Table 2.
Summary of HS-responsive transcript-derived fragments (TDFs) of maturing tomato pollen grains and their homologies to sequences in the databases
| TDF* | Primer combination† | Length (bp)‡ | HS response§ | Exp. pattern¶ | Gene description** | Accesion/AGI no.†† | E-value‡‡ |
| Stress response | |||||||
| HSPs | |||||||
| TDF7 | E-ACA/M-CAA | 320 | + | A | Heat shock protein 81-1 (AtHSP81-1); A. thaliana | AT5G52640p | 3E-48 |
| TDF3.1¶¶ | E-ACA/M-CAT | 390 | + | A | Heat shock protein 81-1 (AtHSP81-1); A. thaliana | AT5G52640p | 5E-44 |
| TDF3.2¶¶ | E-ACA/M-CTT | 299 | + | A | Heat shock protein 90 (LeHSP90); S. Lycopersicum | AF123259 | 3E-13 |
| Oxidative stress | |||||||
| TDF17.1 | E-ACT/M-CTT | 268 | + | A | Glutathione S-transferase T4 (LeGSTT4); S. lycopersicum | AY007561 | 2E-8 |
| TDF6.1 | E-ACT/M-CTT | 523 | – | B | Adenylyl-sulphate reductase; S. lycopersicum | AY568717 | 3E-92 |
| TDF6.2 | E-ACT/M-CTT | 447 | – | B | Adenylyl-sulphate reductase; S. lycopersicum | AY568718 | 8E-68 |
| ER stress response | |||||||
| TDF3.3 | E-ACT/M-CTG | 616 | + | A | Cell division cycle protein 48, putative (AtCDC48); A. thaliana | AT3G53230 | 5E-109 |
| Vesicle fusion | |||||||
| TDF18.1 | E-ACT/M-CTG | 262 | + | C | Vesicle-associated membrane protein 725 (AtVAMP725); A. thaliana | AT2G32670p | 2E-27 |
| TDF17.2 | E-ACA/M-CTT | 298 | + | C | Vesicle-associated membrane protein 726 (AtVAMP726); A. thaliana | AT1G04760p | 8E-24 |
| Ubiquitination | |||||||
| TDF17.3 | E-ACT/M-CTG | 283 | + | D | Kelch repeat-containing F-box family protein; A. thaliana | AT1G15670 | 4E-23 |
| TDF4.1 | E-ACA/M-CTT | 483 | + | A | Protein kinase family protein; A. thaliana | AT5G51270 | 4E-13 |
| TDF32.1¶¶ | E-ACT/M-CTT | 173 | + | C | Ubiquitin-associated (UBA)/TS-N domain-containing protein; A. thaliana | AT1G04850 | 3E-13 |
| Defence | |||||||
| Cell wall modification | |||||||
| TDF10.1 | E-ACT/M-CTG | 334 | – | B | Tomato LAT56 gene for protein P56; S. lycopersicum | X15500 | 3E-57 |
| Metabolism and transport | |||||||
| TDF4.2¶¶ | E-ACT/M-CTG | 565 | + | A | UDP-D-glucuronate 4-epimerase 5 (AtGAE5); A. thaliana | AT4G12250 | 1E-77 |
| TDF27.1 | E-ACA/M-CAG | 143 | + | A | Glucosyl transferase; N. tabacum | AB000623 | 5E-4 |
| TDF27.2¶¶ | E-ACT/M-CTG | 179 | – | B | GL2-expression mudulator (AtGEM); A. thaliana | AT2G22475 | 7E-14 |
| TDF28 | E-ACA/M-CAA | 135 | + | A | Mitochondrial NAD+-dependent malic enzyme (malate dehydrogenase) precursor; S. tuberosum | Z23023 | 4E-5 |
| TDF17.4 | E-ACT/M-CTT | 287 | + | A | FH protein-interacting protein 2/voltage-gated potassium channel (AtFIP2); A. thaliana | AT5G55000 | 4E-13 |
| Signal transduction | |||||||
| TDF31 | E-ACT/M-CAA | 146 | + | A | Calcium and calmodulin-dependent protein kinase 2 (AtCDPK2); A. thaliana | AT1G35670p | 1E-8 |
| TDF26 | E-ACT/M-CAC | 202 | + | A | Calcium-dependent protein kinase 2 (NtCDPK2); N. tabacum | AJ344154 | 1E-8 |
| TDF4.3 | E-ACT/M-CTG | 633 | + | A | Protein kinase; A. thaliana | AT1G68690 | 7E-13 |
| TDF42.1¶¶ | E-ACT/M-CTT | 116 | – | E | AKIN gamma; M. truncatula | AY247269 | 8.6E-9 |
| TDF12.1* | E-ACT/M-CAC | 323 | + | A | 14–3–3 protein (LeTFT9); S. lycopersicum | X98865 | 7E-50 |
| Transcription regulation | |||||||
| TDF14.2 | E-ACA/M-CTC | 301 | + | A | RNA-binding protein RZ-1; N. sylvestris | D83696 | 1E-5 |
| TDF43 | E-ACT/M-CAG | 122 | + | A | DNA-binding protein phosphatase 2C (NtDBP1); N. tabacum | AF520810 | 3E-4 |
| TDF4.4 | E-ACA/M-CTT | 480 | + | A | Argonaute1 (AtAGO1); A. thaliana | AT1G48410 | 8E-76 |
| TDF4.5 | E-ACT/M-CTT | 570 | + | A | Zinc finger protein, putative; G. hirsutum | AY919286 | 6E-27 |
| TDF1 | E-ACT/M-CTG | 811 | + | C | Zinc finger (GATA type) family protein; A. thaliana | AT5G25830 | 8E-46 |
| TDF11.1 | E-ACA/M-CTA | 286 | + | A | TITAN8 (TTN8); ATP binding; A. thaliana | AT3G54670 | 4E-17 |
TDF number, representing the relative location on the acrylamide gel. Fragments in the same location representing different treatment groups or primer combinations are marked.
Primer combination used for the fragment amplification: E indicates EcoRI, M indicates MseI.
Length of the TDF sequence.
Induction (+) or repression (–) of expression by heat stress, detected in at least one of the tested stages of microspore development and one of the cultivars.
Gene expression (Exp.) pattern. Expression patterns were divided into five categories, A–E. A represents HS-induced expression in either or both developmental stages tested, in both cultivars. B represents HS-down-regulated expression in both cultivars in microspores at either the A-3 or A-5 developmental stage. C represents constitutive expression in microspores of cv. 3017 and HS-induced expression in maturing microspores of cv. 3042 at either or both developmental stages. D represents constitutive expression in cv. 3042 and HS-induced expression in microspores of cv. 3017 at either or both developmental stages. E represents constitutive expression in microspores of cv. 3017 and HS-down-regulation in microspores of cv. 3042 at A-3.
Gene description according to NCBI, TAIR, or TIGR database annotation.
Either tomato NCBI accession number (No.), GenBank homologue, or Arabidopsis genome initiative number (AGI) of best hit from TBLASTX search, using a significance threshold of 1.00E-4 are given for each TDF.
E-value indicating the significance level of homology to the corresponding gene.
TDF sequences found on the Affymetrix GeneChip® Tomato Genome Array. All TDF sequences were checked using the NETAFFX program of the Affymetrix service site (http://www.affymetrix.com/analysis/index.affx) for identifying TDFs which are presented on the Affymetrix chip.
Gene sequences found to be highly expressed in Arabidopsis pollen grains (Zimmermann et al., 2004).