Osmotic regulation of root system architecture

Although root system architecture is known to be highly plastic and strongly affected by environmental conditions, we have little understanding of the underlying mechanisms controlling root system development. Here we demonstrate that the formation of a lateral root from a lateral root primordium is repressed as water availability is reduced. This osmotic-responsive regulatory mechanism requires abscisic acid (ABA) and a newly identified gene, LRD2. Mutant analysis also revealed interactions of ABA and LRD2 with auxin signaling. Surprisingly, further examination revealed that both ABA and LRD2 control root system architecture even in the absence of osmotic stress. This suggests that the same molecules that mediate responses to environmental cues can also be regulators of intrinsic developmental programs in the root system.     

植物激素ABA调控植物根系生长的研究进展

脱落酸（abscisic acid,ABA）是一种重要的植物激素,在调控种子发育、种子休眠与萌发、抑制生长、促进落花落果、参与植物应对外界环境胁迫等过程中发挥着重要的生理功能。ABA还能与其他植物激素（如生长素、乙烯等）互作进而精细调控植物根系的生长。本文以模式植物拟南芥（Arabidopsis thaliana（L.）Heynh）为主要对象,对近年来国内外在ABA调控植物根系生长方面的研究成果、ABA与其他植物激素（如GA等）互作调控根系生长及调控非生物逆境下根系发育的机理等进行综述,并对其未来的研究方向进行了展望。     

The soybean root‐specific protein kinase GmWNK1 regulates stress‐responsive ABA signaling on the root system architecture

In humans, members of the WNK protein kinase family are osmosensitive regulators of cell volume homeostasis and epithelial ion transport, and mutation of these proteins causes a rare inherited form of hypertension due to increased renal NaCl re‐absorption. A related class of kinases was recently discovered in plants, but their functions are largely unknown. We have identified a root‐specific WNK kinase homolog, GmWNK1, in soybean (Glycine max). GmWNK1 expression was detected in the root, specifically in root cells associated with lateral root formation, and was down‐regulated by abscisic acid (ABA), as well as by mannitol, sucrose, polyethylene glycol and NaCl. In vitro and in vivo experiments showed that GmWNK1 interacts with another soybean protein, GmCYP707A1, which is a key ABA 8′‐hydroxylase that functions in ABA catabolism. Furthermore, 35S‐GmWNK1 transgenic soybean plants had reduced lateral root number and length compared with wild‐type, suggesting a role of GmWNK1 in the regulation of root system architecture. We propose that GmWNK1 functions to fine‐tune ABA‐dependent ABA homeostasis, thereby mediating the regulation of the root system architecture by ABA and osmotic signals. The study has revealed a new function of a plant WNK1 gene from the important staple crop soybean, and has identified a new component of a regulatory pathway that is involved not only in ABA signaling, but also in the repression of lateral root formation by an ABA‐dependent mechanism distinct from known ABA signaling pathways.     

ABA抑制花生侧根发生

本实验研究了ABA对花生侧根发生的影响。结果表明：用10umol·L-1 ABA浸泡处理花生种子1h或在含ABA的培养基上培养,均抑制侧根的发生,侧根发生率降低,数目减少,长度降低,发生的时间推迟1-2d;用ABA合成抑制剂25umol·L-1 NAPR浸泡后的种子,无论在1／2MS还是在含NAA培养基上培养,侧根发生率、侧根的数目和长度均增加。用NAA的极性运输抑制剂10umol·L-1 TIBA浸泡处理种子后,再在含ABA培养基上培养,侧根不发生,说明ABA抑制花生侧根的发生与种子内源ABA和IAA的水平相关。     

Shoot‐derived abscisic acid promotes root growth

The phytohormone abscisic acid (ABA) plays a major role in regulating root growth. Most work to date has investigated the influence of root‐sourced ABA on root growth during water stress. Here, we tested whether foliage‐derived ABA could be transported to the roots, and whether this foliage‐derived ABA had an influence on root growth under well‐watered conditions. Using both application studies of deuterium‐labelled ABA and reciprocal grafting between wild‐type and ABA‐biosynthetic mutant plants, we show that both ABA levels in the roots and root growth in representative angiosperms are controlled by ABA synthesized in the leaves rather than sourced from the roots. Foliage‐derived ABA was found to promote root growth relative to shoot growth but to inhibit the development of lateral roots. Increased root auxin (IAA) levels in plants with ABA‐deficient scions suggest that foliage‐derived ABA inhibits root growth through the root growth‐inhibitor IAA. These results highlight the physiological and morphological importance, beyond the control of stomata, of foliage‐derived ABA. The use of foliar ABA as a signal for root growth has important implications for regulating root to shoot growth under normal conditions and suggests that leaf rather than root hydration is the main signal for regulating plant responses to moisture.     

Abscisic acid rescues the root meristem defects of the Medicago truncatula latd mutant

The LATD gene of the model legume, Medicago truncatula, is required for the normal function of three meristems, i.e. the primary root, lateral roots and nitrogen-fixing nodules. In latd mutants, primary root growth eventually arrests, resulting in a disorganized root tip lacking a presumptive meristem and root cap columella cells. Lateral root organs are more severely affected; latd lateral roots and nodules arrest immediately after emerging from the primary root, and reveal a lack of organization. Here we show that the plant hormone, abscisic acid (ABA), can rescue the latd root, but not nodule, meristem defects. Growth on ABA is sufficient to restore formation of small, cytoplasm-rich cells in the presumptive meristem region, rescue meristem organization and root growth and formation of root cap columella cells. In contrast, inhibition of ethylene synthesis or signaling fails to restore latd primary root growth. We find that latd mutants have normal levels of ABA, but exhibit reduced sensitivity to the hormone in two other ABA-dependent processes: seed germination and stomatal closure. Together, these observations demonstrate that the latd mutant is defective in the ABA response and indicate a role for LATD-dependent ABA signaling in M. truncatula root meristem function.     

Long-distance signalling of abscisic acid (ABA): the factors regulating the intensity of the ABA signal

Abscisic acid (ABA) is a stress signal, which moves in the xylem from the roots to the aerial parts of the plant, where it regulates stomatal movement and the activity of shoot meristems. Root growth-promoting microorganisms in the rhizosphere, lateral ABA flows in the root cortex across apoplastic barriers, ABA redistribution in the stem, leaf apoplastic pH values, and the action of beta-glucosidases, both in the apoplast and the cytosol of the mesophyll, play an important role in the regulation of signal intensity. The significance of abscisic acid glucose ester as a long-distance stress signal is discussed.     

Response of root branching to abscisic acid is correlated with nodule formation both in legumes and nonlegumes

Legumes are unique among higher plants in forming a symbiosis with Rhizobium. Phylogenetic studies indicate this symbiosis may have evolved as many as three times within the Fabaceae; alternatively, a predisposition for nodulation evolved early in the history of the legume lineage. We have identified a physiological trait-increased lateral root formation in response to abscisic acid (ABA)- that marks all nodulating and non-nodulating legume species in our study set with the exception of Chamaecrista fasciculata and Cercis occidentalis. In contrast, nonlegume species tested decrease lateral root formation in response to ABA. Cercis is not a descendant of any common ancestor hypothesized to have evolved Rhizobium nodulation and has an intermediate response to ABA, partway between that of nonlegumes and legumes. We suggest that acquisition of altered responsiveness of roots to ABA is coincident with the appearance of a predisposition for nodulation within the legumes, followed by a loss in Chamaecrista. In addition, we demonstrate that altered ABA responsiveness of lateral root formation characterizes roots of the actinorhizal nodulator, Casuarina glauca, but not the closely related, nonactinorhizal species, Betula papyrifera. Thus our data provide evidence for a physiological root trait associated with nodulation both in legumes and in an actinorhizal plant.     

A late embryogenesis abundant protein HVA1 regulated by an inducible promoter enhances root growth and abiotic stress tolerance in rice without yield penalty

Regulation of root architecture is essential for maintaining plant growth under adverse environment. A synthetic abscisic acid (ABA)/stress‐inducible promoter was designed to control the expression of a late embryogenesis abundant protein (HVA1) in transgenic rice. The background of HVA1 is low but highly inducible by ABA, salt, dehydration and cold. HVA1 was highly accumulated in root apical meristem (RAM) and lateral root primordia (LRP) after ABA/stress treatments, leading to enhanced root system expansion. Water‐use efficiency (WUE) and biomass also increased in transgenic rice, likely due to the maintenance of normal cell functions and metabolic activities conferred by HVA1 which is capable of stabilizing proteins, under osmotic stress. HVA1 promotes lateral root (LR) initiation, elongation and emergence and primary root (PR) elongation via an auxin‐dependent process, particularly by intensifying asymmetrical accumulation of auxin in LRP founder cells and RAM, even under ABA/stress‐suppressive conditions. We demonstrate a successful application of an inducible promoter in regulating the spatial and temporal expression of HVA1 for improving root architecture and multiple stress tolerance without yield penalty.     

An abscisic acid-sensitive checkpoint in lateral root development of Arabidopsis

Lateral root (LR) formation displays considerable plasticity in response to developmental and environmental signals. The mechanism whereby plants incorporate diverse regulatory signals into the developmental programme of LRs remains to be elucidated. Current concepts of lateral root regulation focus on the role of auxin. In this study, we show that another plant hormone, abscisic acid (ABA), also plays a critical role in the regulation of this post-embryonic developmental event. In the presence of exogenous ABA, LR development is inhibited. This occurs at a specific developmental stage, i.e. immediately after the emergence of the LR primordium (LRP) from the parent root and prior to the activation of the LR meristem, and is reversible. Interestingly, this inhibition requires 10-fold less ABA than the inhibition of seed germination and is only slightly reduced in characterised abi mutants, suggesting that it may involve novel ABA signalling mechanisms. We also present several lines of evidence to support the conclusion that the ABA-induced lateral root inhibition is mediated by an auxin-independent pathway. First, the inhibition could not be rescued by either exogenous auxin application or elevated auxin synthesis. Secondly, a mutation in the ALF3 gene, which is believed to encode an important component in the auxin-dependent regulatory pathway for the post-emergence LR development, does not affect the sensitivity of LRs to ABA. Thirdly, ABA and the alf3-1 mutation do not act at the same developmental point. To summarise, these results demonstrate a novel ABA-sensitive, auxin-independent checkpoint for lateral root development in Arabidopsis at the post-emergence stage. In addition, we also present data indicating that regulation of this developmental checkpoint may require novel ABA signalling mechanisms and that ABA suppresses auxin response in the LRPs.     

Arabidopsis ROP‐interactive CRIB motif‐containing protein 1 (RIC1) positively regulates auxin signalling and negatively regulates abscisic acid (ABA) signalling during root development

Auxin and abscisic acid (ABA) modulate numerous aspects of plant development together, mostly in opposite directions, suggesting that extensive crosstalk occurs between the signalling pathways of the two hormones. However, little is known about the nature of this crosstalk. We demonstrate that ROP‐interactive CRIB motif‐containing protein 1 (RIC1) is involved in the interaction between auxin‐ and ABA‐regulated root growth and lateral root formation. RIC1 expression is highly induced by both hormones, and expressed in the roots of young seedlings. Whereas auxin‐responsive gene induction and the effect of auxin on root growth and lateral root formation were suppressed in the ric1 knockout, ABA‐responsive gene induction and the effect of ABA on seed germination, root growth and lateral root formation were potentiated. Thus, RIC1 positively regulates auxin responses, but negatively regulates ABA responses. Together, our results suggest that RIC1 is a component of the intricate signalling network that underlies auxin and ABA crosstalk.     

Abscisic acid promotes pre-emergence stages of lateral root development in Medicago truncatula

The plant root system is important for plant anchorage and nutrition. Among the different characteristics of the root system, root branching is a major factor of plasticity and adaptation to changing environments. Indeed, many biotic and abiotic stresses, such as drought or symbiotic interactions, influence root branching. Many studies concerning root development and root branching were performed on the model plant Arabidopsis thaliana, but this model plant has a very simplified root structure and is not able to establish any symbiotic interactions. We have recently described 7 stages for lateral root development in the model legume Medicago truncatula and found significant differences in the tissular contribution of root cell layers to the formation of new lateral roots (LR). We have also described 2 transgenic lines expressing the DR5:GUS and DR5:VENUS-N7 reporter genes that are useful to follow LR formation at early developmental stages. Here, we describe the use of these transgenic lines to monitor LR developmental responses of M. truncatula to the phytohormone abscisic acid (ABA) which is a major actor of stress and symbiotic interactions. We show that ABA promotes the formation of new lateral root primordia and their development, mostly at the late, pre-emergence stage.     

Dynamic analysis of ABA accumulation in relation to the rate of ABA catabolism in maize tissues under water deficit

The plant hormone abscisic acid (ABA) accumulates in plant tissues which experience water deficit (stress ABA). This study analysed its accumulation as a function of both synthesis and catabolism in maize tissues. By following the disappearance of the stress ABA when ABA synthesis was blocked by nordihydroguaiaretic acid (NDGA), the rate of the catabolism of stress ABA was determined. When compared with the catabolic rate of baseline (non-stress) ABA, stress ABA showed a catabolic rate >11 times higher. With such an elevated catabolic rate, it is proposed that the xanthophyll precursor pool may not be able to sustain the ABA accumulation, and such a proposition has been substantiated by further experiments where fluridone is used to limit the availability of upstream ABA precursors. When fluridone was used, stress ABA accumulation could only be sustained for a few hours, i.e. approximately 5 h for leaf and 1 h for root tissues. In detached roots, stress ABA accumulation could not be sustained even if fluridone was not used, suggesting that stress ABA accumulation in root systems requires the continuous import of ABA precursors from the shoots. Such an assumption was substantiated by the observation that defoliation or shading significantly reduced ABA accumulation in intact roots. The present study suggests that ABA catabolism is rapid enough to play an important role in the regulation of ABA accumulation.     

Control of root architecture and nodulation by the LATD/NIP transporter

The Medicago truncatula LATD/NIP gene is essential for the development of lateral and primary root and nitrogen-fixing nodule meristems as well as for rhizobial invasion of nodules. LATD/NIP encodes a member of the NRT1(PTR1) nitrate and di-and tri-peptide transporter family, suggesting that its function is to transport one of these or another compound(s). Because latd/nip mutants can have their lateral and primary root defects rescued by ABA, ABA is a potential substrate for transport. LATD/NIP expression in the root meristem was demonstrated to be regulated by auxin, cytokinin and abscisic acid, but not by nitrate. LATD/NIP''s potential function and its role in coordinating root architecture and nodule formation are discussed.Key words: nodule development, lateral root development, root architecture, symbiotic nitrogen fixation, Medicago truncatula, NRT1(PTR) gene familyUnlike most other plants, legumes form two kinds of lateral root organs: lateral roots and nitrogen-fixing root nodules that form in conjunction with compatible symbiotic rhizobium bacteria. Although the morphology and function of these two root organs is distinct, both require the function of the LATD/NIP gene, indicating shared genetic components for these two developmental processes and providing support for a model in which legume nodules evolved from a lateral root blueprint. Both lateral roots and nodules initiate in previously differentiated root cells in response to environmental and developmental cues mediated by hormones. Interestingly, regulation of nodules and lateral roots by hormones is often opposite, allowing formation of one organ or another depending on the conditions.     

The Method of ABA Application Affects Salt Stress Responses in Resistant and Sensitive Potato Lines

The phytohormone abscisic acid (ABA) has been proposed to act as a mediator in plant responses to a range of stresses, including salt stress. Most studies of ABA response apply ABA as a single dose. This may not resemble the prolonged increasing endogenous ABA levels that can occur in association with slowly increasing salinity stresses in nature or field situations. Salt stress response based on method of ABA application was examined in four potato genotypes of varying salt stress resistance: the sensitive ABA-deficient mutant and its normal sibling, a resistant genotype line 9506, and commercial cultivar ‘Norland’ of moderate resistance. ABA was applied by root drench at 0, 50, 75, or 100 μM concentrations through a single dose, or by slowly increasing multiple ABA doses in a sand-based growing system under greenhouse conditions. Salt tolerance was then evaluated after 2 weeks of exposure to 150–180 mM NaCl stress. The method of ABA application had a marked effect on the responses to salt stress. Plant responses to the method of ABA application were differentiated according to (1) growth rate, (2) root water content, and (3) apparent shoot growth response. Under a single dose, growth rate increased in all genotypes under salt stress, whereas slowly increasing multiple ABA applications generally maintained stable growth rates except in the ABA-deficient mutant where there was an upward growth trend. Percent root water content was elevated only under slowly increasing multiple ABA doses in two genotypes, whereas none of the single-dose treatments induced any change. The single ABA dose enhanced vertical growth, whereas the slowly increasing multiple ABA dose applications enhanced lateral shoot growth. Because exogenous application is still an artificial system, endogenous ABA was supplied through grafting of ABA-deficient mutant scions onto rootstocks with known elevated ABA levels. Multiple exogenous ABA applications as low as 50 μM elicited similar shoot water content responses as grafting treatments without ABA application in the mutant genotype but had no effect on the ABA normal sibling. Shoot dry weight was significantly increased through grafting over all exogenous ABA treatments. Our study further indicates that the method of ABA application regime in itself can alter plant responses under salt stress and that certain application regimes may reflect responses to elevated endogenous levels of ABA.     

TTG1-mediated flavonols biosynthesis alleviates root growth inhibition in response to ABA

Key message

Our results demonstrate that the flavonoids biosynthetic pathway can be effectively manipulated to confer enhanced plant root growth under water-stress conditions.

Abstract

Abscisic acid (ABA) is one of most important phytohormones. It functions in various processes during the plant lifecycle. Previous studies indicate that ABA has a negative effect on root growth and branching. Auxin is another key plant growth regulator that plays an essential role in plant growth and development. In contrast to ABA, auxin is a positive regulator of root growth and development at low concentrations. This study was performed to help understand whether flavonoids can suppress the effect of ABA on lateral root growth. The recessive TRANSPARENT TESTA GLABRA 1 (ttg1) mutant was characterized on ABA and sucrose treatments. It was determined that auxin mobilization could be altered by modifying flavonoids biosynthesis, which resulted in alterations of root architecture in response to ABA treatment. Moreover, transgenic TTG1-overexpression (TTG1-OX) seedlings exhibited enhanced root length and lateral root number compared to wild-type seedlings grown under normal or stress conditions. Genetic manipulation of the flavonoids biosynthetic pathway could therefore be employed successfully for the improvement of plant root systems by overcoming the inhibition of ABA and some abiotic stresses.