reviews

Hormones from roots as signals for the shoots of stressed plants

Michael Jackset
Intracellular signalling processes that translate hormone perception into genetic, metabolic and developmental change are the focus of much current plant hormone research. By characterizing these processes it is hoped to establish an understanding of the regulated and integrated behaviour of whole plants at the molecular level. However, for a complete picture, the extracellular signalling processes that coordinate the behaviour and development of the various plant organs must also be uncovered. Roots may use hormones, or their precursors, to provide shoots with early warning of deteriorating soil conditions in ways that increase resilience to stress. Recent progress springs from integrated studies involving sensitive and specific methods of hormone analysis, plant water relations and the use of genetically transformed plants.

he idea that xylem sap flowing into the shoot system from the roots carries not only water and mineral nutrients but also physiologically active substances (hormones) that help to relate root and shoot behaviour has attracted physiologists for many years 1. The concept is bound up with the classic view of hormones as compounds produced in one place and transported to another, where they influence development as part of the mechanism that coordinates the behaviour of differegt organs. Although this is too prescriptive as a definition for plant hormones, the idea is still actively pursued experimentally, such as in the study of flower initiation 2. Currently, there is much interest in how roots inform the shoot of deteriorating soil conditions and, in so doing, help the plant adjust (acclimate) to the altered and more taxing circumstances. Typical reactions to soil stresses - such as drought, flooding, mineral nutrient shortage, salinity or compaction include slower leaf expansion, closure of stomata, epinastic leaf curvature and senescence of older leaves. Treating plants with an appropriate hormone at low doses can often reproduce or overcome these effects, and all the major hormones have been detected in xylem sap. Consequently, these compounds, or their precursors, have received close attention as potential root to shoot signalling molecules. A faltering root system could signal a stress condition simply by a decrease in the supply of water or mineral nutrients. However, analyses using plants with split root systemsa, or placed in specialized pressure vessels4'5 to prevent water shortage in the shoots even when soil is drying or flooded, have shown that this is not necessarily the case. Instead, another chemical signalling mechanism is at work that gives early warning of deteriorating soil conditions and induces changes above ground that may increase stress tolerance. Convincing experimental evidence to support the notion of effective root to shoot signalling in stressed plants is not easily secured. This review summarizes the problems and identifies possible means of overcoming them. The discussion has two main concerns: The hormone abscisic acid (ABA) as a possible signal emanating from roots - ABA has been the focus of much recent research with droughted plants.

The way in which soil flooding promotes epinastic leaf growth by means of the ethylene precursor ACC (l-aminocyclopropane-l-carboxylic acid) as the signalling molecule this is the most convincing example of root to shoot communication involving a hormone. Four ways to signal The ways in which shoots may sense that roots are under stress can be placed into four categories6. The two discussed in this review involve the transport of messenger compounds dissolved in the water of xylem sap, and are termed according to whether they are exported to the shoots in larger ('positive' messages) or smaller ('negative' messages) amounts as the roots become stressed. Water is their solvent, and is either drawn up the plant by internal gradients of water potential generated by transpiration, or propelled more slowly at night, by radial osmotic gradients within the root arising from the active transport of ions into water-filled xylem elements. Other forces may also move water from roots to shoots7, although these remain controversial. There are two less direct means of signalling, by 'accumulation' and 'debit' messages. They are transported in the phloem and are 'mirror images' of xylem-borne positive or negative messages. Thus, when stress at the roots slows the export of phloem-mobile substances from leaves as a result of decreased sink strength, a build-up of a hormone (the accumulation message) can take place in the shoot. For example, foliar concentrations of ABA are increased temporarily by this means when tomato plants are stressed by soil flooding s. The hypothetical debit messages have not yet been examined experimentally. If they do exist, they would be generated when stressed roots increased their demand for phloem-mobile substances, thus depleting the shoot of substances that influence its development. Situations where debit messages are most likely to be found include when root growth is stimulated by shortage of soil nitrogen or by infection with the parasitic weed Striga hermonthica. Root to shoot signalling: a three phase process The model in Box 1 describes stress perception and message (hormone) generation in the roots (phase 1), transit of
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perception in the recipient shoot tissues, and transduc~on of the signal, resulting in actions by receptive cells (e.g. causing stomatal closure) that increase stress tolerance (acclimation).
Phase 1 Phase 2 ,. Phase 3 Shoot system
v

Ir

Root system Stress perception

v

Hormone content may become enriched or depleted as sap flows through the plants. Thus, deliveries or concentrations in sap taken from leaves may not reflect original output from roots below.
A

End of xylem transport system

Partitioning or transport out of xylem sap

v

Signal transduction

After analysis of sap flowing at the rate of whole plant transpiration, the delivery rate is estimated as:
Concentration Sap flow rate

Target cells
v

Altered gene expression and/or metabolism

v

Signal perception
v

Increase or decrease in hormone production

Because of dilution effects, changes in sap flow rate caused by stomatal closure can alter message concentration independently of delivery rate. Thus, concentration is a poor estimate of delivery rate.

Signal transduction

Partitioning or transport into xylem sap ' An increase or decrease in hormone delivery (flux) should be established before the existence of a positive or negative message can be deduced with confidence.

Altered gene expresston and/or metabolism

v

Developmental response

Positive or negative message?

t
Xylem sap transport system to shoot

the message in the xylem sap to targets in the shoot (phase 2), and message decoding in shoot tissue through a second set of perception and intracellular transduction processes (phase 3). These last signals can lead, among other responses, to stomatal closure, epinastic leaf curvature, or senescence by old leaves. The three phase system initiates message formation and redirects it from a localized stress site in the roots to remote unstressed sites in the shoot, where response characteristics to hormones are very different. Knowledge of how stresses and hormones cause sensitive cells to induce their effects in target cells remains rudimentary, but there have recently been exciting advances9'1.

Positive and negative messages in xylem sap Problems of ana/ysis An overriding practical difficulty arises because the xylem sap of transpiring plants is at negative hydrostatic

(subatmospheric) pressure, making direct sampling almost impossible. Indirect methods of sampling are needed, but all are problematic. Xylem sap extracted by pressurizing or centrifuging freshly excised shoot parts has often been used, but the method runs the risk of contaminating sap samples with solutes from the cell wall and symplasm. Furthermore, such samples are often taken from tissues excised some distance from the roots, which could result in the solute content of the sap being changed en route by radial transfer to or from surrounding tissues 11. An alternative approach has been to keep the plant intact and apply pneumatic pressure to the root the pressure is adjusted so that the hydrostatic pressure in the xylem is just above atmospheric. Sap issuing from a cut in the xylem of a cotyledon close to the shoot base is then sampled 12. The solute concentrations and flow rate of this sap are thought to be very close to those of authentic transpiration fluid 1~, although these assumptions have not been
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sap exuding osmotically from the stump of the remaining root (b) Effect on delivery of nitrate ( a ) Dilution of nitrate system. However, the first as sap flow increases as sap flow rate increases droplets can be contaminated with solutes from injured ceUs14. Also, estimates for solute concentrations in this slow-moving sap severely overestimate solute concentrations of faster-moving transpiration fluid, because solutes are ~'E 2 -5 diluted to a much smaller E v extent 6. This dilution effect is (D seen as a curvilinear dilution of z sap solutes when a root system is pressurized pneumatically in gradual steps to accelerate sap flow. The concentration most likely to represent that of xylem sap in intact plants is I I I I I I that given by analysis of sap 1 2 3 4 2 4 flowing at rates of whole plant Sap flow (ram3 s-1) Sap flow (mm a s -1) transpiration (Fig. la and lc). Arithmetic multiplication of solute concentration by sap ((::) Dilution of ACC flow rate generates a delivery (d) Effect on delivery of ACC as sap flow rate increases as sap flow increases rate for that solute. It is a change in the delivery rate that constitutes a chemical message. When a solute becomes more dilute in strict proportion to the 2.0 1.5 speed of sap flow, as shown for nitrate in Fig. la, the computed delivery rate at all or any one 1,5 sap flow rate will give similar delivery values (Fig. lb). These values will all be reasonable estimates of the delivery occur< 0.5 ring in intact plants. Unfortunately, the dilution of hor........ -0 o.5 mones, or their precursors, is often less than proportional to flow (see Fig. lc for an ACC I I I I I I dilution curve in flooded tomato 0 2 4 6 0 2 4 6 plants). This means that, in Sap flow (mm 3 s -1) Sap flow (mm 3 s -1) addition to an effect on concentration, delivery is also influFig. 1. The effect of changes in sap flow rate on solute concentration or delivery in xylem enced by the rate of sap flow sap flowing ~om freshly excised root systems of tomato plants. Sap flow rates were (see Fig. ld for an interaction adjusted by pressurizing the roots pneumatically. Arrows show the rates of transpiration of between flow rate and delivery similar whole plants: concentrations or delivery rates at these flow rates are estimates of of ACC). Thus, deliveries in the values in whole plants. In (a) and (c), the theoretical dilution is shown by the broken intact plants can only be estiline; the actual dilution is shown by the solid line. (a) Nitrate in the xylem sap of mated with the required preciunstressed plants dilutes strictly in proportion to the sap flow rate and thus delivery rates sion by using sap flowing at of nitrate (b) are unaffected by the rate of sap flow. (c) Dilution of ACC (1-aminecyclorates close to those of whole propane-l-carboxylic acid) in flooded plants is less than proportional to the flow rate, and plant transpiration (Fig. ld). consequently delivery is increased by a faster sap flow rate (d). Thus, for ACC, both concentration and delivery rates in flooded plants are only valid for whole plants when the sap The need for care is reinforced analyzed is made to flow at the rate of whole plant transpiration. by the fact that different solutes, and stressed and unstressed plants, behave diffully tested. Unfortunately, estimates of hormone delivery ferently12'15. Delivery calculations can usefully be refined to from roots to the shoot base using this promising method take account of the relative sizes of root and shoot, thereby have yet to be made on stressed plants. A more common generating specific delivery rates (e.g. nanomoles of ABA per approach has been to remove the shoot and analyze xylem square metre of leaf per second), which help interpretation

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where marked differences in size or root : shoot ratios exist between plants 1~. Solutes in the xylem sap of whole plants can also be diluted or concentrated by changes in transpiration rate imposed by environmental conditions and the pattern of stomatal opening. This is an important point, because stress can lead to stomatal closure and thus slow down sap flow. When this happens, an increase in solute concentration in xylem sap passing out of roots into the shoots is inevitable. Thus, no matter how carefully obtained, measurements of increased concentration in the xylem sap of stressed plants may merely be a consequence of stomatal closure. Such a concentration increase does not guarantee that delivery into the shoot is increased by the same amount, if at all, and thus does not demonstrate unequivocally the existence, or magnitude, of a positive message. Unfortunately, many studies have relied upon estimates of concentration to deduce changes in hormone output by roots. Such work runs the risk of mistaking consequence for cause, claiming the existence of a positive message that is not there, or exaggerating the magnitude of the message. and slower transpiration or their cause. Only where increases in ABA concentration are disproportionately greater than any associated slowing of transpiration can an increased ABA input to leaves be safely deduced from concentration data. Where ABA deliveries to leaves have been calculated in droughted plants, they have not necessarily shown a close relationship with stomatal closure, and sometimes relate more closely to changes in ABA concentration2~. This is what would be expected if an increase in ABA concentration in the xylem of shoots were a result and not a cause of stomatal closure. An alternative interpretation is that stomata monitor xylem sap ABA concentration, but not the delivery rate. There is evidence for both points of view 2~'24, and this requires further investigation. However, either way, stressed roots can only be directly responsible for raising the occupancy of ABA receptor sites if they generate a positive message (i.e. increase the mass of ABA supplied to the shoots via the transpiration stream). Much of the ABA from roots is metabolized in leaves23. It seems probable that a balance between leaf metabolism and delivery of the hormone from roots and xylem will determine the local concentration at receptor sites such as stomatal guard cells or expanding leaf cells. The rate of ABA degradation in the shoots may be under environmental control. For example, small decreases in leaf water potential are thought to slow ABA metabolism:5, thereby allowing more hormone from roots to 'survive' passage through shoot tissue to responsive cells.

Delivery results
Despite the problems, there is evidence that hormone output from roots into the shoot base can be altered by soil stresses. Roots undoubtedly synthesize increased amounts of ABA when droughted17 and increased quantities can be delivered into the shoot is, although unambiguous flux data are scarce. In Ricinus communis, a fivefold increase in the supply of ABA in xylem sap from drying roots has been demonstrated. This increase correlates with stomatal closure, and is evident even when precautions are taken to ensure that the ABA has not originated from waterdeficient shoots, and when arithmetic adjustments have been made for effects of soil drying on leaf area and root growth16. In contrast to drought, flooding the soil for 24 h decreases ABA delivery from roots of tomato plants up to eightfold15 (when the hormone has been measured by mass spectrometry). Thus, ABA as a positive message from roots cannot explain the stomatal closure or slow leaf expansion that are symptomatic of flooded plants. Evidence that other soil stresses such as mechanical impedance or physical constriction enhance ABA delivery is also unconvincing ~. The shortage of nitrogen at the roots can reduce delivery of transzeatin and trans-zeatin riboside in Urtica dioica ~9, while drought or soil flooding have the same effect in rice is, poplar and Phaseolus vulgaris 2. Failure to account for effects of sap flow rate has meant that the extent to which cytokinin delivery is depressed by root stresses has been underestimated in most cases. A lessening of cytokinin supply from roots may slow shoot growth and promote leaf senescence.

Do root-sourced messages reach their target? There are many reports of increases in ABA concentration in xylem sap taken from high in the canopy of droughted plants ~'21, while stomata are closing and leaves expanding more slowly than normal. If it is assumed that this ABA was carried from roots and did not enter the xylem sap as it passed through shoot tissue, this xylemborne message appears to be drawn close to potential target cells such as stomatal guard cells and expanding leaf cells. However, in interpreting these data, we are faced with the now familiar problem of deciding whether the increased concentrations are a result of stomatal closure

Are there antitranspirants other than abscisic acid in xylem sap? Experiments with reciprocal grafting of roots and shoots of wild-type and ABA-deficient tomato plants (cv. flacca) helped to rule out the possibility that ABA from roots is responsible for rapid stomatal closure in flooded plants, although ABA from the shoots is required 6. Instead, another as yet unidentified antitranspirant is transported from flooded roots in xylem sap s. Similar grafting experiments with ABA-deficient and wild-type tomato plants under drought stress also point to xylem-borne ABA as being less important than an unknown antitranspirant (N.M. Holbrook and and R. Munns, unpublished). This supports other sporadic reports that xylem sap contains something in addition to ABA that regulates stomatal apertures in droughted plants ls'2~. However, these findings must be set alongside observations that the use of antibodies to deplete xylem sap of ABA results in the loss of stomatalclosing activity27 and that feeding ABA to unstressed leaves at concentrations found in the xylem sap of droughted plants can close stomata2s. The validity of the latter test depends on the accuracy with which ABA in the xylem sap of whole plants is determined; this is not straightforward, and there is a strong risk of overestimating these concentrations. ACC as a positive message that promotes epinasty in flooded plants Flooding the soil with water deprives the roots of oxygen. In tomato plants, this leads to expansion of the adaxial cells of petioles, causing epinastic curvature within 6 h (Box 2b). Epinasty, together with stomatal closure, helps decrease transpiration rates by reducing the intensity and amount of incident radiation at a time when the foliage is liable to become partially dehydrated. This can occur because of a loss of synchrony between the daily rise and fall in root
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estimates for similar-sized plants 34. These improved methods also show that well-drained roots also export ACC (Ref. 35). Thus, the flooding effect is quantitative, and not the qualitative phenomenon it was first thought to be. When account is taken of the areas of leaf into which the ACC is distributed, 72 h flooding can increase this specific delivery rate from under 6 pmol m -2 s-1 to 22 pmol m -2 s-1 (Ref. 36) (Fig. 2).
Phase 3 - shoot responses

3.00

L
2.00 ._> 1.00 o 0 <
{D

..'"

"(~)

Leaf epinasty

Flooded plants

-..."

. . . 0 ......

...... O "

Well-drained plants
i i ,
, , i i

12 Time (h)

24

28

32

36

Fig. 2. Effect of flooding the soil for up to 36 h on the delivery of the ethylene precursor ACC (1-aminocyclopropane-l-carboxylicacid) from the roots in the xylem sap of tomato plants. The analyzed sap was made to flow from freshly decapitated plants at rates close to those of whole plant transpiration. This rate was slower for flooded plants (approximately 1 mm~ s-1) than for well-drained plants (approximately 3.5 mm3 s-l). The black bar on the x axis indicates night.

hydraulic conductivity and opening and closing of stomatal apertures 5, or as a result of root death. Root excision and split root experiments show that a positive message from oxygen-deficient roots is involved which is transported in the xylem and stimulates the formation of additional ethylene in the shoot~9. This extra ethylene is responsible for inducing epinasty. Characterization of the biosynthetic pathway for ethylene by Yang and associates 30 (Box 2a shows the last three steps) led to the message being identified as ACC, the immediate precursor of ethylene. Subsequent work has generated further information on this flooding system for each of the three phases of the signalling model (Box 2; Fig. 2).
Phase 1 - the root response to oxygen shortage

ACC oxidase, the enzyme catalysing ethylene production from ACC, is a nonheme Fe2-ascorbate dioxygenase that requires oxygen. Roots lacking oxygen accumulate ACC, because its synthesis continues anaerobically, at least as long as enough ATP is available for two earlier synthetic steps. Stress perception is thus a simple inhibition of activity of oxygen-requiring ACC oxidase. To this may be added the possibility of inducible expression, under anaerobic conditions, of one member of the family of genes encoding ACC synthase, an unstable pyridoxal phosphate enzyme that generates ACC from S-adenosyl-L-methionine (AdoMet) (Box 2a). This induction is known to occur in rice 31and may be more widely distributed. Studies of anaerobic gene expression in maize roots indicate a role for Ca 2in the signal transduction pathway that regulates transcription 32. Also, the entry of ACC into xylem sap is facilitated by increased solute permeability of root cells caused by oxygen deficiency33and by the entry of some ACC from soil water 1~.
Phase 2 - A C C transport to the shoots

The absolute requirement for ACC from roots to raise ethylene production in the leaves and promote epinastic curvature of flooded plants was demonstrated by the ability of ACC biosynthesis inhibitors (aminoethoxyvinylglycine and amino-oxyacetic acid) to suppress these responses 37. Thus, ACC is transported as a positive message from a site where oxidation, catalysed by the ACC oxidase enzyme, is impossible (anaerobic roots) to a point where the presence of atmospheric oxygen permits oxidation reactions (i.e. in the aerial shoot). However, the existing ACC oxidation capacity of the shoots is inadequate to handle the additional influx of ACC from the anaerobic roots~ Accordingly, the level of ACC oxidase activity in the leaves is raised within 6-12 h of flooding the soil36. This amplification of oxidation capacity raises maximum ethylene production rate and promotes epinastic curvature. This is clear from responses of transgenic tomato plants that contain an antisense construct to one of the three isforms of ACC oxidase present in tomato plants. In the transformed plants, a flooding-induced increase in ACC oxidase activity is suppressed. This, in turn, decreases how much ethylene is made by the leaves of flooded plants thus resulting in less epinastic curvature: The inhibitions occur even though ACC delivery from oxygen-deficient roots is undiminished 36. How oxygen-deficient roots promote ACC oxidase activity in leaves is unknown. It may be induced by the extra ACC or ethylene, because each is known to enhance transcription and activity of ACC oxidase in tomato. Alternatively, an as yet unidentified promoter of ACC oxidase activity or gene expression could be transported from oxygen-deficient roots as a positive message.
Future work

Using an indirect assay, ACC delivery was first shown to occur after 12 h flooding using sap drawn slowly from 'de-topped' tomato plants 34. By using sensitive and chemically specific mass spectrometry, and taking account of the interaction between ACC delivery and sap flow rate (Fig. lc), the effect has now been found to begin after only 6 h of soil flooding and that the peak rate of ACC delivery (2.9 pmol s-~) (Ref. 35) is considerably greater than earlier
26 January1997,Vol.2, No. 1

A theoretical and technical framework is now in place to establish with more certainty the impact of a variety of root stresses on hormone export in the transpiration stream and, in turn, the influences this has on shoot development. The picture is reasonably complete for ACC as a positive message from oxygen-deficient roots that promotes epinastic leaf curvature. There is also a possibility that ACC from drying roots is a positive message that passes to the shoots of citrus, and promotes leaf abscission3s. Evidence points to ABA as a positive message from drying roots that promotes closing of stomata and slows leaf expansion. There is also evidence to indicate that the output of cytokinins from roots is depressed by stress. However, uncertainties and gaps remain in our knowledge of these processes. For example, it is unclear whether putative hormonal messages originating from stressed roots actually arrive at putative target tissues and

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!ii:iiiiiil
(a)
Ethylene biosynthesis 02 + CO2 Methionine + ATP ,- S-adenosylmethionine ~- ACC ~ ' ' ' ~ = Ethylene

I Ac

I I AGO 1

flooded for 24 h, showing reflexed, epinastic leaves. Details

(C)

Phase 1 Stress perception

(e)

Phase 3 Signal transduction

Lack of oxygen in the root system favours ACC accumulation within 4 h as a result of: Arrest of ACC oxidation to ethylene. Increased transcription of the anaerobic ACC synthase gene?

Petioles enriched with ACC from anaerobic roots.

ACC oxidase gene transcription increased in shoots?

ACC oxidase activity increases in petioles. Increased solute permeability of anoxic cells, which promotes entry of ACC into xylem sap. ncreased oxidation of ACC message from anaerobic roots.

(d)

Phase 2 Message transport Ethylene biosynthesis increased sevenfold.

ACC delivery from the roots to the shoot base in the transpiration stream exceeds control rates (0.9 nmol h-1) within 4 h. Maximum delivery rate of 14.4 nmol h-~ reached after 48 h. Rates of delivery exceed those required to support known rates of ethylene biosynthesis in shoots. Unidentified message that promotes ACC oxidase activity (transcription?) in the shoots.

Ethylene promotes cell expansion, resulting in ep~nastic curvature of petioles.

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are not 'lost' during transport up the plant and into leaves. Similarly, more information is required on the hormonal messages that xylem sap may acquire as it passes through the shoot. Such messages merely 'masquerade' as having originated in the roots. Attempts to demonstrate that particular root-sourced hormones really do exert their influence on shoot behaviour as claimed will benefit from the use of mutants that are hormone deficient or hormone insensitive, especially where experiments involve reciprocal root-shoot grafting to insert or remove putative sources of hormonal signals from roots. The isolation of novel substances in xylem sap that close stomata in stressed plants or slow leaf and stem extension can be expected. The discovery of a rapid promotion of ACC oxidase in leaves by oxygen-deprived roots raises the possibility of the remote control of gene expression in shoots being exercised by signals from stressed roots that are transported in the transpiration stream. Xylem sap is far richer in organic compounds than generally realized, and the potential of these solutes for root to shoot communication is only now beginning to be appreciated.
References 1 Went, F.W. (1938) Specific factors other than auxin affecting growth and root formation, Plant Physiol. 13, 55-80 2 McDaniel, C.N. and Hartnett, L.t~ (1993) Floral-stimulus activity in tobacco stem pieces, Planta 189, 577-583 3 Gowing, D.J.G., Davies, W.J. and Jones, H.G. (1993) A positive signal as an indicator of soil drying in apple, Malus domestica Borkh., J. Exp. Bot. 41, 1535-1540 4 Passioura, J.B. (1988) Root signals control leaf expansion in wheat seedlings growing in drying soil, Aust. J. Plant Physiol. 17, 149-157 5 Else, M.A. et al. (1995) A negative hydraulic message from oxygendeficient roots of tomato plants? Influence of soil flooding on leaf water potential, leaf expansion, and the synchrony between stomatal conductance and root hydraulic conductivity, Plant Physiol. 109, 1017-1024 6 Jackson, M.B. (1993) Are plant hormones involved in root to shoot communication?Adv. Bot. Res. 19, 103-187 7 Canny, M.J. (1995) A new theory for the ascent of sap - cohesion supported by tissue pressure, Ann. Bot. 75, 343-357 8 Else, M.A. et al. (1996) Stematal closure in flooded tomato plants involves abscisic acid and a chemically unidentified anti-transpirant in xylem sap, Plant Physiol. 112, 239-247 9 Bleeker, A.B. and Schaller, G.E. (1996) The mechanism of ethylene perception, Plant Physiol. 111, 653-660 16 Allan, A.C. et al. (1994) Two transduction pathways mediate rapid effects of abscisic acid in Commelina guard cells, Plant Cell 6, 1319-1328 11 Schill, V. et al. (1996) The xylem sap of maple (Acerplatanoides) trees sap obtained by a novel method shows changes with season and height, J. Exp. Bot. 47, 123-133 12 Schurr, U. and Schulze, E-D. (1995) The concentration of xylem sap constituents in root exudate and in sap from intact, transpiring castor bean plants (Ricinus communis L.), Plant Cell Environ. 18, 409-420 13 Mtmas, R. (1990) Chemical signals mo~mg from roots to shoots: the case against ABA, in Importance of Root to Shoot Communication in the Responses to Environmental Stress (Monograph 21) (Davies, W.J. and Jeffcoat, B., eds), pp. 175-183, British Society for Plant Growth Regulation 14 Else, M.A. et al. (1994) Concentrations of abscisic acid and other solutes in xylem sap from root systems of tomato and castor-oil plants are distorted by wounding and variable sap flow rates, J. Exp. Bot. 45, 317-323 15 Else, M.A. et al. (1995) Export of abscisic acid, 1-aminocyclopropane-1carboxylic acid, phosphate, and nitrate from roots to shoots of flooded tomato plants. Accounting for effects of xylem sap flow rate on concentration and delivery, Plant Physiol. 107, 377-384 16 Jokhan, A.J., Else, M.A. and Jackson, M.B. Delivery rates of abscisic acid in xylem sap ofRicinus communis L., plants subjected to partial soil drying, J. Exp. Bot. (in press)
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17 Cornish, K. and Zeevaart, J.A.D. (1985) Abscisic acid accumulation by roots of Xanthium strumarium Mill. in relation to water stress, Plant Physiol. 79, 653-658 18 Bano, A. et al. (1993) Abscisic acid and cytokinins as possible root-toshoot signals in xylem sap of rice plants in drying soil, Aust. J. Plant Physiol. 20, 109-115 19 Beck, E. and Wagner, B.M. (1994) Quantification of the daily cytokinin transport from the root to the shoot of Urtica dioica L., Bot. Acta 107, 342-348 26 Smit, B.A., Neumaun D.S. and Stachowiak, M.L. (1990) Root hypoxia reduces leaf growth. Role of factors in the transpiration stream, Plant Physiol. 92, 1021-1028 21 Davis, W.J. and Zhang, J. (1991) Root signals and the regulation of growth and development of plants in drying soil, Annu. Rev. Plant Physiol. Plant Mol. Biol. 42, 55-76 22 Jackson, G.E. et al. (1995) Abscisic acid concentrations and fluxes in droughted conifer saplings, Plant Cell Environ. 18, 13-22 23 Gowing, D.J.G., Jones, H.G. and Davies, W.J. (1993) Xylem-transported abscisic acid: the relative importance of its mass and its concentration in the control of stomatal aperture, Plant Cell Environ. 16, 453-459 24 Trejo, C.L., Clephan, A.L. and Davies, W.J. (1995) How do stomata read abscisic acid signals? Plant Physiol. 109, 803411 25 Tardieu, F. and Davies, W.J. (1992) Stematal response to abscisic acid is a function of current plant water status, Plant Physiol. 98, 540-545 26 Trejo, C.L. and Davies, W.J. (1991) Drought-induced closure of PhaseoIus vulgaris stomata precedes leaf water deficit and any increase in xylem ABA concentration, J. Exp. Bot. 42, 1507-1516 27 Zhang, J. and Davies, W.J. (1991) Antitranspirant activity in the xylem sap of maize plants, J. Exp. Bot. 42, 317-321 28 Tardieu, F., Zhang, J. and Gowing, D.J.G. (1993) A model of stomatal control by both ABA concentration in the xylem sap and leaf water status: test of the model and of alternative mechanisms for droughted and ABA-fedfield-grown maize, Plant Cell Environ. 16, 413-420 29 Jackson, M.B. and Campbell, D.J. (1975) Ethylene and waterlogging effects in tomato, Ann. Appl. Biol. 81, 102-105 30 McKeon, T.A., Fern~dez-Maculet, J.C. and Yang, S.F. (1995) Biosynthesis and metabolism of ethylene, in Plant Hormones: Physiology, Biochemistry and Molecular Biology (Davies, P.J., ed.), pp. 118-139, Kluwer 31 Zarembinski, T.I. and Theologis, A. (1993) Anaerobiosis and plant growth hormones induce two genes encoding 1-a~ainocyclopropane-l-carboxylate synthase in rice (Oryza sativa L.), ~ l . Biol. Cell 4, 363-373 32 Sachs, M.M., Subbaiah, C.C. and Saab, I.N. (1996) Anaerobic gene expression and flooding tolerance in maize, J. Exp. Bot. 47, 1-15 33 Hiatt, A.J. and Lowe, R.H. (1967) Loss of organic acids, K and C1 from barley roots treated anaerobically and with metabolic inhibitors, Plant Physiol. 42, 1275-1278 34 Bradford, I~J. and Yang, S.F. (1980) Xylem transport of 1-aminocyclopropane-l-carboxylic acid, an ethylene precursor, in waterlogged tomato plants, Plant Physiol. 65, 322-326 35 Jackson, M.B. et al. (1994) Hormones and root-shoot relationships in flooded plants - an analysis of methods and results, Plant Soil 167, 99-107 36 English, P.J. et al. (1995) Increased 1-aminocyclopropane-l-carboxylic acid oxidase activity in shoots of flooded tomato plants raises ethylene production to physiologically active levels, Plant Physiol. 109, 1435-1440 37 Bradford, I~J. and Yang, S.F. (1982) Inhibition of ethylene biosynthesis in tomato plants subjected to anaerobic root stress, Plant Physiol. 70, 1503-1507 38 GSmez-Cadenas, A. et al. (1996) Leaf abscission induced by ethylene in water-stressed intact seedings of cleopatra mandarin requires previous abscisic acid accumulation in roots, Plant Physiol. 112, 401-408