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Cloning and expression analysis of LeTIR1 in tomato

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Qiao Y., Feng X.-M., Liu Z.-Z., Wang S.-S., Hao Y.-J., You C.-X.

Acta Biologica Cracoviensia. Series Botanica

2011

tom 53

nr 2

The full-length cDNA of LeTIR1 gene was isolated from tomato with EST-based in silico cloning followed by RACE amplification. LeTIR1 contained an open reading frame (ORF) 1872 bp long, encoding 624 amino acid residues. The predicted protein LeTIR1 had one F-box motif and eleven leucine-rich repeats (LRRs), all of which are highly conserved in TIR1 proteins of other plant species. Phylogenetic analysis showed that the LeTIR1 protein shared high similarity with other known TIR1 proteins. Both sequence and phylogenetic analysis suggested that LeTIR1 is a TIR1 homologue and encodes an F-box protein in tomato. Semi-quantitative RT-PCR indicated that LeTIR1 was expressed constitutively in all organs tested, with higher expression in stem than root, leaf, flower and fruit. Its expression level was positively correlated with the auxin distribution in stem or axillary shoot, and was induced by spraying exogenous IAA.

Ectopic expression of the apple mhgai2 gene brings about GA-intensitive phenotypes in tomatoes

100%

Wang S.-S., Sun C., Liu Z.-Z., Shi Q.-H., Yao Y.-X., You C.-X., Hao Y.-J.

Acta Physiologiae Plantarum

2012

tom 34

nr 6

Bioactive gibberellins (GAs) are phytohormones that regulate plant growth and development. DELLA proteins are highly conserved growth repressors that modulate all aspects of GA responses. In this study, an apple MhGAI2 gene, which encoded a DELLA protein, was isolated from the tea crabapple (Malus hupehensis Redh. Var. pingyiensis). MhGAI2-GFP fusion protein was localized in the nucleus. Anti-MhGAI2 antibody was prepared to test GA sensitivity of the MhGAI2 protein. To elucidate the function of MhGAI2, its GA-insensitive gene mhgai2 was artificially generated via a bridge-PCR approach. Subsequently, mhgai2 was genetically transformed into tomato, while transgenic tomato line pBI containing the empty vector pBI121 was used as control. Transgenic tomato lines TL1 and TL2 ectopically expressed mhgai2 transcripts at high levels, and produced GA-insensitive mhgai2 protein. As a result, the seed germination as well as the growth of roots, hypocotyls and seedlings were much more insensitive to exogenous GA₃ application in transgenic tomato lines TL1 and TL2 than pBI control. TL1 and TL2 exhibited smaller plant statures and produced more compacted inflorescences with smaller flowers, fruits and seeds than pBI control. Therefore, mhgai2 ectopic expression affected not only the vegetative growth but also the reproductive development of transgenic tomatoes. In addition, transgenic seedlings were more resistant to drought and salt stress than pBI. In summary, ectopic expression of apple mhgai2 gene caused GA-insensitive phenotypes in tomatoes.

Ograniczanie wyników

1 Acta Biologica Cracoviensia. Series Botanica

1 Acta Physiologiae Plantarum

2 Hao Y.-J.

2 Liu Z.-Z.

2 Wang S.-S.

2 You C.-X.

1 Feng X.-M.

1 Qiao Y.

1 Shi Q.-H.

1 Sun C.

1 Yao Y.-X.

1 2012

1 2011

Wyniki wyszukiwania

Cloning and expression analysis of LeTIR1 in tomato

Ectopic expression of the apple mhgai2 gene brings about GA-intensitive phenotypes in tomatoes