lanceolate, margin glabrescent, sparsely scabrous or ciliate, midrib (abaxial) glabrescent, ciliolate or pubescent, surface (abaxial and adaxial) glabrous, sparsely scabrous or ciliolate, generally green after drying. Inflorescence single at each prophyll (with a second level of paracladia if panicle-like), head-like (highly reduced panicle-like),; culm 3–6(9) × (0.06)0.08–1.5 cm, compressed, margin glabrous to minutely ciliolate. Paracladia absent or reduced, 1-3 (4) order; peduncle absent or reduced to less than 2 mm, compressed, glabrous. Involucral bract (lowermost) leaf-like: sheath 0-0.2 cm long, mouth, margin and surface white ciliate to pubescent; blade 1–2.5 × –0.2 cm, canaliculated or “M”-shaped at apex, generally green after drying. Male spikelet 4.5–6 × ~1 mm, 5-7 glumes, 2-3 flowers; glume (outermost) 3.5–4.5 mm long, margin glabrous to minute-ciliolate, mucronate (mucro 0.5 mm long); stamen 2-3 anthers, connective projection short conical, ciliate. Female spikelet 4–5 × 1–1.5 mm, generally single, 5-6 glumes; glume (outermost) 3.5–4.5 mm long, margin glabrous to minute-ciliolate, mucronate (mucro –0.75 mm long); stigma 3. Achene 2.5–3.5 × 1.25 mm, oblong, ciliate at apex, light brown to brown, with a constriction zone between main body and beak; hypogynous scale –0.9 mm

112

long, orbicular or elliptic, long white-ciliate, trichomes -0.6 mm long; beak -0.7mm long, short conical, ciliate to pubescent.

Distribution: It is one of the most spread species in the genus, and occurs in most of Venezuelan highlands, and also in the Brazilian Serra do Aracá and Monte Roraima, and at Guyana.

Taxonomic notes: This species was once synonymized under Cephalocarpus rigidus Gilly, but the combination of leaf, involucral bract and external spikelet glumes ornamentation is distinct at each species. While Cephalocarpus rigidus presents them as almost entirely glabrous, C. longibracteatus plants are generally ciliate to pubescent. Additionally, the lowermost glumes of spikelets and achenes have different shapes, and the achene beak is narrower in C. rigidus than in C. longibracteatus.

Examined Material: BRAZIL: Roraima: Uiramutã, Parque Nacional do Monte Roraima, Monte Caburaí, Forzza, R.C. et al. 8204, 7 November 2014 (NY!); GUYANA: Karamand River, Wenamu trail, Maguire, B. 32501 & Fanshawe, D.B., 10 November 1951 (NY!, US!); VENEZUELA: Summit of Mount Duida, Tate, G.H.H. 1035, 20 December 1928 (NY!, US!); Auyan-tepui, Tate, G.H.H. 1345, December 1937 (NY!); summit of Mount Duida, Tate, G.H.H. 720, August 1928 (NY!); Summit of Mount Duida, Tate, G.H.H. 721, August 1929 (NY!); Gran Sabana, Ilu-tepui, Mesa Ridge, Maguire, B. 33393, 14 March 1952 (NY!, US!); Gran Sabana, Ilu-tepui, Maguire, B. 33632, 27 March 1952 (NY!, US!); Amazonas: Serranía Paru, Caño Asísa, Río Ventuari, Cowan, R.S. 31337 & Wurdack, J.J., 7 February 1951 (NY!, US!); Cerro Sipapo (Paráque), Maguire, B. 27592 & Politi, L., 11 December 1948 (NY!, US!); Cerro Sipapo, Maguire, B. 27826 & Politi, L., 20 December 1948 (NY!); Cerro Duida, Culebra Peak, Maguire, B. 29212 & Maguire Jr., B.,April 1949 (NY!, US!); Cerro Huachamacari, Río Cunucunuma, Maguire, B. et al. 30070, 8 December 1950 (NY!, US!); Rio Cunucunuma, Cerro Huachamacari, Maguire, B. et al. 30159, 11 December 1950 (F!, NY!, US!); Serranía Yutaje, Río Manapiare, Maguire, B. 35389 & Maguire, C.K., 23 February 1953 (NY!); Cerro Duida, Steyermark, J.A. 58146, 1 September 1944 (F!); Cerro Duina, Brocchina Hills, Steyermark, J.A. 58148, 1 September 1944 (F!); Atabapo, Parque Nacional Duida-Marahuaca, Fernandez, A. et al. 8154, April 1991 (US!); Atabapo, Liesner, R. 18143, 1 March 1985 (NY!); Atabapo, Liesner, R.L. 18143, 1 March 1985 (NY!); Atures, Valley of Río Coro-Coro, Holst, B.K. 3355 & Liester, R.L., 6 March 1987 (NY!); Atures, Cerro Coro-Coro, Huber, O. 12340, 12 November 1987 (MO!); Sierra Parima, cabeceras del

113

Río Matacuni, Steyermark, J.A. 107511, 19 May 1973 (NY!); Bolivar: , Uaipan-tepui, Koyama, T. 7511 & Agostini, G., 6 March 1967 (NY!, US!); Cerro Guaiquinima, Rio Paragua, Maguire, B. 32764, 25 December 1951 (NY!, US!); Cerro Guaiquinima, Rio Paragua, Maguire, B. 32831, 30 December 1951 (NY!, US!); Cerro Guaiquinima, Rio Paragua, Maguire, B. 32911, 2 January 1952 (NY!, US!); Cerro Guaiquinima, Rio Paragua, Maguire, B. 33054, 10 January 1952 (NY!); Plateau of Ayan tepui, Prance, G.T. 28213 & Huber, O., 26 August 1983 (NY!, US!); Plateau of A[u]yan Tepui, Prance, G.T. 28240 & Huber, O., 27 September 1983 (NY!); Uei-tepui, Steyermark, J.A. 342 & Nilsson, S., 19 April 1960 (NY!, US!); Auyan-tepui, Steyermark, J.A. 93449, 5 May 1964 (NY!); Meseta de Jaua, Cerro Jáua, Steyermark, J.A. 98034, 22 March 1967 (NY!); Meseta de Jaua, Cerro Jáua, Steyermark, J.A. 98035, 22 March 1967 (NY!, US!); Meseta del Jaua, Cerro Sarisariñama, Steyernark, J.A. et al. 108861, 10 February 1974 (NY!); Salto el Toron, Thomas, W.W. 2524, 28 October 1979 (NY!); Auyantepui, E do Bolivar, Vareschi 4877 & Foldats, April 1956 (NY!); Heres, Meseta del Guaiquinima, Huber, O. 10909 & Medina, G., 26 September 1985 (NY!); Heres, macizo del Guaiquinima, Huber, O. 9355, 2 April 1984 (NY!); Heres, Cero Marutani, Steyermark, J.A. et al. 123863, 11 January 1981 (NY!, US!); Piar, Auyan-tepui, Holst, B.K. 3734, 29 March 1987 (NY!); Piar, Altiplanice del Auyan-tepui, Huber, O. et al. 8044, 26 August 1983 (INPA!, NY!); Raul Leoni, 12km al N de Aparaman tepui, Fernandez, A. 3562, September 1986 (NY!); Sifontes, en la ladera media occidental del Cerro Kukenán, Huber, O. 11792, 6 September 1986 (NY!, US!); Bolívar: Sororopan-tepui, Ptari-tepui, Maguire, B. 33916 & Wurdack, J.J., 17 December 1952 (NY!, US!); Meseta Del Jaua, Cerro Sarisariñama, Steyermark, J.A. et al. 108861, 12 February 1974 (F!, NY!, US!); Meseta del Jaua, Cerro Jaua, Steyermark, J.A. et al. 109441, 24 February 1974 (F!, NY!); Cerro Guanacoco, Cumbre, Steyermark, J.A. et al. 109722, 3 March 1974 (NY!); Torono-tepui, Chimantá Massif, Steyermark, J.A. 1120 & Wurdack, J.J., 23 February 1955 (F!, NY!); Ptari- tepui, Steyermark, J.A. 59640, 1 November 1944 (F!, US!); Ptari-tepui, Steyermark, J.A. 59800, 2 November 1944 (F!); Auyan-tepui, Steyermark, J.A. 93483, 7 May 1964 (F!, NY!, US!); Ptari-tepui, first plateau on west side, Thomas, W.W. 2712 ,29 November 1979 (NY!); Piar, Meseta del Auyan-tepui, Huber, O. 9594 & Medina, G., 2 July 1984 (NY!).

13. Cephalocarpus longifolius (Gilly) S.M. Costa, (CHAPTER 2) Basionym: Everardia longifolia Gilly, Brittonia 3(2): 153, f. 2g–h. 1939.

114

Type specimen: VENEZUELA, Bolívar: Mount Auyan-tepui, alt. 2200 m, G.H.H. Tate 1347, December 1937 (NY! – holotype)

Fig. 2 c (Gilly 1941); Fig. 10(Koyama 2003); MAPS 3D

Monoecious herb: sexes mostly at separate paracladia (male basal, female apical), monomorphic. Caudex 5 mm diam. (10–20(30) mm diam. including fibrous sheaths remains and adventitious roots). Leaf: sheath ciliate, dark brown to blackish; contraligule indistinct; blade (15) 20–40 × 0.3–0.6 (0.8) cm, linear-lanceolate, margins and midrib (abaxial) sparse scabrous to scabrous, surface (ad and abaxial) ciliate, concolorous, green. Inflorescence single at each prophyll, panicle-like, (5)6–9 paracladia; culm (20) 30–60 × 0.15–0.4 cm, compressed, glabrous, sparse scabrous or sparse ciliate. Paracladia (lowermost) up to 7(10) cm long, 1(2) order; peduncle up to 5 (7) cm long, compressed, sparse scabrous to sparse ciliate. Involucral bract (lowermost) leaf-like: sheath 1.5–3.5 mm long, sparse ciliate, dark- brown to blackish; blade 2–2.5 × 0.15–0.2 cm, “V”-shaped, sparsely scabrous to scabrous ciliate, green. Male spikelet 8–14 × 1.5–2 mm, 8–9 glumes, 3–4 flowers, 1(2) each cluster; glume (outermost) ~11 mm long, ciliate, mucronate (mucro ~4 mm long), dark-brown; stamen 6–9, connective projection oblong with truncate apex with short trichomes. Female spikelet 7.5–10 × 1–1.5 mm, 5 glumes; glume (outermost) ~11 mm long, ciliate, mucronate (mucro ~4 mm long), dark-brown; stigma 3. Achene 7–9.5 mm × 0.9–1.4 mm, oblong- ellipsoid, smooth, pubescent at the apex, main body continuous to the beak; hypogynous scales ~0.4 mm long, ovoid to ellipsoid, trichome <0.1mm; beak 3.5–4 mm long, narrow- conical, pubescent.

Distribution: It occurs in eastern Venezuelan tepuis.

Taxonomic notes: (vide taxonomic notes under Cephalocarpus angustus).

Examined Material: VENEZUELA: Auyan-tepui, Schnee 1601, 19 April 1956 (F!); Bolivar: Auyan-tepui, Steyermark, J.A. 93703, 11 May 1964 (US!); Piar, Macizo del Chimantá, Huber, O. et al. 10095, 6 February 1985 (US!); Piar, Macizo del Chimantá, Luteyn, J.L. et al. 9513, 16 February 1984 (US!); Piar, Macizo del Chimantá, Steyermark, J.A. et al. 128229, 26 February 1983 (US!); Bolívar: Chimantá massif, Central Section, Steyermark, J.A. 407 & Wurdack, J.J., 4 February 1955 (F!, US!); Chimantá massif, Tonoro-tepui, Steyermark, J.A. 628 & Wurdack, J.J., 9 February 1955 (F!, US!); Chimantá Massif, Apacará-tepuí, Steyermark, J.A. 75765, 20 June 1953 (F!, INPA!); Chimantá Massif, Central Section,

115

Steyermark, J.A. 791 & Wurdack, J.J., 12 February 1955 (F!, US!); Auyan-tepui, Steyermark, J.A. 93482, 7 May 1964 (F!, US!).

14. Cephalocarpus “lowlandia” S.M.Costa, Costa et al. (CHAPTER 3)

Type specimen:— VENEZUELA. Amazonas: Rio Negro, Neblina Massif, 500–700 m, 0°50’N 66°06’W, G. Davidse 27378 & J.S. Miller, 15 July 1984 (NY! – holotype; MO! - isotype)

Figs. 7C, 10C (CHAPTER 3)

(vide Costa et al. CHAPTER 3)

Distribution: its registers are from Cerro de la Neblina (Amazonas, Venezuela)

Taxonomic notes: It is possible to notice that the pubescence and lowermost male glume size increases and the leaf length decreases at higher altitudes at this species. Two collections from Cerro Ariapo (Iparia, Peru), at about 2000 m alt., seem to be part of C. lowlandia and show the abovementioned increase/decrease pattern. With exception of these two collections, the species is only recorded for the Cerro de la Neblina (Amazonas, Venezuela).

Cephalocarpus lowlandia morphologically resembles C. angustus, but it is quite different due to the narrower and less pubescent leaves, the smaller female spikelets, and achene with pubescent beak. Vide “taxonomic notes” under 1. Cephalocarpus angustus for more details.

Examined Material: VENEZUELA: Amazonas: Rio Negro, Neblina Massif, Davidse, G. 27378 & Miller, J.S., 15 July 1984 (US!); Rio Negro, Cerro de la Neblina, Rio Yatua, Maguire, B. et al. 42341, 11 December 1957 (US!); Rio Negro, Cerro de la Neblina, Thomas, W.W. 3033 & Plowman, T., 13 April 1984 (F, US!).

15. Cephalocarpus maguireanus (T.Koyama) S.M. Costa, (CHAPTER 2)

Basionym: Everardia maguireana T. Koyama, Memoirs of the New York Botanical Garden 18: 23. 1969.

116

Type specimen: VENEZUELA, Amazonas: Rio Negro, Serra da Neblina, East trail along escarpment, alt. 8000 ft, B. Maguire 60495 & I.M. Pires, C.K. Maguire, 3 December 1965 (NY00051133 – lectotype; NY00051134!, MG!, K# - isolectotypes)

MAPS 4A

Monoecious herb: sexes mixed or mostly at separate paracladia (male basal, female apical), monomorphic. Caudex 7–15 mm diam. including fibrous sheaths remains and adventitious roots. Leaf: sheath glabrous, brown; contraligule indistinct (difficult to examine); blade 10– 25 × 0.55–0.7 cm, linear-lanceolate, margin and midrib (abaxial) ciliate, concolorous, green. Inflorescence single at each prophyll, panicle-like, (3) 4–6 paracladia; culm 10–25 × 0.12– 0.14 cm, compressed, margin sparsely pubescent to pubescent. Paracladia (lowermost) up to 7 cm long, 2–3(4) order; peduncle up to 4 cm long, compressed, margin sparsely pubescent to pubescent. Involucral bract (lowermost) leaf-like: sheath up to 1 mm long, sparsely pubescent to pubescent, light-brown; blade 0.8–3 × 0.15–0.2 cm, “V”-shaped, sparsely pubescent to pubescent, green. Male spikelet ~5 × ~1.4 mm, 5–7 glumes, 2–3 flowers, solitary or rarely 2– 3(5 – when of both sexes) each cluster; glume (outermost) ~5 mm long, carena sparsely pubescent, mucronate (mucro 1.5–2.5 mm long); stamen 2–4, connective projection not observed. Female spikelet 3–5 × ~1.3 mm, 4–5 glumes; glume (outermost) 3–4.5 mm long, carena sparsely pubescent, mucronate (mucro 1.5–2.5 mm long); stigma 3. Achene ~3 × ~0.75 mm, oblongo-obovoid, smooth, side ridges and apex pubescent, main body continuous to the beak; hypogynous scales ~1.5 mm long, flattened orbicular, trichome up to 1.5 mm long; beak ~1.2 mm long, linear to narrowly conical, pubescent.

Distribution: Only known by the type locality and specimen.

Taxonomic notes: According to Koyama (1969, 2003), this species is distinguishable by its larger female spikelet glumes, almost twice the length of the mature achene, and is closely related to C. vareschii, due to the achene pubescence.

The other diagnostical character pointed out by Koyama (1969, 2003) is not clear-cut, as the relative size of mature achene and glumes can be mistaken and recently described species also have achenes smaller than the glumes. I could not find any additional specimen of Cephalocarpus maguireanus, despite the intensive analysis of collection from the type- locality and vicinity.

117

16. Cephalocarpus martinhae S.M.Costa, (Costa et al. CHAPTER 3)

Type specimen:— VENEZUELA. Amazonas: Atabapo, Cerro Marahuaca, cumbre, sección suroriental, vecinidades dek zanjón, 2685 m, 3°37’N 65°21’W, B. Maguire 65631 & J.A. Steyermark, C. Brewer-Carías, C.K. Maguire, V.C. Espinosa, 15 January 1981 (NY02480927! – holotype, VEN?).

Figs. 7D, 10D (CHAPTER 3)

(vide Costa et al. CHAPTER 3)

Distribution: Only registered to its type-locality and probably restrict to Cerro Marahuaca at altitudes above 2400m. The Mt. Duida, the closest peak and about 25 km far but no specimen of Cephalocarpus martinhae was collected there.

Taxonomic notes: This species is easily distinguishable by its leaves (without midrib) and rounded achene apex with a linear beak without the constriction zone between the beak and the main body.

Examined Material: VENEZUELA: Amazonas, Atabapo: Cerro Marahuaca (cumbre), J.A. Steyermark 65631 & C. Brewer-Carías, C.K. Maguire, V.C. Espinosa, 15 January 1981 (NY! - holotype); Cerro Marahuca (cumbre), J.A. Steyermark 126085 & M. Guariglia, N. Holmgren, J. Luteyn, S. Mori, 3-4 February 1982 (NY!, US!).

16. Cephalocarpus montanus (Ridley) S.M. Costa, (CHAPTER 2) Basionym: Everardia montana Ridley. Timehri 5:210. 1886 Everardia glaucifolia Gilly. Bull. Torrey Club 68: 27, f.2 d-d’. 1941. Type specimen: Venezuela, Amazonas, Mt. Duida, G.H.H. Tate 570 (NY! – holotype, MSC#- spikelets fragments only)] Everardia revoluta Gilly. Bull. Torrey Club 68: 28, f.3 e-e’. 1941. Type specimen: Venezuela, Amazonas, Cerro Duida, 6300 ft alt. G.H.H. Tate 469 (NY00051142!, lectotype; NY00051143!, MSC# - spikelet fragments only - isolectotypes)

118

Everardia guaiquinimae Schnee. Bol. Soc. Venez. Ci. Nat. 9: 83. 1944. Type specimen: Venezuela, Cerro Guaiquinima, F. Cardona 974 (VEN# - holotype; F!, NY!, US!) Everardia surinamensis Gilly. Bull. Torrey Club 76: 292. 1949. Type specimen: Suriname, Tafelberg, B. Maguire 24382 (NY00051145! - lectotype; NY00051146!, U#, VEN#, ISC#- leaf and paracladia fragments, MSC# - leaf and paracladia fragments - isolectotypes). Everardia hitchcockii Lasser & Maguire. Brittonia 7:76. 1950. Type specimen: Venezuela, Amazonas, Cerro Yavi, 1450 m alt. Type specimen: K.D. Phelps & C.B. Hitchcock 15 (NY00051129! – lectotype; NY00051130! - isolectotype) Everardia montana Ridley var. pilosa Gilly. Fieldiana Bot. 28 (1): 54. 1951. Type specimen: Venezuela, Amazonas, Cerro Duida, J.A. Steyermark 58318 (F!, holotype; NY!, photo) Everardia spongiosa M.T. Strong. Novon 24: 403. 2016. Type: Guyana. Cuyuni- Mazaruni Region, Pakaraima Mtns., Mazaruni River, tepui top near Chi-Chi Falls, 5°36’47.5”N, 60°12’59”W, 889 m, 30 Jan. 2004, K. M. Redden 2487A, with M. Lyle, C. Paul & C. Perry (holotype, US; isotype, MO). Syn. Nov. Type specimen: BRITISH GUIANA: Mt. Roraima, 7000-7500 ft., In Thurn 335, October- January 1884-1885 (K# holotype ; US!, BM#, NY photo)

Figs. 2 d–d’, 3 e–f’ (Gilly 1941); Figs. 6, 7C–E (Koyama & Maguire 1965); (Koyama & Maguire 1965); Fig. 9(Koyama 2003); MAPS 5C-D Monoecious herb: sexes intermingled at paracladia, mostly female toward the apex, monomorphic. Caudex 0.5–1 cm (1–2 cm including fibrous sheaths remains and adventitious roots). Leaf: sheath glabrous to densely pubescent, dark-brown to blackish; contraligule indistinct or rounded, white pilose in or shiny under the line; blade 20–60 × 0.5–2 cm, linear- lanceolate to lanceolate, margin, midrib and surface glabrous to densely pubescent, sometimes glaucous and/or with rugose surface, generally green. Inflorescence single at each prophyll, panicle-like, 8-12 paracladia; culms 25–60 × (0.4) 0.17–0.32 (0.4) cm, compressed, glabrous, ciliate or pubescent. Paracladia (lowemost) up to 19 cm long, 3-6 orders; peduncle up to 7 cm long, compressed, glabrous, ciliate or pubescent. Involucral bract (lowermost) leaf-like: sheath 1.5–2.5(3) cm long, glabrous, ciliate or pubescent, brown or dark brown to blackish; blade 2.5–4.5(5.5) × 0.25–0.4(0.47) cm, canaliculated or “V”-shaped, margin, midrib and surface glabrous, ciliate or pubescent. Male spikelet 2.9–7.9 × (0.8) 1–2.4 mm, 6-7 glumes,

119

(4)5–6 flowers, solitary-3/cluster; glume (outermost) 2.4–4.6 mm long, glabrous to densely ciliate, mucronate (mucro 0.95–2.3 mm long); stamen 4-6 (7), connective projection short- conical to linear-cyllindrical, sparse ciliate. Female spikelet 3–6 × 0.75–1.7 mm, 5 glumes; glume (outermost) 2.5–5 mm long, glabrous to densely ciliate, mucronate (mucro 0.7–1.7 mm long); stigma 3. Achene 3.5–6 × 1.1–1.4 mm, ellipsoid, smooth, generally ciliate at apex, main body continuous to the beak; hypogynous scales 0.8–1.3 mm long, orbicular, oblong or spatulate ,trichomes 0.4–0.9 mm long; beak 1.35–3 mm long, conical, sometimes glabrous but normally ciliate.

17.1. Cephalocarpus montanus subsp. montanus This subspecies presents the typical character status of the species. The vegetative and reproductive parts are less pubescent or glabrous, and the leaf surface is frequently smooth. The spikelet sizes are generally bigger too (up to 8 mm long – male – and 6 mm long – female – in the isotype at US). The glumes are darker and more rigid than in other subspecies. Distribution notes:- this typical subspecies is spread through Guiana Shield and vicinity and also occurs at Ecuador and Peru. In the last case, the subspecies occurs in mountains bordering the Andean highlands but with soils of arenisca, as poor as the ones in Guiana Shield highlands.

17.2. Cephalocarpus montanus subsp. glaucifolius (Gilly) S.M. Costa, comb. nov. Basionym: Everardia glaucifolia Gilly, Bull. Torrey Club 68: 27, f.2 d-d’. 1941. This subspecies is quite similar to the typical subspecies, but generally presents glaucous leaf blades and inflorescence. The inflorescences usually have as many paracladia and spikelets as the typical subspecies and the achenes are small to medium sized. Distribution notes:- occurs in almost the same areas of the typical subspecies, except Peru and Ecuador, but less frequently.

17.3. Everardia montanus subsp. guaiquinimae (Schnee) S.M. Costa. Basionym: Everardia guaiquinimae Schnee. Bol. Soc. Venez. Ci. Nat. 9: 83. 1944. Its plants have short, glaucous leaves, just as E. montana subsp. glaucifolia but the leaves are puberulent. At E. montana subsp. guiaiquinimae, the inflorescences have

120

less spikelets and the paracladia are smaller than in the other subspecies. The achenes are small to medium sized. Distribution notes:- this subspecies is restricted to Cerro Guaiquinima and vicinity, in central Venezuela.

17.4. Cephalocarpus montanus subsp. velutinus (T. Koyama) S.M. Costa, comb. nov. Basionym: Everardia montana subsp. velutina T. Koyama, Mem. New York Bot. Gard. 18(2): 26. 1969. Type specimen: NT Silva & U Brazao 60931 (NY!, holotype; NY!, MG!), from Serra Pirapucu, low humid catinga forest alt 1300-1700m, Rio Negro, Amazonas, Brazil. This subspecies is distinguished by the densely vilose leaves, and densely puberulent inflorescences and glumes. The panicle-like inflorescence is more congested than in the typical subspecies. The achenes are small to medium sized. Distribution notes:- All registers are from Cerro de la Neblina (Venezuelan-Brazilian border).

Taxonomic notes:- The “Cephalocarpus montanus” complex is the most intriguing and challenging in the genus. For many years, new species of the complex, formely understood in Everardia Ridley (1886), were described under the paradigm that the isolation of the tepuis should determine groups of different but related species troughout the Guiana Shield. After the collecting efforts and the analysis of the specimens by specialists, this paradigm was questioned and a belief in a more spread distribution of a phenotypic plasticity together with some local subspecies raised.

Along a decade (1941-1951), Mr. Gilly’s described seven species that latter were included as sinonyms of Everardia montana Ridley (=Cephalocarpus montanus) in Mr. Koyama’s works (Koyama & Maguire 1965, Koyama 2003). Gilly had access to only few specimens, and tended to accept scattered and different species, Koyama had access to more specimens, and tended to accept local subspecies immersed in a broadely distributed and morphologically plastic species.

I could examine about 150 specimens in the herbaria and concluded that there is a more spread species with morphological plasticity with many “looks like”, morphologically related taxa mixed together. Some of these related morpho-taxa are restricted to one or two tepuis, but the superficial morphological analysis unattended of statistichal morphological or a

121

molecular analysis is not enough to assert if they are Cephalocarpus montanus subspecies (ecotypes, maybe) or different species (with hibrids, maybe).

To avoid future complications, I recognize Cephalocarpus duidae and C. ptariensis due to the more spread distribution and constant and singular morphology (leaf and achene characters, specially) and maintained the remaining sinonyms under Cephalocarpus montanus. I also recognize three subspecies besides de typical one, as listed above. I maintain Cephalocarpus montanus subsp. glaucifolius, though as spread as C. duidae, due to the remarkable morphological similarities with the typical subspecies. The other two subspecies are quite diferent from the typical one, but due to the low number of specimens available and the observation of intermediary morpho-types, I still recognize them in C. montanus.

I was inclined to include other morphologically related species in “C. montanus” complex (like Cephalocarpus distichous and C. maguireanus), but, again, as I had just a few specimens of these species available and with some structures missing, I prefer to maintain their recognition as independent isolated species.

Examined Material:- BRAZIL: Amazonas: Serra do Aracá, Prance, G.T. et al. 29067, 14 February 1984 (F, INPA, MG!); Serra Aracá ou Serra Natal, Rosa, N.A. 2246 & Lira, S.B., 27 January 1978 (F, INPA, MG, MO!); Rio Negro, Rio Cauaburi, Rio Maturacá, Serra da Neblina, Silva, N.T. 60931 & Brazão, U., 27 January 1966 (MG!); Barcelos, Serra do Aracá, subida pelo Igarapé Pretinho, Costa, S.M. 1115 , 22 August 2014 (UEC!); Barcelos, Serra do Aracá, arredores do acampamento, Costa, S.M. et al. 1121, 22 August 2014 (UEC!); Barcelos, Serra do Aracá, Cachoeira El Dorado, Costa, S.M. et al. 1133,23 August 2014 (UEC!); Barcelos, Serra do Aracá, Cachoeira El Dorado, Costa, S.M. et al. 1135, 23 August 2014 (UEC!); Barcelos, Serra do Aracá, platô, setor norte, Vicentini, A. 1891 & Mesquita, R. de C., 27 August 2001 (INPA!); Barcelos, Serra do Aracá, do Amaral, I.L. 1508, 12 February 1984 (INPA!); COLOMBIA: Vaupés: Rio Kananari y Cerro Isibukuri, García-Barriga, H. 13794, 29 November 1951 (US!); Río Kananari, Cerro Isibukuri, Schultes, R.E. 14502 & Cabrera, I., 29 October 1951 (F!); Rio Kananari (affluent of Rio Apaporis), Schultes, R.E. 14732 & Cabrera, I., 4 December 1951 (US!); Rio Kananari, Cerro Isibukuri, Schultes, R.E. 15050 & Cabrera, I., 23 January 1952 (US!); ECUADOR: Morona-Santiago: Limon Indanza, Región de la Cordillera del Cóndor, Kajekai, C. 204 & Curso de dendrologia, 4 September 2005 (US!); Zamora-Chichipe: Berry, P.E. 7638 & Neill, D., 20 March 2001 (MO!); GUYANA: Pakaraima Mts, Mt Aymatoi, Maas, P.J.M. et al. 5658, 15 October 1981 (MG!); Kamarang

122

river - Wenamu trail, Maguire, B. 32548 & Fanshawe, D.B., 11 November 1951 (F!); Mt. Roraima, in Thurn 335, October 1884 (US!); Cuyuni-Mazaruni: Paruima, 20 km W, on Waukauyengtipu, Clarke, D. 5840 &DAvid, K.; Chin, C.; Perry, C. ,18 July 1997 (US!); Potaro-Siparuni: Mt Ayanhanna, Clarke, H.D. et al. 9500, 23 June 2004 (US!); PERU: Cajamarca: San Ignacio, Distr. Huarango. Cordillera Huarango, Rodriguez, E. et al. 2965, 22 April 2006 (MO!); SURINAM: Tafelberg, Table Moutain, Maguire, B. 24382, 15 August 1944 (F!); VENEZUELA: Rio Guayavaca, Schnee 1471, 16 April 1956 (F!); Auyan tepui, Schnee 1548, 18 April 1956 (F!); Chimantá massif, Central Section, Steyermark, J.A. 902 & Wurdack, J.J., 17 February 1955 (F!); Amazonas: Cerro Sipapo (Paráque), Maguire, B. 27529 & Politi, L., 6 December 1948 (F!, US!); Cerro Huachamacari, Río Cunucunuma, Maguire, B. et al. 29860, 5 December 1950 (F!, US!); Cerro Huachamacari, Rio Cunucunuma, Maguire, B. et al. 3014, 11 December 1950 (F!, US!); Cerro Huachamacari, Rio Cunucunuma, Maguire, B. et al. 30257, 16 December 1950 (US!); Cerro Huachamacari, Río Cunucunuma, Maguire, B. et al. 30290, 17 December 1950 (F!, US!); Caño Guaviarito, Rio Manapiare, Rio Ventuari, Maguire, B. et al. 31708, 2 February 1951 (F!, US!); Serranía Yutaje, Río Manapiare, Maguire, B. 35326 & Maguire, C.K., 18 February 1953 (US!); Serranía Yutaje, Río Manapiare, Maguire, B. 35403 & Maguire, C.K., 25 February 1953 (F, US!); Cerro de la Neblina, Rio Yatua, Maguire, B. 37166 & Wurdack, J.J., 10 January 1954 (F, US!); Cerro de la Neblina, Maguire, B. et al. 37209, 10 January 1954 (F!, US!); Cerro de la Neblina, Rio Yatua, Maguire, B. et al. 42181, 22 November 1957 (US!); Cerro de la Neblina, Nee, M. 31104, 25 February 1985 (US!); Serranía Parú (Asisa), Phelps, K.D. 530 & Hitchcock, C.B., 11 February 1949 (US!); Cerro Autana, Steyermark, J.A. 105108, 21 September 1971 (F!); Cerro Autana, Steyermark, J.A. 105110, 20 September 1971 (F!); Cumbre del Cero Autana, Steyermark, J.A. 105153, 20 September 1971 (F!); Cerro Duida, Caño Negro, Steyermark, J.A. 58043, 26 August 1944 (F!); Cerro Duida, Caño Negro, Steyermark, J.A. 58046, 26 August 1944 (F!); Cerro Duida, Brocchina Hills, Steyermark, J.A. 58136, 1 September 1944 (F!); Cerro Duida, Brocchina Hills, Steyermark, J.A. 58187, 1 September 1944 (F!); Cerro Duida, Caño Negro, Steyermark, J.A. 58213, 2 September 1944 (F!); Atabapo, Plateau of Huachamari, Liesner, R.L. 18070, 1 March 1985 (NY!); Atabapo, Plateau of Duida above Culebra, Liesner, R.L. 18148, 2 March 1985 (NY!); Atabapo, Cerro Duida, cumbre, Steyermark, J.A. et al. 126389, 10 February 1982 (US!); Rio Negro, Cerro de la Neblina, Funk, V.A. 6329, 16 February 1984 (US!); Rio Negro, Cerro de la Neblina, Renner, S.S. 2081, 6 February 1985 (US!); Rio Negro, Cerro de la Neblina, Steyermark, J.A. 129822 & Luteyn,

123

J.L., 7 February 1984 (F!); Rio Negro, vicinity of CAmp VI, Thomas, W.W. 3036 & Plowman, T.,13 April 1984 (F, US!); Río Negro, Neblina and Massif, Camp III, Liesner, R. 16011, 16 February 1984 (US!); Río Negro, Rio Mararinuma, Río Baria, Luteyn, J.L. 9399 & Steyermark, J.A., 7 February 1984 (F, US!); Río Negro, Cerro de la Neblina, Nee, M. 30613, 29 January 1985 (F!, US!); Bolivar: Cerro Guaiquinima, Alto Rio Paragua, Cardona, F. 1119, 15 July 1944 (F, US!); Cerro Guaiquinima, Alto Rio Paragua, Cardona, F. 974 ,October 1943 (F!); Uaipan-tepui, Koyama, T. 7361 & Agostini, G., 1 March 1967 (F, US!); Cerro Guaiquinimae, Rio Paragua, Maguire, B. 32791, 28 December 1951 (F!); Cerro Guaiquinima, Rio Paragua, Maguire, B. 32837, 30 December 1951 (F!); Meseta del Jaua, Cerro Jaua, Steyermark, J.A. et al. 109419, 24 February 1974 (F!); Cerro Guaiquinima, rio Carapo, Steyermark, J.A. et al. 117295, 25 May 1978 (F!); Cerro Guaiquinimae, Steyermark, J.A. et al. 117437, 26 May 1978 (F!); El Cajón, Puente Luis Raúl Vásquez, Steyermark, J.A. et al. 117793, 18 December 1978 (F!); Chimantá massif, Central Section, Steyermark, J.A. 349 & Wurdack, J.J., 2 February 1955 (F!, US!); Ptari-tepui, Steyermark, J.A. 50610, 30 October 1944 (US!); Ptari-tepui, Steyermark, J.A. 59610, 30 October 1944 (F, US!); Ptari-tepui, Steyermark, J.A. 59796, 2 November 1944 (F!); Chimantá Massif, Apácara-tepui, Steyermark, J.A. 75928, 21 June 1953 (F, INPA!); Auyan-tepui, Steyermark, J.A. 93322, 3 May 1964 (US!); Auyan-tepui, Steyermark, J.A. 93323, 3 May 1964 (F, US!); Auyan-tepui, Steyermark, J.A. 93929, 15 May 1964 (F, US!); Meseta Jáua, Cerro Jáua, Steyermark, J.A. 97876, 22 March 1967 (US!); Meseta Jáua, Cerro Jáua, Steyermark, J.A. 97905, 22 March 1967 (US!); Piar, Cerro Venado, Huber, O. 10866 & Huber, M., 24 September 1985 (F!); Piar, Cerro El Venado, Huber, O. et al. 8226, 31 August 1983 (INPA!); Piar, Macizo del Chimantá, Huber, O. 8594 & Dezzeo, N., 31 January 1984 (INPA!); Piar, Macizo del Chimantá, Steyermark, J.A. et al. 127977, 26 January 1983 (US!); Piar/Sifontes, Huber, O. 11320 & Fernandez, A., 18 February 1986 (NY!); Sucre, Meseta de Jáua, cumbre, Steyermark, J.A. et al. 124318, 14 February 1981 (US!).

18. Cephalocarpus neblinensis S.M.Costa, Costa et al. (CHAPTER 3)

Type specimen:— VENEZUELA. Amazonas: Cerro de la Neblina, Rio Yatua, more often in open places about Cumbre Camp, 1800 m alt., B. Maguire 42107 & J.J. Wurdack, C.K. Maguire, 15 November 1957 (F! –holotype; NY!, US! – isotypes)

124

Figs. 7E, 10E (CHAPTER 3)

(vide Costa et al. CHAPTER 3)

Distribution: All collections of Cephalocarpus neblinensis are from Cerro de la Neblina, at the Venezuelan side, but it is highly probable that the species also occurs at the Brazilian side of the area, at the Pico da Neblina National Park.

Taxonomic notes: This is another dioecious species of Cephalocarpus, superficially similar to C. rigidus, but is distinguished due to its longer spikelets (up to ~6 mm long in the female spikelets and up to ~7 mm long in male spikelets) and the dioecism. See the “taxonomic notes” of C. flexifolium to comparisons among dioecious Cephalocarpus spp. and/or Costa et al. (CHAPTER 3).

Examined Material: VENEZUELA: Amazonas: Cerro de la Neblina: Río Yatua, B. Maguire 37021 & J.J. Wurdack, G.S. Bunting, 30 December 1953 (NY!)*; Rio Yatua, B. Maguire 42107 & J.J. Wurdack, C.K. Maguire, 15 November 1957 (NY! – holotype; US!, F! - isotype)*; Rio Yatua, B. Maguire 42455 & J.J.Wurdack, C.K. Maguire, 21 December 1957 (NY!,US!’)*; Rio Negro, B.M. Boom 5808 & A.L.Weitzman, 13 February 1985 (NY!) Rio Negro, Camp III, Neblina and Massif, R. Liesner 16083, 16 February 1984 (NY!, MO!- masc and fem); Rio Negro, Liesner, R. 16651 ,15 March 1984 (NY!, MO!) Rio Negro, J.L. Luteyn 9429 & J.A. Steyermark, 7 February 1984 (F!); Rio Negro, M. Nee 31107[-A], 26 February 1985 (NY!, MO!); Río Negro, río Mawarinumma, Rio Baria, J.L. Luteyn 9429 & J.A. Steyermark, 7 February 1984 (MO!, US!); Cerro de la Neblina, J.A.Steyermark 129827 & J.L. Luteyn, 7-8 February 1984 (MO)

19. Cephalocarpus obovoideus T.Koyama, Memoirs of the New York Botanical Garden 18 (2):22-23, f.4 h-o. 1969. Fig. 2A, 1-7(Koyama 2003); MAPS 4B [Cephalocarpus maguireanus T.Koyama. nomen nudum] [Oreobolus obovoidea T.Koyama. nomen nudum]

Type specimen: BRAZIL: Amazonas: Rio Negro, Rio Cauaburí, Serra da Neblina, east trail along escarpment, 8000 ft. alt., B. Maguire 60496 & J.M. Pires, C.K. Maguire, 3 December 1965 (NY! - holotype)

125

Monoecious herb: male and female spikelets intermingled, monomorphic. Caudex 2–3.6 mm diam. (~6 mm diam. including sheaths remains and adventitious roots). Leaf: sheath dark- brown, pubescent; contraligule indistinct; blade 1.1–2.5 × 0.2–0.25 cm, linear-lanceolate, margin glabrescent, concolorous, greenish. Inflorescence single at each prophyll, head-like; culm 0.8–2.5 × 0.08-0.25 cm, compressed, margin glabrescent or pubescent. Paracladia not developed; peduncle not developed. Involucral bract (lowermost) leaf-like: sheath 0–0.2 mm long; blade 0.6–0.7 × ~0.02 cm, canaliculate, margins glabrous to sparsely scabrous, brown. Male spikelet 3.5–5 × ~1 mm, 5 glumes; glume -~4 mm long, margin glabrous or white- ciliolate, mucronate (mucro –8 mm long); stamen 2, connective projection short ciliate. Female spikelet 3.5–4 × ~1 mm, generally 3 at each cluster, 5-6 glumes; glume -~3 mm long, margin glabrous or white-ciliolate, mucronate (mucro –8 mm long); stigma 3. Achene 2.7–3 × ~1.5 mm, broadly obovoid, smooth, glabrous, olive-brown, with a constriction zone between main body and beak; hypogynous scale –0.2 mm long, orbicular or obtriangular, long ciliate, trichome -1.3 mm long; beak 0.75–1 mm long, round-conical, glabrous or pubescent.

Distribution: The records of C. obovoideus are from Cerro de la Neblina and vicinity, in the Venezuelan-Brazilian border.

Taxonomic notes: This is another Cephalocarpus species known only by few collections. It is distinguished from the other species with the constriction zone in the achene of Cephalocarpus by small and highly congest leaves and broadly obovoid to ellipsoid achenes with a short conical beak.

Examined material: BRAZIL: Amazonas: Rio Negro, Rio Cauaburi, B. Maguire 60496 & J.M. Pires, C.K. Maguire (NY! - holotype); Cerro Neblina, A. Gentry 46697, 17 April 1984 (NY!, MO!).

19. Cephalocarpus ptariensis (Gilly) S.M. Costa, (CHAPTER 2) Basionym: Everardia ptariensis Gilly, Fieldiana Bot. 28(1):55.1961.

Type specimen: VENEZUELA, Bolivar: Ptari-tepui, south-facing slopes overlying stones, on “Cave Rock” above “Cave Camp”, 1810 m alt., J.A. Steyermark 59496, 29 October 1944 (F! - holotype; NY!, ISC#, VEN# - isotypes)

MAPS 4C

126

Monoecious herb: sexes intermingled at paracladia (mostly female at apex), monomorphic. Caudex 3 (5?) mm diam. (8–15 mm diam. including sheaths remains and adventitious roots). Leaf: sheaths glabrous or sparsely ciliate, dark brown to blackish-brown; contraligule indistinct or shiny under a straight line; blades (3)10–15(20) × (0.4)0.6–1.2 cm, lanceolate, margin and midrib (abaxial) ciliate, midrib (ad) densely ciliate, concolorous, green. Inflorescence single at each prophyll, panicle-like, 6-12 paracladia; culm 25–50 × 0.12–0.2 cm, compressed, glabrous or sparsely ciliate. Paracladia up to 15 cm long, 3-5 order; peduncle up to 5.5 cm long, compressed, glabrous or sparsely ciliate. Involucral bract (lowermost) leaf- like: sheath -16 mm long, glabrous or sparsely ciliate at apex, dark brown to blackish-brown; blade –2 × 0.1–0.18 cm, “V”-shaped, blade green, margin and midrib (abaxial) ciliate. Male spikelet (3.6)4–5 × 1–1.5(2) mm, 7–9 glumes, 3–4 flowers; glume (outermost) 3–3.5 mm long, glabrous or ciliate at apex margins, mucronate (mucro 1-1.3 mm long); stamen 4, connective projection short cylindrical with trichomes. Female spikelet 3–4.5 × 0.7–0.95 mm, 5 glumes; glume (outermost) 3–3.5 mm long, glabrous or ciliate at apex margins, mucronate (mucro 1-1.3 mm long); stigma 3. Achene 3–4 × 0.6–0.9 mm, obovoid, smooth, glabrous or sparsely ciliate at apex, main body continuous to the beak; hypogynous scales 0.5–0.7 mm long, elliptic to orbicular, trichome -3 mm long; beak 0.95–1.65 mm long, conical, glabrous or sparsely ciliate.

Distribution: This species occurs at Venezuelan tepuis, especially in the eastern ones.

Taxonomic notes: Cephalocarpus ptariensis was once synonymized under Cephalocarpus montanus at Koyama & Maguire (1965). But, as in the case of Cephalocarpus duidae, this taxon has coherent and constant characters to support its species status. We advocate it due to the following characters: its linear-lanceolate, consistently shiny and smooth, ciliate at margin and midrib (abaxial) leaves. (also see a discussion under 17. Cephalocarpus montanus)

Examined Material: VENEZUELA: Amazonas: Cerro Duida, Steyermark, J.A. 58318, 4 September 1944 (F!); Bolívar: Ptari-tepui, Moore, H.E. et al. 9775, 14 August 1970 (US!); Chimantá Massif, Central section, Steyermark, J.A. 408 & Wurdack, J.J., 4 February 1955 (US!); Chimantá Massif, Central Section, Steyermark, J.A. 409 & Wurdack, J.J., 4 February 1955 (F!); Ptari-tepui, Steyermark, J.A. 59496, 29 October 1941 (F!); Chimantá Massif, Steyermark, J.A. 74847, 13 April 1953 (F!); Chimantá Massif, Steyermark, J.A. 75915, 22 June 1953 (F!); Chimantá Massif, Steyermark, J.A. 75919, 22 June 1953 (F!); Chimantá Massif, Steyermark, J.A. 75921, 22 June 1953 (F!); Auyan-tepui, Steyermark, J.A. 93964, 15

127

May 1964 (US!); Piar, Macizo Chimantá, Chimantá-tepui, Steyermark, J.A. et al. 128130, 26 January 1983 (F!).

20. Cephalocarpus recurviglumis (T.Koyama & Maguire) S.M. Costa, (CHAPTER 2)

Basionym: Everardia recurvigluma T.Koyama & Maguire, Memoirs of the New York Botanical Garden 12: 23, f. 4, F–G. 1965.

Type specimen: VENEZUELA, Bolívar: Ilu-tepui, Gran Sabana, upper escarpment at 2400 m alt., B. Maguire 33525, 20 March 1952 (NY00051140! – lectotype; NY00051141! VEN#- isolectotypes)

Fig. 4 F-H (Koyama & Maguire 1965); Fig. 4 (Koyama 2003); MAPS 4D

Monoecious herb: sexes mostly at separate paracladia (male basal and female apical) , monomorphic. Caudex 3-8 mm diam. ((8) 10–25(30) mm diam. including sheaths remains and adventitious roots). Leaf: sheaths glabrous to ciliolate, brown to dark-brown; contraligule indistinct; blades (10)18–30(40) × (0.34)0.5–1(1.2) cm, linear-lanceolate, margin and midrib (abaxial) ciliolate, concolorous or discolorous, light-green, yellowish-green or green. Inflorescence single at each prophyll, panicle-like, (3)4–8(9) paracladia; culms 20–50(60) × (0.08)0.15–0.2(0.3) cm, compressed, margin glabrous to sparsely ciliate. Paracladia up to 20 cm long, 1-5 (generally 3) orders; peduncle up to 10 cm long, compressed, glabrous or sparsely ciliolate. Involucral bract (lowermost) leaf-like: sheath 1–2.5 mm long, glabrous to sparsely ciliolate, brown to dark-brown; blade 1–2.5(3) × (0.1)0.15–0.23(0.29) cm, “V”- shaped, margin and midrib (abaxial) ciliate, green. Male spikelet 4.9–7.5 × 1–1.8 mm, 7–9 glumes, 5–6 flowers, 1–3 each clusters; glume (outermost) –5 mm long, glabrous, mucronate (mucro 1 mm long); stamen 6, connective projection short truncate/triangular with short trichomes. Female spikelet (2.9)5.7–6.1 × 0.6–1.6 mm, 6 glumes; glume (outermost) –5 mm long, glabrous, mucroate (mucron 1 mm long), recurved at achene maturity; stigma 3. Achene 4.8–6.5 × 0.8–1.2 mm, oblong-ellipsoid, smooth, glabrous, main body continuous to the beak; hypogynous scales 1.5 mm long, truncate, tricomes up to 1.4 mm; beak up to 3.25 mm long, narrow conical, glabrous.

Distribution: It occurs at highlands near the Guyana-Venezuela border.

128

Taxonomic notes: Cephalocarpus recurviglumis is distinguishable from other species by its female spikelet glumes, recurved at achene maturity and after its fall, and by the linear-oblong and completely glabrous achene with long straight scale trichomes.

Examined material: GUYANA: Cuyuni-Mazaruni region: Mt. Maringma, Clarke, H.D. et al. 11750, 15 June 2004 (US!); VENEZUELA: Cerro Apradá, Caroní, Guayana, Cardona, F. 1988, 26 November 1946 (US!); Gran Sabana, Ilu-tepui, Maguire, B. 33526, 20 March 1952 (US!); Bolivar: , Cerro Roraima, Steyermark, J.A. et al. 112456, 16 August 1976 (US!); Bolívar: Chimantá Massif, Torono-tepuí, Steyermark, J.A. 1222 & Wurdack, J.J., 1 March 1955 (F, US!); Mount Roraima, Steyermark, J.A. 58767, 27 September 1944 (F, US!); Chimantá Massif, Central Section, Steyermark, J.A. 608 & Wurdack, J.J., 7 February 1955 (F, US, NY!).

22. Cephalocarpus rigidus Gilly ex Gleason & Killip, Brittonia 3: 152, f.2 a-c. 1939. Cephalocarpus rigidus subsp typicus Gilly. Bull. Torrey Bot. Club 69(4): 296, f. 1 g–i. 1942 Cephalocarpus glabra Strong, Novon 20 (4): 401–403, f.1. Type specimen: Guyana, Cuyuni-Mazaruni region, Pakaraima Mtns., K.M. Redden 2487A M.Lyle, C.Paul & C.Perry (US!-holotype). Syn. Nov.

Type specimen: VENEZUELA, Bolivar: Mount Auyan-tepui, 2200 m alt., G.H.H. Tate 1346, December 1937 (NY! – holotype; MSC#, U#)

Fig. 1 g–i (Gilly 1942); Fig. 7 I–J (Koyama & Maguire 1965); MAPS 5A Monoecious herb: male and female spikelets intermingled, monomorphic. Caudex up to - 8(10) mm diam. including sheaths remains and adventitious roots. Leaf: sheath glabrous (rarely glabrescent), mouth ciliate, reddish-brown to brown; contraligule indistinct; blade 3– 3.5 × 0.2–0.25 mm, linear-lanceolate, glabrous or sparsely scabrous (at apex), reddish-brown or brown. Inflorescence single at each prophyll (with an inconspicuous second level of paracladia if panicle-like), head-like (rarely panicle-like); culm -3 × ~0.1 cm, compressed, glabrous to minutely ciliolate. Paracladia absent or highly reduced, 1 order; peduncle absent or ~1 mm long, compressed, glabrous. Involucral bract (lowermost) leaf-like: sheath -0.2 cm long, glabrous to sparsely ciliolate, reddish-brown after drying; blade 0.5–1 × ~0.1 cm, “V”- shaped, glabrous, reddish-brown. Male spikelet 3.5–3.7 × ~0.7mm, 5-6 glumes, 3-4 flowers;

129

glume (outermost) 3.7–4 mm long, margin minute-ciliolate, mucronate (mucro –1 mm long); stamen 2, connective projection (not observed). Female spikelet ~4 × ~1 mm, generally single, 5 glumes; glume (outermost) 3.7–4 mm long, margin minute-ciliolate, mucronate (mucro –1 mm long); stigma 3. Achene 3–3.3 × ~1.5 mm, broadly ellipsoid, ciliate at apex, paleaceous, with a constriction zone between main body and beak; hypogynous scale ~0.2 mm long, orbicular or elliptic, long white-ciliate, trichomes ~0.8 mm long; beak ~1mm long, round-conical, glabrous or pubescent.

Distribution: This species is also spread in Venezuelan highlands and in Guyana highlands near Venezuela-Guyana border.

Taxonomic notes: Many other taxa, known and unknown, were included under the Cephalocarpus rigidus circumscription probably due to the great variation accepted for the species. A closer look at the type-specimen and obra princeps reveals a narrower variation of the characters and led to the exclusion of almost all synonyms, here accepted as synonyms of Cephalocarpus longibracteatus.

The leaves of Cephalocarpus rigidus, as here accepted, are completely glabrous, except for few specimens with trichomes at the sheath mouth, and up to 3.5 cm long; culms up to 3 cm long; when present, the paracladia never have more than 1 order; and broadly ellipsoid achenes. Strong (2016) recognized the collection from Guyana as belonging to different taxon probably due to the broader circumscription of C. rigidus sensu Koyama & Maguire (1965) and Koyama (2004).

Examined Material: GUYANA: Upper Mazaruni river region, Karowtipu Mountain, Boom, B. 7603 & Gopaul, D., 21 April 1987 (US!); Cuyuni-Mazaruni: top of escarpment above Tihmeri rock paintings, Gillespie, L.J. 2845 & Smart, D.R. ,23 December 1989 (INPA, US!); VENEZUELA: Auyan-tepui, Tate, G.H.H. 1346, December 1937 (NY!); Amazonas: Atabapo, Cerro Marahuaca, Liesner, R. 25266, 23 October 1988 (MO!); Atabapo, Cerro Marahuaca, Steyermark, J.A. 129517, 10 October 1983 (MO!); Atabapo, Cerro Marahuaca, Steyermark, J.A. 129592 ,13 October 1983 (NY!); Atures, Cerro Coro-Coro, Huber, O. 12305, 12 October 1987 (MO!); Atures, Sierra Maigualida, Huber, O. 13088, 24 November 1989 (MO!); Rio Negro, Cerro Aracamuni, Popa Camp., Liesner, R. 22103 & Delascio, F., 18 October 1987 (MO, US!); Bolívar: : Cerro Guanacoco, Steyernark, J.A. et al. 109722, 3 March 1974 (NY!); Chimantá Massif, Steyermark, J.A. 805 & Wurdack, J.J., 1955 (F, NY,

130

US!); Cedeño, Sierra Maigualida, Huber, O. 12819 & Izquierdo, L., 18 November 1988 (MO!); cumbre de Auyan-tepui, Steyermark, J.A. et al. 115984, 26 February 1978 (NY!); cumbre de Auyan-tepui, Steyermark, J.A. et al. 115985, 26 February 1978 (NY!); Gran Sabana, 8km al N de las cabeceras del Rio Waiparu, Fernandez, A. 1934, February 1986 (NY!); Gran Sabana, 8km al N de las cabeceras del Rio Waiparu, Fernandez, A. 2082, February 1986 (NY!); Gran Sabana, Sierra de Lema, Río Carrao alto, Huber, O. 12527, 31 January 1988 (MO!); Piar, Auyan-tepui, Huber, O. 11240, 19 January 1986 (NY!); Piar, cabeceiras, Huber, O. 12206, 10 May 1987 (NY!); Piar, Meseta del Auyantepui, Huber, O. et al. 8110, 27 September 1983 (MO, NY!).

23. Cephalocarpus vareschii (Maguire) S.M. Costa (Chapter 2)

Basionym: Everardia vareschii Maguire, Acta Biologica Venezuelica 2(6): 43. 1957.

Type specimen: VENEZUELA, Bolivar: la parte superior de Auyan-tepui, 2300 m alt., V. Vareschi et E. Fordats 4881, April 1956 (NY! - holotype; G#, VEN# - isotypes)

Fig 1-6 (Maguire 1957); Fig 4A-C (Koyama & Maguire 1965); Fig. 5 (Koyama 2003); MAPS 5B

Monoecious herb: sexes intermingled at paracladia or sometimes in the same cluster, monomorphic. Caudex 3-5 mm diam. (10-18 mm diam. including sheaths remains and adventitious roots). Leaf: sheath ciliate to pubescent, brown; contraligule indistinct; blades 5– 25 × ~0.4– 0.75 cm, linear-lanceolate to lanceolate, pubescent, discolorous, green (ad) and silvery-green (ad). Inflorescence single at each prophyll, panicle-like, 3-4 paracladia; culm (15) 20–40 × 0.12–0.18(0.2) cm, compressed, glabrous to sparsely ciliate. Paracladia up to 15 cm long, 3–5(6) orders; peduncle up to 8 cm long, compressed, sparsely ciliate to pubescent. Involucral bract (lowermost) leaf-like: sheath 1–2.5 cm long, glabrous to ciliate, brown; blade, (1)1.5–3 × 1–1.3 cm, no visible midrib, green, ciliate to pubescent. Male spikelet 4.5– 5.5 × ~1 mm, 7–9 glumes, 5–4 flowers, solitary or rarely 2 each cluster; glume (outermost) ~5 mm long, glabrous or rarely with sparse trichomes, mucronate (mucro 1.6–2 mm long); stamen 3, connective projection compressed, truncate, with short trichomes. Female spikelet 4.6–5 × ~1 mm, 5 glumes, single or rarely 2/cluster; glume (outermost) ~5 mm long, glabrous or rarely with sparse trichomes, mucronate (mucro 1.6–2 mm long); stigma 3. Achene 4.5–5.5

131

× 0.8–1 mm, oblong-elliptic, smooth, pubescent especially at the apex, main body continuous to the beak; hypogynous scales ~2.5 mm long, ellipsoid to ovoid, trichomes up to ~2mm long; beak ~1.5 mm long, narrow conical, pubescent to pilose.

Distribution: The registers of E. vareschii are from central Venezuelan tepuis, at Auyan-tepui and Chimantá massif.

Taxonomic notes: Cephalocarpus vareschii has pubescent leaves, large diffuse inflorescences, and narrow pubescent achenes.

Examined material: VENEZUELA: cumbre del Auyan-tepui, Alto Caroní, Cadorna, F. 2649, February 1949 (US!); Bolívar: Chimantá Massif, Torono-tepui, Steyermark, J.A. 656 & Wurdack, J.J., 9 February 1955 (F!, US!); Auyan-tepui, Steyermark, J.A. 93942, 15 May 1964 (F!,US!).

24. Cephalocarpus “chimanta 2”, Costa et al. (CHAPTER 3)

Type specimen: VENEZUELA, Bolivar: Chimantá Massif, among southwest-facing sandstone bluffs of Chimantá-tepuí (Torono-tepui) (near Southern corner), 1700 m alt., J.A. Steyermark 75480, 19-20 May 1953 (NY!-holotype)

Figs. 7A, 10G (CHAPTER 3)

(see Costa et al. CHAPTER 3)

Distribution: type locality only

Taxonomic notes: This intriguing species resembles a plus-sized Cephalocarpus confertus regarding its vegetative morphology, but additionally to its meaningful bigger size, the achene characters are notably different, without the constriction zone between the body and the beak, lanceolate hypogynous scales and a pyramidal beak. It is only registered from the type- locality and by a single collection).

Examined material: (type collection only)

25. Cephalocarpus “neblina dioica 2” S.M.Costa, Costa et al. (CHAPTER 3)

132

Type specimen: VENEZUELA, Amazonas: Depto Río Negro, Cerro de la Neblina, 0°52- 53’N 66°05’W 1880 m alt., J.A. Steyermark 129812 & J.L. Luteyn, 7-8 February 1984 (MO! Female - holotype);

Fig. 10F (CHAPTER 3)

(see Costa et al. CHAPTER 3)

Distribution:- this species is registered only to Cerro de la Neblina.

Taxonomic notes:- This species resembles a magnified Cephalocarpus confertus as regards its vegetative characters, but besides the bigger caudex and leaf size, it is a dioecious. Another interesting feature is the lateral and terminal placement of the inflorescences of the female plant. All achenes available are immature, but already present the clavate beak typical of Cephalocarpus (sensu Koyama 2003), while Cephalocarpus “chimanta 2” (infl esp séssil) has a triangular beak without the constriction zone beneath.

Examined material (additional): Depto Río Negro, Cerro de la Neblina, 0°52’N 66°05’W 1820 - 1880 m alt., B.M. Boom 5507 & A.L. Weitzman, C. Brewer-Carías, 1 February 1985 (NY! Male - epitype)

Final Comments The reassessment of the circumscriton of the Cryptangieae genera and the discovery of new species buried in herbaria collections increased the number of Cephalocarpus species, from 4 spp sensu Koyama (2003) to 25 species in current understanding.

Also it still is necessary the deeper investigation of groups of morphologically close related species using anatomical, morphometrics and/or molecular tools. For example, the “Cephalocarpus montanus” complex is puzzling, includes four or five species (C. maguireanus (?), C. montanus, C. duidae, C. ptariensis and C. distichous) and four subspecies (C. montanus subsp. montanus, C. montanus subsp. glaucifolius, C. montanus subsp. guiaiquinimae and C. montanus subsp. velutinus). Additional data may help to clarify if it is a single entity or has well-defined boundaries between included species, and especially among the subspecies.

133

Together with the species additions, the morphological variation in the genus increased. Now, Cephalocarpus comprises species with different inflorescence morphology (highly congested, almost head-like, and open, lax panicle-like inflorescences) and achene morphology (Appendix 4A). This new morphological spectrum made it difficult to gather and organize the species in a traditional/dichotomic identification key, especially due to the species that show mixed states of characters of Everardia and Cephalocarpus sensu Koyama (2003) (for example: C. illustrata, C. martinhae, C. “chimanta 2”). An interactive key of the group could ease this problem.

The range of elevation where the species occur also increased, now it goes from 100 m – C. lowlandia- up to 2800 m – C. martinhae, but the species still are restricted to nutrient poor soil, as most of Cryptangieae representatives. Though Cephalocarpus diversity is concentrated at Guiana Shield, some species occur at sub-Andean moutains, for example, the Condor Cordillera in Peru and Ecuadorwhere the habitat is similar to the tepuis (elevation up to 2800 m only on sandstone substrate) (see “Botanical Exploration of the Cordillera del Cóndor” project page, MOBOT. Available at: http://www.mobot.org/MOBOT/Research/ecuador/cordillera/welcome.shtml).

134

MAPS 1 – Geographical distribution of: A-B. Cephalocarpus Nees species and diversity (B – each color represent a diferent species); C. C. angustus; D. C. confertus; E. C. debilis. A B

135

MAPS 2 – Geographical distribution of: A. Cephalocarpus diffusus; B. C. distichus; C. C. dracaenula; D. C. duidae.

136

MAPS 3 – Geographical distribution of: A. Cephalocarpus erecto-laxus; B. E. flexifolium; C. C. longibracteatus; D. C. longifolium.

137

MAPS 4 – Geographical distribution of: A. Cephalocarpus maguireanus; B. C.obovoidea; C. C. ptariensis; D. C. recurvigluma.

138

MAPS 4 – Geographical distribution of: A. Cephalocarpus rigidus; B. C.vareschii; C. C. montanus subsp. montanus; D. C. montanus subsp. glaucifolius (green circle), C. montanus subsp. guaiquinimae (red pentagon) and C. montanus subsp.velutinus (blue triangle).

139

CONCLUSÕES FINAIS O presente estudo enriqueceu de maneira considerável a compreensão sobre Cryptangieae. A partir da investigação de caracteres moleculares e morfológicos, foram realizados estudos taxonômicos e foi possível esclarecer questões sobre a circunscrição tanto das tribos mais basais da subfamília Cyperoideae (Trilepideae, Sclerieae, Bisboeckelereae, Schoeneae e Cryptangieae), como as relações filogenéticas dentro da própria tribo Cryptangieae.

Na primeira etapa da pesquisa, foi reconstruída a hipótese filogenética de Cyperaceae com base em dados de duas regiões de cloroplasto (rbcL e trnL-F) e uma região de DNA ribossomal (ITS) usando análises de Máxima Parcimônia e Inferência Bayesiana (Capítulo 1 – CAP1). Essas análises incluíram de modo suficiente e o quanto possível a diversidade da família. Os resultados apoiaram Cryptangieae como um grupo monofilético em todas as análises (de região única e matriz concatenada) e a manutenção de seu status de tribo tem apoio em suas características morfológicas. Infelizmente não foi possível determinar o grupo-irmão de Cryptangieae, mas provavelmente está em alguns dos grupos hoje aceitos como Schoeneae. Schoeneae é reconstruída como polifilética nos nossos resultados (CAP1) e em outros trabalhos filogenéticos de Cyperaceae com dados moleculares (Muasya et al. 2009, Hinchliff & Roalson 2013); estudos sobre sua circunscrição devem ser publicados em breve (Larridon, comunicação pessoal). Outra lacuna a ser preenchida é a inclusão de sequencias de Koyamaea W.W. Thomas & Davidse em análises com ferramentas moleculares. Este gênero monoespecífico (representado por K. neblinensis) tem poucas coleções disponíveis em herbários, todas provenientes do Pico da Neblina (Venezuela-Brasil) e encontra-se como incertae sedis em Cyperoideae desde o trabalho de Goetghebeur (1998); com indícios morfológicos de proximidade com Sclerieae ou Cryptangieae. Eu (SMC) tentei obter DNA de uma amostra de herbário, usando kits de extração e pelo menos dois protocolos CTAB modificados, mas o DNA total obtido não tinha qualidade para gerar boas sequências, mesmo de 5S-NTS; região com maior número de sequências para amostras de herbário.

Após a apreciação desses primeiros resultados, continuamos tratando Cryptangieae como tribo, sem sua inclusão em Schoeneae como sugerido por Hinchliff & Roalson (2013), e passamos às questões seguintes.

Embora o número de espécies de Cryptangieae utilizadas no CAP1 tenha sido relativamente pequeno (18 spp./ ca. 35%), ficou evidente a necessidade de revisão da circunscrição de seus gêneros. No Capítulo 2 (CAP2) incluímos sequências de

140

aproximadamente 70% das espécies de Cryptangieae e terminais do grupo-externo incluindo representantes de Trilepideae, Sclerieae e Bisboeckelereae. Além das regiões anteriormente utilizadas (rbcL, trnL-F, ITS), adicionamos sequências para mais duas regiões de DNA ribossomal (ETS, 5S-NTS) potencialmente mais informativas.

As regiões de DNA amostradas (rbcL, trnL-F, ITS, ETS e 5S-NTS) foram suficientes para distinguir gêneros mas inadequadas para determinar a relação entre espécies. Talvez estudos utilizando DNA nuclear de baixa cópia ou técnicas de Sequenciamento de Nova Geração possam auxiliar nesta tarefa.

Através das análises de região única e de matriz concatenada, novamente aplicando análises de Máxima Parcimônia e de Inferência Bayesiana, reconhecemos seis grupos coerentes segundo ferramentas moleculares e morfológicas. Defendemos o status de gêneros para os sequintes grupos: Cephalocarpus Nees, Cryptangium Scharder, Didymiandrum Gilly, Exochogyne C.B.Clarke, Krenakia S.M.Costa (nom. nov. para Acrocarpus Nees nom. illeg.), Lagenocarpus Nees. Ainda neste manuscrito, fornecemos chaves de determinação, descrições sucintas e lista de espécies aceitas (incluindo as novas combinações) para os gêneros tal como propostos (CAP2).

Nesta mesma etapa (CAP2) foram realizadas análises de reconstrução de estado de caráter ancestral com os principais caracteres diagnósticos aplicados na taxonomia de Cryptangieae. Ficou evidente a importância de utilizar pelo menos dois caracteres (vegetativos e/ou reprodutivos) para o melhor reconhecimento dos táxons, uma vez que muitos dos caracteres diagnósticos historicamente utilizados para o grupo são variáveis dentro de cada gênero (ornamentação da escama hipógina, por exemplo) e algumas vezes tem surgimento independente (dioicia, por exemplo).

O Capítulo 3 (CAP3) traz as novidades taxonômicas encontradas durante as visitas aos herbários, no Brasil e nos Estados Unidos, durante os anos de doutoramento. As dez espécies novas (sete em Cephalocarpus e três em Lagenocarpus) são adequadamente nomeadas, descritas e ilustradas. Cada uma delas recebeu comentários sobre morfologia, relações com outras espécies e distribuição geográfica. É interessante observar que a maioria dos espécimes destas novas espécies estava depositada há mais de 20 anos nos herbários visitados, principalmente nas décadas de 1950 e 1980. Estas datas coincidem com os esforços de coletas de instituições dos Estados Unidos na região do Escudo das Guianas. Além de demonstrar a importância desse maior esforço de coleta, estas espécies são um bom exemplo

141

de como a biodiversidade pode estar “escondida” em herbários, sendo ainda necessários tratamentos taxonômicos especializados para sua descoberta.

O quarto e último capítulo (CAP4) traz o tratamento taxonômico de Cephalocarpus Nees sob sua circunscrição aqui proposta, incluindo Everardia Ridley. Cephalocarpus passa a ser o gênero com o maior número de espécies da tribo, com 25 spp., e abarca uma maior variedade de morfologia superficial da inflorescência e do aquênio. É fornecida uma descrição mais completa e atualizada para o gênero, assim como chaves de identificação de espécies. As espécies são acompanhadas de sinonímia, descrição sucinta, comentários taxonômicos e geográficos e material examinado. Diversas ilustrações e fotografias são fornecidas; tentando adicionar informações já contempladas por manuscritos anteriores.

Embora este tratamento taxonômico de Cephalocarpus (incluindo Everardia) tenha se baseado na análise de mais espécimes que tratamentos anteriores, não foram visitados os herbários dos demais países de ocorrência do gênero (Colômbia, Peru, Guiana, Venezuela, Equador e Suriname). Tendo em vista o considerável número de novas espécies descobertas durante a identificação de coleções nos herbários dos Estados Unidos, é possível que novas espécies estejam ocultas nessas coleções latino-americanas. Também seria interessante a realização de expedições de coletas as áreas de ocorrência para visualização das espécies em campo. Alguns dos casos de complexos de espécies ou espécies duvidosas poderiam ser reavaliadas com informações de campo, perdidas durante a herborização das plantas. Apenas uma expedição de campo foi realizada para áre de ocorrência de Cephalocarpus, na Serra do Aracá (Manaus - Brasil), onde foram coletados indivíduos de pelo menos três espécies do gênero, uma delas de determinação ainda incerta.

Os tratamentos taxonômicos de Cryptangium, Didymiandrum, Exochogyne, Krenakia e Lagenocarpus segundo circunscrição atualizada e trazendo informações mais recentes sobre sua ocorrência, especialmente no Brasil, devem ser realizados em breve. Dr. Fábio Vitta, sob a orientação do Prof. Dr. George Sheperd e Prof. Dr. W.Wayt Thomas, realizou estudos morfológicos com a maioria das espécies, com exceção das espécies novas, Exochogyne e duas espécies amazônicas de Lagenocarpus durante seu doutoramento. Juntamente com o Prof. Dr. W.W. Thomas, estamos resgatando esses dados e preparando-os para publicação válida, respeitando a propriedade intelectual dos envolvidos.

142

Dada a extensão das questões filogenéticas e taxonômicas estudadas nesta tese, não foi possível tratar outros aspectos interessantes onde Cryptangieae pode ser utilizada como grupo-modelo. Alguns desses temas tratam sobre a evolução dos caracteres morfológicos e anatômicos de suas espécies: desenvolvimento de estruturas caulinares (rizomas e cáudex), desenvolvimento e estrutura de inflorescências (p.ex., lateral vs. terminal, padrões de ramificação), estrutura anatômica dos aquênios nos gêneros, entre outros. A ecofisiologia do grupo, que já foi anteriormente estudada para Lagenocarpus rigidus e adaptações radiculares para o tipo de ambiente onde ocorre a maioria das espécies da tribo (solos pobres no Escudo das Guianas e campos rupestres), também merece ser melhor investigada.

As espécies de Cryptangieae, especialmente pertencentes à Lagenocarpus, são promissoras para estudos confirmando e analisando características sobre anemofilia, por apresentarem flores masculinas e femininas separadas espacialmente e muitas vezes temporalmente; facilitando a montagem de experimentos de polinização. Além de ser um ótimo modelo para estudos biogeográficos de OCBILs neotropicais, uma vez que sua maior diversidade ocorre nas duas regiões mais antigas da América do Sul (Escudo das Guianas e campos rupestres).

Cyperaceae tem importância crucial em muitas das formações vegetacionais abertas onde ocorre e podem servir como modelo em estudos de diferentes áreas da pesquisa botânica. Estes estudos devem ser realizados futuramente; seja por mim (S.M.C.) ou por outros cientistas interessados na complexidade escondida em Cyperaceae.

143

REFERÊNCIAS BIBLIOGRÁFICAS

Ainsworth, C. 2000. Boys and Girls come out to play: the molecular biology of dioecious plants. Annals of Botany 86: 211 –221. DOI 10.1006/anbo.2000.120 Alves, M. & Araujo, C.A.; Prata, A.P.; Vitta, F.; Hefler, S.; Trevisan, R.; Gil, A.S.B.; Martins, S.; Thomas, W. (2009) Diversity of Cyperaceae in Brazil. Rodriguésia. 60(4): 771 – 782. Bebber, D.P.; Carine, M.A.; Wood, J.R.I.; Wortley, A.H.; Harris, D.J.; Prance, G.T.; Davidse, G.; Paige, J.; Pennington, T.D.; Robson, N.K.B. & Scotland, R.W (2010) Herbaria are a major frontier for species discovery. PNAS 107(51): 22169-22171.

Bremer, K. (2002) Gondwanan evolution of the grass alliance of families (Poales). Evolution 56: 1374–1387. https://doi.org /10.1111/j.0014-3820.2002.tb01451.x Bruhl, J.J. (1995) Sedge genera of the world: relationships and a new classification of the Cyperaceae. Australian Systematic Botany 8: 125–305. https://doi.org/10.1071/SB9950125 Chase, M.W., Soltis, D.E., Olmstead, R.G., Morgan, D., Les, D.H., Mishler, B.D., Duvall, M.R., Price, R.A., Hills, H.G., Qiu, Y.-L., Kron, K.A., Rettig, J.H., Conti J, E., Palmer, D., Manhart, J.R., Sytsma, K.J., Michaels, H.J., Kress, W.J., Karol, K.G., Clark, W.D., Hedrén, M., Gaut, B.S., Jansen, R.K., Kim, K.-J., Wimpee, C.F., Smith, J.F., Furnier, G.R., Strauss, S.H., Xiang, Q.-Y., Plunkett, G.M., Soltis,P.S., Swenson, S.M., Williams, S.E., Gadek, P.A., Quinn, C.J., Eguiarte, L.E., Golenberg, E., Learn Jr., G.H., Graham, S.W., Barrett, S.C.H., Dayanandan, S. & Albert., V.A. (1993) Phylogenetics of seed plants: an analysis of nucleotide sequences from the plastid gene rbcL. Annals of the Missouri Botanical Garden 80: 528–586. https://doi.org/10.2307/2399846 Cox, A.V., Bennett, M.D. & Dryer, T.A. 1992. Use of polymerase chain reaction to detect spacer size heterogeneity un plant 5S-rRNA gene clusters and locate such clusters in wheat (Triticum aestivum L.). Theor. Appl. Genet.83: 684--690. Darriba, D., Taboada, G.L., Doallo, R. & Posada, D. (2012) jModelTest 2: more models, new heuristics and parallel computing. Nature Methods 9: 772. https://doi.org/10.1038/nmeth.2109

144

Dhooge, S., Goetghebeur, P. & Muasya, A.M. (2003) Zameioscirpus, a new genus of Cyperaceae from South America. Plant Systematics and Evolution 2431–2: 73–84 https://doi.org/0.1007/s00606-003-0065-3 Elliott, T.L. & Muasya, A.M. (2017) Taxonomic realignment in the southern African Tetraria (Cyperaceae, tribe Schoeneae; Schoenus clade). South African Journal of Botany 112: 354–360. https://doi.org/10.1016/j.sajb.2017.06.011 Esenbeck, N. 1834. Synopsis Generum Cyperacearum. Linnaea 9(3): 273--306. ESRI (2015) ArcGIS Desktop: Release 10.3.1. Environmental Systems Research Institute, California, USA.

Friedman, J. 2011. Gone with the wind: understanding evolutionary transitions between wind and anumal pollination in the angiosperms. New Physiologist 191: 911 –913. Gilly, C. 1941a. The Genus Everardia. Bull. Torrey Bot. Club 68 (1): 20--31. http://www.jstor.org/stable/2481252 Gilly, C. 1941b. The new cyperaceous genus from Northern South America. Bull. Torrey Bot.Club 68 (5): 330--332. http://www.jstor.org/stable/2481606. Gilly, C. 1942. The genus Cephalocarpus Nees (Cyperaceae) Bull. Torrey Bot. Club 69 (4): 290--297. http://www.jstor.org/stable/2481406 . Gilly, C. 1951. Botanical exploration in Venezuela – I. Tribe Cryptangieae (Lagenocarpeae). Fieldiana Bot. 28: 53--60. Goetghebeur, P. (1998) Cyperaceae. In: Kubitzki, K. et al. (eds.) The families and genera of vascular plants. Monocotyledons – Alismatanae and Commelinanae (except Gramineae). Springer, Berlin, pp. 141–190. https://doi.org/10.1007/978-3-662-03531-3_15 Goloboff, P.A., Farris, J.S. & Nixon, K.C. (2008) TNT, a free program for phylogenetic analysis. Cladistics 24: 774–786. https://doi.org/10.1111/j.1096-0031.2008.00217.x Goloboff, P. & Catalano, S. (2016) TNT, version 1.5, with a full implementation of phylogenetic morphometrics. Cladistics 32: 221–232. https://doi.org/10.1111/cla.12160 Govaerts, R., Jimenez-Mejias, P., Koopman, J., Simpson, D., Goetghebeur, P., Wilson, K., Egorova, T. & Bruhl, J. (2017). World Checklist of Cyperaceae. Facilitated by the Royal

145

Botanic Gardens, Kew. Disponível em: http://wcsp.science.kew.org (retrieved 24 October 2017) Guindon, S. & Gascuel, O. (2003) A simple, fast and accurate method to estimate large phylogenies by maximum-likelihood. Systematic Biology 52: 696–704. https://doi.org/10.1080/10635150390235520 Hinchliff, C.E. & Roalson, E.H. (2013) Using supermatrices for phylogenetic inquiry: an example using sedges. Systematic Biology 62: 205–219. http://doi.org/10.1093/sysbio/sys088 Hopper, S.D. 2009. OCBIL theory: towards an integrated understanding of the evolution, ecology and conservation of biodiversity on old, climatically buffered, infertile landscapes. Plant Soil (322): 48—86. Huelsenbeck, J.P. & Ronquist, F. (2001) MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 19: 1572–1574. https://doi.org/10.1093/bioinformatics/btg180 Jung, J. & Choi, H. (2013) Recognition of two major clades and early diverged groups within the subfamily Cyperoideae (Cyperaceae) including Korean sedges. Journal of Plant Research 126: 335–349. https://doi.org/10.1007/s10265-012-0534-2 Katoh, K., Misawa, K., Kuma, K. & Miyata, T. (2002) MAFFT: A novel method for rapid multiple sequence alignment based on fast Fourier transform. Nucleic Acids Research 30: 3059–3066. https://doi.org/10.1093/nar/gkf436 Koyama, T. (2003) A taxonomic account of the genera Cephalocarpus and Everardia (Cyperaceae) – Materials for a Cyperaceae monograph of the Flora Neotropica. Part I. Makinoa 2: 3–38. Koyama, T. (2004) A taxonomic revision of the genus Lagenocarpus (Cyperaceae) – Materials for a Cyperaceae monograph of the Flora Neotropica. Part II. Makinoa 4: 1–73. Koyama, T. & Maguire, B. (1965) Cyperaceae tribe Lagenocarpeae. Botany of the Guayana Highland 12: 8–54. Kress, W.J., Wurdack, K.J., Zimmer, E.A. & Janzen, D.H. (2005) Use of DNA barcodes to identify flowering plants. PNAS 102: 8369–8374. https://doi.org/10.1073/pnas.0503123102

146

Maddison, W. & Maddison, D.R. (2007) Mesquite: A modular system for evolutionary analysis, version 2.01 build j28. Available from:http://www.mesquiteproj.org (accessed 1 April 2018) Muasya, A.M., Simpson, D.A. & Chase, M.W. (2002) Phylogenetic relationships in Cyperus L. s.l. (Cyperaceae) inferred from plastid DNA sequence data. Botanical Journal of the Linnean Society 138: 145–153. https://doi.org/10.1046/j.1095-8339.2002.138002145.x Muasya, A.M., Simpson, D.A., Verboom, G.A., Goetghebeur, P., Naczi, R.F.C., Chase, M.W. & Smets, E. (2009) Phylogeny of Cyperaceae based on DNA sequence data: current progress and future prospects. Botanical Review 75: 2–21. http://doi.org/10.1007/s12229-008-9019-3 Roalson, E.H. & Friar, E.A. (2004) Phylogenetic relationships and biogeographic patterns in North American members of Carex section Acrocystis (Cyperaceae) using nrDNA ITS and ETS sequence data. Plant Systematic and Evolution 243: 175–187. https://doi.org/10.1007/s00606-003-0089-8 Simpson, D.A., Furness, C.A., Hodkinson, T.R., Muasya, M. & Chase, M.W. (2003) Phylogenetic relationships in Cyperaceae subfamily Mapanioideae inferred from pollen and plastid DNA sequence data. American Journal of Botany 90: 1071–1086. https://doi.org/10.3732/ajb.90.7.1071 Simpson, D.A., Muasya, A.M., Alves, M.V., Bruhl, J.J. & Dhooge, S. (2007) Phylogeny of Cyperaceae based on DNA sequence data – a new rbcL analysis. Aliso: A Journal of Systematic and Evolutionary Botany 23 (1): 72–83. http://doi.org/10.5642/aliso.20072301.09 Strong, M. T. 2016. Three new Cyperaceae from the Cuyuni-Mazaruni Region of Guyana (South America). Novon: a Journal for Botanical Nomenclature 24 (4): 401--407. http://dx.doi.org/10.3417/2015032 Thiers, B. (continuously updated) Index Herbariorum: A global directory of public herbaria and associated staff. New York Botanical Garden’s Virtual Herbarium, New York. Available from: http://sweetgum.nybg.org/ih/ (accessed 1 April 2018) Viljoen, J., Muasya, A.M., Barrett, R.L., Bruhl, J.J., Gibbs, A.K., Slingsby, J.A., Wilson, K.L. & Verboom, G.A. (2013) Radiation and repeated transoceanic dispersal of Schoeneae (Cyperaceae) through the southern hemisphere. American Journal of Botany 100: 2494– 2508.

147

https://doi.org/10.3732/ajb.1300105 Vitta, F.A. (2005) Revisão taxonômica e estudos morfológicos e biossistemáticos em Cryptangium Schrad. ex Nees e Lagenocarpus Nees. (Cyperaceae: Cryptangieae). PhD Thesis – Pos-graduate Program on Plant Biology/University of Campinas (UNICAMP), Campinas – São Paulo/Brazil, 276 pp. Disponível em: http://repositorio.unicamp.br/handle/REPOSIP/315028. Wardle, P., Ezcurra, C., Ramirez, C. & Wagstaff, S. (2001) Comparison of the flora and vegetation of the southern Andes and New Zealand. New Zealand Journal of Botany 39: 69–108. Yen, A.C. & Olmstead, R.G. (2000) Molecular systematics of Cyperaceae tribe Cariceae based on two chloroplast DNA regions: ndhF and trnL intron-intergenic spacer. Systematic Botany 25: 479–494. https://doi.org/10.2307/2666691 Zhang, X., Marchant, A., Wilson, K.L. & Bruhl, J.J. (2004) Phylogenetic relationships of Carpha and its relatives (Schoeneae, Cyperaceae) inferred from chloroplast trnL intron and trnL-trnF intergenic spacer sequences. Molecular Phylogenetics and Evolution 31: 647– 657. https://doi.org/10.1016/j.ympev.2003.09.004

148

APPENDIX 1 – Ancestral Character State analysis to each diagnostic morphological character

149

150

151

152

153

154

155

156

157

158

159

160

161

APPENDIX 2 – Phylogenetic hypothesis on Cryptangieae genera based single-marker matrix. [PP values above and BS values below and between brackets. Group/genus color: black – outgroup; light green – Cryptangium; pink – Didymiandrum; brownish-green – Exochogyne; purple – Krenakia; red – Everardia; blue – Cephalocarpus; yellow - Lagenocarpus]

162

163

ITS

164

165

166

APPENDIX 3 – Illustrations of Cephalocarpus from the treatments of Gilly (1942, 1943), Koyama & Maguire (1965) and Koyama (2003)

167

168

169

170

171

172

173

174

175

176

177

178

179

180

181

ANEXO 1- Declaração de bioética e/ou biossegurança

182

ANEXO 2- Declaração referente aos direitos autorais