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RNA quality control using external standard RNA

100%

Hasegawa T., Takahashi J., Iwahashi H.

Polish Journal of Microbiology

2018

tom 67

nr 3

In this paper, we propose a new evaluation method using external standard RNA for quality control of the extracted RNA. RNA Integrity Number and UV absorption are generally used as a basis for RNA quality control; however, these methods do not always reflect the quality of mRNA. While standard RNA is supposedly designed on the basis of mRNA, it has the potential to be used to evaluate the quality of the mRNA. In this study, we took into consideration the three essential factors, viz., yield of mRNA, inhibition to DNA polymerase, and degradation of mRNA for determining the RNA quality using standard RNA. It would be possible to know yield of mRNA and inhibition of the enzyme reaction by adding standard RNA before RNA extraction and looking at standard RNA loss. Degradation was evaluated by comparing the differences in the 3’ and 5’ regions of the RNA. In our study, it was demonstrated that in the crude extract of Saccharomyces cerevisiae, degradation was comparatively higher at the 3’ end of RNA than at the 5’ end. Hence, the degree of RNA degradation can be evaluated by comparing the ratio of degradation from the 3’ and 5’ end.

Identification of dehydrocostus lactone and 4-hydroxy-b-thujone as auxin polar transport inhibitors

51%

Ueda J., Toda Y., Kato K., Kuroda Y., Arai T., Hasegawa T., Shigemori H., Hasegawa K., Kitagawa J., Miyamoto K., Uheda E.

Acta Physiologiae Plantarum

2013

tom 35

nr 07

The survey of naturally occurring of auxin polar transport regulators in Asteraceae was investigated using the radish (Raphanus sativus L.) hypocotyl bioassay established in this study. Significant auxin polar transport was observed when radiolabeled indole-3-acetic acid (IAA) was applied at the apical side of radish hypocotyl segments, but not when it was applied at the basal side of the segments. Almost no auxin polar transport was observed in radish hypocotyl segments treated with synthetic auxin polar transport inhibitors of N-(1- naphthyl)phthalamic acid (NPA) and 9-hydroxyfluorene-9- carboxylic acid (HFCA) at 0.5 lg/plant. 2,3,5-Triiodobenzoic acid (TIBA) at 0.5 μg/plant was less effective than NPA and HFCA, and p-chlorophenoxyisobutyric acid (PCIB) at 0.5 μg/ plant had almost no effect on auxin polar transport in the radish hypocotyl bioassay. These results strongly suggest that the radish hypocotyl bioassay is suitable for the detection of bioassay-derived auxin polar transport regulators. Using the radish hypocotyl bioassay and physicochemical analyses, dehydrocostus lactone (decahydro-3,6,9-tris-methylene-azulenol (4,5-b)furan-2(3H)-one) and 4-hydroxy-β-thujone (4-hydroxy- 4-methyl-1-(1-methylethyl)-bicyclo[3.1.0]hexan-3-one) were successfully identified as auxin polar transport inhibitors from Saussurea costus and Arctium lappa, and Artemisia absinthium, respectively. About 50 and 40 % inhibitions of auxin polar transport in radish hypocotyl segments were observed at 2.5 μg/plant pre-treatment (see ‘‘Materials and methods’’) of dehydrocostus lactone and 4-hydroxy-β-thujone, respectively. Although the mode of action of these compounds in inhibiting auxin polar transport has not been clear yet, their possible mechanisms are discussed.

Ograniczanie wyników

1 Acta Physiologiae Plantarum

1 Polish Journal of Microbiology

2 Hasegawa T.

1 Arai T.

1 Hasegawa K.

1 Iwahashi H.

1 Kato K.

1 Kitagawa J.

1 Kuroda Y.

1 Miyamoto K.

1 Shigemori H.

1 Takahashi J.

1 Toda Y.

1 Ueda J.

1 Uheda E.

1 2018

1 2013

Wyniki wyszukiwania

RNA quality control using external standard RNA

Identification of dehydrocostus lactone and 4-hydroxy-b-thujone as auxin polar transport inhibitors