Drought and Abscisic Acid Effects on Aquaporin Content Translate into Changes in Hydraulic Conductivity and Leaf Growth Rate: A Trans-Scale Approach

The effects of abscisic acid (ABA) on aquaporin content, root hydraulic conductivity (Lp_r), whole plant hydraulic conductance, and leaf growth are controversial. We addressed these effects via a combination of experiments at different scales of plant organization and tested their consistency via a model. We analyzed under moderate water deficit a series of transformed maize (Zea mays) lines, one sense and three antisense, affected in NCED (for 9-cis-epoxycarotenoid dioxygenase) gene expression and that differed in the concentration of ABA in the xylem sap. In roots, the mRNA expression of most aquaporin PIP (for plasma membrane intrinsic protein) genes was increased in sense plants and decreased in antisense plants. The same pattern was observed for the protein contents of four PIPs. This resulted in more than 6-fold differences between lines in Lp_r under both hydrostatic and osmotic gradients of water potential. This effect was probably due to differences in aquaporin activity, because it was nearly abolished by a hydrogen peroxide treatment, which blocks the water channel activity of aquaporins. The hydraulic conductance of intact whole plants was affected in the same way when measured either in steady-state conditions or via the rate of recovery of leaf water potential after rewatering. The recoveries of leaf water potential and elongation upon rehydration differed between lines and were accounted for by the experimentally measured Lp_r in a model of water transfer. Overall, these results suggest that ABA has long-lasting effects on plant hydraulic properties via aquaporin activity, which contributes to the maintenance of a favorable plant water status.     

Cold Stress-Induced Acclimation in Rice is Mediated by Root-Specific Aquaporins

Cold acclimation process plays a vital role in the survival of chilling- and freezing-tolerant plants subjected to cold temperature stress. However, it remains elusive whether a cold acclimation process enhances root water uptake (a component of chilling tolerance) in chilling-sensitive crops such as rice. By analyzing the root hydraulic conductivity under cold stress for a prolonged time, we found that cold stress induced a gradual increase in root osmotic hydraulic conductivity [Lp(r(os))]. Compared with the control treatment (roots and shoots at 25°C), low root temperature (LRT) treatment (roots at 10°C; shoots at 25°C) dramatically reduced Lp(r(os)) within 1 h. However, Lp(r(os)) gradually increased during prolonged LRT treatment and it reached 10-fold higher values at day 5. Moreover, a coordinated up-regulation of root aquaporin gene expression, particularly OsPIP2;5, was observed during LRT treatment. Further, comparison of aquaporin gene expression under root-only chilling (LRT) and whole-plant chilling conditions, and in the roots of intact plants vs. shootless plants, suggests that a shoot to root signal is necessary for inducing the expression of aquaporin genes in the root. Collectively, these results demonstrate that a cold acclimation process for root water uptake functions in rice and is possibly regulated through aquaporins.     

Transpiration difference under high evaporative demand in chickpea (Cicer arietinum L.) may be explained by differences in the water transport pathway in the root cylinder

• Terminal drought substantially reduces chickpea yield. Reducing water use at vegetative stage by reducing transpiration under high vapor pressure deficit (VPD), i.e. under dry/hot conditions, contributes to drought adaptation. We hypothesized that this trait could relate to differences in a genotype's dependence on root water transport pathways and hydraulics.
• Transpiration rate responses in conservative and profligate chickpea genotypes were evaluated under increasing VPD in the presence/absence of apoplastic and cell‐to‐cell transport inhibitors.
• Conservative genotypes ICC 4958 and ICC 8058 restricted transpiration under high VPD compared to the profligate genotypes ICC 14799 and ICC 867. Profligate genotypes were more affected by aquaporin inhibition of the cell‐to‐cell pathway than conservative genotypes, as measured by the root hydraulic conductance and transpiration under high VPD. Aquaporin inhibitor treatment also led to a larger reduction in root hydraulic conductivity in profligate than in conservative genotypes. In contrast, blockage of the apoplastic pathway in roots decreased transpiration more in conservative than in profligate genotypes. Interestingly, conservative genotypes had high early vigour, whereas profligate genotypes had low early vigour.
• In conclusion, profligate genotypes depend more on the cell‐to‐cell pathway, which might explain their higher root hydraulic conductivity, whereas water‐saving by restricting transpiration led to higher dependence on the apoplastic pathway. This opens the possibility to screen for conservative or profligate chickpea phenotypes using inhibitors, itself opening to the search of the genetic basis of these differences.

     

Differential responses of plasma membrane aquaporins in mediating water transport of cucumber seedlings under osmotic and salt stresses

It has long been recognized that inhibition of plant water transport by either osmotic stress or salinity is mediated by aquaporins (AQPs), but the function and regulation of AQPs are highly variable among distinct isoforms and across different species. In this study, cucumber seedlings were subjected to polyethylene glycol (PEG) or NaCl stress for duration of 2 h or 24 h. The 2 h treatment with PEG or NaCl had non‐significant effect on the expression of plasma membrane AQP (CsPIPs) in roots, indicating the decrease in hydraulic conductivity of roots (Lp_r) and root cells (Lp_rc) measured in these conditions were due to changes in AQP activity. After both 2 h and 24 h PEG or NaCl exposure, the decrease in hydraulic conductivity of leaves (K_leaf) and leaf cells (Lp_lc) could be attributed to a down‐regulation of the two most highly expressed isoforms, CsPIP1;2 and CsPIP2;4. In roots, both Lp_r and Lp_rc were further reduced after 24 h PEG exposure, but partially recovered after 24 h NaCl treatment, which were consistent with changes in the expression of CsPIP genes. Overall, the results demonstrated differential responses of CsPIPs in mediating water transport of cucumber seedlings, and the regulatory mechanisms differed according to applied stresses, stress durations and specific organs.     

Quantifying relationships between rooting traits and water uptake under drought in Mediterranean barley and durum wheat

In Mediterranean regions drought is the major factor limiting spring barley and durum wheat grain yields. This study aimed to compare spring barley and durum wheat root and shoot responses to drought and quantify relationships between root traits and water uptake under terminal drought.One spring barley（Hordeum vulgare L. cv. Rum） and two durum wheat Mediterranean cultivars（Triticum turgidum L. var durum cvs Hourani and Karim） were examined in soil‐column experiments under well watered and drought conditions. Root system architecture traits, water uptake, and plant growth were measured. Barley aerial biomass and grain yields were higher than for durum wheat cultivars in well watered conditions. Drought decreased grain yield more for barley（47%） than durum wheat（30%, Hourani）. Root‐to‐shoot dry matter ratio increased for durum wheat under drought but not for barley, and root weight increased for wheat in response todrought but decreased for barley. The critical root length density（RLD） and root volume density（RVD） for 90% available water capture for wheat were similar to（cv. Hourani） or lower than（cv. Karim） for barley depending on wheat cultivar. For both species, RVD accounted for a slightly higher proportion of phenotypic variation in water uptake under drought than RLD.     

Arbuscular mycorrhizal symbiosis and methyl jasmonate avoid the inhibition of root hydraulic conductivity caused by drought

Hormonal regulation and symbiotic relationships provide benefits for plants to overcome stress conditions. The aim of this study was to elucidate the effects of exogenous methyl jasmonate (MeJA) application on root hydraulic conductivity (L) of Phaseolus vulgaris plants which established arbuscular mycorrhizal (AM) symbiosis under two water regimes (well-watered and drought conditions). The variation in endogenous contents of several hormones (MeJA, JA, abscisic acid (ABA), indol-3-acetic acid (IAA), salicylic acid (SA)) and the changes in aquaporin gene expression, protein abundance and phosphorylation state were analyzed. AM symbiosis decreased L under well-watered conditions, which was partially reverted by the MeJA treatment, apparently by a drop in root IAA contents. Also, AM symbiosis and MeJA prevented inhibition of L under drought conditions, most probably by a reduction in root SA contents. Additionally, the gene expression of two fungal aquaporins was upregulated under drought conditions, independently of the MeJA treatment. Plant aquaporin gene expression could not explain the behaviour of L. Conversely, evidence was found for the control of L by phosphorylation of aquaporins. Hence, MeJA addition modified the response of L to both AM symbiosis and drought, presumably by regulating the root contents of IAA and SA and the phosphorylation state of aquaporins.     

Transgenic <Emphasis Type="Italic">Arabidopsis</Emphasis> and tobacco plants overexpressing an aquaporin respond differently to various abiotic stresses

Despite the high isoform multiplicity of aquaporins in plants, with 35 homologues including 13 plasma membrane intrinsic proteins (PIPs) in Arabidosis thaliana, the individual and integrated functions of aquaporins under various physiological conditions remain unclear. To better understand aquaporin functions in plants under various stress conditions, we examined transgenic Arabidopsis and tobacco plants that constitutively overexpress Arabidopsis PIP1;4 or PIP2;5 under various abiotic stress conditions. No significant differences in growth rates and water transport were found between the transgenic and wild-type plants when grown under favorable growth conditions. The transgenic plants overexpressing PIP1;4 or PIP2;5 displayed a rapid water loss under dehydration stress, which resulted in retarded germination and seedling growth under drought stress. In contrast, the transgenic plants overexpressing PIP1;4 or PIP2;5 showed enhanced water flow and facilitated germination under cold stress. The expression of several PIPs was noticeably affected by the overexpression of PIP1;4 or PIP2;5 in Arabidopsis under dehydration stress, suggesting that the expression of one aquaporin isoform influences the expression levels of other aquaporins under stress conditions. Taken together, our results demonstrate that overexpression of an aquaporin affects the expression of endogenous aquaporin genes and thereby impacts on seed germination, seedling growth, and stress responses of the plants under various stress conditions. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Effects of different arbuscular mycorrhizal fungal backgrounds and soils on olive plants growth and water relation properties under well‐watered and drought conditions

The adaptation capacity of olive trees to different environments is well recognized. However, the presence of microorganisms in the soil is also a key factor in the response of these trees to drought. The objective of the present study was to elucidate the effects of different arbuscular mycorrhizal (AM) fungi coming from diverse soils on olive plant growth and water relations. Olive plants were inoculated with native AM fungal populations from two contrasting environments, that is, semi‐arid – Freila (FL) and humid – Grazalema (GZ) regions, and subjected to drought stress. Results showed that plants grew better on GZ soil inoculated with GZ fungi, indicating a preference of AM fungi for their corresponding soil. Furthermore, under these conditions, the highest AM fungal diversity was found. However, the highest root hydraulic conductivity (Lp_r) value was achieved by plants inoculated with GZ fungi and growing in FL soil under drought conditions. So, this AM inoculum also functioned in soils from different origins. Nine novel aquaporin genes were also cloned from olive roots. Diverse correlation and association values were found among different aquaporin expressions and abundances and Lp_r, indicating how the interaction of different aquaporins may render diverse Lp_r values.     

水分亏缺下紫花苜蓿和高粱根系水力学导度 与水分利用效率的关系

利用聚乙二醇(PEG-6000)模拟水分亏缺条件(胁迫水势-0.2MPa,胁迫48h),研究了变水条件下紫花苜蓿(品种:阿尔冈金和陇东)和高粱(品种:抗四)根系水力学导度(Lp_r)、根系活力、根叶相对含水量、水分利用效率等参数的动态变化,以期进一步明确植物水分吸收及散失过程调控的生理生态学基础。结果表明:水分亏缺限制了紫花苜蓿和高粱根系吸水,表现在Lp_r的下降和根系活力的降低;继而调控了其地上部反应,引起气孔导度、光合速率、叶片相对含水量和蒸腾速率等的下降,但限制性的提高了其水分利用效率,尤其在胁迫初期。恢复到正常供水条件后,Lp_r、根系活性、气孔导度等水分利用参数逐渐部分或完全恢复到了胁迫前水平,但恢复程度存在种间和品种间差异,并且根系吸水能力的恢复对于是植株地上部生长状态的恢复至关重要,尤其是水分恢复初期。紫花苜蓿根系中检测到水通道蛋白(AQPs)的存在,水分亏缺对紫花苜蓿Lp_r的影响认为主要是通过影响AQPs的活性实现的。比较紫花苜蓿和高粱水分吸收与利用状况在变水条件下的动态变化,认为紫花苜蓿幼苗对干旱逆境的适应能力相对弱于高粱,品种间陇东适应能力更强。     

Influence of water stress on the glucosinolate profile of Brassica oleracea var. italica and the performance of Brevicoryne brassicae and Myzus persicae

Drought stress alters the chemical composition of plants, which can influence their tolerance to insect herbivory. To evaluate plant chemical responses to drought stress, broccoli, Brassica oleracea L. var. italica Plenck (Brassicaceae), was grown under well‐watered, drought, and water‐logged conditions. The glucosinolate (GS) levels and the performance of two aphid species, the specialist Brevicoryne brassicae (L.) and the generalist Myzus persicae (Sulzer) (both Hemiptera: Aphididae), in relation to water stress conditions were studied. High Performance Liquid Chromatography analysis showed that water stress changed the levels of GS in broccoli plants. Plants grown for 2 weeks under drought stress were significantly smaller and showed decreased levels of total GS when compared with GS contents of well‐watered plants, whereas water‐logged conditions led to a slight increase in the GS contents. A substantial decrease in indolyl GS was detected in water‐deficient plants, whereas aliphatic GS decreased slightly. Analysis of sugar levels in phloem sap of broccoli plants revealed that plants under water‐logged conditions contained the highest amounts of sugars followed by drought‐stressed and well‐watered plants. The two aphid species responded differently to water stress‐induced changes in their host plants. Significantly larger populations of M. persicae were recorded on plants with a limited water supply than on plants grown under well‐watered or water‐logged conditions. Brevicoryne brassicae was less affected by water stress, and similar population sizes were found on plants that were subject to different treatments. Analysis of covariance showed a significant effect of the plants’ water condition but no significant effect of GS content on the performance of M. persicae. However, the specialist B. brassicae remained unaffected by changes induced under water stress conditions.     

Root hydraulic conductivity and whole-plant water balance in tropical saplings following a shade-to-sun transfer

We hypothesized that pioneer and late successional species show different morphological and physiological responses in water use after gap formation. The magnitude of the responses was compared between two pioneer species (Macaranga gigantea and Trema orientalis) and four late successional species (Shorea sp.), in an experiment in which saplings were transferred from shade to sun. Although transpiration demand increased following the transfer, root hydraulic conductivity (Lp_r) decreased. Lp_r was sensitive to brief treatments with HgCl₂ (a specific inhibitor of aquaporins). This allows Lp_r to be divided into two components: cell-to-cell and apoplastic pathways. The Lp_r of cell-to-cell pathway decreased in all species following the transfer, relating to aquaporin depression in roots. Following the transfer, leaf osmotic potentials at full hydration decreased and both leaf mass per area [leaf mass/leaf area (LMA)] and fine-root surface area/leaf surface area (root SA/leaf SA) increased in almost all species, allowing saplings to compensate for the decrease in Lp_r. Physiologically, pioneer species showed larger decreases in Lp_r and more effective osmotic adjustment than late successional species, and morphologically, pioneer species showed larger increases in root SA/leaf SA and LMA. Water balance at the whole-plant level should be regulated by coupled responses between the aboveground and the belowground parts. Interspecific differences in responses after gap formation suggest niche differentiation in water use between pioneer and late successional species in accordance with canopy-gap size.     

Aquaporin regulation in roots controls plant hydraulic conductance,stomatal conductance,and leaf water potential in Pinus radiata under water stress

Stomatal regulation is crucial for forest species performance and survival on drought‐prone sites. We investigated the regulation of root and shoot hydraulics in three Pinus radiata clones exposed to drought stress and its coordination with stomatal conductance (g_s) and leaf water potential (Ψ_leaf). All clones experienced a substantial decrease in root‐specific root hydraulic conductance (K_root‐r) in response to the water stress, but leaf‐specific shoot hydraulic conductance (K_shoot‐l) did not change in any of the clones. The reduction in K_root‐r caused a decrease in leaf‐specific whole‐plant hydraulic conductance (K_plant‐l). Among clones, the larger the decrease in K_plant‐l, the more stomata closed in response to drought. Rewatering resulted in a quick recovery of K_root‐r and g_s. Our results demonstrated that the reduction in K_plant‐l, attributed to a down regulation of aquaporin activity in roots, was linked to the isohydric stomatal behaviour, resulting in a nearly constant Ψ_leaf as water stress started. We concluded that higher K_plant‐l is associated with water stress resistance by sustaining a less negative Ψ_leaf and delaying stomatal closure.     

Aquaporin expression in response to different water stress intensities and recovery in Richter-110 (<Emphasis Type="Italic">Vitis</Emphasis> sp.): relationship with ecophysiological status

Aquaporins seem essential for the regulation of plant water status and expenses. Richter-110 is a Vitis hybrid (Vitis berlandieri × rupestris) reputed to be strongly drought-tolerant. Three irrigation treatments were established in Richter-110 plants growing outdoors defined by the resulting maximum stomatal conductance (g _s), and ensuring water stress situations not severe enough as to stop photosynthesis and growth: well-watered plants (g _s about 250 mmol H₂O m⁻² s⁻¹), moderate water stress (g _s about 150 mmol H₂O m⁻² s⁻¹) and severe water stress (g _s about 50 mmol H₂O m⁻² s⁻¹). Plants under water stress were kept at constant water availability for 7 days to check for possible acclimation. Finally, plants were re-watered, and allowed to recover, for 3 days. Stomatal conductance, leaf water potential, xylem abscisic acid (ABA) content and root and stem hydraulic conductivity were determined. The relative amounts of expression of mRNA encoding seven putative aquaporins were determined in roots and leaves by RT-PCR. The decrease in stomatal conductance with moderate and severe water stress was associated with increasing ABA contents, but not with the leaf water potential and hydraulic conductivities, which remained unchanged during the entire experiment. Aquaporin gene expression varied depending on which aquaporin, water stress level and the plant organ. We suggest that aquaporin expression was responsive to water stress as part of the homeostasis, which resulted in constant leaf water potential and hydraulic conductivity.     

Phosphorus deprivation effects on water relations of <Emphasis Type="Italic">Nicotiana tabacum</Emphasis> plant via reducing plasma membrane permeability

Plants grown in phosphorus-deprived solutions often exhibit disruption of water transport due to reduction in root hydraulic conductivity (Lp_r). To uncover the relationship between root Lp_r and water permeability coefficient (P_f) of plasma membrane and the role of aquaporins, we evaluated P_f of plasma membrane and also PIP-type aquaporin gene expression in tobacco (Nicotiana tabacum L.) plant roots after seven days P-deprivation. The results showed significant reduction in sap flow rate (J_v) and osmotic root hydraulic conductivity (L_pr-o) in P-deprived roots. These effects were reversed 24 h after P-resupplying. Interestingly, the P_f of root protoplasts was 57% lower in P-deprived plants compared with P-sufficient ones. The expression of NtPIP1;1 and NtPIP2;1 aquaporins did not change significantly in P-deprived plants compared with P-sufficient ones, but the copy number of NtAQP1 increased significantly in P-deprived plants. P-deprivation did not change Lpr-o significantly in antisense NtAQP1 plants. Taken together, these findings suggest that P-deprivation may play an important role in modulation of root hydraulic conductivity by affecting P_f in transcellular pathway of water flow across roots and aquaporins. Finally, we concluded that dominant water transport pathway under P-deprivation was transcellular one.     

Reciprocal grafting separates the roles of the root and shoot in sex‐related drought responses in Populus cathayana males and females

Drought stress responses and sensitivity of dioecious plants, such as Populus cathayana Rehd., are determined by different mechanisms in each sex. In general, males tend to be more resistant while females are more sensitive. Here, we used reciprocal grafting between males and females to determine the relative importance of roots and shoots when plants are exposed to drought stress. Total dry matter accumulation (DMA), photosynthetic capacity, long‐term water‐use efficiency (Δ), water potential and ultrastructure of mesophyll cells were evaluated to determine the different roles of root and shoot in sex‐related drought responses. Plants with male roots were found to be more resistant and less sensitive to water stress than those with female roots under drought conditions. On the contrary, plants with female shoots grew better than those with male shoots under well‐watered conditions. These results indicated that the sensitivity of males and females to water stress is primarily influenced by root processes, while under well‐watered conditions sexual differences in growth are primarily driven by shoot processes. Furthermore, grafting female shoot scion onto male rootstock was proved to be an effective mean to improve resistance to water stress in P. cathayana females.     

Hydraulic properties of rice and the response of gas exchange to water stress

We investigated the role of xylem cavitation, plant hydraulic conductance, and root pressure in the response of rice (Oryza sativa) gas exchange to water stress. In the field (Philippines), the percentage loss of xylem conductivity (PLC) from cavitation exceeded 60% in leaves even in watered controls. The PLC versus leaf water potential relationship indicated diurnal refilling of cavitated xylem. The leaf water potential causing 50 PLC (P(50)) was -1.6 MPa and did not differ between upland versus lowland rice varieties. Greenhouse-grown varieties (Utah) were more resistant to cavitation with a 50 PLC of -1.9 MPa but also showed no difference between varieties. Six-day droughts caused concomitant reductions in leaf-specific photosynthetic rate, leaf diffusive conductance, and soil-leaf hydraulic conductance that were associated with cavitation-inducing water potentials and the disappearance of nightly root pressure. The return of root pressure after drought was associated with the complete recovery of leaf diffusive conductance, leaf-specific photosynthetic rate, and soil-leaf hydraulic conductance. Root pressure after the 6-d drought (61.2 +/- 8.8 kPa) was stimulated 7-fold compared with well-watered plants before drought (8.5 +/- 3.8 kPa). The results indicate: (a) that xylem cavitation plays a major role in the reduction of plant hydraulic conductance during drought, and (b) that rice can readily reverse cavitation, possibly aided by nocturnal root pressure.     

Review article. New aspects of plant aquaporin regulation and specificity

Recent findings concerning the sensitivity of aquaporins to heavy metal ions and their possible consequences to plant physiology are discussed. Furthermore, results obtained from heterologous expression of plant aquaporin cRNAs in Xenopus oocytes are related to the significance of water flux regulation by modification of the aquaporin protein or change of gene activity is given and new data concerning the selectivity of plasma membrane aquaporins are presented.Keywords: Aquaporin, regulation, specificity, sensitivity to heavy metal ions.      

Over-expression of a barley aquaporin increased the shoot/root ratio and raised salt sensitivity in transgenic rice plants

Barley HvPIP2;1 is a plasma membrane aquaporin and its expression was down-regulated after salt stress in barley [Katsuhara et al. (2002) Plant Cell Physiol. 43: 885]. We produced and analyzed transgenic rice plants over-expressing barley HvPIP2;1 in the present study. Over-expression of HvPIP2;1 increased (1) radial hydraulic conductivity of roots (Lp(r)) to 140%, and (2) the mass ratio of shoot to root up to 150%. In these transgenic rice plants under salt stress of 100 mM NaCl, growth reduction was greater than in non-transgenic plants. A decrease in shoot water content (from 79% to 61%) and reduction of root mass or shoot mass (both less than 40% of non-stressed plants) were observed in transgenic plants under salt stress for 2 weeks. These results indicated that over-expression of HvPIP2;1 makes rice plants sensitive to 100 mM NaCl. The possible involvement of aquaporins in salt tolerance is discussed.