Rational design of novel fluorescent enzyme biosensors for direct detection of strigolactones

We developed genetically encoded fluorescent strigolactone biosensors based on the strigolactone receptors DAD2 from Petunia hybrida, and HTL7 from Striga hermonthica via domain insertion of circularly permuted GFP

Chesterfield

2020

Scholarcy highlights

  • Strigolactones are plant hormones involved in nutrient distribution, environmental adaptation, and plant development
  • Generation of an SL biosensor by FP domain insertion into DAD2 To develop a single-component SL biosensor, we applied the approach of fluorescent protein domain insertion into the P. hybrida SL receptor, DAD2
  • D14/DAD2 is composed of two major domains denoted here as the catalytic domain and lid domain, the latter of which comprises α helices 5-8
  • Two new high-sensitivity biosensors were developed based on DAD2 and ShHTL7
  • The sensitivity is on par with current mass spectrometry limits of detection: a recent SL quantification method development paper reported a limit of detection of 4.94 μg/L for rac-GR24 49
  • The biosensors cannot identify specific strigolactones; given they report on receptor binding, they may be able to distinguish functionally active strigolactones from inactive precursors and other molecules
  • Plasmid construction DAD2, ShHTL7, circularly permuted green fluorescent protein and LSSmOrange were codon optimised and synthesised as gene fragments, where DAD2 was synthesised in fusion to LexA DNA binding domain for use in the Y2H-based sensor

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