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Heat-induced proteomes in tomato pollen mother cells

ABSTRACT: Tomato pollen production and viability is highly vulnerable to higher temperature. Hot summers with temperature reaching above 32°C can disrupt production of viable pollens and fruit set, resulting in yield loss. In recent years, temperature above 35-38oC has become a norm during mid-summer with potential adverse impacts on the production of tomatoes and many other crop species. Pollens are developed through the microsporogenesis and micro-gametogenesis stages. The most heat sensitive period is from the meiotic process of the microsporocytes, at the young microspore stage (uninucleate stage of microspore) to during late pollen development (pollen mitosis). This project studied the heat-induced proteomes in microsporocyte, also called pollen mother cells (PMC). Homogenous PMC samples were collected from cross-sectioned frozen fresh anther tissues of tomato ‘Maxifort’ using laser capture microdissection (LCM). Tandem mass tag (TMT) proteomics analysis was conducted to identify proteomics changes related to heat tolerance during pollen development.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Solanum Lycopersicum

TISSUE(S): Plant Cell, Flower

SUBMITTER: Ted Thannhauser

LAB HEAD: Theodore Thannhauser

PROVIDER: PXD018915 | Pride | 2022-04-07

REPOSITORIES: pride

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			Action	DRS
	10July2018_Suping_10plexTMT_Exp3_F1.raw	Raw
	10July2018_Suping_10plexTMT_Exp3_F10.raw	Raw
	10July2018_Suping_10plexTMT_Exp3_F11.raw	Raw
	10July2018_Suping_10plexTMT_Exp3_F12.raw	Raw
	10July2018_Suping_10plexTMT_Exp3_F2.raw	Raw

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Project description:Bolting is a key process in the growth and development of lettuce (Lactuca sativa L.). High temperature can induce earlier bolting which decreases in both quality and production of lettuce. However, knowledge underlying lettuce bolting is still lacking. To better understand the molecular basis of bolting, a comparative proteomics analysis was conducted on lettuce stems in the bolting period induced by high temperature (33 °C) compared with a control (20 °C) using iTRAQ-based proteomics, phenotypic measures, and biological verifications. High temperature induced lettuce bolting, while control temperature did not. Of the 6656 proteins identified, 758 proteins significantly altered their expression level induced by high-temperature relative to the control, of which 409 were up-regulated and 349 down-regulated. Proteins with abundance level change were mainly involved in photosynthesis, carbohydrate metabolism, stress response, hormone synthesis, and signal transduction. These differential proteins were mainly enriched in pathways associated with photosynthesis and tryptophan metabolism involving in auxin (IAA) biosynthesis. Among the differentially expressed proteins associated with photosynthesis and tryptophan metabolism were up-regulated. Moreover, in gibberellin (GA) biosynthesis pathway, 10 of main enzymes of P450 were up-regulated. Proteins related to SAUR and GRP, implicated in IAA and GA signal transduction were up-regulated, and the phosphorylation and ubiquitination related proteins regulating IAA and GA signal transduction were also induced. These findings indicate that a high temperature enhances the function of photosynthesis, IAA and GA synthesis and signal transduction to promote the process of bolting, which is in line with the physiology and transcription levels of IAA and GA metabolism. Our data provide a first comprehensive dataset for gaining novel understanding of the molecular basis underlying lettuce bolting induced by high temperature. It is potentially important for further functional analysis and genetic manipulation for molecular breeding to breed new cultivar of lettuce to restrain early bolting, which is vital for improving vegetable quality.

Project description:In Arabidopsis, jasmonate is required for stamen and pollen maturation. Mutants deficient in jasmonate synthesis, such as opr3, are male-sterile but become fertile when jasmonate is applied to developing flower buds. We have used ATH1 oligonucleotide arrays to follow gene expression in opr3 stamens for 22 hours following jasmonate treatment. In these experiments, a total of 821 genes were specifically induced by jasmonate and 480 repressed. Comparisons with data from previous studies indicate that these genes constitute a stamen-specific jasmonate transcriptome, with a large proportion (70%) of the genes expressed in the sporophytic tissue but not in the pollen. Bioinformatics tools allowed us to associate many of the induced genes with metabolic pathways that are likely up-regulated during jasmonate-induced maturation. Our pathway analysis led to the identification of specific genes within larger families of homologues that apparently encode stamen-specific isozymes. Extensive additional analysis of our dataset identified 13 transcription factors that may be key regulators of the stamen maturation processes triggered by jasmonate. Two of these transcription factors, MYB21 and MYB24, are the only members of subgroup 19 of the R2R3 family of MYB proteins. A myb21 mutant obtained by reverse genetics exhibited shorter anther filaments, delayed anther dehiscence and greatly reduced male fertility. A myb24 mutant was phenotypically wild type, but production of a myb21Â myb24 double mutant indicated that introduction of the myb24 mutation exacerbated all three aspects of the myb21 phenotype. Exogenous jasmonate could not restore fertility to myb21 or myb21Â myb24 mutant plants. Together with the data from transcriptional profiling, these results indicate that MYB21 and MYB24 are induced by jasmonate and mediate important aspects of the jasmonate response during stamen development. Experiment Overall Design: Three replicates for opr3 at each time point of 0 hour, 30 minutes, 2 hours, 8 hours and 22 hourss of either JA or OPDA treatment. One replicate of wild-type stamens. <br><br>Note that data files GSM106833.txt and GSM106907.txt as downloaded from GEO are identical.

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Heat-induced proteomes in tomato pollen mother cells

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