Although mulberry cultivars Wubu, Yu711, and 7307 display distinct anatomical, morphological, and agronomic characteristics under natural conditions, it remains unclear if they differ in drought tolerance. To address this question and elucidate the underlying regulatory mechanisms at the whole-plant level, 2-month old saplings of the three mulberry cultivars were exposed to progressive soil water deficit for 5 days. The physiological responses and transcriptional changes of
PIPs
in different plant tissues were analyzed. Drought stress led to reduced leaf relative water content (RWC) and tissue water contents, differentially expressed
PIPs
, decreased chlorophyll and starch, increased soluble sugars and free proline, and enhanced activities of antioxidant enzymes in all plant parts of the three cultivars. Concentrations of hydrogen peroxide (H
2
O
2
), superoxide anion (O
2
•−
), and malonaldehyde (MDA) were significantly declined in roots, stimulated in leaves but unaltered in wood and bark. In contrast, except the roots of 7307, soluble proteins were repressed in roots and leaves but induced in wood and bark of the three cultivars in response to progressive water deficit. These results revealed tissue-specific drought stress responses in mulberry. Comparing to cultivar Yu711 and 7307, Wubu showed generally slighter changes in leaf RWC and tissue water contents at day 2, corresponding well to the steady
PIP
transcript levels, foliar concentrations of chlorophyll, O
2
•−
, MDA, and free proline. At day 5, Wubu sustained higher tissue water contents in green tissues, displayed stronger responsiveness of
PIP
transcription, lower concentrations of soluble sugars and starch, lower foliar MDA, higher proline and soluble proteins, higher ROS accumulation and enhanced activities of several antioxidant enzymes. Our results indicate that whole-plant level responses of
PIP
transcription, osmoregulation through proline and soluble proteins and antioxidative protection are important mechanisms for mulberry to cope with drought stress. These traits play significant roles in conferring the relatively higher drought tolerance of cultivar Wubu and could be potentially useful for future mulberry improvement programmes.
