METHOD FOR INDIRECT ASSESSMENT OF INTENSITY OF OXIDATIVE PHOSPHORYLATION IN GRAPE TISSUES USING RELATIVE COPY INDEX OF MITOCHONDRIAL DNA Russian patent published in 2025 - IPC A01H1/04 

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology and is a method for indirectly assessing the intensity of oxidative phosphorylation in grape tissues. The method includes an assessment of photosynthesis of plant seedlings, germination of a Vitis vinifera L. plant of the variety 'Chardonnay', an assessment of the effect of temperature treatment of plant seedlings in vitro at 45 °C 120 min on the intensity of oxidative phosphorylation processes in grape leaves, based on the relative copy of mitochondrial DNA, while grape explants were grown on a Murashige base medium-Borax with the addition of 30 g/l sucrose, 0.2 mg/l indolyl-3-butyric acid (IBA) and 2 mg/l 6-benzylaminopurine (6-BAP), experimental vessels with nutrient medium and explants, were divided into 2 groups: the experimental group of plants was subjected to heat treatment at 45 °C, 120 min (exp) in a dry-air thermostat, after the end of the heat treatment of grape leaves, the experimental group and the control group of plants that were not subjected to heat treatment were combined and the plants were incubated at a temperature of 23 ± 1With a 16-hour photoperiod and illumination with 40-watt fluorescent lamps of cold white light with an intensity of 105-115 mmol PPFD/m²/c for 30 days, samples of leaf plates were randomly selected from each group of plants for subsequent extraction of total DNA, and a reaction was carried out with preparations of the resulting total DNA quantitative RT-PCR in 20 µl of a reaction mixture consisting of 10 µl of LightCycler 480 SYBR Green I Master Mix, 5 ng of DNA, 5 µl, 3 µl of water and 1 µl each of the corresponding primers forward and reverse, 0.33 µm, RT-PCR was performed using an automatic analyzer at initial denaturation at 95 °C in for 5 min - 1 cycle, followed by 45 cycles of denaturation at 95 °C for 10 s, annealing at 58 °C for 25 s and elongation at 72 °C for 25 s, primary data were obtained in Ct format, the relative copyicity of the mitochondrial DNA NAD1 gene was determined using the chromosomal DNA gene GAPDH as a reference, the relative copyicity of the mitochondrial NAD1 gene was calculated separately for experimental (exp) and control samples.

EFFECT: invention makes it possible to simplify the indirect assessment of the intensity of oxidative phosphorylation processes in grape leaves.

! cl, 2 dwg