Molecular and morphological analysis of subfamily ... · huv dqg haworthia zlwk yhu\ pxfk vpdoohu...
Transcript of Molecular and morphological analysis of subfamily ... · huv dqg haworthia zlwk yhu\ pxfk vpdoohu...
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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\
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MATERIALS AND METHODS
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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�$ORRLGHDH�JURXSLQJV��)LJ��6���ZLWK�PRGHUDWH�WR�VWURQJ�VXSSRUW�EXW�SRRU�UHVROXWLRQ�ZLWKLQ�HDFK�JURXS��Haworthia VXEJ��Haworthia��Aloe VHFW��Macrifoliae��A. VHFW. Kumara��D�SRO\WRP\�LQFOXGLQJ WKH�µWUHH�Aloe¶�VSHFLHV�SOXV�Aloe V�VWU���LQFOXGLQJ�Chortolirion�� DQG�D�FODGH�FRPSULVLQJ�Astroloba�
Table .�6WDWLVWLFV�IURP�0RGHO7HVW�DQG�03�DQDO\VHV�REWDLQHG�IURP�VHSDUDWH�DQG�FRPELQHG�GDWDVHWV�
matK rbcLa trnH-psbA&RPELQHG�SODVWLG� ,76�
&RPELQHG� SODVWLG�,76�
1R��RI�WD[D�LQFOXGHG ��� ��� ��� ��� ��� ���1R��RI�LQFOXGHG�FKDUDFWHUV ��� ��� ��� ���� ��� ����1R��RI�FRQVWDQW�FKDUDFWHUV ��� ��� ��� ���� ��� ����1R��RI�YDULDEOH�VLWHV ��������� �������� ��������� ��������� ��������� ���������1R��RI�SDUVLPRQ\�LQIRUPDWLYH�VLWHV ��������� �������� ������� ��������� �������� ���������1R��RI�WUHHV��)LWFK� ��� ��� ��� 661 ��� ����1R��RI�VWHSV��7UHH�OHQJWK� ��� ��� ��� ��� ��� ����&RQVLVWHQF\�LQGH[ ���� ���� ���� ���� ���� ����5HWHQWLRQ�LQGH[ ���� ���� ���� ���� ���� ����$YHUDJH�QXPEHU�RI�FKDQJHV�SHU�YDULDEOH�VLWH� �QXPEHU�RI�VWHSV�QXPEHU�RI�YDULDEOH�VLWHV� ��� ��� ��� ��� ��� ���0RGHO�VHOHFWHG�E\�$NDLNH�LQIRUPDWLRQ�FULWHULRQ 790�* +.<�* 790�,�* 790�,�* 751�* *75�,�*
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TAXON����������)HEUXDU\���������±��'DUX��DO����Phylogeny of Alooideae
�� 9HUVLRQ�RI�5HFRUG��LGHQWLFDO�WR�SULQW�YHUVLRQ��
Gasteria� Haworthia�VXEJ��Hexangulares�DQG�VXEJ��Robusti-pedunculares� DQG Aloe aristata�
Combined plastid and nuclear dataset. —�$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�
VLVWHU�WR�D�ZHOO�VXSSRUWHG��%6� �������33� �������EXW�LQWHUQDOO\�XQUHVROYHG�FODGH�LQFOXGLQJ�WKH�RWKHU�PHPEHUV�RI�WKH�VXEJHQXV�
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)
��
'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
1.00
1.00 1.00
0.99
1.00
0.92
0.51
1.00
0.60
0.591.00
0.77
1.00
1.00
0.93
1.00
0.68
1.00
0.95
1.00
0.74
0.99
0.97
1.00
1.00
0.67
1.00
0.64
0.75
1.00
1.00
0.88
1.00
0.78
1.00
0.55
0.98
0.55
0.99
0.74
1.00
1.00
0.99
0.93
1.00
0.57
0.95
0.83
0.98
0.77
56(3)
í(3)
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
85(2)
89(3)
61(1)
92(6)
58(1)
99(6)
77(1)
100(11)
í(1)
79(2)75(2)
100(1)
100(10)
í(0)
80(1)
í(1)
í(1)82(2)
81(5)67(1)
57(2)í(1)
78(2)í
54(3)
í
í(3)
97(4)
88
70(2)
92(3)
61(1)
63(1)
58(1)
í
Gasteria
Haworthia subg.Hexangulares I
Haworthia subg.Robustipedunculares
Astroloba
Aloe sect. Aristatae
Haworthia sect. Hexangulares II
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�����
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TAXON����������)HEUXDU\���������±��'DUX��DO����Phylogeny of Alooideae
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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]
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0.92
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0.51
0.90
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1.00
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1.00
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0.53
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1.00
0.94
0.96
0.99
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0.740.82
0.88
0.82
0.76
0.79
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Haworthia subg.Haworthia
Aloe sect. Macrifoliae
Aloe sect. Kumara
Tree Aloe spp.
Outgroups
AB
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'DUX��DO����Phylogeny of AlooideaeTAXON����������)HEUXDU\���������±��
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5HODWLRQVKLSV�DPRQJ�H. attenuata��WKH�UHVW�RI�H. VXEJ��Hexan-gulares�DQG�Gasteria�UHPDLQ�XQUHVROYHG�
Gasteria. — Gasteria�VSHFLHV�IRUP�D�ZHOO�VXSSRUWHG�FODGH�LQ�ERWK�DQDO\VHV��%6� �����33� �������)LJ���$���7KH�FODGH�LV�UHFRYHUHG�LQ�DOO�WUHHV�DV�RQH�HOHPHQW�RI�D�PRGHUDWHO\�WR�ZHOO�VXSSRUWHG�WULFKRWRP\��%6� �����33� �������WKDW�LQFOXGHV�WKH�VSHFLHV�RI�Haworthia VXEJ��Hexangulares.
Evolution of morphological traits. —�$OO����PRUSKR-ORJLFDO�WUDLWV�WKDW�ZHUH�DQDO\VHG�DUH�VKRZQ�WR�EH�KRPRSODVLRXV��QDPHO\�KDELW��OHDI�LQVHUWLRQ��OHDI�PDUJLQV��OHDI�PDFXODWLRQ�DQG�WXEHUFXODWLRQ��OHDI�DSH[��LQIORUHVFHQFH�EUDQFKLQJ�DQG�RULHQWD-WLRQ��LQIORUHVFHQFH�OHQJWK�UHODWLYH�WR�SHGLFHO��LQIORUHVFHQFH�VKDSH�DQG�FRORXU��SHULDQWK�RULHQWDWLRQ�DW�DQWKHVLV��IORZHU�DUUDQJHPHQW�RQ�SHGXQFOH��SHULDQWK�V\PPHWU\�DQG�FRORXU��SHULDQWK�VKDSH�DQG�FXUYDWXUH��SHGLFHO�OHQJWK��WHSDO�FRQQDWLRQ��DQG�VWDPHQ�OHQJWK��$SSHQGL[�6����7KH�GLVWULEXWLRQ�RI�QLQH�FKDUDFWHUV�RI�SDUWLFX-ODU�LPSRUWDQFH�LQ�JHQHULF�DQG�LQIUDJHQHULF�FODVVLILFDWLRQ�ZLWKLQ�$ORRLGHDH�LV�VXPPDULVHG�LQ�7DEOH����VHH�DOVR�)LJV��6�±6���
Habit. —�$FDXOHVFHQFH�LV�SOHVLRPRUSKLF�ZLWKLQ�$ORRLGHDH��ZLWK�FHUWDLQ�OLQHDJHV�FKDUDFWHULVHG�E\�VHFRQGDU\�FDXOHVFHQFH�DQG� DUERUHVFHQFH� �)LJ�� 6�$��� &DXOHVFHQFH� KDV� GHYHORSHG�VHYHUDO� WLPHV� LQ� WKH�VXEIDPLO\�� LQFOXGLQJ�Aloe� LWVHOI� �QRWD-EO\�A. VHFW��Macrifoliae��DQG�LQ�VRPH�RI�WKH�VHJUHJDWH�JHQHUD��QRWDEO\�Astroloba DQG�VRPH�VSHFLHV�RI�Gasteria�DQG�Haworthia VXEJ��Hexangulares��$UERUHVFHQFH�LV�XQFRPPRQ�LQ�WKH�VXE-IDPLO\�DQG�LV�FKDUDFWHULVWLF�RI�VSHFLHV�LQ�Aloe VHFW��Aloidendron DQG�A. VHFW��Dracoaloe� LQFOXGLQJ�A. eminens �)LJ��6�$��
Leaf insertion. — 3RO\VWLFKRXV�OHDI�LQVHUWLRQ�LV�FRPPRQ�ZLWKLQ�WKH�VXEIDPLO\�EXW�GLVWLFKRXV�OHDI�LQVHUWLRQ�LV�UDUH��RF-FXUULQJ�VHYHUDO�WLPHV�LQ�Aloe��RQFH�LQ�Haworthia�DQG�SRVVLEO\�RQFH�LQ�Gasteria��)LJ��6�%��
Leaf tuberculation. — 7KH�SUHVHQFH�RI�ZKLWH�WXEHUFOHV�RQ�WKH�OHDYHV�LV�ODUJHO\�UHVWULFWHG�WR�Gasteria DQG�Haworthia�VXEJ��Hexangulares�DQG�VXEJ��Robustipedunculares�EXW�WXEHUFXOHV�DUH�DOVR�GHYHORSHG�LQ�VRPH�Aloe VSHFLHV��H�J���Aloe aristata� A. haworthioides��A. verecunda��DQG�LQ�Chortolirion��)LJ��6�&��
Table .�&KDUDFWHU�VWDWHV�IRU�QLQH�VHOHFWHG�WUDLWV�ZKLFK�KDYH�EHHQ�HPSOR\HG�LQ�SUHYLRXV�$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)
$UERUHVFHQW Aloe plicatilis (A. VHFW��Kumara���A. VHFW��Pachydendron DQG� A. VHFW��Dracoaloe
/HDI�LQVHUWLRQ6SLUDO�SRO\VWLFKRXV� 0RVW�$ORRLGHDH�
'LVWLFKRXV Gasteria obliqua� Aloe nubigena� A. plicatilis� A. verecunda DQG�Haworthia truncata
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0DFXODWH $OO Gasteria� VHYHUDO�Aloe VSHFLHV��H�J���A. aristata DQG A. VHFW�� Leptoaloe
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6SUHDGLQJ Chortolirion� Astroloba��Haworthia DQG�D�IHZ�Aloe
3HQGXORXV 0RVW�Aloe DQG�Gasteria VSHFLHV
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$FWLQRPRUSKLF� Haworthia�VXEJ��Robustipedunculares DQG�VHYHUDO�Aloe� H�J���A. albida
:HDNO\�ELODELDWH� Gasteria VSHFLHV
6WURQJO\�ELODELDWH Haworthia VXEJ��Haworthia DQG�VXEJ��Hexangulares SOXV�Chortolirion angolense
3HULDQWK�FXUYDWXUH6WUDLJKW� Astroloba��Chortolirion��Haworthia VXEJ��Hexangulares DQG�VXEJ��
Robustipedunculares DQG�PDMRULW\�RI�Aloe
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3HULDQWK&\OLQGULFDO�DQG�WDSHULQJ�LQWR�SHGLFHO� 0RVW�Aloe� Astroloba� Chortolirion� Haworthia VXEJ��Haworthia DQG�
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,QVHUWHG Chortolirion� Gasteria DQG�WKH�WKUHH�Haworthia VXEJHQHUD
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%RWK�ZKRUOV�FRQQDWH�����RU�PRUH�RI�OHQJWK� Gasteria�DQG�VRPH�Aloe VSHFLHV�
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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�
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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����������$OWKRXJK�VWLOO�UHFRJQLVDEO\�µAloe-OLNH¶��WKH�IORZHUV�RI�EHH�SROOLQDWHG�Aloe VSHFLHV��VHQVX�%RWHV��DO���������VXFK�DV�A. linearifolia DQG�A. minima�GLV-SOD\�VHYHUDO�FKDUDFWHULVWLFV�RI�WKH�Haworthia�W\SH�IORZHU�DSDUW�IURP�UHGXFHG�VL]H��QDPHO\�WKHLU�QHDUO\�KRUL]RQWDO�RULHQWDWLRQ��ZKLWLVK�DQG�ZHDNO\�ELODELDWH�SHULDQWK��DQG�VRPHWLPHV�LQFOXGHG�VWDPHQV��6PLWK��9DQ�:\N��������VXJJHVWHG�WKDW�IORUDO�]\JR-PRUSK\�LQ�Astroloba, Chortolirion DQG�Haworthia�UHSUHVHQWV�DQ�DGYDQFHG�VWDWH�GHULYHG�IURP�WKH�SOHVLRPRUSKLF�DFWLQRPRU-SKLF�SDWWHUQ��ZKLFK�LV�VXSSRUWHG�E\�RXU�DQDO\VLV�
The Gasteria-type flower. —�7KH�VR�FDOOHG�JDVWHULIRUP�IORZHU��FXUYHG�DQG�IODVN�VKDSHG�ZLWK�DQ�RYRLG��LQIODWHG�WXEH�DW�OHDVW�KDOI�DV�ORQJ�DV�WKH�SHULDQWK��DQG�LQFOXGHG�RU�VKRUWO\�
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Tepal connation. —�)ORUDO�V\QGURPHV�LQGLFDWH�WKDW�Astro-loba��Ł�Poellnitzia) rubriflora, Gasteria�DQG�VHYHUDO�Aloe VSH-FLHV�DUH�ELUG�SROOLQDWHG�DQG�RXU�UHFRQVWUXFWLRQ�VXJJHVWV�WKDW�RUQLWKRSKLO\�LQ�Gasteria�DQG�Astroloba�DUH�VHFRQGDU\�DGDSWD-WLRQV�IURP�DQ�DQFHVWUDO�HQWRPRSKLORXV��Haworthia�W\SH�IORZHU�UDWKHU�WKDQ�GHULYHG�IURP�D�PRUH�W\SLFDO�Aloe�W\SH�IORZHU�
,W� LV� LQFUHDVLQJO\�HYLGHQW� WKDW� WKH�GLIIHUHQFHV� LQ� IORUDO�PRUSKRORJ\�WKDW�ZHUH�XVHG�DV�WKH�SULPDU\�FKDUDFWHULVWLF�MXV-WLI\LQJ�WKH�UHFRJQLWLRQ�RI�WKHVH�YDULRXV�OLQHDJHV�DV�VHJUHJDWH�JHQHUD�UHSUHVHQW�V\QGURPHV�DVVRFLDWHG�ZLWK�VKLIWV�LQ�SROOLQD-WLRQ�V\VWHPV�IURP�ELUG�WR�LQVHFW�DQG�EDFN��+LVWRULFDOO\��PRVW�RI�WKH�VHJUHJDWH�JHQHUD�ZHUH�HUHFWHG�EHIRUH�LQIRUPDWLRQ�RQ�WKH�IXOO�YDULDWLRQ�LQ�Aloe�ZDV�NQRZQ��DQG�FHUWDLQO\�EHIRUH�WKH�VLJQLILFDQFH�RI�IORUDO�V\QGURPHV�LQ�SROOLQDWLRQ�ZDV�DSSUHFLDWHG��,Q�$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 1. – 7KH�SK\ORJHQHWLF�WUHH�UHSUHVHQWHG�D�JHQH�WUHH��7KLV�LV�KLJKO\�XQOLNHO\�WR�DSSO\�WR�RXU�FRPELQHG�DQDO\VLV��ZKLFK�LV�EDVHG�RQ�IRXU�JHQHV��WKUHH�SODVWLGV�DQG�RQH�QXFOHDU�UHJLRQ��
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��$��%HUJHU� [= Aloe subspicata� �%DNHU��%RDWZU���-�&��0DQQLQJ@
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'DUX��DO����Phylogeny of AlooideaeTAXON����������)HEUXDU\���������±��
��9HUVLRQ�RI�5HFRUG��LGHQWLFDO�WR�SULQW�YHUVLRQ��
Adams, S.P., Leitch, I.J. & Bennett, M.D. ������5LERVRPDO�'1$�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.��������±����
Devey, D.S., Leitch, I., Rudall, P.J., Pires, J.C., Pillon, Y. & Chase, M.W. ������6\VWHPDWLFV�RI�;DQWKRUUKRHDFHDH�VHQVX�ODWR��ZLWK�HPSKDVLV�RQ�Bulbine��Aliso��������±����
Doyle, J.J. & Doyle, J.L. ������$�UDSLG�'1$�LVRODWLRQ�SURFHGXUH�IRU�VPDOO�TXDQWLWLHV�RI�IUHVK�OHDI�WLVVXH��Phytochem. Bull.�������±���
Duval, H.A. ������Plantae succulentae in horto Alenconio��3DULV�Edgar, R.C. ������086&/(��0XOWLSOH�VHTXHQFH�DOLJQPHQW�ZLWK�KLJK�
DFFXUDF\�DQG�KLJK�WKURXJKSXW��Nucl. Acids Res.���������±�����Felsenstein, J. ������&RQILGHQFH�OHYHOV�RQ�SK\ORJHQLHV��$Q�DSSURDFK�
XVLQJ�WKH�ERRWVWUDS��Evolution��������±����Fritz, G.P.J. ������5HYLHZ�RI�WKH�WKUHH�VSHFLHV�DFFHSWHG�LQ�Chortolirion
$��%HUJHU��;DQWKRUUKRHDFHDH��$VSKRGHORLGHDH���Aloe������±���Germishuizen, G., Meyer, N.L., Steenkamp, Y. & Keith, M. (eds.)
������A checklist of South African plants. 6RXWKHUQ�$IULFDQ�%R-WDQLFDO�'LYHUVLW\�1HWZRUN�5HSRUW�1R������3UHWRULD��6$%21(7�
Gildenhuys, S.D. ������Haworthia marxii �$VSKRGHODFHDH�$ORDFHDH���D�QHZ�VSHFLHV�IURP�WKH�/LWWOH�.DURR��6RXWK�$IULFD��Aloe �����±��
Glen, H.F. & Hardy, D.S. ������$ORDFHDH��Aloe��3S���±���LQ��*HUPLV�KXL]HQ��*���HG����Flora of Southern Africa��YRO�����SW�����IDVF���� Aloaceae (First part): Aloe��3UHWRULD��1DWLRQDO�%RWDQLFDO�,QVWLWXWH�
Glen, H.F. & Smith, G.F. ������1RWHV�RQ�WKH�W\SLILFDWLRQ�RI�VRPH�VSHFLHV�RI�Aloe �$VSKRGHODFHDH�$ORDFHDH���Bothalia ������±���
Goldblatt, P. & Manning, J.C. ������5DGLDWLRQ�RI�SROOLQDWLRQ�V\VWHPV�LQ�WKH�,ULGDFHDH�RI�VXE�6DKDUDQ�$IULFD��Ann. Bot. (Oxford) �������±����
Hargreaves, A.L., Harder, L.D. & Johnson, S.D. ������Aloe incon-spicua��7KH�ILUVW�UHFRUG�RI�DQ�H[FOXVLYHO\�LQVHFW�SROOLQDWHG�DORH��S. African J. Bot. �������±����
Hillis, D.M. & Dixon, M.T. ������5LERVRPDO�'1$��0ROHFXODU�HYROX-WLRQ�DQG�SK\ORJHQHWLF�LQIHUHQFH��Quart. Rev. Biol. �������±����
Holland, P.G. ������$Q�HYROXWLRQDU\�ELRJHRJUDSK\�RI�WKH�JHQXV�Aloe. J. Biogeogr. ������±����
Huelsenbeck, J.P. & Ronquist, F. ������0U%D\HV��%D\HVLDQ�LQIHUHQFH�RI�SK\ORJHQHWLF�WUHHV��Bioinformatics��������±����
Jeppe, B. ������South African aloes��&DSH�7RZQ��3XUQHOO�Jombart, T. & Dray, S. ������DGHSK\OR��([SORUDWRU\�DQDO\VHV�IRU�WKH�
SK\ORJHQHWLF�FRPSDUDWLYH�PHWKRG��Bioinformatics ��������±�����Klopper, R.R. & Smith, G.F. ������7KH�JHQXV�Aloe /���$SVKRGHODFHDH��
$ORRLGHDH��LQ�1DPDTXDODQG��6RXWK�$IULFD��Haseltonia �����±���Klopper, R.R., Van Wyk, A.E. & Smith, G.F. ����� 3K\ORJHQHWLF�
UHODWLRQVKLSV�LQ�WKH�IDPLO\�$VSKRGHODFHDH��$VSDUDJDOHV���Biodiv-ers. & Ecol.�����±���
Linnaeus, C. ������Species plantarum���QG�HG��6WRFNKROP�Maddison, W.P. & Maddison, D.R. ������0HVTXLWH��$�PRGXODU�V\VWHP�
IRU�HYROXWLRQDU\�DQDO\VLV��YHUVLRQ�������KWWS���PHVTXLWHSURMHFW�RUJManning, J.C. & Smith, G.F. ������$VSKRGHODFHDH��$ORRLGHDH��7KH�
JHQXV�Poellnitzia�LQFOXGHG�LQ�Astroloba��Bothalia��������
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TAXON����������)HEUXDU\���������±��'DUX��DO����Phylogeny of Alooideae
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Meyer, N.L. & Smith, G.F. ������Astroloba corrugata��'HVFULSWLRQ�RI�D�ORQJ�NQRZQ�VSHFLHV�LQ�D�VRXWKHUQ�$IULFDQ�HQGHPLF�$ORRLG�JHQXV��Bothalia ������±���
Meyer, N.L. & Smith, G.F. ������$ORDFHDH��3S�����±����LQ��(JJOL��8���HG����Illustrated handbook of succulent plants: Monocotyledons. %HUOLQ��6SULQJHU�
Mössmer, M., Smith, G.F., Van Wyk, B.E. & Condy, G. ������Hawor-thia fasciata ID��fasciata��Fl. Pl. Africa ������±���
Obermeyer, A.A. ������$ORDFHDH��Haworthia koelmaniorum�2EHUP���+DUG\. Fl. Pl. Africa ���������
Obermeyer, A.A. ������/LOLDFHDH��Aloe, Chamaealoe� Haworthia� Astroloba� Poellnitzia DQG�Chortolirion. Bothalia ��������
Palumbi, S.R. ������1XFOHLF�DFLGV�,,��7KH�SRO\PHUDVH�FKDLQ�UHDFWLRQ��3S�����±����LQ��+LOOLV��'�0���0RULW]��&���0DEOH��%�.���HGV����Molecular systematics���QG�HG��6XQGHUODQG��6LQDXHU�
Paradis, E., Claude, J. & Strimmer, K. ������$3(��$QDO\VHV�RI�SK\OR-JHQHWLFV�DQG�HYROXWLRQ�LQ�5�ODQJXDJH��Bioinformatics��������±����
Posada, D. & Crandall, K.A. ������02'(/7(67��7HVWLQJ�WKH�PRGHO�RI�'1$�VXEVWLWXWLRQ��Bioinformatics �������±����
R Development Core Team ������5��$�ODQJXDJH DQG�HQYLURQPHQW�IRU�VWDWLVWLFDO�FRPSXWLQJ��5�)RXQGDWLRQ�IRU�6WDWLVWLFDO�&RPSXWLQJ��9LHQQD��$XVWULD��KWWS���ZZZ�5�SURMHFW�RUJ�
Ramdhani, S., Barker, N.P. & Cowling, R.M. ������5HYLVLWLQJ�PRQR-SK\O\�LQ�Haworthia 'XYDO��$VSKRGHODFHDH���,QFRQJUXHQFH��K\-EULGL]DWLRQ�DQG�FRQWHPSRUDU\�VSHFLDWLRQ��Taxon ��������±�����
Ratnasingham, S. & Herbert, P.D. ����� %2/'��7KH�%DUFRGH�RI�/LIH�'DWD�6\VWHP��KWWS���ZZZ�EDUFRGLQJOLIH�RUJ���Molec. Ecol. Notes �� ���±����
Reeves, G., Chase, M.W., Goldblatt, P., Fay, M.F., Cox, A.V., Lejeune, B. & Souza-Chies, T. ������0ROHFXODU�V\VWHPDWLFV�RI�,ULGDFHDH��(YLGHQFH�IURP�IRXU�SODVWLG�'1$�UHJLRQV��Amer. J. Bot. ��������±�����
Reynolds, G.W. ������The aloes of tropical Africa and Madagascar. 0EDEDQH��6ZD]LODQG��7KH�$ORHV�%RRN�)XQG�
Reynolds, G.W. ������The aloes of South Africa.�&DSH�7RZQ��$�$��%DONHPD�
Ronquist, F. & Huelsenbeck, J.P. ������0U%D\HV����%D\HVLDQ�SK\ORJH-QHWLF�LQIHUHQFH�XQGHU�PL[HG�PRGHOV��Bioinformatics���������±�����
Salm-Dyck, J.D. ����±������Monographia generum Aloe et Mesem-bryanthemi����YROV��%RQQ�
Sang, T., Crawford, D.J. & Steussy, T.F. ������&KORURSODVW�'1$�SK\-ORJHQ\��UHWLFXODWH�HYROXWLRQ�DQG�ELRJHRJUDSK\�RI�Paeonia��3DHRQL�DFHDH���Amer. J. Bot.���������±�����
Seelanan, T., Schnabel, A. & Wendel, J.F. ������&RQJUXHQFH�DQG�FRQVHQVXV�LQ�WKH�FRWWRQ�WULEH��0DOYDFHDH���Syst. Bot. �������±����
Small, R.L., Cronn, R.C. & Wendel, J.F. ������8VH�RI�QXFOHDU�JHQHV�IRU� SK\ORJHQ\� UHFRQVWUXFWLRQ� LQ�SODQWV��Austral. Syst. Bot. �������±����
Smith, G.F. ������+LVWRULFDO�UHYLHZ�RI�WKH�WD[RQRP\�RI�Chortolirion %HUJHU��$VSKRGHODFHDH��$ORRLGHDH�. Aloe ������±���
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Viljoen, A.M. ������A chemotaxonomic study of phenolic leaf com-pounds in the genus Aloe. 3K�'��WKHVLV��5DQG�$IULNDDQV�8QLYHUVLW\��-RKDQQHVEXUJ��6RXWK�$IULFD�
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