Mitochondrial pathway of programmed cell death in Paeonia lactiflora pollen cryopreservation

Programmed cell death (PCD) is an important factor to reduces the viability of plant germplasm after cryopreservation. However, the pathways by which PCD occurs is not fully understood. To investigate whether there is a mitochondrial pathway for pollen PCD after cryopreservation, the pollen of Paeonia lactiflora two cultivars with different PCD levels after cryopreservation was used as test material and the changes of mitochondrial calcium ions (Ca2+), structure, function and their relationship with PCD were compared. The results showed that compared with fresh pollen, the PCD of ‘Feng Huang Nie Pan’ was significantly reduced after cryopreservation. Their mitochondrial Ca2+ content decreased by 74.27%, mitochondrial permeability transition pore (MPTP) opening reduced by 25.41%, mitochondrial membrane potential slightly decreased by 5.02%, cardiolipin oxidation decreased by 65.31%, and oxygen consumption remained stable, with a slightly ATP production increase. On the contrary, compared with fresh pollen, ‘Zi Feng Chao Yang’ showed severe PCD after cryopreservation. The decline in mitochondrial Ca2+-ATPase activity led to an accumulation of excessive Ca2+ within mitochondria, triggering widespread opening of MPTP, significantly affecting mitochondrial respiration and energy synthesis. These results suggest the mitochondrial pathway of PCD exists in pollen cryopreservation. •After cryopreservation, the imbalance of mitochondrial homeostasis leads to the occurrence of PCD.•The opening of MPTP plays an indispensable role in the occurrence of PCD during pollen cryopreservation.•Mitochondrial Ca2+ signaling is a pivotal regulatory factor in PCD occurrence.•For the first time established the measurement method for respiratory metabolism (OCR and ECAR) of pollen.