Most plants use the C₃ pathway for carbon fixation. However, some plants, such as sugar cane, corn, and cacti that grow in hot conditions, use alternative pathways to fix carbon and conserve energy loss due to photorespiration. Photorespiration is the process that occurs when the oxygen concentration is high. Under such conditions, the rubisco enzyme in the Calvin cycle binds O₂ instead of CO₂, which halts photosynthesis and consumes energy.

C₄ Pathway

The C₄ pathway is used by plants such as corn and sugarcane. In the first step, CO₂ enters the mesophyll cells, and the enzyme phosphoenolpyruvate carboxylase (PEP carboxylase) adds it to the 3-carbon compound PEP to form the 4-carbon compound oxaloacetate. Oxaloacetate is then converted into organic acids, predominantly malate. Malate is then transported into the bundle sheath cells deep in the leaf, where the oxygen concentration is low. It is here that malate is broken down to release a molecule of CO₂. CO₂ then enters the Calvin cycle and is converted into sugar with the help of the enzyme RuBisCo.

In hot, arid conditions, the physical separation of carbon fixing and the Calvin cycle is advantageous, as the plants can close their stomata to conserve water. As a result, they can keep the oxygen concentration low and therefore, favor the binding of CO₂ to RuBisCo rather than O₂.

CAM Pathway

Other plants, such as cacti and pineapple, use the crassulacean acid metabolism (CAM) pathway to fix carbon. CAM plants open their stomata at night to prevent water loss during hot days. Once the CO₂ enters the mesophyll cells, it combines with PEP to form oxaloacetate and eventually malate.

Malate is then stored in vacuoles until the next day when it is released to enter the Calvin cycle. So, the carbon fixing occurs at night, and the Calvin cycle, fuelled by the light-dependent reactions, proceeds during the day. In this manner, CAM plants separate carbon fixation and sugar synthesis across different times of the day.