The demand for new sources of bioactive plant secondary metabolites (SMs) with importance as pharmaceutical and food additives, cosmetics and flavors, have emerged as rising scientific topic in parallel to the protection of natural habitats due to overexploitation and decreasing of biodiversity. Cowslip (Primula veris L. subsp. veris) is a perennial medicinal plant in which pharmaceutically important SMs such as phenolic acids glycosides (primulaverin, primeverin) and triterpene saponins (primulic acid I and II) are produced in the roots and aerial parts. These metabolites serve as quality markers for the evaluation of raw materials used for the production of expectorant and diuretic drugs. Alternatively to traditional soil-cultivation of whole plants, in vitro adventitious root production systems have the potential for elevated and stable production of plant SMs through elicitation processes. Additionally, these systems have an advantageous perspective from ecological and economical aspect comparing with conventional method for mass production of valuable compounds. In the present study eight abiotic elicitors [salicylic acid (SA), cazeine hydrolysate (CH), L-proline, silver nitrate (AgNO3), cobalt chloride (CoCl2), sucrose, jasmonic acid (JA) and putrescine], under a 60-day culture period into 250 mL flasks were tested. Afterwards, the following three elicitors applied in single (100 mg/L CH, 100 mg/L L-proline, 25 μM JA) or combinations (JA + CH, JA + L-proline), under two exposure times (30 and 60 days). The results showed that primeverin [35.7 mg/g dry weight (DW)], and cumulative phenolic acids glycosides (52.9 mg/g) content, yield index (YI) of primeverin (97.4 mg of pure dry extract), cumulative phenolic acids glycosides (144.4 mg), primulic acid I (37.2 mg), primulic acid II (9.1 mg) and cumulative saponins (45.4 mg), as well as root fresh biomass growth rate (× 10.2-fold), and root DW (2.7 g) were promoted after elicitation with 100 mg/L L-proline for 60 days. Primulaverin (23.6 mg/g), primeverin (14.4 mg/g) and the cumulative phenolic acids glycosides (38.1 mg/g) content was considerably maximized after treatment with 25 μM JA for 30 days. Combined elicitation of roots with 25 μM JA + 100 mg/L L-proline for 30 days, indicated the highest content in primulic acid II (5.9 mg/g), cumulative saponins (22 mg/g), while primulic acid I represented the highest content under 100 μM JA for 60 days (27 mg/g). Among the eight elicitors tested, L-proline, JA, and L-proline + JA proved to be the most beneficial treatments for SMs production. Following that, root fresh biomass growth rate (× 8.1), content, and YI of primulaverin (14.3 mg/g, 149.1 mg), primeverin (0.3 mg/g, 3.2 mg), cumulative phenolic acids glycosides (14.6 mg/g, 152.3 mg), primulic acid I (10.3 mg/g, 93.3 mg), primulic acid II (2.5 mg/g, 19.2 mg), and cumulative saponins (12.8 mg/g, 112.5 mg) were considerably higher after 90 days of culture in a 2 L balloon bubble type bioreactor system than after 60 days. Elicitor type, elicitor concentration, and elicitation exposure duration are factors that play a key role in the production of SMs. In the present study, the importance of abiotic elicitation strategies, and bioreactor systems which are novel in the scale-up process, on adventitious root in vitro cultures of P. veris subsp. veris have been discussed in detail and are highlighted for the first time. The production systems applied herein lay the possibilities for the large-scale manufacturing of specialized plant-derived SMs including triterpene saponins and phenolic acid glycosides, presenting commercial applications due to higher root biomass and bioactive compounds productivity obtained.