Molecular phylogeny of Glyphochloa (Poaceae, Panicoideae), an endemic grass genus from the Western Ghats, India

The genus Glyphochloa (Poaceae: Panicoideae: Andropogoneae: Rottboellinae) is endemic to peninsular India and is distributed on lateritic plateaus of low and high altitude in and around Western Ghats and the Malabar Coast. The genus presumably originated and diversified in the Western Ghats. Species relationships in the genus Glyphochloa were deduced here based on molecular phylogenies inferred using nuclear ribosomal ITS sequences and plastid intergenic spacer regions (atpB-rbcL, trnT-trnL, trnL-trnF), and new observations were made of spikelet morphology, caryopsis morphology and meiotic chromosome counts. We observed two distinct clades of Glyphochloa s.l. One of these (group I') includes Ophiuros bombaiensis, and is characterized by a single-awned lower glume and a base chromosome number of 6; it grows in low elevation coastal areas. The other clade (group II') has a double-awned lower glume, a base chromosome number of 7, and is restricted to higher elevation lateritic plateaus; G. ratnagirica may belong to the group II clade, or may be a third distinct lineage in the genus. A sister-group relationship between group I and II taxa (with or without G. ratnagirica) is not well supported, although the genus is recovered as monophyletic in shortest trees inferred using ITS or concatenated plastid data. We present a key to species of Glyphochloa and make a new combination for O. bombaiensis.

Cucumber chloroplast trnL(CAA) gene: nucleotide sequence and in vivo expression analysis in etiolated cucumber seedlings treated with benzyladenine and light.

The nucleotide sequence of a 714 bp BamHI-EcoRI fragment of cucumber chloroplast DNA was determined. The fragment contained a gene for tRNA(Leu) together with its flanking regions. The trnL(CAA) gene sequence is about 99% in similarity to broad bean, cauliflower, maize, spinach and tobacco corresponding genes. The relative expression level of the gene was determined by Northern (tRNA) gel blot and Northern (total cellular RNA) slot-blot analyses using the trnL gene probe in 6-day old etiolated cucumber seedlings and the seedlings that had been kept in the dark (dark-grown), treated with benzyladenine (BA) and kept in the dark (BA-treated dark-grown), illuminated (light-grown), and treated with BA and illuminated (BA-treated light-grown), for additional 4, 8 or 12 hr. The trnL transcripts and tRNA(Leu) levels in BA-treated dark-grown seedlings were 5 and 3 times higher, respectively after 4 hr BA treatment, while in the BA treated light-grown seedlings the level of trnL transcripts was only 3 times higher and had no detectable effect on mature tRNA(Leu) when compared to the time-4 hr dark-grown seedlings. However, the level of mature tRNA(Leu) did not show marked changes in the light-grown seedlings, whereas the level of trnL transcripts increases 3 times after 8 hr illumination of dark-grown seedlings. These data indicate that both light and cytokinin can signal changes in plastid tRNA gene expression. The possible regulatory mechanisms for such changes are discussed.

Cucumber chloroplast trnL(CAA) gene: nucleotide sequence and in vivo expression analysis in etiolated cucumber seedlings treated with benzyladenine and light

The nucleotide sequence of a 714 bp BamHI-EcoRI fragment of cucumber chloroplast DNA was determined. The fragment contained a gene for tRNA(Leu) together with its flanking regions. The trnL(CAA) gene sequence is about 99% in similarity to broad bean, cauliflower, maize, spinach and tobacco corresponding genes. The relative expression level of the gene was determined by Northern (tRNA) gel blot and Northern (total cellular RNA) slot-blot analyses using the trnL gene probe in 6-day old etiolated cucumber seedlings and the seedlings that had been kept in the dark (dark-grown), treated with benzyladenine (BA) and kept in the dark (BA-treated dark-grown), illuminated (light-grown), and treated with BA and illuminated (BA- treated light-grown), for additional 4, 8 or 12 hr. The trnL transcripts and tRNA(Leu) levels in BA-treated dark-grown seedlings were 5 and 3 times higher, respectively after 4 hr BA treatment, while in the BA treated light-grown seedlings the level of trnL transcripts was only 3 times higher and had not detectable effect on mature tRNA(Leu) when compared to the time-4 hr dark-grown seedlings. However, the level of mature tRNA(Leu) did not show marked changes in the light-grown seedlings, whereas the level of trnL transcripts increases 3 times after 8 hr illumination of dark-grown seedlings. These date indicate that both light and cytokinin can signal changes in plastid tRNA gene expression. The possible regulatory mechanisms for such changes are discussed.

On the systematic position of some Asian enigmatic genera of Asclepiadoideae (Apocynaceae)

The phylogenetic structure of Asclepiadoideae (Apocynaceae) has been elucidated at the tribal and subtribal levels in the last two decades. However, to date, the systematic positions of seven Asian genera, Cosmostigma, Graphistemma, Holostemma, Pentasachme, Raphistemma, Seshagiria and Treutlera, have not been investigated. In this study, we examine the evolutionary relationships among these seven small enigmatic Asian genera and clarify their positions in Asclepiadoideae, using a combination of plastid sequences of rbcL, rps16, trnL and trnL- F regions. Cosmostigma and Treutlera are resolved as members of the non-Hoya clade of Marsdenieae with strong support (maximum parsimony bootstrap support value BSMP = 96, maximum likelihood bootstrap support value BSML = 98, Bayesian-inferred posterior probability PP = 1.0). Pentasachme is resolved as sister of Stapeliinae to Ceropegieae with moderate support (BSMP = 64, BSML = 66, PP = 0.94). Graphistemma, Holostemma, Raphistemma and Seshagiria are all nested in the Asclepiadeae-Cynanchinae clade (BSMP = 97, BSML = 100, PP = 1.0). The study confirms the generally accepted tribal and subtribal structure of the subfamily. One exception is Eustegia minuta, which is placed here as sister to all Asclepiadeae (BSMP = 58, BSML = 76, PP = 0.99) and not as sister to the Marsdenieae + Ceropegieae clade. The weak support and conflicting position indicate the need for a placement of Eustegia as an independent tribe. In Asclepiadeae, a sister group position of Cynanchinae to the Asclepiadinae + Tylophorinae clade is favoured (BSMP = 84, BSML = 88, PP = 1.0), whereas Schizostephanus is retrieved as unresolved. Oxystelma appears as an early-branching member of Asclepiadinae with weak support (BSMP = 52, BSML = 74, PP = 0.69). Calciphila and Solenostemma are also associated with Asclepiadinae with weak support (BSMP = 37, BSML = 45, PP = 0.79), but all alternative positions are essentially without support. The position of Indian Asclepiadoideae in the family phylogeny is discussed. (c) 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 174, 601-619.

Generic identity of Camptorrhiza indica (Colchicaceae) based on cytogenetics and molecular phylogenetics

The tribe Iphigenieae (Colchicaceace, Liliales) includes two genera, viz. Camptorrhiza and Iphigenia, which are distributed in Africa, India, and Australasia. Iphigenia is represented by 12 species, of which six occur in India while Camptorrhiza comprises one species each in Africa (C. strumosa) and India (C. indica). The genus Camptorrhiza possesses a knee-shaped tuber attached to the corms, filaments with a thick bulge in the middle and styles with single stigma. Iphigenia on the other hand lacks knee-shaped tuber, bears linear filaments and has styles with three stigmas. Camptorrhiza indica possesses ovoid corms, linear filaments and styles with a single stigma. These characters are intermediate between Iphigenia and Camptorrhiza and hence we studied the cytogenetics and phylogenetic placement of this species to ascertain its generic identity. Somatic chromosome count (2n = 22) and karyotypic features of C. indica are very similar to that of Iphigenia species. Molecular phylogenetic studies based on atpB-rbcL, rps16, trnL, and trnL-F regions showed that C. indica is nested within a lineage of Indian Iphigenia species. Thus, C. indica was reduced to a species of Iphigenia, i.e., I. ratnagirica. Camptorrhiza is now a monotypic genus restricted only to southern Africa. A key to the Indian Iphigenia species is provided. In addition, a new combination Wurmbea novae-zelandiae is proposed for Iphigenia novae-zelandiae.