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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

Fold-change differences with standard errors for copy number of mitochondrial genes among wild-type line B and MSC lines. Normalization was performed using the nuclear gene gadph as compared to wild-type B as described by Livak and Schmittgen (2001). Significant difference between an MSC mutant for a given mitochondrial gene as compared to wild-type B was established at α = 0.05 using pairwise t-test. Lower and higher copy number comparison between an MSC mutant and wild-type line B are shown as “a” and “b,” respectively.
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fig2: Fold-change differences with standard errors for copy number of mitochondrial genes among wild-type line B and MSC lines. Normalization was performed using the nuclear gene gadph as compared to wild-type B as described by Livak and Schmittgen (2001). Significant difference between an MSC mutant for a given mitochondrial gene as compared to wild-type B was established at α = 0.05 using pairwise t-test. Lower and higher copy number comparison between an MSC mutant and wild-type line B are shown as “a” and “b,” respectively.

Mentions: We focused on mitochondrial genes that appeared in lower copy number in MSC3, MSC12, and MSC16 relative to wild-type B (Table 1 and Table S1). The polycistronic region nad5ex4-atp4-nad5ex5 in MSC3 and rps7 in MSC12 and MSC16 were hypothesized to be likely candidates for their respective MSC phenotypes and their relative amounts were assessed using quantitative (q) PCR (Figure 2 and Table S1). Normalization was performed using the nuclear gene gadph, because it has been previously validated as an appropriate reference gene for leaf tissues (Hruz et al. 2011), and we did not detect copy number differences among the MSC lines and wild-type B (Table S2).


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)

Fold-change differences with standard errors for copy number of mitochondrial genes among wild-type line B and MSC lines. Normalization was performed using the nuclear gene gadph as compared to wild-type B as described by Livak and Schmittgen (2001). Significant difference between an MSC mutant for a given mitochondrial gene as compared to wild-type B was established at α = 0.05 using pairwise t-test. Lower and higher copy number comparison between an MSC mutant and wild-type line B are shown as “a” and “b,” respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig2: Fold-change differences with standard errors for copy number of mitochondrial genes among wild-type line B and MSC lines. Normalization was performed using the nuclear gene gadph as compared to wild-type B as described by Livak and Schmittgen (2001). Significant difference between an MSC mutant for a given mitochondrial gene as compared to wild-type B was established at α = 0.05 using pairwise t-test. Lower and higher copy number comparison between an MSC mutant and wild-type line B are shown as “a” and “b,” respectively.
Mentions: We focused on mitochondrial genes that appeared in lower copy number in MSC3, MSC12, and MSC16 relative to wild-type B (Table 1 and Table S1). The polycistronic region nad5ex4-atp4-nad5ex5 in MSC3 and rps7 in MSC12 and MSC16 were hypothesized to be likely candidates for their respective MSC phenotypes and their relative amounts were assessed using quantitative (q) PCR (Figure 2 and Table S1). Normalization was performed using the nuclear gene gadph, because it has been previously validated as an appropriate reference gene for leaf tissues (Hruz et al. 2011), and we did not detect copy number differences among the MSC lines and wild-type B (Table S2).

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