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Flower development of Lilium longiflorum - The Lilium information ...

Flower development of Lilium longiflorum - The Lilium information ...

Chapter 2 7 4 4 17 20 30

Chapter 2 7 4 4 17 20 30 13 50 26 32 36 52 74 78 27 60 33 21 89 89 0.1 37 68 99 69 100 59 84 79 93 54 99 73 100 100 97 AG Arabidopsis thaliana BAG1 Brassica napus LMADS2 Lilium longiflorum STK Arabidopsis thaliana CUM10 Cucumis sativus 100 FBP11 Petunia x hybrida FBP7 Petunia x hybrida GGM3 Gnetum gnemon DAL2 Picea abies 100 SAG1 Picea mariana LLAG1 Lilium longiflorum HAG1 Hyacinthus orientalis 100 PeMADS1 Phalaenopsis equestris HvAG1 Hordeum vulgare OsMADS3 Oryza sativa 100 CaMADS1 Corylus avellana PTAG1 Populus balsamifera PTAG2 Populus balsamifera 100 MdMADS15 Malus domestica RAG1 Rosa hybrida STAG1 Fragaria x ananassa CUS1 Cucumis sativus CUM1 Cucumis sativus FAR Antirrhinum majus TAG1 Lycopersicon esculentum FBP14 (pMADS3) Petunia x hybrida NAG1 Nicotiana tabacum FBP6 Petunia x hybrida PLE Antirrhinum majus LAG Liquidambar styraciflua VvMADS1 Vitis vinifera GAG1 Panax ginseng GAGA2 Gerbera hybrida GAGA1 Gerbera hybrida HaAG Helianthus annuus RAP1 Rumex acetosa SLM1 Silene latifolia ZAG1 Zea mays HvAG2 Hordeum vulgare WAG Triticum aestivum OsMADS13 Oryza sativa ZMM1 / ZAG2 Zea mays

Characterisation of LLAG1 in Arabidopsis Figure 2. Phylogram of MADS-box gene members with C and D functions of the AG subfamily. The tree was generated by ClustalX version 1.8 program (Thompson et al., 1997) using a Phylip distance matrix with 1,000 bootstrap trials and the graphic representation was given by TREEVIEW software (Page, 1996). LLAG1 is indicated in bold text. The species of origin are given after the abbreviated name of the genes. Bar represents 10% amino acid substitution per site along the 185-domain. Within the MADS domain, LLAG1 shares 100% (56/56) amino acid similarity with AG from Arabidopsis thaliana, HAG from Hyacinthus orientalis, PeMADS1 from Phalaenopsis equestris, OsMADS3 from Oryza sativa, WAG from Triticum aestivum, pMADS3 from Petunia hybrida and CaMADS1 from Corylus avellana while there is only one non-conserved amino acid substitution in PLE from Antirrhinum majus, leading to 98% similarity. The C-terminus is the least conserved portion of the AG homologues. LLAG1 shares 81% (58/72) amino acid similarity with HAG1, 92% (67/73) with PeMADS1, 59% (47/80) with OsMADS3, 82% (64/78) with WAG, 93% (65/70) with CaMADS1, 89% (62/70) with pMADS3, 84% (59/70) with PLE, and 60% (48/80) with AG. As a whole, the similarities in the predicted primary structure of LLAG1 range from 89% with HAG1 (202/228) to 67% with AG (191/284) among the sequences shown in Figure 1. Phylogenetic analyses indicate that MADS-box sub-families representing monophyletic clades tend to show similar sequences, expression patterns and related functions (Purugganan, 1997). Multiple alignment with LLAG1 and other members of the monophyletic AG clade is presented as a phylogram in Figure 2. It shows that LLAG1 is closely related to the monocot AG orthologues, specially HAG1 and PeMADS1, the latter being from an orchid species. LLAG1 expression pattern. The spatial expression pattern of LLAG1 in floral organs of lily was investigated by RT-PCR using gene specific primers designed to amplify the 3’ portion, which is its least conserved section. Amplification of a GAPDH fragment was used as a constitutive control. A fragment of approximately 500 bp corresponding to LLAG1 transcripts could only be detected in stamens and carpels while it was not detectable in tepals or leaves (Figure 3). This expression pattern suggests that this gene is involved in the development of reproductive floral organs since it is expressed in stamens and carpels but remains 21

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