||Species of Phalaenopsis Blume (Orchidaceae) are found throughout tropical Asia, namely South China, Indochina, India, Southeast Asia, and Australia. This genus is comprised of approximately 66 species according to the latest classification. Most of them possess commercial value. Thousands of Phalaenopsis cultivars have been grown for commercial goals. Although this orchid is very beautiful and popular throughout the world, studies on the molecular systematics and phylogenetic relationships among these orchids are still deficient. |
Phylogenetic trees inferred from the internal transcribed spacers 1 and 2 (ITS1+ITS2) region of nuclear ribosomal DNA (nrDNA) and chloroplast DNAs (cpDNAs), including the intron of trnL, the IGS of trnL-trnF, and the IGS of atpB-rbcL, were used to clarify the phylogenetics and evolutionary trends of the genus Phalaenopsis (Orchidaceae). Molecular data are provided to clarify the latest systematics of the genus Phalaenopsis as suggested by Christenson (2001). He treated the genera of Doritis and Kingidium as synonyms of the genus Phalaenopsis and divided it into the five subgenera of Proboscidioides, Aphyllae, Parishianae, Polychilos, and Phalaenopsis. The results concurred that the genera Doritis and Kingidium should be treated as synonyms of the genus Phalaenopsis as suggested by Christenson (2001). The subgenera of Aphyllae and Parishianae were both shown to be monophyletic groups, and to be highly clustered with the subgenus Proboscidioides and two sections (including sections Esmeralda and Deliciosae) of the subgenus Phalaenopsis, which have the same morphological characters of four pollinia as well as similar biogeographies. Furthermore, neither the subgenus Phalaenopsis nor Polychilos was found to be a monophyletic group in this study. In addition, the phylogenetic tree indicates that Phalaenopsis is monophyletic and does not support the existing subgeneric and sectional classification.
The phylogenetic tree of the genus Phalaenopsis is basically congruent with the geographical distributions of this genus. Based on the tree, two major clades were separated within the genus Phalaenopsis. The first clade, having four pollinia, included sections Proboscidiodes, Parishianae, and Esmeralda, of which are distributed in South China, India, and Indochina. The second clade, bearing two pollinia, included the sections Phalaenopsis, Polychilos, and Fuscatae, of which are distributed in Malaysia, Indonesia, and the Philippines. In addition, the biogeography of the genus Phalaenopsis is congruent with the historical geology of the distribution regions of this genus as well. According to molecular evidences, biogeography, historical geology, and the evolutionary trend of pollinia number of orchid, evolutionary trends of the genus Phalaenopsis were deduced. The subgenus Aphyllae was suggested to be the origin of Phalaenopsis and South China was suggested to be the origin center of Phalaenopsis. In addition, there were two dispersal pathways of Phalaenopsis from the origin center to Southeast Asia. In one pathway, Phalaenopsis species dispersed from South China to Southeast Asia, in particular the Philippines, using Indochina, older lands of the Philippines (Mindoro, Palawan, Zamboanga, etc.) as steppingstones, from which the subgenus Phalaenopsis developed. In the other pathway, Phalaenopsis species dispersed from South China to Southeast Asia, in particular Indonesia and Malaysia, using the Malay Peninsula as a steppingstone, from which the subgenus Polychilos developed.
Furthermore, molecular data and geological dating were used to estimate the substitution rates of DNA from the genus Phalaenopsis based on the hypothesis of the molecular clock. The substitution rates of both ITS and cpDNA data from the genus Phalaenopsis were 2.4~4.7 x 10-9 and 3.9~7.8 x 10–10 substitutions/site/year, respectively. The substitution rates of ITS data of the genus Phalaenopsis are approximately six times those of cpDNA. Based on the substitution rates, the divergence time among most of the P. lueddemanniana complex was estimated to have been during the Pleistocene. The section Deliciosae separated from the section Stauroglottis at 21~10.5 Mya.
Furthermore, the phylogenetics of the close species of Phalaenopsis will be evaluated based on molecular data, involving three groups of close Phalaenopsis species, namely the P. amabilis complex, P. sumatrana complex, and P. violacea complex. For the first complex, the internal transcribed spacer 1 and 2 (ITS1+ITS2) regions of nuclear ribosomal DNA (nrDNA) were applied to evaluate the phylogenetics of the P. amabilis complex, namely P. amabilis, P. amabilis subsp. moluccana, P. amabilis subsp. rosenstromii, P. aphrodite, P. aphrodite subsp. formosana, and P. sanderiana. Based on molecular data, each of species/subspecies from the P. amabilis complex with the exception of P. aphrodite and its subspecies could be separated from each other. Phalaenopsis aphrodite from different locations and its subspecies could not be separated from each other, but all of them were separable from different populations/subspecies of P. amabilis. In addition, P. sanderiana was nested within both P. amabilis and its subspecies. These results do not support P. sanderiana being treated as a separate species from P. amabilis. In addition, I suggest that P. aphrodite is the origin of the P. amabilis complex and originated in the Philippines. Phalaenopsis amabilis and P. sanderiana descended from P. aphrodite (or its ancestor). Based on the phylogenetic tree, evolutionary trends of the P. amabilis complex were suggested. Within evolutionary trends of P. amabilis complex, two different lineages with different dispersal pathways were suggested. First, P. aphrodite, dispersed into Palawan and evolved to be P. amabilis, thereafter further dispersing into Borneo and Sumatra. Second, P. aphrodite dispersed into southern Mindanao and evolved into P. sanderiana, thereafter further dispersing into Sulawesi and New Guinea, from which P. amabilis subsp. moluccana and P. amabilis subsp. rosenstromii developed, respectively.
For the second complex, the phylogenetic relationship of the P. sumatrana complex, namely P. sumatrana, P. corningiana, and P. zebrina, was detected based on the ITS1 and ITS2 regions of nrDNA, the intron of trnL, and the IGS of atpB-rbcL of cpDNA. The P. sumatrana complex includes the two species of P. sumatrana and P. corningiana, as well as a problem species, P. zebrina, according to the concepts of Sweet (1980) and Christenson (2001). Based on the phylogenetic tree inferred from the ITS sequence, accessions of P. sumatrana cannot be separated from those of P. corningiana. Furthermore, accessions of P. zebrina can be separated from those of both P. sumatrana and P. corningiana. In addition, analyses of both sequences of the trnL intron and atpB-rbcL IGS of cpDNA apparently cannot discriminate among these three species of the P. sumatrana complex. Based on the molecular data of this study, plants of P. zebrina might be treated as a separate species from both P. sumatrana and P. corningiana. In the evolutionary trend of the P. sumatrana complex, plants of P. zebrina were deduced to be the relative origin group of the P. sumatrana complex based on the phylogenetic tree and biogeography. In addition, plants of both P. sumatrana and P. corningiana might have descended from plants of P. zebrina.
For the third complex, the phylogenetic trees inferred from the internal transcribed spacer 1 and 2 (ITS1+ITS2) regions of nuclear ribosomal DNA (nrDNA), the intron of trnL, and the intergenic spacer of atpB-rbcL of chloroplast DNA (cpDNA) were used to clarify the phylogenetic relationships of the P. violacea complex. The complex includes the two species of P. violacea and P. bellina, according to the concept of Christenson (2001). Based on the phylogenetic tree inferred from the ITS sequence, P. bellina could not be separated from most populations from P. violacea with the exception of the population distributed on Mentawai Is., Indonesia. In addition, analyses of both the intron of trnL and the IGS of atpB-rbcL of cpDNA apparently could not discriminate among the three species of the P. sumatrana complex. Based on the morphological characters, P. violacea from Mentawai Is. bears a long floral rachis and was separable from the other groups of the P. violacea complex. Therefore, the results in this study have a trend to treat the population of Mentawai Is. of the P. violacea complex as a separate species from P. violacea. In the evolutionary trend of the P. violacea complex, Mentawai plants of this complex might be descended from those of Sumatra/the Malay Peninsula according to the phylogenetic analysis and biogeography.