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Background and aims The stable carbon isotope ratio of leaf dry matter (δ 13Cp) is generally a reliable recorder of intrinsic water-use efficiency in C3 plants. Here, we investigated a previously reported pattern of developmental change in leaf δ 13Cp during leaf expansion, whereby emerging leaves are initially 13C-enriched compared to mature leaves on the same plant, with their δ 13Cp decreasing during leaf expansion until they eventually take on the δ 13Cp of other mature leaves. Methods We compiled data to test whether the difference between mature and young leaf δ 13Cp differs between temperate and tropical species, or between deciduous and evergreen species. We also tested whether the developmental change in δ 13Cp is indicative of a concomitant change in intrinsic water-use efficiency. To gain further insight, we made online measurements of 13C discrimination (∆ 13C) in young and mature leaves. Key Results We found that the δ 13Cp difference between mature and young leaves was significantly larger for deciduous than for evergreen species (−2.1 ‰ vs. −1.4 ‰, respectively). Counter to expectation based on the change in δ 13Cp, intrinsic water-use efficiency did not decrease between young and mature leaves; rather, it did the opposite. The ratio of intercellular to ambient CO2 concentrations (ci/ca) was significantly higher in young than in mature leaves (0.86 vs. 0.72, respectively), corresponding to lower intrinsic water-use efficiency. Accordingly, instantaneous ∆ 13C was also higher in young than in mature leaves. Elevated ci/ca and ∆ 13C in young leaves resulted from a combination of low photosynthetic capacity and high day respiration rates. Conclusion The decline in leaf δ 13Cp during leaf expansion appears to reflect the addition of the expanding leaf’s own 13C-depleted photosynthetic carbon to that imported from outside the leaf as the leaf develops. This mixing of carbon sources results in an unusual case of isotopic deception: less negative δ 13Cp in young leaves belies their low intrinsic water-use efficiency.

Directional change in leaf dry matter δ 13C during leaf development is widespread in C3 plants