Exogenous glycine betaine alleviates water-deficit stress in Indian pennywort (Centella asiatica) under greenhouse conditions.

Centella asiatica (Indian pennywort) is a green leafy vegetable containing centelloside' (triterpenoid), a key phytochemical component in traditional medicine. Being a glycophytic species, they exhibit decline in growth performance and yield traits when subjected to water-deficit (WD) conditions. Glycine betaine (GB) is a low molecular-weight organic metabolite that plays a crucial role in abiotic stress conditions in higher plants. The objective of this study was to investigate the potential of GB in alleviating water-deficit stress (in terms of morphological and physiological responses) in two different genotypes of Indian pennywort, "Nakhon Pathom" (NP; high centelloside-yielding genotype) and "Pathum Thani" (PT; low centelloside-yielding genotype). The genotypes of Indian pennywort were propagated by stolon cutting and transplanted into plastic bags containing 2 kg of garden soil. At the flower-initiation stage (30 days after transplantation), uniform plant material was treated exogenously with 0 (control), 25, and 50 mM GB at 100 mL per plant (one-time foliar spray) and then divided into two groups, 1) well watered (WW; irrigated daily with 400 mL fresh water; 98% field capacity) and 2) water deficit (WD; withheld water for 14 days; 72% field capacity). Foliar application of GB (25 mM) significantly improved leaf osmotic potential in NP under WD conditions via osmotic adjustment by free proline and fructose. Differences in leaf temperature (T_leaf) between WD and WW in NP were maximized (+ 1.93 °C) and the gap of T_leaf was reduced in the case of 25-50 mM GB application. Similarly, crop water stress index (CWSI) in NP and PT plants under WD condition was significantly increased by 1.95- and 1.86-fold over the control, respectively; however, it was significantly decreased by exogenous GB application. Increasing T_leaf and CWSI in drought-stressed plants was closely related to stomatal closure, leading to reduced gas exchange parameters, i.e., stomatal conductance (g_s), transpiration rate (E), net photosynthetic rate (P_n), and intercellular CO₂ concentration (C_i), and consequently decreased plant biomass and total centelloside yield. Overall physiological, morphological, and secondary metabolite traits were enhanced in NP under WD conditions using 25 mM GB exogenous application compared with the control. The study highlights the significance of GB in Indian pennywort production under limited water irrigation (water deficit) with higher vegetable yield and phytochemical stabilization.