Guizhou Province is one of the regions in China where macadamia is cultivated. The area is characterized by prominent karst landforms, with uneven distribution of precipitation and utilizable water resources, which poses significant challenges to macadamia production. To explore the effects of different drought levels on the anatomical structure and physiological characteristics of macadamia seedlings, and to reveal their adaptation mechanisms and regulatory responses to drought stress, this study established a drought stress experiment on O.C (Own Choice) macadamia seedlings. The seedlings were subjected to stress in a 25% PEG-6000 solution for 0 h (CK), 24 h, 36 h, 48 h, and 72 h, and cellular structural features of stems and leaves were measured, as well as physiological and biochemical indices. The results indicated that macadamia seedlings gradually exhibited dehydration and chlorosis with prolonged drought stress. At 72 h of drought stress, root water potential, leaf water potential, chlorophyll content, relative water content, and root activity decreased by 353%, 98%, 44%, 72%, and 79%, respectively. Leaf thickness, palisade tissue thickness, and spongy tissue thickness were reduced by 19%, 33%, and 29%, respectively. Stomatal density increased by 50%, while stomatal aperture, vessel diameter, and cell wall thickness significantly decreased. Photosynthesis was markedly impaired: Pn, Tr, Gs, WUE, Fv/Fm, qP, and ΦPSII declined by 73%, 25%, 67%, 64%, 0.23, 60%, and 84%, respectively, whereas Ci and qN increased by 107% and 11%, respectively. Cell membranes began to sustain damage after 24 h of drought stress, with electrolyte leakage and MDA content rising by 266% and 672%, respectively, at 72 h. Prolonged drought stress reduced IAA, CTK, and GA levels by 37%, 33%, and 16%, respectively, while ABA content increased by 48%. To counteract drought stress, seedlings activated osmotic adjustment and reactive oxygen species (ROS) scavenging mechanisms. Osmolyte content significantly increased with stress duration, reaching 61%, 73%, 697%, and 107% increments in SS, SP, Pro, and betaine at 72 h. Antioxidant enzyme activities initially rose, peaking at 24 h (SOD, POD, CAT, and APX increased by 132%, 288%, 110%, and 46%, respectively), then gradually declined. By 72 h, SOD and APX activities fell below control levels, while POD and CAT remained elevated. These findings demonstrate that under PEG-6000-simulated drought stress, macadamia seedlings alleviate damage by modifying leaf and stem cellular structures and activating antioxidant and osmotic adjustment mechanisms. This study provides a theoretical basis for understanding the physiological mechanisms of macadamia drought stress response.
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