<title>Abstract</title>Gravity is a critical environmental factor affecting the morphology and function of plants on Earth. Gravistimulation triggered by changes in the gravity vector induces an increase in the cytoplasmic free calcium ion concentration ([Ca²⁺]_c) as an early process of gravity sensing; however, its role and molecular mechanism are still unclear. When seedlings of <italic>Arabidopsis thaliana</italic> expressing apoaequorin were rotated from the upright position to the upside-down position, a biphasic [Ca²⁺]_c-increase composed of a fast-transient [Ca²⁺]_c-increase followed by a slow [Ca²⁺]_c-increase was observed. We find here a novel type [Ca²⁺]_c-increase, designated a very slow [Ca²⁺]_c-increase that is observed when the seedlings were rotated back to the upright position from the upside-down position. The very slow [Ca²⁺]_c-increase was strongly attenuated in knockout seedlings defective in MCA1, a mechanosensitive Ca²⁺-permeable channel (MSCC), and was partially restored in <italic>MCA1</italic>-complemented seedlings. The mechanosensitive ion channel blocker, gadolinium, blocked the very slow [Ca²⁺]_c-increase. This is the first report suggesting the possible involvement of MCA1 in an early event related to gravity sensing in Arabidopsis seedlings.