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The Mosaic Mutants of Cucumber: A Method to Produce Knock-Downs of Mitochondrial Transcripts.

Del Valle-Echevarria AR, Kiełkowska A, Bartoszewski G, Havey MJ - G3 (Bethesda) (2015)

Bottom Line: Cytoplasmic effects on plant performance are well-documented and result from the intimate interaction between organellar and nuclear gene products.Mass spectrometry and Western blots did not corroborate transcriptional differences in the mitochondrial proteome of the MSC mutant lines, indicating that post-transcriptional events, such as protein longevity, may compensate for reduced transcription in MSC mitochondria.Our results support cucumber as a model system to produce transcriptional "knock-downs" of mitochondrial genes useful to study mitochondrial responses and nuclear interactions important for plant performance.

View Article: PubMed Central - PubMed

Affiliation: Department of Horticulture, University of Wisconsin, Madison, Wisconsin 53706.

No MeSH data available.


Related in: MedlinePlus

Western blot analyses (A) of amounts of COX2 and ATP4 (arrow) proteins in three replicated samples of protein from plants of wild-type B and MSC3, MSC12, and MSC16. Relative amounts of ATP4 and COX2 (B and C) were estimated after normalization to amounts of nuclear-encoded mitochondrially targeted PORIN. Statistical analyses were performed using log10 of the ratio, and the asterisk indicates significantly greater amounts of COX2 protein in the MSC3 and MSC12 as compared to B at P < 0.05.
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fig5: Western blot analyses (A) of amounts of COX2 and ATP4 (arrow) proteins in three replicated samples of protein from plants of wild-type B and MSC3, MSC12, and MSC16. Relative amounts of ATP4 and COX2 (B and C) were estimated after normalization to amounts of nuclear-encoded mitochondrially targeted PORIN. Statistical analyses were performed using log10 of the ratio, and the asterisk indicates significantly greater amounts of COX2 protein in the MSC3 and MSC12 as compared to B at P < 0.05.

Mentions: Western blots were performed using whole leaf protein extracted from wild-type B, MSC3, MSC12, and MSC16 to evaluate the quantity of ATP4 as the potential genetic basis of MSC3, and using COX2 as a representative mitochondrially encoded protein for assessing potentially compromised mitochondrial translation in MSC12 and MSC16 (Figure 5A). The results revealed that amounts of the ATP4 protein in the MSC lines were similar to wild-type B (Figure 5B). For COX2, MSC12 had a significantly (P < 0.05) increased amount of the COX2 protein as compared to wild-type B, and MSC16 was not significantly different (Figure 5C).


The Mosaic Mutants of Cucumber: A Method to Produce Knock-Downs of Mitochondrial Transcripts.

Del Valle-Echevarria AR, Kiełkowska A, Bartoszewski G, Havey MJ - G3 (Bethesda) (2015)

Western blot analyses (A) of amounts of COX2 and ATP4 (arrow) proteins in three replicated samples of protein from plants of wild-type B and MSC3, MSC12, and MSC16. Relative amounts of ATP4 and COX2 (B and C) were estimated after normalization to amounts of nuclear-encoded mitochondrially targeted PORIN. Statistical analyses were performed using log10 of the ratio, and the asterisk indicates significantly greater amounts of COX2 protein in the MSC3 and MSC12 as compared to B at P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4478549&req=5

fig5: Western blot analyses (A) of amounts of COX2 and ATP4 (arrow) proteins in three replicated samples of protein from plants of wild-type B and MSC3, MSC12, and MSC16. Relative amounts of ATP4 and COX2 (B and C) were estimated after normalization to amounts of nuclear-encoded mitochondrially targeted PORIN. Statistical analyses were performed using log10 of the ratio, and the asterisk indicates significantly greater amounts of COX2 protein in the MSC3 and MSC12 as compared to B at P < 0.05.
Mentions: Western blots were performed using whole leaf protein extracted from wild-type B, MSC3, MSC12, and MSC16 to evaluate the quantity of ATP4 as the potential genetic basis of MSC3, and using COX2 as a representative mitochondrially encoded protein for assessing potentially compromised mitochondrial translation in MSC12 and MSC16 (Figure 5A). The results revealed that amounts of the ATP4 protein in the MSC lines were similar to wild-type B (Figure 5B). For COX2, MSC12 had a significantly (P < 0.05) increased amount of the COX2 protein as compared to wild-type B, and MSC16 was not significantly different (Figure 5C).

Bottom Line: Cytoplasmic effects on plant performance are well-documented and result from the intimate interaction between organellar and nuclear gene products.Mass spectrometry and Western blots did not corroborate transcriptional differences in the mitochondrial proteome of the MSC mutant lines, indicating that post-transcriptional events, such as protein longevity, may compensate for reduced transcription in MSC mitochondria.Our results support cucumber as a model system to produce transcriptional "knock-downs" of mitochondrial genes useful to study mitochondrial responses and nuclear interactions important for plant performance.

View Article: PubMed Central - PubMed

Affiliation: Department of Horticulture, University of Wisconsin, Madison, Wisconsin 53706.

No MeSH data available.


Related in: MedlinePlus