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62 TAXON )HEUXDU\ ± 'DUX DO Phylogeny of Alooideae 62 9HUVLRQ RI 5HFRUG LGHQWLFDO WR SULQW YHUVLRQ INTRODUCTION $VSKRGHODFHDH VXEIDP $ORRLGHDH VHQVX $3* ,, $ORDFHDH VHQVX 6PLWK 6WH\Q DUH DQ 2OG :RUOG JURXS FRPSULVLQJ VRPH VSHFLHV FKDUDFWHULVHG E\ PRUH RU OHVV GLV- WLQFWO\ VXFFXOHQW OHDYHV RIWHQ ZLWK SULFNO\ RU WRRWKHG PDUJLQV DQG D PDUNHGO\ ELPRGDO NDU\RW\SH ZLWK WKH EDVLF FKURPR- VRPH QXPEHU x 7D\ORU 6PLWK 9DQ :\N 5HSUHVHQWDWLYHV RI WKH VXEIDPLO\ DOVR VKDUH VRPH FKHPLFDO FKDUDFWHUV QRWDEO\ WKH SUHVHQFH RI DQWKURQH&JO\FRVLGHV LQ WKHLU OHDYHV DQG PHWK\OK\GUR[\DQWKUDTXLQRQHV LQ WKHLU URRWV 6PLWK 9DQ :\N 0RGHUQ WD[RQRP\ RI WKH JURXS EHJLQV ZLWK /LQQDHXV ZKRVH UDWKHU KHWHURJHQHRXV FRQFHSW RI Aloe / HQFRP- SDVVHG VHYHUDO $VSDUDJDOLDQ WD[D ZLWK PRUH RU OHVV WXEXODU IORZHUV DQG OHDWKHU\ RU VXFFXOHQW OHDYHV 2I WKH QDPHV LQ- FOXGHG E\ KLP LQ WKH JHQXV IRXU DUH QRW PHPEHUV RI $ORRLGHDH WKUHH DUH QRZ LQ Sansevieria 7KXQE DQG RQH LQ Kniphofia 0RHQFK 7KH UHPDLQGHU JURXSHG E\ IORZHU VL]H DQG VKDSH DUH FXUUHQWO\ VHJUHJDWHG DPRQJ Aloe IRXU VSHFLHV Gasteria 'XYDO RQH VSHFLHV Astroloba 8LWHZDDO RQH VSHFLHV DQG Haworthia 'XYDO ILYH VSHFLHV /LQQDHXV¶V SUHOLPLQDU\ JURXSLQJ RI WKH DORRLG VSHFLHV ZDV VXEVHTXHQWO\ IRUPDOLVHG ILUVW DW VHF- WLRQDO OHYHO ZLWKLQ Aloe E\ 6DOP'\FN ± DQG ODWHU E\ WKH UHFRJQLWLRQ RI WKH VHJUHJDWH JHQHUD Gasteria GHILQHG E\ WKH PRGHUDWHO\ ODUJH FXUYHG DQG RIWHQ JDVWHULIRUP IORZ- HUV DQG Haworthia ZLWK YHU\ PXFK VPDOOHU ZKLWLVK IORZHUV 'XYDO Haworthia ZDV IXUWKHU VSOLW ZKHQ WKRVH VSHFLHV ZLWK PRUH RU OHVV DFWLQRPRUSKLF IORZHUV ZHUH VHSDUDWHG IURP WKRVH ZLWK ELODELDWH IORZHUV LQWR WKH VPDOO JHQXV Astroloba 8LWHZDDO 7KUHH DGGLWLRQDO VPDOO JHQHUD KDYH VLQFH EHHQ UHFRJQLVHG QDPHO\ Chortolirion $ %HUJHU Lomato- phyllum :LOOG DQG Poellnitzia 8LWHZDDO EXW WKH FXUUHQW FODV- VLILFDWLRQ RI WKH VXEIDPLO\ UHYLHZHG LQ .ORSSHU DO LQFOXGHV Lomatophyllum LQ Aloe DQG Poellnitzia LQ Astroloba WKXV UHWDLQLQJ WKH ILYH JHQHUD Aloe Astroloba Chortolirion Molecular and morphological analysis of subfamily Alooideae (Asphodelaceae) and the inclusion of Chortolirion in Aloe Barnabas H. Daru, John C. Manning, James S. Boatwright, Olivier Maurin, Norman Maclean, Hanno Schaefer, Maria Kuzmina & Michelle van der Bank 1 African Centre for DNA Barcoding, University of Johannesburg, P.O. Box 524 Auckland Park 2006, Johannesburg, South Africa 2 Compton Herbarium, South African National Biodiversity Institute, Private Bag X7, Claremont 7735, South Africa Research Centre for Plant Growth and Development, School of Biological and Conservation Sciences, University of KwaZulu-Natal, Pietermaritzburg, Private Bag X01, Scottsville 3209, South Africa Department of Biodiversity and Conservation Biology, University of the Western Cape, Private Bag X17, Bellville 7535, Cape Town, South Africa School of Biological Sciences, University of Southampton, Highfield, Southampton, Hants, SO16 7PX, U.K. 6 Technische Universitaet Muenchen, Biodiversitaet der Pflanzen, Maximus-von-Imhof Forum 2, 85354 Freising, Germany International Barcode of Life Project, Biodiversity Institute of Ontario, University of Guelph, Guelph, Ontario N1G 2W1, Canada $XWKRU IRU FRUUHVSRQGHQFH Barnabas H. Daru, [email protected] Abstract $VSKRGHODFHDH VXEIDP $ORRLGHDH $VSDUDJDOHV FXUUHQWO\ FRPSULVHV ILYH JHQHUD IRXU RI ZKLFK DUH HQGHPLF WR VRXWKHUQ $IULFD 'HVSLWH WKHLU LPSRUWDQFH LQ FRPPHUFLDO KRUWLFXOWXUH WKH HYROXWLRQDU\ UHODWLRQVKLSV DPRQJ WKH JHQHUD DUH VWLOO LQFRPSOHWHO\ XQGHUVWRRG 7KLV VWXG\ H[DPLQHV SK\ORJHQHWLF UHODWLRQVKLSV LQ WKH VXEIDPLO\ XVLQJ DQ H[SDQGHG PROHFXODU VHTXHQFH GDWDVHW IURP WKUHH SODVWLG UHJLRQV matK rbcLa trnH-psbA DQG WKH ILUVW VXEXQLW RI WKH QXFOHDU ULERVRPDO LQWHUQDO WUDQVFULEHG VSDFHU ,76 6HTXHQFH GDWD ZHUH DQDO\VHG XVLQJ PD[LPXP SDUVLPRQ\ DQG %D\HVLDQ VWDWLVWLFV DQG VHOHFWHG PRUSKRORJLFDO WUDLWV ZHUH PDSSHG RQWR WKH PROHFXODU SK\ORJHQ\ Haworthia LV FRQILUPHG DV EHLQJ SRO\SK\OHWLF FRPSULVLQJ WKUHH PDLQ FODGHV WKDW ODUJHO\ FRUUHODWH ZLWK FXUUHQW VXEJHQHULF FLUFXPVFULSWLRQV Astroloba DQG Gasteria DUH HYLGHQWO\ HDFK PRQRSK\OHWLF DQG VLVWHU UHVSHFWLYHO\ WR Astroloba DQG H. VXEJ Robustipedunculares Chortolirion LV VKRZQ WR EH GHHSO\ QHVWHG ZLWKLQ Aloe DQG LV IRUPDOO\ LQFOXGHG LQ WKDW JHQXV Aloe LWVHOI LV FOHDUO\ SRO\SK\OHWLF ZLWK WKH GZDUI VSHFLHV A. aristata DOOLHG WR Haworthia VXEJ Robustipedunculares 7KH WD[RQRPLF LPSOLFDWLRQV RI WKHVH ILQGLQJV DUH H[DPLQHG EXW EUDQFK VXSSRUW DW FULWLFDO ORZHU QRGHV LV LQVXIILFLHQW DW WKLV VWDJH WR MXVWLI\ LPSOHPHQWLQJ PDMRU WD[RQRPLF FKDQJHV Keywords $VSKRGHODFHDH ,76 matK PROHFXODU SK\ORJHQ\ PRUSKRORJ\ QRPHQFODWXUH rbcLa trnH-psbA WD[RQRP\ Supplementary Material 7KH (OHFWURQLF 6XSSOHPHQW )LJV 6±6 $SSHQGLFHV 6±6 DQG WKH DOLJQPHQW DUH DYDLODEOH LQ WKH 6XSSOHPHQWDU\ 'DWD VHFWLRQ RI WKH RQOLQH YHUVLRQ RI WKLV DUWLFOH KWWSZZZLQJHQWDFRQQHFWFRPFRQWHQWLDSWWD[ 5HFHLYHG 0D\ UHYLVLRQ UHFHLYHG 2FW DFFHSWHG 1RY

Molecular and morphological analysis of subfamily ... · huv dqg haworthia zlwk yhu\ pxfk vpdoohu zklwlvk iorzhuv 'xydo haworthia zdv ixuwkhu vsolw zkhq wkrvh vshflhv zlwk pruh ru

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Page 1: Molecular and morphological analysis of subfamily ... · huv dqg haworthia zlwk yhu\ pxfk vpdoohu zklwlvk iorzhuv 'xydo haworthia zdv ixuwkhu vsolw zkhq wkrvh vshflhv zlwk pruh ru

62

TAXON����������)HEUXDU\���������±��'DUX��DO����Phylogeny of Alooideae

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INTRODUCTION

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Molecular and morphological analysis of subfamily Alooideae (Asphodelaceae) and the inclusion of Chortolirion in Aloe

Barnabas H. Daru,! John C. Manning,"#$ James S. Boatwright,% Olivier Maurin,! Norman Maclean,& Hanno Schaefer,' Maria Kuzmina( & Michelle van der Bank!

1 African Centre for DNA Barcoding, University of Johannesburg, P.O. Box 524 Auckland Park 2006, Johannesburg, South Africa2 Compton Herbarium, South African National Biodiversity Institute, Private Bag X7, Claremont 7735, South Africa�� Research Centre for Plant Growth and Development, School of Biological and Conservation Sciences, University of

KwaZulu-Natal, Pietermaritzburg, Private Bag X01, Scottsville 3209, South Africa�� Department of Biodiversity and Conservation Biology, University of the Western Cape, Private Bag X17, Bellville 7535, Cape Town,

South Africa�� School of Biological Sciences, University of Southampton, Highfield, Southampton, Hants, SO16 7PX, U.K.6 Technische Universitaet Muenchen, Biodiversitaet der Pflanzen, Maximus-von-Imhof Forum 2, 85354 Freising, Germany�� International Barcode of Life Project, Biodiversity Institute of Ontario, University of Guelph, Guelph, Ontario N1G 2W1, Canada$XWKRU�IRU�FRUUHVSRQGHQFH� Barnabas H. Daru, [email protected]

Abstract $VSKRGHODFHDH�VXEIDP��$ORRLGHDH��$VSDUDJDOHV��FXUUHQWO\�FRPSULVHV�ILYH�JHQHUD��IRXU�RI�ZKLFK�DUH�HQGHPLF�WR�VRXWKHUQ�$IULFD��'HVSLWH�WKHLU�LPSRUWDQFH�LQ�FRPPHUFLDO�KRUWLFXOWXUH�WKH�HYROXWLRQDU\�UHODWLRQVKLSV�DPRQJ�WKH�JHQHUD�DUH�VWLOO�LQFRPSOHWHO\�XQGHUVWRRG��7KLV�VWXG\�H[DPLQHV�SK\ORJHQHWLF�UHODWLRQVKLSV�LQ�WKH�VXEIDPLO\�XVLQJ�DQ�H[SDQGHG�PROHFXODU�VHTXHQFH�GDWDVHW�IURP�WKUHH�SODVWLG�UHJLRQV��matK��rbcLa� trnH-psbA��DQG�WKH�ILUVW�VXEXQLW�RI�WKH�QXFOHDU�ULERVRPDO�LQWHUQDO�WUDQVFULEHG�VSDFHU��,76����6HTXHQFH�GDWD�ZHUH�DQDO\VHG�XVLQJ�PD[LPXP�SDUVLPRQ\�DQG�%D\HVLDQ�VWDWLVWLFV��DQG�VHOHFWHG�PRUSKRORJLFDO�WUDLWV�ZHUH�PDSSHG�RQWR�WKH�PROHFXODU�SK\ORJHQ\��Haworthia LV�FRQILUPHG�DV�EHLQJ�SRO\SK\OHWLF��FRPSULVLQJ�WKUHH�PDLQ�FODGHV�WKDW�ODUJHO\�FRUUHODWH�ZLWK�FXUUHQW�VXEJHQHULF�FLUFXPVFULSWLRQV��Astroloba DQG�Gasteria DUH�HYLGHQWO\�HDFK�PRQRSK\OHWLF�DQG�VLVWHU�UHVSHFWLYHO\�WR�Astroloba DQG�H. VXEJ� Robustipedunculares��Chortolirion LV�VKRZQ�WR�EH�GHHSO\�QHVWHG�ZLWKLQ�Aloe DQG�LV�IRUPDOO\�LQFOXGHG�LQ�WKDW�JHQXV��Aloe LWVHOI�LV�FOHDUO\�SRO\SK\OHWLF��ZLWK�WKH�GZDUI�VSHFLHV�A. aristata DOOLHG�WR�Haworthia VXEJ��Robustipedunculares��7KH�WD[RQRPLF�LPSOLFDWLRQV�RI�WKHVH�ILQGLQJV�DUH�H[DPLQHG�EXW�EUDQFK�VXSSRUW�DW�FULWLFDO�ORZHU�QRGHV�LV�LQVXIILFLHQW�DW�WKLV�VWDJH�WR�MXVWLI\�LPSOHPHQWLQJ�PDMRU�WD[RQRPLF�FKDQJHV�

Keywords $VSKRGHODFHDH� ,76���matK� PROHFXODU SK\ORJHQ\��PRUSKRORJ\��QRPHQFODWXUH� rbcLa� trnH-psbA��WD[RQRP\

Supplementary Material 7KH�(OHFWURQLF�6XSSOHPHQW��)LJV��6�±6���$SSHQGLFHV�6�±6���DQG�WKH�DOLJQPHQW�DUH�DYDLODEOH�LQ�WKH�6XSSOHPHQWDU\�'DWD�VHFWLRQ�RI�WKH�RQOLQH�YHUVLRQ�RI�WKLV�DUWLFOH��KWWS���ZZZ�LQJHQWDFRQQHFW�FRP�FRQWHQW�LDSW�WD[��

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MATERIALS AND METHODS

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03�DQDO\VHV�ZHUH�SHUIRUPHG�RQ�WKH�,76���FRPELQHG�SODV-WLG��DQG�WRWDO�FRPELQHG�GDWDVHWV�ZKHUHDV�%,�ZDV�HPSOR\HG�RQO\�RQ�WKH�WRWDO�FRPELQHG�GDWDVHW��03�DQDO\VHV�ZHUH�SHUIRUPHG�XVLQJ�3$83 �Y����E�����6ZRIIRUG���������7UHH�VHDUFKHV�ZHUH�FRQGXFWHG�XVLQJ������UDQGRP�VHTXHQFH�DGGLWLRQV��UHWDLQLQJ�WHQ�WUHHV�DW�HDFK�VWHS��ZLWK�WUHH�ELVHFWLRQ�UHFRQQHFWLRQ��7%5��EUDQFK�VZDSSLQJ�DQG�0XO7UHHV�LQ�HIIHFW��7KH�UHVXOWLQJ�WUHHV�ZHUH�WKHQ�XVHG�DV�VWDUWLQJ�WUHHV�LQ�D�VHFRQG�VHDUFK�ZLWK�WKH�VDPH�SDUDPHWHUV�EXW�ZLWKRXW�D�OLPLW�IRU�WKH�QXPEHU�RI�WUHHV�SHU�UHSOLFDWH��VZDSSLQJ�WR�FRPSOHWLRQ���LQ�RUGHU�WR�VHH�LI�WKH�VKRUWHVW�WUHHV�ZHUH�IRXQG�LQ�WKH�SUHYLRXV�DQDO\VLV��'HOD\HG�WUDQVIRUPDWLRQ��'(/75$1��FKDUDFWHU�RSWLPLVDWLRQ�ZDV�XVHG�LQVWHDG�RI�DFFHOHUDWLRQ�RI�WUDQVIRUPDWLRQ��$&&75$1��IRU�FDO-FXODWLQJ�EUDQFK�OHQJWKV�EHFDXVH�RI�UHSRUWHG�HUURUV�ZLWK�YHU-VLRQ����E����RI�3$83 ��KWWS���SDXS�FVLW�IVX�HGX�SUREOHPV�KWPO���%UDQFK�VXSSRUW�ZDV�HVWLPDWHG�XVLQJ�ERRWVWUDS�DQDO\VLV��)HOVHQ-VWHLQ��������ZLWK������UHSOLFDWHV��VLPSOH�VHTXHQFH�DGGLWLRQ��7%5�VZDSSLQJ��ZLWK�0XO7UHHV�LQ�HIIHFW�VDYLQJ�WHQ�WUHHV�SHU�UHSOLFDWH��2QO\�JURXSV�RI�JUHDWHU�WKDQ�����ERRWVWUDS�VXSSRUW��%6��ZHUH�UHSRUWHG��7KH�IROORZLQJ�DUELWUDU\�VFDOH�IRU�HYDOXDW-LQJ�%6�ZDV�DSSOLHG��ZHDN�����±������PRGHUDWH�����±�����RU�VWURQJ�����±�������%,��+XHOVHQEHFN��5RQTXLVW��������5RQTXLVW��+XHOVHQEHFN��������ZDV�SHUIRUPHG�XVLQJ�0U%D\HV�Y��������)RU�HDFK�PDWUL[��,76���matK��rbcLa��trnH-psbA��WKH�PRVW�DSSURSULDWH�PRGHO�ZDV�VHOHFWHG�EDVHG�RQ�$NDLNH�LQIRU-PDWLRQ�FULWHULRQ��$,&��LPSOHPHQWHG�LQ�02'(/7(67�Y�������3RVDGD��&UDQGDOO���������7DEOH����

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Page 6: Molecular and morphological analysis of subfamily ... · huv dqg haworthia zlwk yhu\ pxfk vpdoohu zklwlvk iorzhuv 'xydo haworthia zdv ixuwkhu vsolw zkhq wkrvh vshflhv zlwk pruh ru

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XVHG�WR�SURGXFH�D�����PDMRULW\�UXOH�FRQVHQVXV�WUHH�VKRZLQJ�WKH�IUHTXHQFLHV��SRVWHULRU�SUREDELOLWLHV�RU�33��RI�DOO�REVHUYHG�EL�SDUWLWLRQV��7KH�IROORZLQJ�VFDOH�ZDV�XVHG�WR�HYDOXDWH�WKH�33V��EHORZ�������ZHDNO\�VXSSRUWHG������±�����VWURQJO\�VXSSRUWHG��7R�PDS�WKH�ERRWVWUDS�YDOXHV��%6��DQG�33�YDOXHV��0U%D\HV�WUHH��RQWR�WKH�WUHH��WKH�QH[XV�WUHH�ILOH�IURP�WKH�%,�DQDO\VLV�ZDV�UHVFDOHG�XVLQJ�³DSH´�Y��������3DUDGLV��DO���������DQG�³DGH-SK\OR´�Y������-RPEDUW��'UD\��������SDFNDJHV�LPSOHPHQWHG�LQ�5��5�'HYHORSPHQW�&RUH�7HDP��������

Coding of morphological characters. —� $�PDWUL[� RI����PRUSKRORJLFDO�FKDUDFWHUV�ZDV�SUHSDUHG�IRU�WKH���� WD[D RI�$ORRLGHDH�DQG�RXWJURXSV�LQFOXGHG�LQ�WKH�DQDO\VHV��0RVW�WD[RQRPLF�VWXGLHV�LQ�$ORRLGHDH�LGHQWLILHG�WKHVH�GLDJQRVWLF�FKDUDFWHUV��RIWHQ�DW�VSHFLHV�OHYHO��WR�LQIHU�UHODWLRQVKLSV�ZLWKLQ�WKH�VXEIDPLO\��5H\QROGV��������������-HSSH��������%D\HU��������������������������9DQ�-DDUVYHOG��������������������������9DQ�-DDUVYHOG��DO���������*OHQ��6PLWK��������0|VVPHU��DO���������6PLWK������D��E��6PLWK��DO.��������0H\HU��6PLWK��������������*OHQ��+DUG\��������9DQ�:\N��6PLWK��������9DQ�-DDUVYHOG��9DQ�:\N��������������������6PLWK��6WH\Q��������*HUPLVKXL]HQ��DO���������*LOGHQKX\V��������.ORSSHU��6PLWK���������7KHVH�FKDUDFWHUV�ZHUH�VFRUHG�DV�SUHVHQW�RU�DEVHQW�LQ�D�����PDWUL[�DV�LQGLFDWHG�LQ�$SSHQGL[�6���7KH�SDWWHUQV�RI�HYROXWLRQ�RI�WKHVH�FKDUDFWHUV�ZHUH�H[DPLQHG�E\�UHFRQVWUXFW-LQJ�WKHP�RQWR�WKH�PDMRULW\�UXOH�FRQVHQVXV�WUHH�SURGXFHG�E\�WKH�%,�DQDO\VLV�XVLQJ�0HVTXLWH�Y�������0DGGLVRQ��0DGGLVRQ���������0RUSKRORJLFDO�FKDUDFWHUV�DQG�FKDUDFWHU�VWDWHV�DUH�GH-ILQHG�LQ�$SSHQGL[�6��DQG�WKH�GDWD�PDWUL[�XVHG�IRU�FKDUDFWHU�UHFRQVWUXFWLRQV�LV�SUHVHQWHG�LQ�$SSHQGL[�6��

RESULTS

Comparison of sequence partitions. —�7KH�VWDWLVWLFV�IURP�WKH�03�DQDO\VHV�IRU�WKH�VLQJOH�SODVWLG�DQDO\VLV��FRPELQHG�SODV-WLG��PDW.࣠�UEF/D࣠�trnH-psbA���,76��DQG�WKH�WRWDO�FRPELQHG�GDWDVHW�DUH�VKRZQ�LQ�7DEOH����,76��KDG�D�VLJQLILFDQWO\�KLJKHU�

QXPEHU�RI�YDULDEOH�VLWHV�������FRPSDUHG�WR�WKH�SODVWLG�UHJLRQV�FRPELQHG��KHQFHIRUWK�FDOOHG�SODVWLG�GDWDVHW���7KH�QXPEHU�RI�SRWHQWLDOO\�SDUVLPRQ\�LQIRUPDWLYH�FKDUDFWHUV�IRU�WKH�SODVWLG�GDWDVHW�ZLWKLQ�$ORRLGHDH�������LV�PXFK�ORZHU�WKDQ�IRU�,76���������7KH�DYHUDJH�QXPEHU�RI�FKDQJHV�SHU�YDULDEOH�VLWH�IRU�,76�������FKDQJHV�SHU�YDULDEOH�VLWH��UHWHQWLRQ�LQGH[�5,� �������DQG�FRQVLVWHQF\�LQGH[�&,� �������LV�KLJKHU�WKDQ�LQ�WKH�SODVWLG�GDWDVHW������FKDQJHV�SHU�YDULDEOH�VLWH��5,� �������&,� ��������$QDO\VLV�RI�HDFK�RI�WKH�WKUHH�SODVWLG�UHJLRQV��matK� rbcLa� trnH-psbA) UHVXOWHG�LQ�WUHHV�WKDW�ZHUH�VLPLODU�LQ�WRSRORJ\��7UHHV�UHVXOWLQJ�IURP�WKH�IROORZLQJ�DQDO\VHV�DUH�SUHVHQWHG��VLPSOLILHG�WUHH�WR-SRORJLHV�RI�WKH�FRPELQHG�SODVWLG�UHJLRQV��PDW.࣠�UEF/D࣠+࣠WUQ+�psbA��)LJ���$���,76���)LJ���%��DQG�FRPELQHG�SODVWLG�DQG�QXFOHDU�(PDW.࣠�UEF/D࣠+࣠WUQ+�SVE$�,76���)LJ���&���FRPELQHG�SODVWLG�UHJLRQV��PDW.࣠�UEF/D࣠+࣠WUQ+�SVE$���)LJ��6����,76���)LJ��6���DQG�FRPELQHG�SODVWLG�DQG�QXFOHDU�UHJLRQV��PDW.࣠�UEF/D࣠+࣠WUQ+�psbA�,76���)LJ�����

Combined plastid data. —�,QGLYLGXDO�SODVWLG�QXFOHRWLGH�VHTXHQFH�DQDO\VHV��UHVXOWV�QRW�VKRZQ��ZHUH�WRSRORJLFDOO\�FRQ-VLVWHQW��QHJOLJLEOH�WR�]HUR�LQFRQJUXHQFH���DQG�IRU�WKH�SXUSRVH�RI�WKH�UHVXOWV�DQG�GLVFXVVLRQ�ZHUH�FRPELQHG�DQG�WUHDWHG�DV�D�VLQJOH�GDWDVHW��7KH�VWDWLVWLFV�IRU�03�DQDO\VLV�IRU�WKH�FRPELQHG�SODVWLG�GDWD�LV�SUHVHQWHG�LQ�7DEOH����)URP�WKH�KHXULVWLF�VHDUFK��ZH�IRXQG�����PRVW�SDUVLPRQLRXV�SK\ORJHQHWLF�WUHHV�RI�ZKLFK�RQH�LV�SUHVHQWHG�LQ�)LJXUH�6���$ORRLGHDH�DUH�VXSSRUWHG�DV�D�PRQRSK\OHWLF�JURXS��EXW�UHVROXWLRQ�LQ�WKH�UHVW�RI�WKH�WUHH�LV�ORZ��6RPH�JURXSV�LQ�Aloe��Haworthia�VXEJ��Haworthia��H��VXEJ��Robustipedunculares�DQG�Gasteria�ZHUH�PRGHUDWHO\�VXSSRUWHG�

ITS data. —�6XPPDU\�VWDWLVWLFV�IRU�WKH�,76��GDWD�PDWUL[�LV�SUHVHQWHG�LQ�7DEOH����$QDO\VLV�RI�WKH�QXFOHDU�GDWDVHW�DOVR�UHWULHYHG�D�PRQRSK\OHWLF�$ORRLGHDH��DOWKRXJK�WKHUH�LV�VRPH�LQFRQJUXHQFH�ZLWK�WKH�SODVWLG�GDWD��LW�DOORZV�XV�WR�LGHQWLI\�ILYH�PDMRU�$ORRLGHDH�JURXSLQJV��)LJ��6���ZLWK�PRGHUDWH�WR�VWURQJ�VXSSRUW�EXW�SRRU�UHVROXWLRQ�ZLWKLQ�HDFK�JURXS��Haworthia VXEJ��Haworthia��Aloe VHFW��Macrifoliae��A. VHFW. Kumara��D�SRO\WRP\�LQFOXGLQJ WKH�µWUHH�Aloe¶�VSHFLHV�SOXV�Aloe V�VWU���LQFOXGLQJ�Chortolirion�� DQG�D�FODGH�FRPSULVLQJ�Astroloba�

Table .�6WDWLVWLFV�IURP�0RGHO7HVW�DQG�03�DQDO\VHV�REWDLQHG�IURP�VHSDUDWH�DQG�FRPELQHG�GDWDVHWV�

matK rbcLa trnH-psbA&RPELQHG�SODVWLG� ,76�

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1R��RI�WD[D�LQFOXGHG ��� ��� ��� ��� ��� ���1R��RI�LQFOXGHG�FKDUDFWHUV ��� ��� ��� ���� ��� ����1R��RI�FRQVWDQW�FKDUDFWHUV ��� ��� ��� ���� ��� ����1R��RI�YDULDEOH�VLWHV ��������� �������� ��������� ��������� ��������� ���������1R��RI�SDUVLPRQ\�LQIRUPDWLYH�VLWHV ��������� �������� ������� ��������� �������� ���������1R��RI�WUHHV��)LWFK� ��� ��� ��� 661 ��� ����1R��RI�VWHSV��7UHH�OHQJWK� ��� ��� ��� ��� ��� ����&RQVLVWHQF\�LQGH[ ���� ���� ���� ���� ���� ����5HWHQWLRQ�LQGH[ ���� ���� ���� ���� ���� ����$YHUDJH�QXPEHU�RI�FKDQJHV�SHU�YDULDEOH�VLWH� �QXPEHU�RI�VWHSV�QXPEHU�RI�YDULDEOH�VLWHV� ��� ��� ��� ��� ��� ���0RGHO�VHOHFWHG�E\�$NDLNH�LQIRUPDWLRQ�FULWHULRQ 790�* +.<�* 790�,�* 790�,�* 751�* *75�,�*

Page 7: Molecular and morphological analysis of subfamily ... · huv dqg haworthia zlwk yhu\ pxfk vpdoohu zklwlvk iorzhuv 'xydo haworthia zdv ixuwkhu vsolw zkhq wkrvh vshflhv zlwk pruh ru

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Gasteria� Haworthia�VXEJ��Hexangulares�DQG�VXEJ��Robusti-pedunculares� DQG Aloe aristata�

Combined plastid and nuclear dataset. —�$OWKRXJK�WKH�SDUWLWLRQHG�%UHPHU�VXSSRUW�WHVW�LQGLFDWHG�VRPH�LQFRQJUXHQ-FLHV�EHWZHHQ�WKH�QXFOHDU�DQG�SODVWLG�GDWDVHWV��YLVXDO�LQVSHF-WLRQ�RI�WKH�VHSDUDWH�DQDO\VHV��)LJ�����VKRZV�WKDW�QRQH�RI�WKH�VWURQJO\�VXSSRUWHG�FODGHV�ZHUH�PXWXDOO\�LQFRPSDWLEOH��DQG�WHVWV�IRU�LQFRQJUXHQFH�KDYH�EHHQ�VKRZQ�WR�EH�RI�YDULDEOH�UHOLDELOLW\��5HHYHV��DO���������<RGHU��DO����������%DVHG�RQ�WKH�HYLGHQW�FRQJUXHQFH�EHWZHHQ�WKH�WZR�GDWDVHWV��ZH�WKHUHIRUH�FRPELQHG�DOO�GDWD��6HHODQDQ��DO���������:LHQV���������)LJ������6WDWLVWLFV�IRU�WKH�SK\ORJHQHWLF�IUDPHZRUN�RI�WKH�FRQFDWHQDWHG�GDWDVHW�LV�VXP-PDULVHG�LQ�7DEOH����$�PRQRSK\OHWLF�$ORRLGHDH�ZDV�UHFRYHUHG�ZLWK�KLJK�VXSSRUW��)LJ�����%6� �����33� ��������7KH�FRPELQHG�GDWDVHW�UHVROYHG�HLJKW�PDMRU�JURXSLQJV�UHSUHVHQWHG�LQ�GLIIHUHQW�FRORXUV�DQG�FDSLWDO�OHWWHUV�LQ�)LJXUH�����$��WKH�WUHH�Aloe VSHFLHV���%��Aloe plicatilis� �&��Aloe VHFW��Macrifoliae���'��Haworthia VXEJ��Haworthia� �(��WKH�µWUXH¶�Aloe VSHFLHV���)��WKH�µ+DZRU-WKLRLG¶ FODGH��H. VXEJ��5REXVWLSHGXQFXODUHV࣠+࣠$VWUR�ORED࣠+࣠$ORH�aristata�� �*��Haworthia VXEJ��Hexangulares DQG��+��Gasteria�

The ‘tree aloes’. —�7KH�DQDO\VLV�UHWULHYHV�PRVW�RI�WKH�WUHH�Aloe�VSHFLHV��A. barberae��A. dichotoma��A. eminens��DV�D�FODGH�ZLWK�VWURQJ�VXSSRUW�LQ�WKH�%,�DQDO\VLV��%6� �����33� ��������7KLV�OLQHDJH�FRQVWLWXWHV�WKH�HDUOLHVW�GLYHUJLQJ�HOHPHQWV�LQ�VXEIDPLO\�$ORRLGHDH��)LJ���%�. 7KH�UHPDLQLQJ�WUHH�VSHFLHV��A. plicatilis� RFFXSLHV�DQ�LVRODWHG�SRVLWLRQ�LQ�DQ�XQUHVROYHG�SRO\WRP\�ZLWK�A. VHFW��Macrifoliae DQG�Haworthia VXEJ��Haworthia�

Haworthia subg. Haworthia. —�7KLV�FODGH�LV�ZHOO�VXS-SRUWHG� LQ� ERWK�03� DQG�%,� DQDO\VHV� �%6� � ���� 33� � ������)LJ���%�.�7KH�JUDVV�OLNH�Haworthia blackburniae LV�UHVROYHG�DV�

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The ‘rambling aloes’. —�7KLV�JURXS��UHSUHVHQWLQJ�Aloe VHFW��Macrifoliae (A. ciliaris� A. commixta��A. gracilis� A. stria-tula��A. tenuior��ZDV�UHFRYHUHG�LQ�ERWK�DQDO\VHV�DV�D�VWURQJO\�VXSSRUWHG�FODGH��%6� �����33� �������)LJ���%��

The ‘true aloes’. —�7KLV�JURXS��ZKLFK�FRPSULVHV�WKH�PD-MRULW\�RI�Aloe VSHFLHV�SOXV�Chortolirion �)LJ���$��ZDV�UHWULHYHG�ZLWK�VWURQJ�VXSSRUW�LQ�WKH�%,��33� ��������6HYHUDO�LQWHUQDO�FODGHV��QRQH�RI�ZKLFK�FRUUHVSRQG�H[DFWO\�WR�FXUUHQW�VHFWLRQV�UHFRJQLVHG�LQ�WKH�WD[RQRP\�RI�WKH�JHQXV��ZHUH�ZHOO�VXSSRUWHG�LQ�WKH�%,�DQDO\VLV��$PRQJ�WKHVH��WKH�PDMRULW\�RI�WKH�µJUDVV�DORHV¶��A. VHFW��Leptoaloe� JURXSHG�WRJHWKHU��%6� �����33� �������LQ�D�FODGH�WKDW�LQFOXGHG�Chortolirion angolense �%DNHU��$��%HUJHU� A. kouebok-keveldensis� A. buhrii� A. lutescens� A. reynoldsii� A. spicata��A. striata VXEVS. komaggasensis DQG�A. striata VXEVS��striata DOEHLW�ZLWK�ZHDN�%,�VXSSRUW��33� ������� ,Q�WKH�03�WUHH�WKLV�FODGH�ZDV�UHFRYHUHG�DV�SDUW�RI�D�ODUJHU�SRO\WRP\�ZLWK�QR�ERRW-VWUDS�VXSSRUW��$�VHFRQG�FODGH�FRPSULVHG�VHYHUDO�RI�WKH�µVLQJOH�VWHPPHG�DORHV¶��Aloe VHFW��Pachydendron��SOXV�RWKHU�VSHFLHV��ZLWK�ZHDN�03�VXSSRUW��%6� �����EXW�VWURQJ�VXSSRUW�LQ�WKH�%,�DQDO\VLV��33� ��������$�WKLUG�FODGH�FRPSULVLQJ�D�KHWHURJHQRXV�DVVHPEODJH�RI�Aloe VSHFLHV��A. excelsa� A. petricola� A. munchii� A. chabaudii��A. vryheidensis��ZDV�ZHOO�VXSSRUWHG�LQ�WKH�%,��%6� �����33� ��������$�IRXUWK�FODGH�UHFRYHUHG�DQRWKHU�KHWHURJHQRXV�DVVHPEODJH�RI�Aloe VSHFLHV�FRQWDLQLQJ�WKH�W\SH�RI�WKH�JHQXV��A. perfoliata� ZLWK�ZHDN�WR�PRGHUDWH�VXSSRUW��%6� �����33� ��������7ZR�DGGLWLRQDO�µJUDVV�DORHV¶��A. albida DQG�A. chortoliri-oides� ZHUH�UHFRYHUHG�DV�VLVWHUV�ZLWK�VWURQJ�VXSSRUW��%6� �����33� ��������7KH�UHPDLQLQJ�µWUXH�DORHV¶�DUH�ODUJHO\�XQUHVROYHG�

Fig. .�&ODGRJUDPV�VXPPDULVLQJ�UHODWLRQVKLSV�RI�PDMRU�WD[RQRPLF�JURXSLQJV�ZLWKLQ�$ORRLGHDH��1XPEHUV�DERYH�WKH�EUDQFKHV�DUH�03�ERRWVWUDS�VXSSRUW�YDOXHV�DQG�RQHV�EHORZ�DUH�%,�SRVWHULRU�SUREDELOLWLHV�

Gasteria (n = 20)

Haworthia subg. Hexangulares (n = 15)52

6 2

65

92

Aloe haworthioides

H. subg. Robustipedunculares (n = 5)

Astroloba (n = 4)

Aloe sect. Aristatae (n = 3)

Haworthia koelmaniorum (n = 2)

H. subg. Haworthia (n = 49)

True Aloe spp .

Chortolirion angolense

T ree Aloe spp. (n = 5)

Out g rou p s ( n = 13 ) Out g rou p ( Bulbine s p.)

Gasteria (n = 20)

H. subg. Hexangulares (n = 16)

80

0.74

92

-

H. subg. Robusti-

pedunculares (n = 5)

Astroloba (n = 4)

Haworthia koelmaniorum (n = 2)

Chortolirion angolense

Aloe sect. Macrifoliae (n = 5)

T ree Aloe spp. (n = 5)

Out g rou p s (n = 13 )

d e n i b m o C ) C ( 1 S T I ) B ( d i t s a l P ) A (

Aloe sect. Aristatae (n = 3)

H. subg. Haworthia (n = 49)

Aloe sect. Kumara (n = 2)

True Aloe spp. (n = 40)

1.00

1.00

58 0.68

77 0.95

-

56 0.97

1.00

-

0.53

93 1.00

77

99

59

Aloe sect. Macrifoliae (n = 5)

57 88

95

T rue Aloe spp .

Chortolirion angolense

Gasteria (n = 20)

H. subg. Robusti-

pedunculares (n = 5)

H. subg. Hexangulares

Aloe sect. Aristatae (n = 3)

57

61

Astroloba (n = 4)

85 85

61

99

T ree Aloe spp. (n = 5)69

Aloe sect. Kumara (n = 2)

Aloe sect. Kumara (n = 2)

H. subg. Haworthia (n = 44)

Page 8: Molecular and morphological analysis of subfamily ... · huv dqg haworthia zlwk yhu\ pxfk vpdoohu zklwlvk iorzhuv 'xydo haworthia zdv ixuwkhu vsolw zkhq wkrvh vshflhv zlwk pruh ru

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'DUX��DO����Phylogeny of AlooideaeTAXON����������)HEUXDU\���������±��

��9HUVLRQ�RI�5HFRUG��LGHQWLFDO�WR�SULQW�YHUVLRQ��

Aloe succotrina

Aloe hereroensis

Aloe haworthioides

Aloe anivoranoensis

Aloe dewinteri

Aloe propagulifera

Aloe greatheadii subsp. davyana

Aloe striata subsp. karasbergensis

Aloe glauca

Aloe brevifolia

Aloe albida

Aloe chortolirioides

Aloe melanacantha

Aloe pearsonii

Aloe arenicola

Aloe perfoliata

Aloe vryheidensis

Aloe chabaudii

Aloe munchii

Aloe petricola

Aloe excelsa

Aloe angelica

Aloe alooides

Aloe comosa

Aloe rupestris

Aloe thraskii

Aloe lineata

Aloe lineata var. muirii

Aloe lineata var. muirii

Aloe pictifolia

Aloe microstigma

Aloe arborescens

Aloe ferox

Aloe spicata

Chortolirion angolense

Aloe lutescens

Aloe reynoldsii

Aloe striata

Aloe striata subsp. kommagasensis

Aloe buhrii

Aloe kouebokkeveldensis

Aloe saundersiae

Aloe verecunda

Aloe vossii

Aloe challisii

Aloe fouriei

Aloe ecklonis

Aloe nubigena

H. koelmaniorum var. mcmutryi

H. koelmaniorum var. mcmutryi

Aloe aristata

Aloe aristata

Aloe aristata

Astroloba rubriflora

Astroloba foliolosa

Astroloba herrei

Astroloba corrugata

Haworthia marginata

Haworthia kingiana

Haworthia pumila

Haworthia marginata

Haworthia kingiana

Haworthia attenuata

Haworthia venosa

Haworthia venosa var. granulata

Haworthia limifolia

Haworthia limifolia

Haworthia limifolia var. ubomboensis

Haworthia fasciata

Haworthia longiana

Haworthia sordida

Haworthia bruynsii

Haworthia glauca

Haworthia nigra

Haworthia reinwardtii

Haworthia coarctata var. adelaidensis

Haworthia reinwardtii var. brevicula

Haworthia coarctata

G. pillansii var. pillansii

G. pillansii var. ernesti-ruschii

G. croucheri

G. carinata var. carinata

G. carinata var. retusa

G. acinacifolia

G. nitida var. armstrongii

G. batesiana var. dolomitica

G. ellaphieae

G. excelsa

G. vlokii

G. obliqua

G. rawlinsonii

G. glauca

G. glomerata

G. pulchra

G. tukhelensis

G. disticha

G. doreeniae

G. polita

[Fig. 3B, next page]

0.97

1.00

0.73

1.00

0.60

1.00

0.92

0.73

0.99

0.93

0.90

0.89

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1.00 1.00

0.99

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0.92

0.51

1.00

0.60

0.591.00

0.77

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0.93

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0.74

0.99

0.97

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1.00

0.67

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0.64

0.75

1.00

1.00

0.88

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0.78

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0.55

0.98

0.55

0.99

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0.93

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0.57

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56(3)

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80(5)

80

í(0)

90(3)

53(1)í(0)

53(1)84(1)

51(1)66(1)

61(2)

71(4) 70(1)

55(0)

86(2)

í(0)í(0)

91(2)

í(0)

í(0) 95(2)í

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Gasteria

Haworthia subg.Hexangulares I

Haworthia subg.Robustipedunculares

Astroloba

Aloe sect. Aristatae

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Aloe sect.Leptoaloe

True Aloe spp.

E

F

G

H

(1)

(1)

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Fig. . 7KH�����PDMRULW\�UXOH�FRQVHQVXV�WUHH�REWDLQHG�IURP�WKH�%,�DQDO\VLV�RI�WKH�FRPELQHG�SODV-WLG�DQG�,76��GDWDVHW�IRU�$OR�RLGHDH�SOXV�RXWJURXSV��:H�PDSSHG�WKH�ERRWVWUDS�YDOXHV�IURP�WKH�03�WUHH�RQWR�WKH�%,�PDMRULW\�UXOH�FRQVHQVXV�WUHH�EHFDXVH�WKH�03�DQG�%,�DQDO\VHV�SURGXFHG�VLPLODU�WRSRORJLHV��1XPEHUV�DERYH�QRGHV�DUH�QRQSDUDPHWULF�03�ERRWVWUDS�VXSSRUW�YDOXHV��WKRVH�DERYH�QRGHV�EXW�ZLWKLQ�SDUHQWKHVHV�DUH�GHFD\�LQGLFHV�FDOFXODWHG�XVLQJ�7UHH5RW��6RUHQVHQ��)UDQ]RVD���������WKRVH�EHORZ�QRGHV�DUH�%,�SRVWH-ULRU�SUREDELOLWLHV�DERYH�����

Page 9: Molecular and morphological analysis of subfamily ... · huv dqg haworthia zlwk yhu\ pxfk vpdoohu zklwlvk iorzhuv 'xydo haworthia zdv ixuwkhu vsolw zkhq wkrvh vshflhv zlwk pruh ru

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‘Haworthioid’ clade (Haworthia subg. Robustipeduncu-lares, Astroloba, Aloe aristata, H. koelmaniorum). —�$�PRG-HUDWHO\�VXSSRUWHG�µKDZRUWKLRLG¶�FODGH��%6� �����33� �������ZDV�UHWULHYHG�DV�VLVWHU�WR�WKH�Gasteria-Hexangulares FODGH�ZLWK�VWURQJ�VXSSRUW��%6� �����33� ��������Haworthia koelmaniorum YDU��mcmurtryi (H. VXEJ��Hexangulares��ZDV�UHWULHYHG�ZLWK�ZHDN�03�DQG�%,�VXSSRUW��%6� �����33� �������DV�WKH�HDUOLHVW�GLYHUJLQJ�OLQHDJH�LQ�WKH�FODGH��7KHUH�LV�PRGHUDWH�WR�VWURQJ�VXSSRUW��%6� �����33� �������IRU�D�VLVWHU�UHODWLRQVKLS�EHWZHHQ�

A. aristata DQG�H. VXEJ��5REXVWLSHGXQFXODUHV࣠�Astroloba��7KH�UHPDLQLQJ�VSHFLHV�RI�H. VXEJ��Robustipedunculares ZHUH�UH-FRYHUHG�ZLWK�VWURQJ�VXSSRUW��%6� ������33� �������EXW�DUH�RQO\�ZHDNO\�VXSSRUWHG�LQ�WKH�%,��33� �������DV�VLVWHU�WR�D�PRGHUDWH�WR�ZHOO�VXSSRUWHG��%6� �����33� �������Astroloba.

Haworthia subg. Hexangulares. — Haworthia� VXEJ��Hexangulares� �EXW� H[FOXGLQJ�H. koelmaniorum�� )LJ�� �$��ZDV�UHFRYHUHG�LQ�ERWK�03�DQG�%,�DQDO\VHV�ZLWK�PRGHUDWH�WR�VWURQJ�VXSSRUW��%6� �����33� �������LQ�D�FODGH�ZLWK�Gasteria��

Anthericum liliago

Tecophilaea cyanocrocus

Zephyra elegans

Xanthorrhoea resinosa

Xanthorrhoea sp.

Xanthorrhoea sp.

Asphodeline lutea

Eremurus spectabilis

Kniphofia galpinii

Kniphofia uvaria

Bulbine fistulosa

Bulbine semibarbata

Bulbine frutescens

Aloe barberae

Aloe eminens

Aloe dichotoma subsp. ramosissima

Aloe dichotoma

Aloe pillansii

Aloe plicatilis

Aloe plicatilis

Aloe commixta

Aloe gracilis

Aloe striatula

Aloe ciliaris

Aloe tenuior

Haworthia blackburniae

Haworthia blackburniae var. derustensis

Haworthia blackburniae var. graminifolia

Haworthia mirabilis

Haworthia bayeri

Haworthia zantneriana

Haworthia angustifolia

Haworthia emelyae var. comptoniana

Haworthia truncata

Haworthia variegata

Haworthia pygmaea

Haworthia chloracantha

Haworthia outeniquensis

Haworthia decipiens

Haworthia arachnoidea

Haworthia arachnoidea

Haworthia mucronata var. bijliana

Haworthia mucronata var. morrisiae

Haworthia mucronata var. rycroftiana

Haworthia monticola var. asema

Haworthia pulchella var. globifera

Haworthia lockwoodii

Haworthia mucronata var. habdomadis

Haworthia cooperi

Haworthia cymbiformis var. setulifera

Haworthia cymbiformis var. ramosa

Haworthia decipiens var. virella

Haworthia decipiens var. pringlei

Haworthia cooperi var. doldii

Haworthia cymbiformis

Haworthia cymbiformis var. setulifera

Haworthia vlokii

Haworthia wittebergensis

Haworthia emelyae

Haworthia floribunda

Haworthia marxii

Haworthia semiviva

Haworthia marumiana var. archeri

Haworthia mutica

Haworthia mutica var. nigra

Haworthia retusa

Haworthia mirabilis var. triebneriana

Haworthia mirabilis var. calcarea

Haworthia mirabilis var. paradoxa

Haworthia magnifica var. dekenahii

Haworthia herbacea var. flaccida

Haworthia maculata

Haworthia reticulata

Haworthia herbacea var. paynei

[Fig. 3A, previous page]

1.00

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Haworthia subg.Haworthia

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Tree Aloe spp.

Outgroups

AB

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Page 10: Molecular and morphological analysis of subfamily ... · huv dqg haworthia zlwk yhu\ pxfk vpdoohu zklwlvk iorzhuv 'xydo haworthia zdv ixuwkhu vsolw zkhq wkrvh vshflhv zlwk pruh ru

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Leaf tuberculation. — 7KH�SUHVHQFH�RI�ZKLWH�WXEHUFOHV�RQ�WKH�OHDYHV�LV�ODUJHO\�UHVWULFWHG�WR�Gasteria DQG�Haworthia�VXEJ��Hexangulares�DQG�VXEJ��Robustipedunculares�EXW�WXEHUFXOHV�DUH�DOVR�GHYHORSHG�LQ�VRPH�Aloe VSHFLHV��H�J���Aloe aristata� A. haworthioides��A. verecunda��DQG�LQ�Chortolirion��)LJ��6�&��

Table .�&KDUDFWHU�VWDWHV�IRU�QLQH�VHOHFWHG�WUDLWV�ZKLFK�KDYH�EHHQ�HPSOR\HG�LQ�SUHYLRXV�$ORRLGHDH�V\VWHPDWLFV�

&KDUDFWHU &KDUDFWHU�VWDWH 6SHFLHV

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6WHPOHVV 0RVW�Gasteria� KDOI�RI�Haworthia�VXEJ��Hexangulares� H. VXEJ��Robustipedunculares

&DXOHVFHQW Gasteria obliqua��KDOI�RI�Haworthia�VXEJ��Hexangulares� DOO�Astroloba �LQFOXGLQJ�Poellnitzia)

$UERUHVFHQW Aloe plicatilis (A. VHFW��Kumara���A. VHFW��Pachydendron DQG� A. VHFW��Dracoaloe

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/HDI�PDFXODWLRQ�,PPDFXODWH� 0RVW�$ORRLGHDH

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6SUHDGLQJ Chortolirion� Astroloba��Haworthia DQG�D�IHZ�Aloe

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Robustipedunculares DQG�PDMRULW\�RI�Aloe

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Page 11: Molecular and morphological analysis of subfamily ... · huv dqg haworthia zlwk yhu\ pxfk vpdoohu zklwlvk iorzhuv 'xydo haworthia zdv ixuwkhu vsolw zkhq wkrvh vshflhv zlwk pruh ru

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TAXON����������)HEUXDU\���������±��'DUX��DO����Phylogeny of Alooideae

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Perianth colour. —�)URP�RXU�DQDO\VLV��QRW�VKRZQ���YLYLG�\HOORZ��RUDQJH�RU�UHGGLVK�IORZHUV�DUH�FOHDUO\�SOHVLRPRUSKLF�ZLWKLQ� $ORRLGHDH�� +RZHYHU��Chortolirion��Haworthia DQG�Astro loba �H[FOXGLQJ�Poellnitzia��DQG�VHYHUDO�VSHFLHV�RI�Aloe��HVSHFLDOO\�LQ�A. VHFW��Leptoaloe��DUH�FKDUDFWHULVHG�E\�ZKLWLVK�RU�JUHHQLVK�SHULDQWK�

The Haworthia-type flower. —�8VXDOO\�ZKLWH�DQG�VRPH-WLPHV�ZLWK�VWURQJO\�]\JRPRUSKLF�SHULDQWKV��DQG�LQVHUWHG�DQ-WKHUV�DUH�FKDUDFWHULVWLF�RI�WKH�JHQXV�Haworthia DQG�Chorto-lirion��&KDUDFWHU�UHFRQVWUXFWLRQ�VXJJHVWV�WKDW�WKLV�IORZHU�W\SH�LV�KRPRSODVLRXV��DQG�WKDW�LW�KDV�HYROYHG�LQGHSHQGHQWO\�DW�OHDVW�WKUHH�DQG�SRVVLEO\�IRXU�WLPHV��RQFH�HDFK�LQ�Chortolirion DQG�H. VXEJ� Haworthia�DQG�RQFH�RU�WZLFH�LQ�H. VXEJ��Hexangu-lares�VXEJ��Robustipedunculares ZLWK�SRVVLEOH�UHYHUVDOV� LQ�A. aristata��A. haworthioides DQG�Gasteria���)LJ��6�$±&��

The Gasteria-type flower. —�)ODVN�VKDSHG�IORZHUV�ZLWK�LQIODWHG�EDVHV�RFFXU�QRW�RQO\�LQ�Gasteria� EXW�DOVR�LQ�PDQ\�Aloe VSHFLHV�DQG�KDYH�HYLGHQWO\�HYROYHG�VHYHUDO�WLPHV��)LJ��6�$���DOWKRXJK�ZLWKLQ�Aloe�� WKH�VWDPHQV�DUH�RIWHQ�EXW�QRW�DOZD\V�H[VHUWHG��)LJ��6�%���6LPLODU�IORZHUV�DUH�FKDUDFWHULVWLF�DOVR�RI�Astro loba �Ł�Poellnitzia) rubriflora�EXW�KHUH�DUH�XQLTXHO\�KHOG�HUHFW�RQ�DQ�LQFOLQHG�UDFHPH��$�ZHOO�GHYHORSHG�SHULDQWK�WXEH�IRUPHG�E\�WKH�IXVLRQ�RI�ERWK�WHSDO�ZKRUOV�KDV�HYROYHG�LQGH-SHQGHQWO\�LQ�Gasteria��Astroloba�DQG�Aloe kouebokkeveldensis �)LJ��6�&��

Tepal connation. —�7KH�SHULDQWK�LQ�$ORRLGHDH�FRPSULVHV�VL[�WHSDOV�LQ�WZR�ZKRUOV��YDULRXVO\�FRQQDWH�LQWR�D�VKRUW�RU�SURP-LQHQW� WXEH��%DVDOO\�FRQQDWH�WHSDOV�UHSUHVHQW� WKH�SOHVLRPRU-SKLF�FRQGLWLRQ��,Q�VHYHUDO�RI�WKH�RUQLWKRSKLORXV�Aloe�VSHFLHV��KRZHYHU��WKH�RXWHU�WHSDO�ZKRUO�LV�FRQQDWH�LQ�WKH�EDVDO�KDOI��IRUPLQJ�D�GLVWLQFW�SHULDQWK�WXEH��)LJ� 6�&���)ORZHUV�ZLWK�ERWK�ZKRUOV�FRQQDWH�IRU�KDOI�RU�PRUH�RI�WKHLU�OHQJWK�DUH�GLDJQRVWLF�IRU�Gasteria��Astroloba (Poellnitzia) rubriflora��DQG�A. koue-bokkeveldensis�DQG�DSSHDU�WR�KDYH�HYROYHG�LQGHSHQGHQWO\�LQ�WKHVH�WKUHH�OLQHDJHV�IURP�DQFHVWUDO�W\SHV�ZLWK�WHSDOV�FRQQDWH�DW�WKH�EDVH�RQO\��)LJ��6�&)�

DISCUSSION

Relationships within Alooideae. —�7KH�PRQRSK\O\�RI�$ORRLGHDH�ZDV�ILUVW�GHPRQVWUDWHG�E\�&KDVH��DO���������XVLQJ�WKH�WZR�SODVWLG�UHJLRQV�rbcL�DQG�trnL-F� DQG�PRUH�UHFHQWO\�E\�7UHXWOHLQ��DO� �����D��E���XVLQJ�matK�DQG�JHQRPLF�ILQJHU-SULQWLQJ��7KLV�ILQGLQJ�LV�VWURQJO\�VXSSRUWHG�E\�RXU�VWXG\��XVLQJ�VLJQLILFDQWO\�JUHDWHU�WD[RQ�VDPSOLQJ�DQG�D�FRPELQHG�DQDO\VLV�RI�WKUHH�SODVWLG�UHJLRQV��matK� rbcLa� trnH-psbA��SOXV�WKH�QXFOHDU�UHJLRQ�,76���$OWKRXJK�WKH�PRQRSK\O\�RI�WKH�VXEIDP-LO\�LV�QR�ORQJHU�LQ�TXHVWLRQ��WKLV�FDQQRW�EH�DFFHSWHG�IRU�DOO�RI�WKH�JHQHUD��,W�KDV�EHFRPH�LQFUHDVLQJO\�FOHDU�WKDW�QHLWKHU�Aloe QRU�Haworthia DUH�PRQRSK\OHWLF�DV�FXUUHQWO\�FLUFXPVFULEHG��7UHXWOHLQ��DO�������D��E��5DPGKDQL��DO����������2XU�DQDO\VLV�RI�WKH�FRPELQHG�SODVWLG�DQG�QXFOHDU�'1$�GDWDVHWV��)LJ�����FRQ-ILUPV�WKDW�ERWK�Aloe DQG�Haworthia DUH�SRO\SK\OHWLF�DV�FXU-UHQWO\�FLUFXPVFULEHG��:H�LGHQWLI\�HLJKW�SULPDU\�PRQRSK\OHWLF�OLQHDJHV�LQ�$ORRLGHDH�WKDW�ODUJHO\�FRUUHVSRQG�WR�WKH�IROORZLQJ�FXUUHQWO\�UHFRJQLVHG�JHQHULF�DQG�LQIUDJHQHULF�JURXSV������Aloe

VHFW��Dracoaloe�A. VHFW��Aloidendron������A. VHFW��Kumara������A��VHFW��Macrifoliae������Haworthia�VXEJ��Haworthia������A. VHFW� Aristatae�H.�VXEJ��Robustipedunculares�Astroloba��(6) H��VXEJ��Hexangulares������Gasteria������UHPDLQLQJ�VSH-FLHV�RI�Aloe��7KLV�WRSRORJ\�LV�FRQJUXHQW�ZLWK�WKDW�RI�7UHXWOHLQ��DO�������E���ZKLFK�KDG�PXFK�UHGXFHG�VDPSOLQJ��$PRQJ�WKH�VPDOOHU�VHJUHJDWH�JHQHUD��ERWK�Astroloba DQG�Gasteria�DUH�PRQRSK\OHWLF�EXW�Chortolirion� LV�GHHSO\�HPEHGGHG�DPRQJ�WUXH�Aloe�VSHFLHV�

6HYHQ�RI�WKH�HLJKW�SULPDU\�OLQHDJHV�DUH�HQGHPLF�WR�VRXWK-HUQ�$IULFD��ZLWK�RQO\�Aloe V�VWU��H[WHQGLQJ�EH\RQG�WKH�UHJLRQ��7KLV�VXJJHVWV�WKDW�WKH�HDUO\�GLYHUVLILFDWLRQ�RI�WKH�VXEIDPLO\�WRRN�SODFH�LQ�WKH�VXEFRQWLQHQW��ZKLFK�LV�DOVR�WKH�PDLQ�FHQWUH�RI�GLVWULEXWLRQ�IRU�WKH�VXEIDPLO\��6PLWK��9DQ�:\N��������

Relationships of the ‘tree aloes’. —�2XU�DQDO\VLV�GRHV�QRW�UHWUHLYH�WKH�µWUHH�DORHV¶�VHQVX�9DQ�:\N��6PLWK��������DV�D�PRQRSK\OHWLF�JURXS�EXW�DV�WZR�VHSDUDWH�OLQHDJHV��Aloe VHFW��$ORLGHQGURQ࣠�A. VHFW��Dracoaloe��LQFOXGLQJ�A. eminens IURP�6RPDOLD��HPHUJHV�DV�SRVVLEO\�RQH�RI�WKH�HDUO\�GLYHUJLQJ�OLQH-DJHV�ZLWKLQ�WKH�VXEIDPLO\��)LJ���%���7KH�LPPHGLDWH�UHODWLRQ-VKLSV�RI�WKH�UHPDLQLQJ�WUHH�Aloe��A. plicatilis (A. VHFW��Kumara) DUH�XQUHVROYHG�

Relationships of Aloe sect. Macrifoliae. — Aloe VHFW��Macri foliae� WKH�µUDPEOLQJ�DORHV¶��VHQVX�9DQ�:\N��6PLWK��������� FRPSULVLQJ� ILYH� FORVHO\� UHODWHG� VSHFLHV� �A. ciliaris� A. commixta��A. gracilis� A. striatula��A. tenuior���LV�UHFRYHUHG�DV�D�VWURQJO\�VXSSRUWHG�FODGH�EXW�LWV�UHODWLRQVKLS�WR�WKH�UHPDLQ-LQJ�VSHFLHV�LQ�WKH�VXEIDPLO\�UHPDLQV�XQUHVROYHG��7KH�VHFWLRQ�LV�GHILQHG�YHJHWDWLYHO\�E\�LWV�FDQH�OLNH�VWHPV��DQG�VOHQGHU��VKHDWK-LQJ��XQVSRWWHG�DQG�PHVRSK\WLF�OHDYHV�ZLWK�PLQXWH�PDUJLQDO�WHHWK��DQG�IORUDOO\�E\�WKH�PRUH�RU�OHVV�HQWLUHO\�FRQQDWH�RXWHU�WHSDOV��*OHQ��+DUG\��������

Relationships within the ‘true aloes’. —�7KH�UHPDLQLQJ�VSHFLHV�RI�Aloe �H[FOXGLQJ�A. aristata��DUH�UHWULHYHG�DV�D�FODGH�EXW�UHODWLRQVKLSV�DPRQJ�WKHP�DUH�SRRUO\�UHVROYHG��$OWKRXJK�VRPH�RI�WKH�FXUUHQWO\�UHFRJQLVHG�VHFWLRQV�PD\�EH�PRQRSK\-OHWLF��RWKHUV�DUH�QRW��DQG�D�PXFK�PRUH�H[WHQVLYH�VDPSOLQJ�RI�VSHFLHV�DQG�DGGLWLRQDO�JHQH�UHJLRQV�LV�UHTXLUHG�WR�HYDOXDWH�WD[R-QRPLF�DQG�HYROXWLRQDU\�UHODWLRQVKLSV�DPRQJ�WKHP�

$OWKRXJK�Chortolirion� LV�GHHSO\�HPEHGGHG�ZLWKLQ�WKLV�JURXS��DV�SDUW�RI�D�FODGH�LQFOXGLQJ�PRVW�RI�WKH�JUDVV�Aloe VSH-FLHV��LWV�SUHFLVH�UHODWLRQVKLSV�DUH�VWLOO�XQFOHDU��&ORVH�PRUSKRORJ-LFDO�VLPLODULW\�WR�VSHFLHV�VXFK�DV�A. bowiea�DQG�A. inconspicua LQ�YHJHWDWLYH�SDUWV��QDPHO\�WKH�JUDVV�OLNH�OHDYHV�ZLWK�EXOE�OLNH�VZHOOLQJ��DQG�LQ�WKH�VPDOO��ELODELDWH�IORZHUV��VXJJHVW�D�FORVH�UHODWLRQVKLS�WR�SDUW�RI�A. VHFW��Leptoaloe�

Relationships of Gasteria. — Gasteria�IRUPV�D�VWURQJO\�VXSSRUWHG�FODGH�VLVWHU�WR�Haworthia�VXEJ��Hexangulares. ,W�LV�GHILQHG�E\�VHYHUDO�PRUSKRORJLFDO�V\QDSRPRUSKLHV��QRWDEO\�WKH�XQDUPHG��YHUUXFRVH�OHDYHV�DQG�LQFOLQHG�VHFXQG�LQIORUHVFHQFHV�RI�SHQGXORXV��JDVWHULIRUP�IORZHUV�ZLWK�D�ZHOO�GHYHORSHG�IORUDO�WXEH��9DQ�-DDUVYHOG���������DQG�ZDV�XQVXUSULVLQJO\�RQH�RI�WKH�HDUOLHVW�VHJUHJDWHV�RI�Aloe WR�EH�UHFRJQLVHG��2XU�SK\OR-JHQHWLF�DQDO\VLV�SODFHV�WKH�JHQXV�VLVWHU�WR�Haworthia VXEJ��Hexangulares��ZKLFK�LQFOXGHV�VSHFLHV�ZLWK�UHPDUNDEO\�VLPLODU�OHDYHV��7KH�XQLTXH�Gasteria�W\SH�IORZHUV�DUH�PRVW�SDULVPRQL-RXVO\�LQWHUSUHWHG�DV�D�UHYHUVLRQ�WR�ELUG�SROOLQDWLRQ�IURP�WKH�

Page 12: Molecular and morphological analysis of subfamily ... · huv dqg haworthia zlwk yhu\ pxfk vpdoohu zklwlvk iorzhuv 'xydo haworthia zdv ixuwkhu vsolw zkhq wkrvh vshflhv zlwk pruh ru

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'DUX��DO����Phylogeny of AlooideaeTAXON����������)HEUXDU\���������±��

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HQWRPRSKLORXV�Haworthia�W\SH�IORZHU�ZLWK�LQFOXGHG�VWDPHQV��,W�LV�KDUGO\�VXUSULVLQJ�WKHQ�WKDW�Gasteria IORZHUV�DUH�QRW�SUH-FLVHO\�PDWFKHG�E\�DQ\�ELUG�SROOLQDWHG�IORZHUV�LQ�Aloe�

Relationships of Astroloba. —�6SHFLHV�RI�Astroloba DUH�UHWULHYHG�DV�D�FODGH�VLVWHU�WR�Haworthia VXEJ��Robustipeduncu-lares��7KH�JHQXV�LV�PRUSKRORJLFDOO\�GHILQHG�E\�LWV�FDXOHVFHQW�KDELW�ZLWK�VWLII�� LPEULFDWH�OHDYHV��DQG�VPDOO��DFWLQRPRUSKLF�IORZHUV�ZLWK�LQFOXGHG�VWDPHQV�

Relationships within Haworthia. —�2XU�DQDO\VLV�DFFRUGV�ZLWK�SUHYLRXV�VWXGLHV�LQGLFDWLQJ�WKDW�Haworthia�LV�QRW�PRQR-SK\OHWLF�EXW�UDWKHU�UHSUHVHQWV�WKUHH�OLQHDJHV�FRUUHVSRQGLQJ�WR�WKH�WKUHH�VXEJHQHUD�SURSRVHG�E\�%D\HU���������������6SHFLHV�RI�H. VXEJ��Haworthia�FRPSULVH�D�VWURQJO\�VXSSRUWHG�FODGH��GHILQHG�PRUSKRORJLFDOO\�E\�WKH�EDVDOO\�WULDQJXODU�SHULDQWK��REFODYDWH� IORZHUV�DQG�XSFXUYHG�VW\OH� �%D\HU���������������$GGLWLRQDO�VXSSRUW�IRU�WKLV�DOOLDQFH�FRPHV�IURP�6PLWK��DO����������ZKR�UHSRUWHG�WKH�RFFXUUHQFH�RI�KH[RVH�SRRU�QHFWDU��OHVV�WKDQ�����VXFURVH�HTXLYDOHQWV��LQ�H. VXEJ��Haworthia LQ� FRQWUDVW� WR� KH[RVH�ULFK� QHFWDU� �PRUH� WKDQ� ���� VXFURVH�HTXLYDOHQWV��LQ�H. VXEJ��Hexangulares DQG�H. VXEJ��Robusti-pedunculares. 7KH�ODWWHU�LV�D�VPDOO�JURXS�RI�IRXU�VSHFLHV�WKDW�LV�ZHOO�VXSSRUWHG�DV�PRQRSK\OHWLF�DQG�VLVWHU�WR�Astroloba��,W�LV�GHILQHG�PRUSKRORJLFDOO\�E\�LWV�PRUH�RU�OHVV�VWUDLJKW�SHULDQWK�DEUXSWO\�MRLQHG�WR�WKH�SHGLFHO��%D\HU���������������7KH�IORZHU�W\SH�IRXQG�LQ�H. VXEJ��Robustipedunculares LV�QRW�GLVVLPLODU�WR�WKDW�LQ�Astroloba��GLIIHULQJ�HVVHQWLDOO\�E\�LWV�VOLJKW�]\JRPRUSK\��0HPEHUV�RI�H. VXEJ��Robustipedunculares�DUH�RIWHQ�UREXVW�ZLWK�DWWHQXDWH�OHDYHV��RIWHQ�VFDEULG�DQG�SDWWHUQHG�ZLWK�ZKLWH�WXEHUFOHV��%D\HU���������$�VLVWHU�UHODWLRQVKLS�EHWZHHQ�Astroloba DQG�H. VXEJ��Robustipedunculares�LV�VXSSRUWHG�E\�VLPLODULWLHV�LQ�QHFWDU�VXFURVH�FRQFHQWUDWLRQV��9DQ�:\N��DO�� ������

7KH�GZDUI�Aloe aristata LV VLVWHU�WR�H. VXEJ��Robustipe-dunculares�Astroloba��7KLV�PRUSKRORJLFDOO\�XQXVXDO�VSH-FLHV�LV�XQLTXH�LQ�Aloe LQ�KDYLQJ�³Haworthia-OLNH´ OHDYHV�ZLWK�GU\��DZQ�WLSSHG�DSLFHV�DQG�ZKLWH�WXEHUFOHV�DQG�GLVWLQFWLYH��GRZQFXUYHG�IORZHUV�ZLWK�EDVDO�VZHOOLQJ��*OHQ��+DUG\���������7KH�FORVH�YHJHWDWLYH�VLPLODULW\�EHWZHHQ�WKLV�VSHFLHV�DQG�VRPH�PHPEHUV�RI�H. VXEJ��Robustipedunculares LV�FRQVLVWHQW�ZLWK�D�FORVH�UHODWLRQVKLS�EHWZHHQ�WKHP��DQG�WKH�SULPDU\�GLIIHUHQFH�EHWZHHQ�WKH�WZR�JURXSV�LV�HYLGHQWO\�WKH�ODUJH��RUDQJH�IORZHUV�RI�A. aristata��7KLV�IORZHU�W\SH�LV�DVVRFLDWHG�ZLWK�ELUG�SROOLQD-WLRQ�DQG�KDV�DULVHQ�VHYHUDO�WLPHV�ZLWKLQ�WKH�VXEIDPLO\��$ORQH��LW�LV�WKHUHIRUH�QRW�QHFHVVDULO\�DQ�LQGLFDWLRQ�RI�UHODWLRQVKLSV�

8QOLNH� WKH� RWKHU� WZR� VXEJHQHUD� RI�Haworthia�� VXEJ��Hexangulares LV�SRVVLEO\�SRO\SK\OHWLF�ZLWK�WKH�LQFOXVLRQ�RI�H. koelmaniorum��ZKLFK�RFFXSLHV�DQ�LVRODWHG�SRVLWLRQ�VLVWHU�WR�H. VXEJ��Robustipedunculares�$VWURORED࣠�Aloe aristata EXW�ZLWK�RQO\�ZHDN�VXSSRUW.�7KH�JHRJUDSKLFDO�GLVWULEXWLRQ�RI�WKLV�VSHFLHV�LV�ZHOO�QRUWK�RI�PRVW�RWKHU�VSHFLHV�RI�Haworthia ZLWK�WKH�H[FHSWLRQ�RI�H. limifolia��:KHQ�ILUVW�GHVFULEHG��2EHUPH\HU���������H. koelmaniorum ZDV�WUHDWHG�LQ�H. VHFW��Margariti-fera �QRZ�H. VXEJ��Robustipedunculares��EXW�ODWHU�WUDQVIHUUHG�WR�H. VXEJ��Hexangulares��ZKHUH�LWV�UHODWLRQVKLSV�DSSHDU�WR�OLH�ZLWK�H. limifolia �%D\HU�������. )XUWKHU�HYLGHQFH�IRU�D�ILQDO�GHFLVLRQ�RQ�LWV�SRVLWLRQ�LV�UHTXLUHG��6SHFLHV�RI�H. VXEJ��Hex-angulares�GLVSOD\�WKH�ODUJHVW�YHJHWDWLYH�GLYHUVLW\�LQ�Haworthia��ZLWK�VRPH�VSHFLHV�FORVHO\�UHVHPEOLQJ�PHPEHUV�RI�Astroloba

DQG�H. VXEJ��Robustipedunculares LQ�WKHLU�YHJHWDWLYH�PRUSKRO-RJ\��HVSHFLDOO\�WKH�SUHVHQFH�RI�WXEHUFOHV�RQ�WKH�OHDYHV�

Evolution of selected characters in Alooideae. —�2XU�UHFRQVWUXFWLRQ�RI�WKH�QLQH�PRUSKRORJLFDO�FKDUDFWHUV�RQWR�WKH�%,�PDMRULW\�UXOH�FRQVHQVXV�WUHH�UHYHDOHG�WKDW�QRQH�LV�XQLTXH�WR�D�VLQJOH�FODGH�LGHQWLILHG�LQ�WKLV�VWXG\�

Habit. —�3UHYLRXV FODVVLILFDWLRQV��H�J���5H\QROGV��������+ROODQG��������DUH�LPSOLFLW�LQ�WUHDWLQJ�DUERUHVFHQFH�LQ�Aloe DV�D�GHULYHG�VWDWH��%UDQGKDP��������SURSRVHG�WKDW�VFDQGHQW�Aloe VSHFLHV�ZLWK�XVXDOO\�UHODWLYHO\�PHVRSK\WLF�OHDYHV��H�J���A��tenu-ior�DQG�A. ciliaris��$��VHFW��Macrifoliae���UHSUHVHQW�WKH�SULPLWLYH�VWDWH�LQ�Aloe VSHFLHV�EXW�6PLWK��9DQ�:\N��������DUJXHG�WKDW�ERWK�VPDOO��KLJKO\�VXFFXOHQW�WD[D�DQG�DUERUHVFHQW�IRUPV�ZHUH�GHULYHG�IURP�D�PHVRSK\WLF��FRPSDUDWLYHO\�DFDXOHVFHQW�WD[RQ��2XU�DQDO\VLV�VXSSRUWV�WKLV�K\SRWKHVLV��LQGLFDWLQJ�WKDW�DUERUHV-FHQFH�LQ�Aloe LV�IRXQG�QRW�RQO\�LQ�HDUO\�GLYHUJLQJ�OLQHDJHV�EXW�DOVR�LQ�RWKHUV�GHHSO\�HPEHGGHG�ZLWKLQ�Aloe��DQG�WKDW�WKH�VPDOO��VWHPOHVV�JUDVV�Aloe �$��VHFW��Leptoaloe��DUH�GHULYHG�

Leaf insertion. —�'LVWLFK\�LV�HYLGHQWO\�WKH�MXYHQLOH�FRQGL-WLRQ��SUHVHQW�LQ�DOO�Aloe�DQG�Gasteria�VHHGOLQJV��DQG�LWV�SHUVLV-WHQFH�LQ�DGXOW�SODQWV�LV�EHVW�LQWHUSUHWHG�DV�QHRWHQ\�

Leaf tuberculation. — 7XEHUFXODWLRQ�LV�FHUWDLQO\�D�GHULYHG�FRQGLWLRQ��DV�K\SRWKHVLVHG�E\�6PLWK��9DQ�:\N���������,Q�Gasteria��9DQ�-DDUVYHOG��������KDV�SURSRVHG�WKDW�LWV�HYROXWLRQ�ZDV�GULYHQ�E\�WKH�DEVHQFH�RI�WKH�ELWWHU�FRQVWLWXHQW�W\SLFDO�RI�Aloe VSHFLHV��LPSO\LQJ�WKDW�WKH�ULJLG�WXEHUFOHV�PD\�PDNH�WKH�OHDYHV�OHVV�SDODWDEOH�

Perianth colour. —�7KH�EULJKW�\HOORZ��RUDQJH�RU�UHGGLVK�IORZHUV�W\SLFDO�RI�PRVW�$ORRLGHDH�DUH�VWURQJO\�DVVRFLDWHG�ZLWK�RUQLWKRSKLO\��7KH�ZKLWLVK�RU�JUHHQLVK�SHULDQWK�FKDUDFWHULVWLF�RI�Chortolirion��Haworthia, Astroloba �H[FOXGLQJ�Poellnitzia) DQG�Aloe�VHFW��Leptoaloe��DSSHDUV�WR�EH�D�GHULYHG�DGDSWDWLRQ�WR�HQWRPRSKLO\��%RWHV��DO���������+DUJUHDYHV��DO���������

The Haworthia-type flower. —�6PDOO��VSUHDGLQJ�IORZHUV�ZLWK�D�ZKLWLVK��PRUH�RU�OHVV�ELODELDWH�SHULDQWK�DQG�LQFOXGHG�DQWKHUV�DUH�GLDJQRVWLF�RI�Haworthia DQG�Chortolirion �%D\HU���������%D\HU��������LGHQWLILHG�VPDOO�IORUDO�GLIIHUHQFHV�LQ�WKH�WKUHH�VXEJHQHUD�RI�Haworthia��ZKLFK�LV�FRQVLVWHQW�ZLWK�WKH�LQGHSHQGHQW�HYROXWLRQ�RI�WKLV�IORZHU�W\SH�LQ�WKLV�JHQXV��7KH�FRQYHUJHQFH�LQ�WKLV�IORZHU�V\QGURPH�LQ�Chortolirion SURYLGHV�FOHDU�HYLGHQFH�WKDW�VXFK�D�IORZHU�W\SH�FDQ�HYROYH�LQGHSHQGHQWO\�IURP�DQ�RUQLWKRSKLORXV�DQFHVWRU� 5HODWLYHO\�VKRUW�WXEHG��ZKLW-LVK�RU�FUHDP�FRORXUHG�IORZHUV�LQ�VRPH�Aloe�VSHFLHV�VXFK�DV�A. inconspicua KDYH�EHHQ�VKRZQ�WR�EH�DQ�DGDSWDWLRQ�WR�LQVHFW�SROOLQDWLRQ��%RWHV��DO����������$OWKRXJK�VWLOO�UHFRJQLVDEO\�µAloe-OLNH¶��WKH�IORZHUV�RI�EHH�SROOLQDWHG�Aloe VSHFLHV��VHQVX�%RWHV��DO���������VXFK�DV�A. linearifolia DQG�A. minima�GLV-SOD\�VHYHUDO�FKDUDFWHULVWLFV�RI�WKH�Haworthia�W\SH�IORZHU�DSDUW�IURP�UHGXFHG�VL]H��QDPHO\�WKHLU�QHDUO\�KRUL]RQWDO�RULHQWDWLRQ��ZKLWLVK�DQG�ZHDNO\�ELODELDWH�SHULDQWK��DQG�VRPHWLPHV�LQFOXGHG�VWDPHQV��6PLWK��9DQ�:\N��������VXJJHVWHG�WKDW�IORUDO�]\JR-PRUSK\�LQ�Astroloba, Chortolirion DQG�Haworthia�UHSUHVHQWV�DQ�DGYDQFHG�VWDWH�GHULYHG�IURP�WKH�SOHVLRPRUSKLF�DFWLQRPRU-SKLF�SDWWHUQ��ZKLFK�LV�VXSSRUWHG�E\�RXU�DQDO\VLV�

The Gasteria-type flower. —�7KH�VR�FDOOHG�JDVWHULIRUP�IORZHU��FXUYHG�DQG�IODVN�VKDSHG�ZLWK�DQ�RYRLG��LQIODWHG�WXEH�DW�OHDVW�KDOI�DV�ORQJ�DV�WKH�SHULDQWK��DQG�LQFOXGHG�RU�VKRUWO\�

Page 13: Molecular and morphological analysis of subfamily ... · huv dqg haworthia zlwk yhu\ pxfk vpdoohu zklwlvk iorzhuv 'xydo haworthia zdv ixuwkhu vsolw zkhq wkrvh vshflhv zlwk pruh ru

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TAXON����������)HEUXDU\���������±��'DUX��DO����Phylogeny of Alooideae

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Tepal connation. —�)ORUDO�V\QGURPHV�LQGLFDWH�WKDW�Astro-loba��Ł�Poellnitzia) rubriflora, Gasteria�DQG�VHYHUDO�Aloe VSH-FLHV�DUH�ELUG�SROOLQDWHG�DQG�RXU�UHFRQVWUXFWLRQ�VXJJHVWV�WKDW�RUQLWKRSKLO\�LQ�Gasteria�DQG�Astroloba�DUH�VHFRQGDU\�DGDSWD-WLRQV�IURP�DQ�DQFHVWUDO�HQWRPRSKLORXV��Haworthia�W\SH�IORZHU�UDWKHU�WKDQ�GHULYHG�IURP�D�PRUH�W\SLFDO�Aloe�W\SH�IORZHU�

,W� LV� LQFUHDVLQJO\�HYLGHQW� WKDW� WKH�GLIIHUHQFHV� LQ� IORUDO�PRUSKRORJ\�WKDW�ZHUH�XVHG�DV�WKH�SULPDU\�FKDUDFWHULVWLF�MXV-WLI\LQJ�WKH�UHFRJQLWLRQ�RI�WKHVH�YDULRXV�OLQHDJHV�DV�VHJUHJDWH�JHQHUD�UHSUHVHQW�V\QGURPHV�DVVRFLDWHG�ZLWK�VKLIWV�LQ�SROOLQD-WLRQ�V\VWHPV�IURP�ELUG�WR�LQVHFW�DQG�EDFN��+LVWRULFDOO\��PRVW�RI�WKH�VHJUHJDWH�JHQHUD�ZHUH�HUHFWHG�EHIRUH�LQIRUPDWLRQ�RQ�WKH�IXOO�YDULDWLRQ�LQ�Aloe�ZDV�NQRZQ��DQG�FHUWDLQO\�EHIRUH�WKH�VLJQLILFDQFH�RI�IORUDO�V\QGURPHV�LQ�SROOLQDWLRQ�ZDV�DSSUHFLDWHG��,Q�$IULFDQ�,ULGDFHDH�LQ�SDUWLFXODU��ZKHUH�QXPHURXV�VSHFLDOLVW�SROOLQDWLRQ�V\VWHPV�KDYH�EHHQ�GRFXPHQWHG��VHYHUDO�HUVWZKLOH�µJHQHUD¶�KDYH�EHHQ�VKRZQ�WR�UHSUHVHQW�DUWLILFLDO�DVVRFLDWLRQV�RI�VSHFLHV�EDVHG�RQ�IORUDO�FKDUDFWHUV�DVVRFLDWHG�ZLWK�SROOLQDWLRQ�V\VWHPV��*ROGEODWW��0DQQLQJ��������

Implications for taxonomy. —� 2XU� UHVXOWV� DUH� HVVHQ-WLDOO\�FRQJUXHQW�ZLWK�WKRVH�RI�HDUOLHU�V\VWHPDWLF�VWXGLHV��H�J���7UHXWOHLQ��DO�������D��E��5DPGKDQL��DO����������7KH�FXUUHQW�FODVVLILFDWLRQ�RI�WKH�VXEIDPLO\�LQWR�ILYH�JHQHUD��VHH�UHYLHZ�E\�.ORSSHU��DO���������KDV�EHHQ�LQIRUPHG�E\�HYLGHQFH�IURP�FODGLVWLF�VWXGLHV�RI�PRUSKRORJLFDO�WUDLWV��6PLWK��9DQ�:\N��������6PLWK��6WH\Q��������.ORSSHU��DO����������DV�ZHOO�DV�F\WRORJLFDO��7D\ORU��������DQG�FKHPLFDO�GDWD��9LOMRHQ��DO�� ������9LOMRHQ���������7KHVH�VWXGLHV��DOWKRXJK�XQDEOH�WR�UHVROYH�WKH�UHODWLRQVKLSV�ZLWKLQ�WKH�VXEIDPLO\��SURYLGHG�D�ZRUNLQJ�K\-SRWKHVLV�IRU�WKLV�VWXG\�

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Ɣ� Scenario 2. –�5HWDLQ�WKH�VWDWXV�TXR��7KLV�VFHQDULR�UH-TXLUHV�WKH�DFFHSWDQFH�RI�SDUDSK\O\�LQ�ERWK�Aloe DQG�Hawor-thia. $V�7UHXWOHLQ��DO�������E��ULJKWO\�SRLQW�RXW� WKH�FXUUHQW�VXEIDPLOLDO�FODVVLILFDWLRQ�GRHV�QRW�UHIOHFW�DYDLODEOH�HYLGHQFH�IURP�SK\ORJHQHWLF�DQDO\VHV��DQG�WKH�IXUWKHU�LW�GHSDUWV�IURP�WKH�SK\ORJHQHWLF�HYLGHQFH�WKH�PRUH�GLIILFXOW�LW�ZLOO�EH�WR�LQWHJUDWH�SUDFWLFH�DQG�WKHRU\�

Ɣ� Scenario 3. –�$�WD[RQRPLF�µVSOLWWHU¶V¶�DSSURDFK�WKURXJK�WKH�UHFRJQLWLRQ�RI�DGGLWLRQDO�VPDOOHU�JHQHUD�ZLWKLQ�$ORRLGHDH�LQ�RUGHU�WR�UHWDLQ�DOO�RU�PRVW�RI�WKH�FXUUHQWO\�DFFHSWHG�JHQHUD�DV�PRQRSK\OHWLF��$OWKRXJK�ZH�DUH�QRZ�LQ�SRVVHVVLRQ�RI�D�ZHOO�VDPSOHG�WUHH��WKH�SRRU�VXSSRUW�DORQJ�WKH�EDFNERQH�SODFHV�WKH�VDPH�FRQVWUDLQWV�XSRQ�XV��8QWLO�WKHVH�EDVDO�QRGHV�DUH�EHWWHU�VXSSRUWHG�ZH�UHPDLQ�XQDEOH�WR�SURSRVH�DQ�DOWHUQDWLYH�FODVVLIL-FDWLRQ��Aloe aristata LV�FOHDUO\�PLVSODFHG�ZLWK�WKH�RWKHU�DORHV�

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2Q�PRUSKRORJLFDO�JURXQGV�WKHUH�LV�OLWWOH�WR�SUHFOXGH�LP-SOHPHQWLQJ�WKLV�RSWLRQ�VLQFH�Aloe�DOUHDG\�LQFOXGHV�PRVW�RI�WKH�YDULDWLRQ�HYLGHQW�LQ�WKH�VXEIDPLO\��%UDQFK�VXSSRUW�DW�WKH�ORZHU�QRGHV�LQ�RXU�DQDO\VLV�LV�QRW�KLJK�HQRXJK�WR�SUHGLFDWH�WKH�DGRS-WLRQ�RI�WKLV�RSWLRQ��DQG�VHTXHQFLQJ�RI�DGGLWLRQDO�JHQH�UHJLRQV�PLJKW�VWLOO�JURXS�WKH�WUHH�DQG�UDPEOLQJ�DORHV�ZLWK�WKH�WUXH�DORHV��8QWLO�D�VXLWDEO\�ZHOO�VXSSRUWHG�WRSRORJ\�LV�DYDLODEOH�RQ�ZKLFK�WR�EDVH�D�SK\ORJHQHWLF�FODVVLILFDWLRQ�LW�VHHPV�EHVW�WR�UHIUDLQ�IURP�PDMRU�IRUPDO�WD[RQRPLF�DQG�QRPHQFODWXUDO�DGMXVWPHQWV�

NOMENCLATURE

Aloe VHFW� Chortolirion��$��%HUJHU��%RDWZU���-�&��0DQQLQJ��comb. & stat. nov.�Ł�Chortolirion�$��%HUJHU�LQ�(QJOHU��3IODQ]HQU��,9�����,,,������������±�7\SH��Chortolirion an-golense (%DNHU��$��%HUJHU� [= Aloe subspicata� �%DNHU��%RDWZU���-�&��0DQQLQJ@

Page 14: Molecular and morphological analysis of subfamily ... · huv dqg haworthia zlwk yhu\ pxfk vpdoohu zklwlvk iorzhuv 'xydo haworthia zdv ixuwkhu vsolw zkhq wkrvh vshflhv zlwk pruh ru

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'DUX��DO����Phylogeny of AlooideaeTAXON����������)HEUXDU\���������±��

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Adams, S.P., Leitch, I.J. & Bennett, M.D. ������5LERVRPDO�'1$�HYROXWLRQ�DQG�SK\ORJHQ\�LQ�Aloe �$VSKRGHODFHDH���Amer. J. Bot. ��������±�����

APG II ������$Q�XSGDWH�RI�WKH�$QJLRVSHUP�3K\ORJHQ\�*URXS�FODV-VLILFDWLRQ�IRU�WKH�RUGHUV�DQG�IDPLOLHV�RI�IORZHULQJ�SODQWV��$3*�,,��Bot. J. Linn. Soc.���������±����

Baldwin, B.G., Sanderson, M.J., Porter, J.M., Wojciechowski, M.F., Campbell, C.S. & Donoghue, M.J. ������7KH�,76�UHJLRQ�RI�QX-FOHDU�ULERVRPDO�'1$�$�YDOXDEOH�VRXUFH�RI�HYLGHQFH�RQ�$QJLR-VSHUP�SK\ORJHQ\��Ann. Missouri Bot. Gard.��������±����

Bayer, M.B. ������Haworthia handbook. .LUVWHQERVFK��1DWLRQDO�%R-WDQLF�*DUGHQV�RI�6RXWK�$IULFD�

Bayer, M.B. ������The new Haworthia handbook��.LUVWHQERVFK��&DSH�7RZQ��1DWLRQDO�%RWDQLF�*DUGHQV�RI�6RXWK�$IULFD�

Bayer, M.B. ������Haworthia revisited: A revision of the genus��+DW-ILHOG��8PGDXV�3UHVV�

Bayer, M.B. ������Haworthia update: Essays on Haworthia��YRO�����+DWILHOG��8PGDXV�3UHVV�

Bayer, M.B. ������Haworthia supplement��/RQGRQ��7KH�+DZRUWKLD�6RFLHW\�

���Aloe aestivalis %RDWZU���-�&��0DQQLQJ��nom. nov.�Ł�Chor-tolirion latifolium�=RQQ���*�3�-��)ULW]�LQ�%UDGOH\D���������ILJV���±����������QRQ�Aloe latifolia��+DZ���+DZ��±�+ROR-W\SH��6RXWK�$IULFD��>)UHH�6WDWH@��%ORHPIRQWHLQ�QHDU�DLUSRUW��������Fritz 1025��35(��

���Aloe subspicata �%DNHU��%RDWZU���-�&��0DQQLQJ��comb. nov.� Ł� Haworthia subspicata� %DNHU� LQ� %XOO�� +HUE��%RLVVLHU��VHU������������������±�+RORW\SH��6RXWK�$IULFD��>*DXWHQJ@��0RGGHUIRQWHLQ����6HS�������Conrath 645��=��LVRW\SH��.��

= Haworthia angolensis %DNHU�LQ�7UDQV��/LQQ��6RF��/RQGRQ��%RW���������������Ł�Chortolirion angolense (%DNHU��$��%HUJHU�LQ�(QJOHU��3IODQ]HQU��,9�����,,,�������������QRQ�Aloe angolen-sis %DNHU�±�+RORW\SH��$QJROD��+XLOOD��1RY�������Welwitsch 3756��%0��

���Aloe tenuifolia��(QJO���%RDWZU���-�&��0DQQLQJ��comb. nov. Ł�Haworthia tenuifolia�(QJO��LQ�%RW��-DKUE��6\VW���������W����������Ł�Chortolirion tenuifolium��(QJO���$��%HUJHU�LQ�(QJOHU��3IODQ]HQU��,9�����,,,������������±�+RORW\SH��6RXWK�$IULFD��>1RUWKHUQ�&DSH@��QHDU�.XUXPDQ��)HE�������Marloth 1049��%��LVRW\SH��35(��

ACKNOWLEDGEMENTS

:H�ZRXOG�OLNH�WR�WKDQN�%UXFH�0��%D\HU��6HDQ�*LOGHQKX\V��(UQVW�YDQ�-DDUVYHOG��DQG�WKH�0ROHFXODU�6\VWHPDWLFV�VHFWLRQ�RI�-RGUHOO�/DER-UDWRU\�DW� WKH�5R\DO�%RWDQLF�*DUGHQV��.HZ��/DV]OR�&VLED�DQG�)HOL[�)RUHVW��IRU�VDPSOHV��7KH�1DWLRQDO�5HVHDUFK�)RXQGDWLRQ��15)��6RXWK�$IULFD�DQG�WKH�5R\DO�6RFLHW\�RI�/RQGRQ�DUH�JUHDWO\�DFNQRZOHGJHG�IRU�IXQGLQJ��7KLV�SURMHFW�ZDV�DOVR�SDUWO\�IXQGHG�E\�WKH�*RYHUQPHQW�RI�&DQDGD�WKURXJK�*HQRPH�&DQDGD�DQG�WKH�2QWDULR�*HQRPLFV�,QVWLWXWH�������2*,�,&,������:H�DSSUHFLDWH�WZR�DQRQ\PRXV�UHIHUHHV�ZKR�SUR-YLGHG�XVHIXO�FRPPHQWV�RQ�DQ�HDUOLHU�GUDIW�RI�WKH�PDQXVFULSW�

LITERATURE CITED

Botes, C., Johnson, S.D. & Cowling, R.M. ������&RH[LVWHQFH�RI�VXF-FXOHQW�WUHH�DORHV��3DUWLWLRQLQJ�RI�ELUG�SROOLQDWRUV�E\�IORUDO�WUDLWV�DQG�IORZHULQJ�SKHQRORJ\��Oikos ��������±����

Botes, C., Wragg, P.D. & Johnson, S.D. ������1HZ�HYLGHQFH�IRU�EHH�SROOLQDWLRQ�V\VWHPV�LQ�Aloe �$VSKRGHODFHDH��$ORLGHDH���D�SUHGRPL-QDQWO\�ELUG�SROOLQDWHG�JHQXV��S. African J. Bot. �������±����

Brandham, P.E. ������(YROXWLRQ�LQ�D�VWDEOH�FKURPRVRPH�V\VWHP��,Q��%UDQGKDP��3�(���%HQQHWW��0�'���HGV����Kew Chromosome Confer-ence II. /RQGRQ��*HRUJH�$OOHQ��8QZLQ�

Bremer, K. ������7KH�OLPLWV�RI�DPLQR�DFLG�VHTXHQFH�GDWD�LQ�DQJLRVSHUP�SK\ORJHQHWLF�UHFRQVWUXFWLRQ��Evolution��������±����

CBOL Plant Working Group ������$�'1$�EDUFRGH�IRU�ODQG�SODQWV��Proc. Natl. Acad. Sci. U.S.A.�����������±������

Chase, M.W., De Bruijn, A.Y., Cox, A.V., Reeves, G., Rudall, P.J., Johnson, M.A.T. & Eguiarte, L.E. ������3K\ORJHQHWLFV�RI�$VSKR-GHODFHDH��$VSDUDJDOHV���$Q�DQDO\VLV�RI�SODVWLG�rbcL�DQG�trnL-F '1$�VHTXHQFHV��Ann. Bot. (Oxford)��������±����

DeSalle, R. & Brower A.V.Z. ������3URFHVV�SDUWLWLRQV��FRQJUXHQFH�DQG�WKH�LQGHSHQGHQFH�RI�FKDUDFWHUV��,QIHUULQJ�UHODWLRQVKLSV�DPRQJ�FORVHO\�UHODWHG�+DZDLLDQ�Drosophila�IURP�PXOWLSOH�JHQH�UHJLRQV��Syst. Biol.��������±����

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