Loss and recovery dynamics of mitochondria in laticifer vessels of the rubber tree under repeated latex harvesting

Natural rubber is harvested by periodically incising the laticifer vessels in the bark of rubber trees to release latex, the cytoplasm of laticifers. Although mitochondria are suspended in the cytoplasm and were expected to leak out, some earlier studies failed to detect their presence. In this study, we identified mitochondria and plastids in expelled latex using molecular methods and confocal microscopy, quantifying their abundance at 13 850 ± 800 mitochondria per microliter latex. Each tapping event released approximately 746 ± 35 mtDNA copies and 113 ± 7 mitochondria per laticifer cell (mean cell volume: 0.008134 μl). Individual mitochondria contained 6.7 ± 0.6 genome copies (mean ± SD), a value significantly higher than the 1.5 ± 0.2 mtDNA copies per mitochondrion observed in leaves. This suggests that laticifer mitochondria are primed for proliferation. We further investigated mitochondrial dynamics during tapping cycles by measuring temporal changes in concentration. Initial latex flow exhibited the highest mitochondrial concentration (18 749 ± 954/μl), which progressively decreased to 40% of the initial level (7542 ± 940/μl) within 30 min, likely due to dilution from water influx. Following latex vessel plugging, the mitochondrial population rebounded rapidly, surpassing the initial concentration by 1.4-fold within 3 days. Subsequent tapping cycles (second and third) exhibited similar mitochondrial loss and recovery trends, though recovery kinetics shifted from a linear (first cycle) to a logarithmic pattern. These results indicate that tapping stimulates mitochondrial proliferation and that laticifer mitochondria lack protective mechanisms comparable to those of the nucleus, resulting in their expulsion with latex during harvesting.