The Monophyly of Diplazium

166 parsimony-informative characters. This result showed that gene rbcL sequence is a good enough marker for inferring phylogenetic hiphothesis of Diplazium.

8.3.4. Phylogenetic Analysis

The 721-bp region from the pair number 1 to 721, numbered from the initial methionin codon of Nicotiana tabacum was used for analysis. As a references, species outsite of Malesia are included in the analysis: D. lonchophyllum from Central America and D. cavalerianum, D. mesosorum, D. rhachidosorus , D. sibiricum, D. squamigerum and D. wichurae from Japan and Table 8.1.. The aligned matrix of the 77- taxon contained 544 75 of parsimony- informative characters, 35 5 variable of parsimony-uninformative characters and 142 20 constant characters. The consistency index CI and retention index RI; Farris 1989 are 0.79 and 0.94, respectively. The topology of Most Parsimony MP of 8 trees and those strict concensus trees are generally similar Figure 8.1..

8.3.4.1. The Monophyly of Diplazium

Recent molecular phylogenetic analysis of Diplazium and closely related genera revealed that Diplazium is monophyletic. Preliminary phylogenetic study of Diplazium conducted by Sano et al 2000a based on chloroplast rbcL gene sequences showed the monophyletic of Diplazium clade supported by Bootstrap value 86. This clade includes Monomelangium pullingeri Bak. Tagawa. By using evidence from chloroplast trnL-F region sequences Wang et al 2003 showed the monophyletic of Diplazium clade that include Callipteris Bory, Allantoidea R. Br. Emend. Ching, and Diplaziopsis C. Chr. with supported Bootstrap value 98. All species studied in this study form a monophyletic group. The monophyly of Diplazium is strongly support in the tree with 100 bootstrap value. It is revealed that separation of Diplazium from Athyrium are strongly resolved. Thus, the monophyly of West Malesian Diplazium has been verified based on this molecular study. 167 Morphologically, Diplazium can be distinguished from other close related genera including from Athyrium by characters combination as follow Kato, 1977: 1 stipe bases neither swollen nor bearing pneumatophores; 2 Frond axes U-shaped with flat base in most species; 3 Acroscopic basal pinnules equal or smaller; lamina margin not cartilaginous, spines absent; 4 Vein free, or goniopetrid or sagenoid – anstomousing, ending near the margin; 5 Sori dorsal on the vein and linear, either single Asplenoid or double Diplazioid; generally Asplenoid sori along the acroscopic side of a vein and Diplazioid sori are confined to both sides of the basal acroscopic; both parts of double sori of about equal length; 6 Scales entire or toothed with consisting of two upturned ends of adjacent marginal cells. This morphological study see Chapter 9 supported the previous workers Ching 1940; van Alderwerelt van Rosenburgh 1908; Alston 1956; Sledge 1962; Holttum 1940, 1966; Kato 1977, 1995; Edie 1978; Tagawa Iwatsuki 1988; Andrews 1990; Kramer et al. 1990. in separating Diplazium from Athyrium .

8.3.4.2. Relationships among species within Diplazium