Characterization of inhibitor(s) of β-glucuronidase enzyme activity in GUS-transgenic wheat

Ramadan, Ahmed M.; Eissa, Hala F.; El-Domyati, Fotouh M.; Saleh, O. M.; Nasser, Ibrahim; Salama, M.; Mahfouz, Magdy M.; Bahieldin, Ahmed

doi:10.1007/s11240-011-9988-5

Cited by 7 publications

(3 citation statements)

References 41 publications

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“…This probably indicated that the cells with transformation competency could not regenerate after co-cultivation or that gus gene expression was either lost, silenced or inhibited in regenerated plantlets. A similar failure in detecting GUS activity in tissues of transgenic wheat was reported (Ramadan et al 2011), which could result from the presence of some potential GUS enzyme inhibitors in those tissues (Bahieldin et al 2005;Ramadan et al 2011). Kumar et al (2011) found that the CaMV 35S promoter was ineffective in early stages of transgenic plant development in sorghum.…”

Section: Discussionsupporting

confidence: 55%

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Regeneration and Agrobacterium-mediated transformation of multiple lily cultivars

Wang

Kronenburg

Menzel

et al. 2012

Plant Cell Tiss Organ Cult

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To pursue genetic improvement of lily, efficiency of both regeneration and transformation from callus cultures induced from different explants were evaluated in multiple cultivars. Thirty-five callus lines induced from filaments or styles and one control callus line derived from bulb scales of in total twenty lily cultivars representing Lilium longiflorum, Oriental 9 Trumpet and Longiflorum 9 Asiatic hybrids were maintained on a medium with 8.3 lM picloram (PIC). In this study, they were tested for their regeneration potential by transferring them onto a regeneration medium supplemented with 0.4 lM PIC and 0.044 lM 6-benzyladenine. Regeneration was obtained in all cultivars examined and the percentage varied from zero to 89 % in the 36 callus lines. Regeneration frequency was significantly influenced by the genotype (cultivar). Subculturing the calli every 4 weeks by refreshing the regeneration medium contributed positively to bulblet formation, when compared to an eight week subculture frequency. It was found that the regeneration ability generally decreased with an increasing age of the callus cultures for all cultivars. The origin of the callus (style or filament) did not lead to significant differences in regeneration frequency, but there was an interaction between callus origin and genotype. Calli of eight randomly chosen cultivars were co-cultivated with Agrobacterium tumefaciens strain AGL0 carrying binary vectors with the gus gene as reporter and putative transgenic plants were produced. GUS histochemical assays demonstrated transient and stable expression of the gus gene in both calli and regenerated lily plants. Transient expression frequencies ranged from 0.3 to 20.6 % while stable transformation was much lower, only 1.4 % as the maximum.

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Section: Discussionsupporting

confidence: 55%

“…Nontransgenic controls never showed blue staining, while sometimes GUS expression and staining can be lacking despite the fact that T-DNA has been integrated (Ramadan et al 2011;Kumar et al 2011). In the first transformants obtained with this protocol PCR showed the presence of gus gene sequences in totally blue-stained lily plants.…”

Section: Discussionmentioning

confidence: 99%

Regeneration and Agrobacterium-mediated transformation of multiple lily cultivars

Wang

Kronenburg

Menzel

et al. 2012

Plant Cell Tiss Organ Cult

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“…This result indicates that the low GUS activity in root extracts of hydroponically grown plants is largely responsible for this discrepancy in GUS performance between substrates. Low GUS activity, which has often been reported in transgenic wheat harboring the uidA gene, is attributed to non-proteinaceous inhibitors of the β-glucuronidase that are released once cell membranes are disrupted (Bahieldin et al 2005, Ramadan et al 2011. Therefore, it was hypothesized that inhibitors of this enzyme were synthesized preferentially in hydroponically grown plants, which resulted in root extracts with very low GUS activity.…”

Section: Discussionmentioning

confidence: 99%

Characterization of the high-affinity phosphate transporter PHT1;4 gene promoter of Arabidopsis thaliana in transgenic wheat

et al. 2017

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The root specificity and phosphate (Pi) deficiency responsiveness of high-affinity phosphate transporter (PHT1) genes point to their promoters as a sustainable system to drive Pi acquisition-related transgenes in plants. In this study, a 3-kb promoter of the AtPHT1;4 gene from Arabidopsis thaliana fused to the -glucuronidase (GUS) reporter gene was biolistically introduced into wheat (Triticum aestivum L.) and functionally characterized in transgenic plants grown in hydroponics and in pots with soil under various Pi supply rates. From among 27 T 1 progeny derived from 250 T 0 , four transgenic lines reached T 3 , with two of them showing detectable GUS activity in the roots of T 4 plants. An unusually high number of transgene insertions characterized these transgenic lines, along with an irregular pattern of histochemical GUS staining and weak GUS activity. GUS expression driven by AtPHT1;4 was consistently higher under most assay conditions, as it was unaffected by 0 to 0.5 mM Pi in hydroponically grown plants, as well as by 16 to 20 mg(P) kg -1 (soil) in potted plants. Raising the soil P up to or above 40 mg kg -1 significantly down-regulated the quantity of GUS transcripts. These results show that the responsiveness of the AtPHT1;4 promoter to Pi availability in transgenic wheat was restricted to soil-grown plants, which highlighted the relevance of the substrate and Pi supply rates in assessing molecular responses to Pi deficiency.

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The ethylene response factor SmERF6 co-regulates the transcription of SmCPS1 and SmKSL1 and is involved in tanshinone biosynthesis in Salvia miltiorrhiza hairy roots

Bai

Jia

et al. 2018

Planta

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The SmERF6, which recognizes the GCC-box of SmCPS1 and SmKSL1 promoter in nucleus, regulates the tanshinone biosynthesis in Salvia miltiorrhiza hairy roots. Tanshinone, an important medicinal ingredient in Salvia miltiorrhiza, is best known for its use in medicine. However, the transcription factor regulation of tanshinone biosynthesis is unclear. Here, we isolated and identified a transcription factor in the ERF family of S. miltiorrhiza, SmERF6, which was screened from an S. miltiorrhiza cDNA library by the promoters of two key tanshinone synthesis genes (SmKSL1 and SmCPS1); this factor regulated tanshinone biosynthesis. The gene was highly expressed in the root and responded to ethylene treatment. SmERF6 modulated tanshinone biosynthesis by directly binding to an ethylene-responsive element (GCC-box) of the SmKSL1 and SmCPS1 promoters and activating their transcription. Overexpression of SmERF6 in the hairy roots increased their tanshinone accumulation, and SmERF6 silencing by RNAi led to a lower tanshinone content. Furthermore, tanshinone accumulation maintained homeostasis with the total phenolic acid and flavonoid contents in S. miltiorrhiza. These findings elucidated how SmERF6 directly co-regulates the transcription of SmCPS1 and SmKSL1 and modulates tanshinone synthesis to accelerate the metabolic flux of tanshinone accumulation in S. miltiorrhiza.

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Characterization of inhibitor(s) of β-glucuronidase enzyme activity in GUS-transgenic wheat

Cited by 7 publications

References 41 publications

Regeneration and Agrobacterium-mediated transformation of multiple lily cultivars

Regeneration and Agrobacterium-mediated transformation of multiple lily cultivars

Characterization of the high-affinity phosphate transporter PHT1;4 gene promoter of Arabidopsis thaliana in transgenic wheat

The ethylene response factor SmERF6 co-regulates the transcription of SmCPS1 and SmKSL1 and is involved in tanshinone biosynthesis in Salvia miltiorrhiza hairy roots

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