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

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

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Chapter 2 inactive in the perianth and vegetative tissues. These findings are consistent with the hypothesis that LLAG1 has a similar function as AG in lily floral development. L T S C LLAG1 GAPDH Figure 3. Expression of LLAG1 in different floral and vegetative organs. RT-PCR using primers for the 3'-terminal portion of LLAG1 was used for specific amplification. This result indicated that LLAG1 transcripts are present only in stamen (S) and carpels (C) of the flower and not in leaves (L) or tepals (T). Loading and RNA quality control were verified by amplification of the constitutive lily GAPDH. Ectopic expression of LLAG1 in Arabidopsis. Functional analysis of LLAG1 cDNA was done by ectopic expression in Arabidopsis to understand whether the sequence and expression similarities between LLAG1 and AG also point to a functional relationship. A binary vector carrying 35S::LLAG1 and a kanamycin resistance gene was introduced into Arabidopsis via Agrobacterium transformation and the phenotypic alterations of the transformed plants were analysed in the T1 and T2 generations. According to the ABC model of flower development, transgenic plants overexpressing AG are expected to show homeotic modifications in the first and second whorls of the flower. This results in the formation of carpelloid organs in the first whorl and petals replaced by organs with a staminoid identity, acquiring characteristics of the ap2 mutant phenotype (Bowman et al., 1991), due to the negative interaction between A- and C-functions. Out of 60 independent kanamycin-resistant plants analysed in the T1, 26 exhibited phenotypic alterations. Plants showing homeotic changes were divided into strong and weak ap2-like phenotypes. In general, strong ap2-like plants displayed reduced height, small and curled leaves, loss of inflorescence indeterminacy, bumpy siliques and also flowered earlier than wild type plants (Figure 4), whereas the weak ap2-like group showed normal vegetative growth with less pronounced floral homeotic modifications. 22

A C E 4 3 1 2 1 B D F Characterisation of LLAG1 in Arabidopsis Figure 4. Floral and vegetative morphology of Arabidopsis. A, Wild-type flower consisting of 4 sepals, 4 petals, 6 stamens and a pistil. B, ag-1 mutant flower in which stamens are converted to petals and the pistil to a new flower in a reiterated manner. C and D, Transgenic plants overexpressing LLAG1 under 35S promoter show homeotic mutations in the first and second whorls similar to those found in ap2 mutants. Arrow indicates a complete conversion of a petal into a stamen in the second whorl, which is visualised in between first whorl organs. Whorl numbers are indicated on the organs. E, Rosette leaves of a wild type plant. F, Rosette leaves of a transgenic plant with a strong ap2-like phenotype. 23

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