In this study, a system to measure leaf internal CO
2 (
Ci) was incorporated into an open gas exchange system (LI-COR, Lincoln, NE, USA). The
Ci was directly measured with a cup attached to the abaxial surface of sunflower (
Helianthus annuus L.) leaves with open stomata while normal CO
2 and water vapor exchange through the same section of adaxial surface was simultaneously detected. The potential problems in the system, namely bulk air flow through the leaf, diffusion leaks, and change in the CO
2 gradient inside the leaf, were examined with the aim to apply the system to measure net photosynthesis at various
Ci (i.e.
A−
Ci curves). A micro blower constantly circulated the air in a loop without pressure pulses or bulk air movement through the leaf. The measured
Ci (
Ci(m)) generally followed the external CO
2 as much as the calculated one (
Ci(c)). There was close agreement between the
Ci(m) and the
Ci(c) particularly at low
Ci, and the diffusion leak hardly affected the relationship between the two. Despite possible alterations of leaf properties by cup attachment, the direct measurement is expected to cast a new light on leaf gas exchange.
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