Aim of this study was to develop a method to investigate the impact of root-gulling phylloxera (Daktulosphaira vitifoliae (Fitch)) (PY) on grapevine (Vitis vinifera L., Zweigelt), grafted onto rootstock Kober 5BB, within two growth chambers (GC). The underlying background hypothesis says that PY is able to cause an additional sink on roots, also of common used rootstocks. As a consequence carbohydrate partitioning might be negatively influenced and fruit quality decreases. The first objective was to cultivate one year old potted grapevines within the GC, each containing one treatment (“control” or “PY”) with two variants (non-fructifying and fructifying vines). Trial plants were cultivated successfully within the GC under a low light regime. Measurements of chlorophyll fluorescence (CF) were taken with different types of fluorimeters. Various CF parameters were calculated and three (Fv/Fm, PSII or Y(II) and qP) are presented. Significant differences between the GC were rarer with ongoing development of plants and with closer time periods between measurements. Less differences and thus more comparable conditions were detected when reducing trial plants. The second objective was to inoculate roots of the treatment PY with eggs of grape PY at four different dates, differing either in inoculation-technique, material, or both. Infestation by PY was checked visually in both GC and was not detected after the third inoculation. A control after the fourth inoculation verified hatching of larvae for each plant of the treatment PY. Further several adaptions, optimisations and suggestions, which can be implemented in future trials, are found. An effect by each GC, thus unequal growth conditions, cannot be excluded. Measurements on sugar physiology, pattern of assimilate transport and distribution, agronomic indicators and grape quality parameters, result in a high and valuable amount of reproducible data. Then an impact of PY on fruit and wine quality can be quantified.