Karst regions are characterized by heterogeneous soil habitats, with shallow wide soil (SW) on hilly slopes and deep narrow soil (DN) in rocky trenches. To make full use of limited water and nutrients, plants have therefore developed a number of root extension strategies. This study investigated the effect of soil moisture on horizontal root extension in SW and vertical root extension in DN by assessing root growth responses, biomass allocation, and root distribution. A full two-way factorial blocked design of soil dimensions by water availability was followed. The perennial grass
Lolium perenne
L. was grown in SW and DN under high (W100%), moderate (W50%), and low (W30%) water availability, respectively. The main results were as follows: (1) The total biomass of
L. perenne
was not influenced either by soil habitat or by water application. Root length, root surface area, root biomass and root to shoot ratio all decreased with decreasing water application in SW, but not in DN soil. (2) With decreasing water application, the cumulative percentage of root length, root surface area and root biomass in 4 rings from the center out to 12 cm of SW soil showed a trend of W50% > W30% > W100% in SW, however, the cumulative percentage of root biomass in 4 layers from the surface to a depth of 36 cm was not significantly different between different water treatments in DN. (3) Under all three water treatments, specific root length showed an increase but root length density showed a decreasing trend from the center outward in SW soil or from the surface to bottom in DN soil. Overall, these results suggest that in SW habitat, soil moisture determines horizontal expansion of the roots in
L. perenne
, although the overall expansion ability was limited in severe drought. However, due to the relatively strong water retention ability, soil moisture changes were less obvious in DN, resulting in no significant vertical extension of the root system. The root response of
L. perenne
helps our understanding of how herbaceous plants can adjust their belowground morphology to support their growth in harsh karst soil environments.
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