A Test for Molecular Identification of Japanese Archaeological Beans and Phylogenetic Relationship of Wild and Cultivated Species of Subgenus Ceratotropis (Genus Vigna, Papilionaceae) using Sequence Variation in Two Non-Coding Regions of theTrnl andTrnf Genes

Phylogenetic Relationship of Wild and Cultivated Vigna (Subgenus Ceratotropis, Fabaceae) from Myanmar Based on Sequence Variations in Non-coding Regions of trnT-F

“…Using 25 µl of a reaction mixture containing 2.5 µl of 10 × reaction buffer, 2.5 µl of 25 mM MgCl 2 , 2.5 µl of 1.25 Table 1. Species or species groups reported by Tateishi (1996) and Tomooka et al (2002a) Tateishi ( Okinawa, Iriomote/Japan AB304025/AB304075 1) Same accessions as those used by Yano et al (2004). mM dNTPs, 1.25 µl of each 10 µM primer, 0.1 µl of Taq (5 units/µl), 1.0 µl of 10.0 ng/µl genomic DNA and 13.9 µl of dH 2 O, the trnT-L region was amplified by polymerase chain reactions in a thermocycler (Gene Amp PCR System 2700, Applied Biosystems) as follows: initial denaturation for 2 min at 96°C, followed by 35 cycles of denaturation (94°C for 1 min), annealing (57°C for 45 sec) and extension (72°C for 2 min), with final extension at 72°C for 5 min in the thermocycler.…”

“…Rich. of the subgenus Plectotropis were selected as outgroups based on their relationship with the ingroup described in the previous studies (Fatokun et al 1993, Goel et al 2002, Maréchal et al 1978, Yano et al 2004, Yasuda and Yamaguchi 1996. Wild, weedy and cultivated races were assigned in reference to the reports of Yamaguchi (1992) and Tomooka et al (2002b).…”

“…mM dNTPs, 1.25 µl of each 10 µM primer, 0.1 µl of Taq (5 units/µl), 1.0 µl of 10.0 ng/µl genomic DNA and 13.9 µl of dH 2 O, the trnT-L region was amplified by polymerase chain reactions in a thermocycler (Gene Amp PCR System 2700, Applied Biosystems) as follows: initial denaturation for 2 min at 96°C, followed by 35 cycles of denaturation (94°C for 1 min), annealing (57°C for 45 sec) and extension (72°C for 2 min), with final extension at 72°C for 5 min in the thermocycler. The trnL intron and trnL-F spacer regions were amplified by using the same protocol as that described by Yano et al (2004). Amplified products were purified according to the instructions included in the manuals of the QIA quick Spin PCR Purification Kit (QIAgen, Valencia, Califonia, USA).…”

“…*Corresponding author (e-mail: hyama102@plant.osakafu-u.ac.jp) on the analysis of the ITS sequences, recognized two groups in Ceratotropis. A phylogenetic analysis based on the trnL and trnL-F sequences (Yano et al 2004) revealed the presence of two major clades, the azuki bean group and the mung bean group in Ceratotropis. Furthermore, a molecular phylogenetic tree constructed by Yano et al (2004) was incongruent with the other trees (Doi et al 2002, Goel et al 2002 in the grouping of V. reflexo-pilosa, which was classified into the mung bean group rather than the azuki bean group.…”

“…A phylogenetic analysis based on the trnL and trnL-F sequences (Yano et al 2004) revealed the presence of two major clades, the azuki bean group and the mung bean group in Ceratotropis. Furthermore, a molecular phylogenetic tree constructed by Yano et al (2004) was incongruent with the other trees (Doi et al 2002, Goel et al 2002 in the grouping of V. reflexo-pilosa, which was classified into the mung bean group rather than the azuki bean group. Despite repeated taxonomic revisions based on field studies and molecular analyses on the subgenus Ceratotropis, the phylogenetic relationship of several wild Vigna species occuring in South and South East Asia, in particular, V. minima (Roxb.)…”