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Profiling Gene Expression in Germinating Brassica Roots.

Park MR, Wang YH, Hasenstein KH - Plant Mol. Biol. Rep. (2014)

Bottom Line: The expression level of all genes differed significantly at each sample position.SPGE needles can be used within two weeks when stored at 4 °C.Our data indicate that gene expression studies that are based on the entire root miss important differences in gene expression that SPGE is able to resolve for example growth adjustments during gravitropism.

View Article: PubMed Central - PubMed

ABSTRACT
Based on previously developed solid-phase gene extraction (SPGE) we examined the mRNA profile in primary roots of Brassica rapa seedlings for highly expressed genes like ACT7 (actin7), TUB (tubulin1), UBQ (ubiquitin), and low expressed GLK (glucokinase) during the first day post-germination. The assessment was based on the mRNA load of the SPGE probe of about 2.1 ng. The number of copies of the investigated genes changed spatially along the length of primary roots. The expression level of all genes differed significantly at each sample position. Among the examined genes ACT7 expression was most even along the root. UBQ was highest at the tip and root-shoot junction (RS). TUB and GLK showed a basipetal gradient. The temporal expression of UBQ was highest in the MZ 9 h after primary root emergence and higher than at any other sample position. Expressions of GLK in EZ and RS increased gradually over time. SPGE extraction is the result of oligo-dT and oligo-dA hybridization and the results illustrate that SPGE can be used for gene expression profiling at high spatial and temporal resolution. SPGE needles can be used within two weeks when stored at 4 °C. Our data indicate that gene expression studies that are based on the entire root miss important differences in gene expression that SPGE is able to resolve for example growth adjustments during gravitropism.

No MeSH data available.


Related in: MedlinePlus

Relative expression of highly expressed genes (ACT7, UBQ; a) or low expressed genes (TUB, GLK; b) along 10-mm-long primary Brassica roots. Root were sampled about 36 h after germination at five positions (a, insert) and the copy number of ACT7, UBQ, TUB, and GLK determined at each sample position [1 (tip) to 5 (root–shoot junction); average ± SE, n = 9]. Least significance differences at p < 0.05 for copy numbers of ACT7, UBQ, TUB, and GLK are 61, 229, 32, and 21 copies, respectively
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Fig6: Relative expression of highly expressed genes (ACT7, UBQ; a) or low expressed genes (TUB, GLK; b) along 10-mm-long primary Brassica roots. Root were sampled about 36 h after germination at five positions (a, insert) and the copy number of ACT7, UBQ, TUB, and GLK determined at each sample position [1 (tip) to 5 (root–shoot junction); average ± SE, n = 9]. Least significance differences at p < 0.05 for copy numbers of ACT7, UBQ, TUB, and GLK are 61, 229, 32, and 21 copies, respectively

Mentions: The number of copies of highly expressed genes (ACT7, UBQ; Fig. 6a) at five positions in 10-mm-long primary roots of Brassica varied with the sampling position. UBQ in the root–shoot junction (RS; Fig. 6a, sample position 5) showed the highest copy number, about 4,800. Low expressed genes (GLK, Tub, Fig. 6b) had the lowest copy number in the meristematic zone (sample position 1). The highest copy number for ACT7, TUB, and GLK was found at sample positions 2 and 3, respectively (Fig. 6). This data set illustrates that the expression of all examined gene differed significantly along the root.Fig. 6


Profiling Gene Expression in Germinating Brassica Roots.

Park MR, Wang YH, Hasenstein KH - Plant Mol. Biol. Rep. (2014)

Relative expression of highly expressed genes (ACT7, UBQ; a) or low expressed genes (TUB, GLK; b) along 10-mm-long primary Brassica roots. Root were sampled about 36 h after germination at five positions (a, insert) and the copy number of ACT7, UBQ, TUB, and GLK determined at each sample position [1 (tip) to 5 (root–shoot junction); average ± SE, n = 9]. Least significance differences at p < 0.05 for copy numbers of ACT7, UBQ, TUB, and GLK are 61, 229, 32, and 21 copies, respectively
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3926982&req=5

Fig6: Relative expression of highly expressed genes (ACT7, UBQ; a) or low expressed genes (TUB, GLK; b) along 10-mm-long primary Brassica roots. Root were sampled about 36 h after germination at five positions (a, insert) and the copy number of ACT7, UBQ, TUB, and GLK determined at each sample position [1 (tip) to 5 (root–shoot junction); average ± SE, n = 9]. Least significance differences at p < 0.05 for copy numbers of ACT7, UBQ, TUB, and GLK are 61, 229, 32, and 21 copies, respectively
Mentions: The number of copies of highly expressed genes (ACT7, UBQ; Fig. 6a) at five positions in 10-mm-long primary roots of Brassica varied with the sampling position. UBQ in the root–shoot junction (RS; Fig. 6a, sample position 5) showed the highest copy number, about 4,800. Low expressed genes (GLK, Tub, Fig. 6b) had the lowest copy number in the meristematic zone (sample position 1). The highest copy number for ACT7, TUB, and GLK was found at sample positions 2 and 3, respectively (Fig. 6). This data set illustrates that the expression of all examined gene differed significantly along the root.Fig. 6

Bottom Line: The expression level of all genes differed significantly at each sample position.SPGE needles can be used within two weeks when stored at 4 °C.Our data indicate that gene expression studies that are based on the entire root miss important differences in gene expression that SPGE is able to resolve for example growth adjustments during gravitropism.

View Article: PubMed Central - PubMed

ABSTRACT
Based on previously developed solid-phase gene extraction (SPGE) we examined the mRNA profile in primary roots of Brassica rapa seedlings for highly expressed genes like ACT7 (actin7), TUB (tubulin1), UBQ (ubiquitin), and low expressed GLK (glucokinase) during the first day post-germination. The assessment was based on the mRNA load of the SPGE probe of about 2.1 ng. The number of copies of the investigated genes changed spatially along the length of primary roots. The expression level of all genes differed significantly at each sample position. Among the examined genes ACT7 expression was most even along the root. UBQ was highest at the tip and root-shoot junction (RS). TUB and GLK showed a basipetal gradient. The temporal expression of UBQ was highest in the MZ 9 h after primary root emergence and higher than at any other sample position. Expressions of GLK in EZ and RS increased gradually over time. SPGE extraction is the result of oligo-dT and oligo-dA hybridization and the results illustrate that SPGE can be used for gene expression profiling at high spatial and temporal resolution. SPGE needles can be used within two weeks when stored at 4 °C. Our data indicate that gene expression studies that are based on the entire root miss important differences in gene expression that SPGE is able to resolve for example growth adjustments during gravitropism.

No MeSH data available.


Related in: MedlinePlus