HighlightHere, a function for SlARF9, of the tomato ARF gene family, is defined and new insight provided into the mechanism by which auxin controls cell division and early fruit development.

Homogeneous populations of developing microspores and pollen from anthers of lily (Lilium longiflorum Thumb.) and tobacco (Nicotiana tabacum L.) show a continuous production of biomass, reaching a maximum in young pollen. The rate of RNA synthesis was 460 fg · h(-1) in young binucleate cells, 138 fg · h(-1) in late binucleate cells and 56 fg · h(-1) in microspores. The mRNA population in developing pollen can be separated into three groups. In the first group, certain types of mRNAs are present at a constant level during all stages of development. A second group is characteristic of young pollen and increases quantitatively until anthesis. A third group is seen transiently; to this belong mRNAs present only before mitosis or at a distinct cell stage after mitosis. Some of the translation products of this latter group of mRNAs showed similarities between lily and tobacco on two-dimensional gels in respect of molecular weight and isolectric point, indicating that those mRNAs and proteins play a role in the regulation of pollen development.

Transcripts of the ntp303 gene accumulate abundantly throughout pollen development, whereas the protein only accumulates to detectable levels after pollen germination. In an attempt to explain the divergence in the accumulation profiles of the mRNA and the protein, we investigated the role of the untranslated regions (UTRs) in enhancing ntp303 translation during the transition from developing to germinating pollen. Luciferase reporter gene fusion constructs containing the ntp3035′-UTR gave rise to luciferase activity that was up to 60-fold higher during pollen tube growth than that of constructs containing different 5′-UTRs. No apparent differences in the luciferase activity of these constructs were observed during pollen development. Thentp303 5′-UTR-mediated increase in luciferase activity was not significantly influenced by coding region or 3′-UTR sequences. Furthermore, enhanced luciferase activity directed by thentp303 5′-UTR occurred predominantly at the post-transcriptional level. A series of 5′-UTR deletion constructs was created to identify putative regulatory sequences required for the high level of translation during pollen tube growth. Two predicted stem loop structures (H-I and H-II) caused a complete inhibition of the enhanced translation after their total or partial deletion. A (GAA)8repeat within the H-I stem loop structure was demonstrated to be important for the modulation of translation efficiency. The H-II stem loop structure was found to be essential for the determination of mRNA stability.

Sexual reproduction is a critical process in the life-cycle of plants and very sensitive to environmental perturbations. To better understand the effect of high temperature on plant reproduction, we cultivated tomato (Solanum lycopersicum) plants in continuous mild heat. Under this condition we observed a simultaneous reduction in pollen viability and appearance of anthers with pistil-like structures, while in a more thermotolerant genotype, both traits were improved. Ectopic expression of two pistil-specific genes, TRANSMITTING TISSUE SPECIFIC and TOMATO AGAMOUS LIKE11, in the anthers confirmed that the anthers had gained partial pistil identity. Concomitantly, expression of the B-class genes TOMATO APETALA3, TOMATO MADS BOX GENE6 (TM6) and LePISTILLATA was reduced in anthers under continuous mild heat. Plants in which TM6 was partially silenced reacted hypersensitively to temperature elevation with regard to the frequency of pistilloid anthers, pollen viability and pollen quantity. Taken together, these results suggest that high-temperature-induced down-regulation of tomato B-class genes contributes to anther deformations and reduced male fertility. Improving our understanding of how temperature perturbs the molecular mechanisms of anther and pollen development will be important in the view of maintaining agricultural output under current climate changes.

The characterization of a gene with a unique microspore-specific expression pattern is reported. Isolated microspores from tobacco were used to synthesize a cDNA library. Clones that did not hybridize to leaf cDNA were further characterized by northern analysis. One clone proved to be a microspore-specific cDNA, representing a transcript of 650 nt. The corresponding gene, NTM19 (Nicotiana tabacum microspore-specific), was isolated and its sequence analysed. The gene encodes a protein of 10.8 kDa with a pI of 6.92 and a putative signal sequence at the N-terminus. A localization study revealed a unique spatial and temporal distribution. The transcript was only detected in the unicellular microspore. No hybridization signals were observed in other pollen developmental stages, nor in the surrounding anther tissues or other vegetative tissues of the plant. Therefore it can be concluded that NTM 19 is a gene with a highly microspore-specific character according to both localization and stage of expression. Southern blot analysis demonstrated the presence of a small gene family. The occurrence of TNM 19 was investigated in a range of closely and distantly related species and was found to be present in other solanaceous species, including the ancestors of tobacco and in a monocot species.

No abstract

No abstract

No abstract