Germination of photoblastic lettuce seeds is regulated via the control of endogenous physiologically active gibberellin content, rather than of gibberellin responsiveness

Phytochrome regulates lettuce (Lactuca sativa L. cv. Grand Rapids) seed germination via the control of the endogenous level of bioactive gibberellin (GA). In addition to the previously identified LsGA20ox1, LsGA20ox2, LsGA3ox1, LsGA3ox2, LsGA2ox1, and LsGA2ox2, five cDNAs were isolated from lettuce seeds: LsCPS, LsKS, LsKO1, LsKO2, and LsKAO. Using an Escherichia coli expression system and functional assays, it is shown that LsCPS and LsKS encode ent-copalyl diphosphate synthase and ent-kaurene synthase, respectively. Using a Pichia pastoris system, it was found that LsKO1 and LsKO2 encode ent-kaurene oxidases and LsKAO encodes ent-kaurenoic acid oxidase. A comprehensive expression analysis of GA metabolism genes using the quantitative reverse transcription polymerase chain reaction suggested that transcripts of LsGA3ox1 and LsGA3ox2, both of which encode GA 3-oxidase for GA activation, were primarily expressed in the hypocotyl end of lettuce seeds, were expressed at much lower levels than the other genes tested, and were potently up-regulated by phytochrome. Furthermore, LsDELLA1 and LsDELLA2 cDNAs that encode DELLA proteins, which act as negative regulators in the GA signalling pathway, were isolated from lettuce seeds. The transcript levels of these two genes were little affected by light. Lettuce seeds in which de novo GA biosynthesis was suppressed responded almost identically to exogenously applied GA, irrespective of the light conditions, suggesting that GA responsiveness is not significantly affected by light in lettuce seeds. It is proposed that lettuce seed germination is regulated mainly via the control of the endogenous content of bioactive GA, rather than the control of GA responsiveness

Elobixibat improves rectal sensation in patients with chronic constipation aged ≥60 years: a randomised placebo-controlled study

Objective High rectal sensory thresholds (RSTs) are associated with chronic constipation (CC), especially in older patients. Bile acids (BAs) affect the RSTs of healthy individuals. Here, we aimed to investigate the effects of the BA transporter inhibitor elobixibat in patients with CC aged ≥60 years.Design We prospectively compared the RSTs of 17 patients with CC aged ≥60 years with those of 9 healthy individuals of the same age range. We next performed a prospective, randomised, parallel-group, double-blind, placebo-controlled clinical trial of 17 patients with CC who administered elobixibat or placebo daily for 1 week. Using barostat methodology, their first constant sensation volume (FCSV), defaecatory desire volume (DDV), and maximum tolerable volume (MTV) thresholds; their rectal compliance; and their faecal BA concentrations were measured before and after treatment.Results There were no significant differences in the RSTs of healthy individuals and patients with CC, but all of these tended to be higher in the latter group. Elobixibat increased the desire to defaecate, significantly reduced the threshold for FCSV (p=0.0018), and tended to reduce the threshold for DDV (p=0.0899) versus placebo. However, there were no differences in the MTV or rectal compliance of the two groups. The total faecal BA concentration increased, and particularly that of secondary BAs in the elobixibat group. Elobixibat was most efficacious in participants with a longer duration of CC and a history of treatment for CC.Conclusion Elobixibat reduces the RSTs of patients with CC aged ≥60 years, which may be important for its therapeutic effects.Trial registration number jRCTs061200030

Interlaboratory validation data on real-time polymerase chain reaction detection for unauthorized genetically modified papaya line PRSV-YK

This article is referred to research article entitled “Whole genome sequence analysis of unidentified genetically modified papaya for development of a specific detection method” (Nakamura et al., 2016) [1].Real-time polymerase chain reaction (PCR) detection method for unauthorized genetically modified (GM) papaya (Carica papaya L.) line PRSV-YK (PRSV-YK detection method) was developed using whole genome sequence data (DDBJ Sequenced Read Archive under accession No. PRJDB3976). Interlaboratory validation datasets for PRSV-YK detection method were provided. Data indicating homogeneity of samples prepared for interlaboratory validation were included. Specificity and sensitivity test data for PRSV-YK detection method were also provided. Keywords: Genetically modified, Real-time PCR, Carica papaya L., Validation dat

Changes in the transcript levels of gibberellin (GA) metabolism genes in the cotyledon end and hypocotyl end of lettuce seeds

(A) Frozen seeds were divided into two parts: cotyledon end, including the cotyledons (Cot), fruit wall (FW), seed coat (SC), and endosperm (ES); and the hypocotyl end, including the hypocotyl (Hyp), root apical meristem (RAM), shoot apical meristem (SAM), and part of the Cot, FW, SC, and ES. (B) Expression levels of GA metabolism genes after light treatment, determined using QRT-PCR. See for light treatments. The results were normalized to the expression of 18S rRNA (internal control), and the highest value was set to 100. Two independent experiments were performed, and means with standard errors are shown.<p><b>Copyright information:</b></p><p>Taken from "Germination of photoblastic lettuce seeds is regulated via the control of endogenous physiologically active gibberellin content, rather than of gibberellin responsiveness"</p><p></p><p>Journal of Experimental Botany 2008;59(12):3383-3393.</p><p>Published online 24 Jul 2008</p><p>PMCID:PMC2529229.</p><p></p

Effects of abscisic acid (ABA) treatment on the expression levels of gibberellin (GA) metabolism genes in imbibed lettuce seeds

(A) The expression levels of the genes were analysed by QRT-PCR. The results were normalized to the expression of 18S rRNA (internal control), and the expression levels of all genes examined are given relative to the reference value of the transcript level of at 0 h, set to 1. Three independent experiments were performed, and means with standard errors are shown. (B) Expression analysis using seeds that had been cut in half. The results were normalized to the expression of 18S rRNA (internal control), and the highest value was set to 100. Two independent experiments were performed, and means with standard errors are shown.<p><b>Copyright information:</b></p><p>Taken from "Germination of photoblastic lettuce seeds is regulated via the control of endogenous physiologically active gibberellin content, rather than of gibberellin responsiveness"</p><p></p><p>Journal of Experimental Botany 2008;59(12):3383-3393.</p><p>Published online 24 Jul 2008</p><p>PMCID:PMC2529229.</p><p></p

Changes in the transcript levels of gibberellin (GA) metabolism genes in imbibed lettuce seeds after various light treatments

(A) Time-course of lettuce seed germination after light treatment. Zero (0) h indicates seeds imbibed for 3 h in the dark that received no light treatment. FR, FR/R, and FR/R/FR indicate seeds treated with far-red light, far-red followed by red light, and far-red followed by red and then far-red light, respectively. FR/R+ABA indicates seeds treated with far-red followed by red light and 0.1 mM ABA. Triplicate experiments were performed, and means with standard errors are shown. (B) Expression levels of GA metabolism genes after light treatment. The expression levels of these genes were analysed by QRT-PCR. The results were normalized to the expression of 18S rRNA (internal control); the expression levels of all genes examined are given relative to the reference value of the transcript level of at 0 h, set to 1. Three independent experiments were performed, and means with standard errors are shown.<p><b>Copyright information:</b></p><p>Taken from "Germination of photoblastic lettuce seeds is regulated via the control of endogenous physiologically active gibberellin content, rather than of gibberellin responsiveness"</p><p></p><p>Journal of Experimental Botany 2008;59(12):3383-3393.</p><p>Published online 24 Jul 2008</p><p>PMCID:PMC2529229.</p><p></p

Expression of and during germination, and gibberellin (GA) responsiveness in lettuce seeds

(A) Expression levels of and after light treatment. The expression analysis was carried out by QRT-PCR. See for light treatments. The results were normalized to the expression of 18S rRNA (internal control), and the expression levels of all genes examined are given relative to the reference value of the transcript level of at 0 h, set to 1. Three independent experiments were performed, and means with standard errors are shown. (B) Expression levels of and in the cotyledon end and the hypocotyl end of lettuce seeds after light treatment. The results were normalized to the expression of 18S rRNA (internal control), and the highest value was set to 100. Two independent experiments were performed, and means with standard errors are shown. (C) Germination frequency of lettuce seeds in the presence of a GA biosynthesis inhibitor and various concentrations of GA. Five sets of 20 decoated lettuce seeds were incubated in the dark at 25 °C in medium containing 50 μM uniconazol-P and various concentrations of GA. After red light treatment, seeds were incubated at 25 °C in the dark for 24 h and the germination frequency was recorded. Means with standard errors are shown.<p><b>Copyright information:</b></p><p>Taken from "Germination of photoblastic lettuce seeds is regulated via the control of endogenous physiologically active gibberellin content, rather than of gibberellin responsiveness"</p><p></p><p>Journal of Experimental Botany 2008;59(12):3383-3393.</p><p>Published online 24 Jul 2008</p><p>PMCID:PMC2529229.</p><p></p