Strawberry and wild strawberry are among the most popular horticultural crops. Due to the development of soilless cultivation systems, the whole-year production of these economically important fruit crops is achievable even in countries with temperate climate. However, the responses of strawberry (Fragaria × ananassa Duch.) and wild strawberry (Fragaria vesca L.) to microclimate conditions in greenhouse gutter cultivation in the autumn–winter season in Poland have not been yet determined. The aim of this study was to analyze the physiological, biochemical, and biometrical responses of two cultivars of strawberry ‘Ostara’ and ‘San Andreas’ and two cultivars of wild strawberry ‘Baron von Solemacher’ and ‘Regina’ grown for 20 weeks, starting from September 17th, in controlled greenhouse conditions on coconut mats in gutters in the autumn–winter season and irradiated with sodium lamps. Strawberry ‘San Andreas’ produced 30% larger leaves and almost three-fold higher fresh and dry weight of biomass than ‘Ostara’. The strawberry plants ‘San Andreas’ had a higher content (20%) of chlorophyll a and 30% of chlorophyll b than ‘Ostara’ plants. Generally, ‘San Andreas’ displayed an overall higher concentration of intercellular CO2 (about 14%) than ‘Ostara’ plants providing higher gas exchange processes. Photosynthetic rate amounted to 13.0 μmol·m−2·s−1 for ‘San Andreas’ that was almost two-fold higher than for ‘Ostara’. ‘San Andreas’ flower and fruit productions were uniform and the six-fold higher individual fruit yield proved the excellent attributes of this cultivar to the greenhouse cultivation. Even though the productivity of the two studied wild strawberry cultivars was similar, ‘Regina’ showed higher values of some parameters than ‘Baron von Solemacher’ (40% larger leaves, 25% higher photosynthetic rate, 10% higher concentration of intercellular CO2). A high nutritional value of fruits is maintained compared to traditional open-air cultivation.