Acta Manilana 59 (2011), pp. 49–55 Printed in the Philippines ISSN: 0065–1370 Acta Manilana • Volume 59 (2011) Polyphyly of Bikkia Reinw. (Rubiaceae) based on multi- locus sequence analysis (ITS, rps16, trnL-F ), with emphasis on the endemic Bikkia philippinensis Val. including its conservation status Grecebio Jonathan D. Alejandro 1,2 *, Ian Kevin C. Balete 1 , Julian Fabian C. Caagbay 1 , Jana May Marie B. Cruz 1 , Christine Joy C. Narciso 1 , Dominic E. Nazareno 1 , Cecilia I. Banag 1,2 , and Millard M. Uy 1,3 1 College of Science, 2 Research Center for Natural and Applied Sciences, University of Santo Tomas, España Boulevard, 1015 Manila, Philippines, 3 School of Science and Engineering, Ateneo de Manila University, Philippines Bikkia Reinw. is a shrubby genus of the tribe Chiococceae s.s. (Rubiaceae) with ca. 20 species. A recent molecular phylogenetic study of the tribe using two markers showed that Bikkia is polyphyletic forming two separate clades, the bases of which are habitat and corolla shape. The endemic Bikkia philippinensis was excluded in their analysis due to the lack of plant material. The present study utilized three molecular markers (ITS, rps16, and trnL-F) to test the polyphyly of Bikkia as currently circumscribed, determine the phylogenetic position of the B. philippinensis in the previously identified two groups, and assess its conservation status. Our combined data tree showed that Bikkia is highly polyphyletic. The B. philippinensis is nested within the coastal group, together with the type species Bikkia tetranda. The result of the molecular analysis is also congruent with the habitat data and morphology wherein B. philippinensis was found to thrive in coastal areas and possess infundibular corolla. Finally, B. philippinensis was found to be a critically endangered species based on population size. Keywords: Bikkia, conservation, endemic, ITS, Philippines, rps16, trnL-F *To whom correspondence should be addressed Email: gdalejandro@mnl.ust.edu.ph / balejan@yahoo.com INTRODUCTION The Rubiaceae (coffee family) is the fourth most diverse families among Angiosperms with ca. 13,000 species worldwide. Rubiaceous members are found mainly in the tropics and aside from being speciose, the family is known for its high endemicity. Such narrow or restricted distributions can be attributed to the ecologic sensitivity of its species [1]. In the case of the Philippine species, which constitute about 13% of total Rubiaceae worldwide [2], they are very much understudied taxonom- ically, and previously excluded in molecular phylogenetic studies due to lack