David A. J. McArthur and N. Richard Knowles*. Department of Plant ... of root nitrate uptake is diminished within 1 d of P depriva- tion and well before ...... Plant SoillO4 71-78. Lambert DH, Baker DE, Cole H (1979) The role of mycorrhizae in.
Plant Physiol. (1993) 102: 771-782
lnfluence of Species of Vesicular-Arbuscular Mycorrhizal Fungi and Phosphorus Nutrition on Growth, Development, and Mineral Nutrition of Potato (Solanum tuberosum 1.)' David A. J. McArthur and N. Richard Knowles* Department of Plant Science, 4-1 O Agriculture/Forestry Center, University of Alberta, Edmonton, Alberta, Canada, T6G 2P5
area (Woolley and Wareing, 1972; Fredeen et al., 1989). Photosynthesis and carbohydrate utilization for shoot growth are hindered when P nutrition is inadequate (Qiu and Israel, 1992), resulting in a shift in photoassimilate and P partitioning to favor root growth (Cogliatti and Clarkson, 1983). For the potato (Solanum tuberosum L.), adequate P nutrition is critica1for tuber development and simultaneously to maintain a high photosynthetic rate during tuber bulking (Nelson et al., 1947). Because the potato has an inherently low root density and restricted ability to recover fertilizer P (Pursglove and Sanders, 1981), P deficiency can be a limiting factor to yield in commercial potato production (MacKay et al., 1988). Many P-deficiency responses of plants have been described, some of which are suggested to be adaptive and to improve plant P nutrition. These include the induction in roots of a P-transport system that exhibits enhanced affinity and transport capacity for P (Cogliatti and Clarkson, 1983), enhanced extracellular and intracellular acid phosphatase activity (Goldstein et al., 1988; Lefebvre et al., 1990), and modifications to glycolysis that alleviate the effects of a P starvation-induced decrease in ATP availability (Duff et al., 1989). Although these adaptations may improve P nutrition specifically, P starvation has a rapid negative influence on the overall mineral nutrition of plants. For example, the rate of root nitrate uptake is diminished within 1 d of P deprivation and well before root growth is inhibited (Lee, 1982; Rufty et al., 1990). Similar results have been reported for sulfate (Lee, 1982) and Mg (Skinner and Matthews, 1990). Such changes in root selectivity of ion uptake are at least partially responsible for reductions in general mineral nutrient accumulation in P-deficient plants. Because P nutrition is linked to uptake and assimilation of other mineral nutrients, it is possible that some of the growth and physiological responses attributed to P deficiency may actually reflect P starvation-induced deficiencies in other essential elements (Skinner and Matthews, 1990). Interpretation of the direct influence of P nutrition on plant mor-
Crowth, development, and mineral physiology of potato (Solanum tuberosum l. plants ) in response to infection by three species of vesicular-arbuscular mycorrhizal (VAM) fungi and different levels of P nutrition were characterized. P deficiency in no-P and low-P (0.5 mM) nonmycorrhizal plants developed between 28 and 84 d after planting. By 84 d after planting, P deficiency decreased plant relative growth rate such that no-P and low-P plants had, respectively, 65 and 45% less dry mass and 76 and 55% less total P than plants grown with high P (2.5 mM). A severe reduction in leaf area was also evident, because P deficiency induced a restriction of lateral bud growth and leaf expansion and, also, decreased the relative plant allocation of dry matter to leaf growth. Root growth was less influenced by P deficiency than either leaf or stem growth. Moreover, P-deficient plants accumulated a higher proportion of total available P than high-P plants, indicating that P stress had enhanced root efficiency of P acquisition. Plant P deficiency did not alter the shoot concentration of N, K, Mg, or Fe; however, the total accumulation of these mineral nutrients in shoots of P-stressed plants was substantially less than that of highP plants. P uptake by roots was enhanced by each of the VAM symbionts by 56 d after planting and at all levels of abiotic P supply. Species differed in their ability to colonize roots and similarly to produce a plant growth response. In this regard, Glomus intraradices (Schenck and Smith) enhanced plant growth the most, whereas Glomus dimorphicum (Boyetchko and Tewari) was least effective, and Glomus mosseae ([Nicol. and Cerd.] Cerd. and lrappe) produced an intermediate growth response. l h e partia1 alleviation of P deficiency in no-P and low-P plants by VAM fungi stimulated uptake of N, K, Mg, Fe, and Zn. VAM fungi enhanced shoot concentrations of P, N, and Mg by 28 d after planting and, through a general improvement of overall plant mineral nutrition, promoted plant growth and development.
P nutrition exerts a significant influence on plant growth and development, as is readily evident when deficiency occurs. P deprivation induces a rapid decline in plant hydraulic conductivity that results in an appreciable inhibition of leaf expansion (Radin and Eidenbock, 1984). Axillary bud growth is also responsive to P nutrition, and P deficiency can restrict development of the shoot canopy and photosynthetic surface This research was supported by grants from Agriculture Canada and the Natural Sciences and Engineering Research Council of Canada to N.R.K.Graduate student support to D.A.J.M. was provided by the Simonet Endowment to Horticulture. * Corresponding author; fax 1-403-492-4265. -.
/71
Abbreviations: ANOVA, analysis of variance; DAI', days after planting; Lw, leaf weight; LA, leaf area; LAR, leaf area ratio; Lw/W, leafplant dry weight ratio; NAR, net assimilation rate; NM, nonmycorrhizal; Rw,root weight; RGR, relative growth rate; Rw/W, root:plant dry weight ratio; SAR, specific absorption rate; SWIW, shoot:plant dry weight ratio; SLA, specific leaf area; Tw/W, tuber:plant dry weight ratio; VAM, vesicular-arbuscularmycorrhizal; W, plant weight.
McArthur and Knowles
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phology and physiology may be further confounded by involvement of a mycorrhizal symbiosis. Uptake of P by plants in P-deficient soils is substantially improved by mycorrhizal symbioses (Harley and Smith, 1983). Furthermore, a VAM symbiosis can also increase uptake of other mineral nutrients (Lambert et al., 1979; Kucey and Janzen, 1987; Smith and Gianinazzi-Pearson, 1988), although this latter effect may be through the alleviation of plant P deficiency (Lambert et al., 1979; Lee, 1982). On the other hand, some studies indicate that enhanced Zn and Cu uptake by VAM fungi, and subsequent transfer of these minerals to the host, may specifically improve plant growth under Zn- or Cu-deficient conditions (Kucey and Janzen, 1987). Other influences of a VAM symbiosis on plant development may be totally unrelated to plant mineral physiology (Smith and Gianinazzi-Pearson, 1988). In previous studies, we demonstrated detrimental effects of P deficiency on growth and physiology of potato grown under tuber-inducing conditions and characterized some adaptive responses of plants to P stress (McArthur and Knowles, 1992, 1993). Inoculation of roots with a VAM fungus (Glomusfasciculatum) produced a substantial infection in low-P plants relative to high-P plants, thus indicating enhanced plant compatibility to VAM fungi due to P deficiency. Under tuber-inducing conditions, however, the VAM symbiosis alleviated P deficiency to only a minor extent. Because it is well established that source-sink relationships and demand for mineral nutrients by potato plants are different after tuber induction has occurred (Dwelle, 1985), symbiont-host interactions may have been influenced by tuber growth. Colonization of roots by VAM fungi is known to be affected by the symbiont-host combination and nutritional state of the host (Anderson, 1992). The study reported in this paper extends our previous work to provide an integrated view of changes in the potato plant's growth, development, and mineral physiology that occur under non-tuber-inducing conditions in response to P-deficiency stress and VAM fungi. By accounting for the effects of 'I nutrition and the VAM symbiosis on plant P status, and simultaneously describing plant N, K, Mg, Fe, and Zn status, we have characterized both direct and indirect effects of these treatments on the mineral physiology of potato at the whole-plant level. Different VAM species were compared for the development of root colonization of plants grown under different P levels and the response of the host to each species.
MATERIALS A N D METHODS
Inocula
Inocula for NM and VAM (Glomus dimorphicum Boyetchko and Tewari, Glomus intraradices Schenck and Smith, and Glomus mosseae [Nicol. and Gerd.] Gerd. and Trappe) treatments consisted of soil and roots of clover (Trifolium repens L. cv Altaswede) plants grown in an autoclaved sand:soil medium (3:1, v/v) for 120 d. Pot culture of G. dimorphicum was obtained as reported by Boyetchko and Tewari (1986), and G. intraradices and G. mosseae were obtained from the Intemational Culture Collection of VA Mycorrhizal Fungi at the University of Florida, Gainesville. Plants were grown in 2-L pots and were fertilized weekly with 50 mL of a modified
Plant Physiol. Vol. 102, 1993
(minus P) HoagJandsolution. Roots of NM clover plants were uninfected, whereas those of VAM plants were 75% infected and had many vesicles and chlamydospores. To quaniify infection, we ciit roots to I-cm lengths and stored them in a solution consisting of 5% formalin, 5% acetic acid, and 45% ethanol (v/v) until further processed by clearing and stainmg (Phillips and Hayman, 1970). Percentage of infection was assessed using the gridline intersect method (Giovanetti and Mosse, 1980) with 100 counts slide-' and 10 clover or 10 potato (Solanu,rll tuberosum L. cv Russet Burbank) slides plant-'. Plant Crowth Conditions and Experimental Design
Certified potato seed tubers, taken from 4OC (95% RH) storage, were surface sterilized with 0.1% (w/v) NaOCl for 10 min and rinsed thoroughly with distilled H20. Single-eye seedpieces (6.0 g) were cut from the midregion of the tribers and rinsed. After air drying for 1 h, seedpieces were planted in vermiculite and were sprouted in the dark for 12 d at 27OC (95% RH). Excised sprouts with attached roots were blocked for sprout length before transplanting onto NM or VAM soil inociilum (100 g fresh weight pot-') in 25-cm diameter pots (1 sprout pot-I). Each pot contained 7.0 kg (air dry) of an autoclaved sand:soil medium (3:1, v/v) (8 pg g-' of NaHC03-extractable P). An equal mix of fine and coarse sand was used in the medium, and the soil was a silty clay loam with a cation exchange capacity of 0.5 millimhos cm-'. A water extract equivalent to 2 volumes of this soil had a pH of 62!, no detectable P, 33 pg of total N, 40 pg of Ca, 11 pg of Mg, and 5 pg of K (g-' of soil). Pots were placed in a growth chainber set at 25/18OC (day/night) with a 16-h photoperiod. A F'PFD of 300 pmol m-' s-' was provided by fluorescent and incandescent lights rnaintained at 10 cm from the shoot tips. In each of the first 3 weeks, each pot received 100 mL of nutrient solution (40 mM KNO,; 20 mM Ca(N03)2;20 m MgS04. 185 p~ H3B03; 36.5 p~ MnC13; 0.3 p~ ZnS04; 1.3 p~ CuS04; 0.065 W M H2M004;10 mg L-' of ethylenediamine di(o-hydroxyphenyl) acetic acid [pH 6.01) with added P. In the 4th week and thereafter, each pot received 100 mL three times per week (30 applications total). Thus, treatments consisted of three levels of P (0.0, 0.5, or 2.5 m KHzPO4),four inocula (NM or one of three VAM species), and four harvest dates (O, 28,56, and 84 DAP) arranged factorially in a randomized complete block design (four blocks). At the time of harvest, leaf number, leaf area, and stem number were recorded. Plant tissues were divided into stems, leaves, tubers, and roots and lyophilized for dry weight and mineral analyses. From these vanables, Lw/W, Rw/W, Sw/W, Tw/W, LAR (LA/ W ) , RGR (1/W X dW/dt), SAR (1/Rw X dP/dt), SLA (LA,IL,), and NAR (l/LA X dW/dt) were calculated (Hunt, 1982). P accumulated/P available was calculated as follows: (the cumulative total of plant P at each harvest date minus iriitial plant P content at transplanting)/(cumulative total of P supplied at each harvest date, including total NaHC03-extractable soil P pot-I). For the determination of P, K, Mg, Fe, and Zn concentrations, lyophilized plant material was ground through a Mriley mil1 (40 mesh), and 200 mg were dry ashed at 5OOOC for 8 h. Ash was dissolved in 1.0 mL of HCl and made up to 10
Potato, P Deficiency, and Mycorrhizal Fungi mL using 0.1 N H2S04.Tissue P was detennined by the method of Serrano et al. (1976). Shoot Mg, Fe, and Zn concentrations were determined with a Perkin-Elmer model 4000 atomic absorption spectrophotometer at A ~ o z .A248.3, ~, and A213.9, respectively. Flame emission spectroscopywith an A766.5was used to determine shoot K levels. To decrease interferences by P and ionization in Mg and K determinations, samples were diluted to produce solutions containing O. 1 % La (LaC13.6H20)and 0.1% KCl (for Mg) or 0.1% La (for K) (Anonymous, 1979). For analysis of shoot N, 50 mg of ground tissue was digested in H2S04 containing a catalyst (Se02:CuS04:K2S04,1:10:100), and N content was determined with an ,4630 by the method of Fawcett and Scott (1960). Soluble N and Soluble Carbohydrate Assays
For determination of plant N compounds and carbohydrate content, 50 mg of ground, lyophilized plant material was extracted (4OC) for 3 min (mortar and pestle) in 5 mL of 50 mM Hepes buffer (pH 7.4). The homogenate was centrifuged (16408, 4OC) for 30 min, and free amino N, soluble protein N, and nitrate N were detennined from 100, 200, and 50 pL of cold supernatant, respectively. Soluble protein N was measured by a modified Lowry method (Bensadoun and Weinstein, 1976) with BSA (15.6% N) as the standard. Ninhydrin was used to assay free amino N (Rosen, 1956) with a Leu (10.7% N) standard. Nitrate N was determined by the methods of Cataldo et al. (1975), with a KN03 standard. Reducing sugars were assayed colorimetrically by the methods of Nelson (1944) and Somogyi (1952) using a 100-pL aliquot of extract and a Glc standard. Total soluble carbohydrates were detennined from a 50-pL aliquot of supernatant with the phenol-sulfuric acid reagent (Dubois et al., 1956) and a Glc standard. Statistical Analyses
Growth and physiological data were subjected to ANOVA and, where appropriate, sums of squares were partitioned into individual degree of freedom components of both main effects and interactions. Percentage of infection data were arc-sin transformed to achieve homogeneity of variance before ANOVA was calculated. Based on the results of the ANOVA, regression analysis was used to derive polynomial models for describing the various relationships. In this study, Pdeviation indicates a significant residual variance that is not explained by a linear model and with no further trend analysis possible.
RESULTS Development of Root infection
Root colonization by G. intraradices and G. mosseae was rapid, as evidenced by a 43 and 40% level of root infection, respectively, at 28 DAP (Fig. 1). At 28 DAP, infected roots showed many penetration points, hyphal coils, and arbusa l e s , whereas vesicles were absent. By 84 DAP, the percentage of root infection had doubled, and root vesicles and externa1 clusters of chlamydospores were common. Roots
O
773
20
40
60
80
DAYSAFTERPLANnNG
effect of time on percentage of root infection by C. dimorphicum (A),C. intraradices (O),or C. mosseae (O) for potato plants (averaged for plants grown with 0.0, 0.5, or 2.5 m M P) inoculated with VAM clover pot culture. F values for C. dimorphicum versus (average of C. intraradices and C. mosseae) and C. mosseae versus G. intraradices were significant at the 0.01 level. The interaction of Timequadrafic x C. dimorphicum versus (averageof C. intraradices and C. mosseae) was significant at the 0.01 level. Inset, Main effect of increasing abiotic P supply on percentage of root infection, averaged for VAM species and during an 84-d growth period (O).The F value for the linear trend was significant at the 0.01 level. Figure 1. Main
inoculated with G. dimorphicum initially had a low rate of infection, but this increased later in the experiment so that differences in level of infection between G. dimorphicum and other species averaged only 9% at 84 DAP. Differences among species’ ability to colonize roots were evident throughout this study, however, with G. intraradices producing the most root infection and G. dimorphicum producing the least. The main effect of increasing abiotic P supply was to decrease the percentage of root infection throughout the entire growth period by 8 and 17% for low-P (0.5 mM) and high-P (2.5 mM) plants, respectively, relative to plants without supplemental P (no-P) (Fig. 1, inset). No significant interaction between VAM species and abiotic P level was evident for root infection, indicating that high P diminished host compatibility with VAM fungi regardless of differences among funga1 species. Analysis of Plant Crowth, Development, and Morphology
Figure 2 shows that plant growth during the first 28 DAP was unaffected by differential P nutrition, probably reflecting the initial small size of plants relative to the P available. By 56 DAP, however, NM plants fertilized with no P (0.0 m ~ ) or low P had accumulated 39 and 32% less dry matter, respectively, than plants grown with high P. By 84 DAP, noP plants and low-P NM plants had 65 and 45% less dry weight, respectively, than high-P plants. Inoculation of roots with a VAM fungus provided a distinct advantage to plants grown with no P or low P, as demonstrated by an average 73 and 43% greater plant dry weight, respectively, at 84 DAP, compared to NM plants with the same abiotic P supply. The amount by which plant growth was stimulated by VAM species depended on the rate at which root infection was established (Abbott and Robson, 1982), and, hence, G. intraradices and G. mosseae, which more rapidly colonized roots, increased plant dry weight at 84 DAP by an average of 10%
Plant Physiol. Vol. 102, 1993
McArthur and Knowles
774
-
Growth and shoot canopy development were restrictetl by inadequate P nutrition by 56 DAP and substantially so between 56 and 84 DAP. A primary reason for the diminished total leaf area was a significant restriction of lateral bud growth associated with low P nutrition (Fig. 3). Expansion of photosynthetic surface area for Russet Burbank plants depends on lateral branch growth after the primary slnoot flowers (between 28 and 56 DAP in our study). Between 56 and 84 DAP, no-P NM plants increased their branch nurnber by 34%, whereas high-P NM plants nearly doubled their branch number. Leaf number for no-P NM plants also increased by 32% from 56 to 84 DAP; however, this potential contribution to total leaf area was partially offset by an 18% reduction in area per leaf for the same period (P < 0.01). As a result of this inhibition of growth and development, no-P NM plants had a 75 and 80% lower total leaf area and leaf dry weight, resyectively, than high-P NM plants at 84 DAP. Stem (including tubers) and root growth were somewhat less restricted by inadequate P nutrition, as indicated by the 60
60
c
0 3
5
2 40 3
2 20
a O 60
s
s
f> 40 K
n
5
20
2 O
-
60
o)
I-
I
40
> K
2 z
*
O.OmMP
20
O
O
20 40 60 DA% AFTER PLANTING
80
Figure 2. Time course of dry matter accumulation of NM (O) and VAM plants (C. dimorphicum, A; C. intraradices, O, C. mosseae, O ) grown with 0.0, 0.5, or 2.5 mM P for 84 d. F values for t h e main effects of VAM, C. dimorphicum versus (average of C. intraradices and C. mosseae), Timequadratlc, and the interaction of VAM X P,,,,,, x Timequadratlc were significant at the 0.01 level. Inset, Main effect of abiotic P supply on dry matter accumulation for NM plants and the average of VAM plants (O).F values for PI,,,,, and the interaction of VAM x PI,.,,, were significant at the 0.01 level.
more than did G. dimorphicum (P < 0.01). When P was relatively sufficient (Fig. 2, inset), no significant influence of a VAM symbiosis on dry mass accumulation was observed, in spite of an average 53% level of root infection (Fig. 1, inset). P nutrition and the VAM symbiosis exerted strong influentes over plant development and morphology. The polygonal diagrams in Figure 3 summarize the effects of VAM fungi and abiotic P supply on plant yield components at 28 (inner polygon), 56, and 84 DAP. Results of ANOVA for treatment effects on each yield component depicted in Figure 3 appear in Table 1. A positive response to increasing abiotic P supply, time, or the VAM symbiosis was evident for each plant yield component. Significant interactions between P nutrition and the VAM fungi were indicated by ANOVA and characterized a relatively large benefit from the VAM symbiosis for no-P and low-P plants but a relatively minor effect of the VAM symbiosis on high-P plants.
i
$ 2
30
T 4M
30
T
0.5 mM P
Non-mycorrhizal
Leaf
VA-mycorrhizal
Figure 3. Polygonal diagrams describing growth, development, and overall plant morphology of NM and VAM potato plants grown with 0.0, 0.5, or 2.5 mM P and harvested at 28 (inner polygon), 56, and 84 DAP (outerpolygon).Axes are defined on the 2.5 mM P polygonal diagrams, and stem dry weight includes tuber dry weight. ,4 summary of the statistical analysis for each yield component is presented in Table I. Data are pooled for three VAM species.
Potato, P Deficiency, and Mycorrhizal Fungi
775
Table 1. Partia/ summary of ANOVAs for yield components of NM and VAM potato plants grown with various levels of P Plants were inoculated with NM or VAM (C. dimorphicum [Cd], C. intraradices [Gil, or C. mosseae [Cml) clover pot cultures and fertilized with a complete nutrient solution containing 0.0, 0.5, or 2.5 mM P. Plants were harvested 28, 56, and 84 DAP. Data are Dresented in Figure 3. No.
Dry wt
Source of Variation
VAM P VAM X PL VAM X Timeg PL X T h e Q VAM X PL x Timeq Ci and Cm > Cd Ci > Cm
Branch
Leaf
O.Olb L
0.01
0.01
L
ns
Leaf
Stem'
Root
Plant
L, D
0.01 L, D
0.01 L
0.01 L, D
0.01 L, D
0.01 0.01 0.01 0.01 0.01
0.01 0.01 0.01 0.01 0.01
0.01 0.01
ns
0.01
0.10 0.05 0.01 0.01 0.01
0.01
0.01 0.01 0.01 0.01 0.01
ns
ns
ns
ns
ns
ns
0.01 0.01
ns
0.01
0.01 0.01 0.05
0.01 0.01
ns
lncludes tuber dry weight. Significance levels or significant trend (L, linear; Q, quadratic; D, deviations with P c 0.05) for indicated sources of variation (ns, not significant). a
and 36% lower dry weights, respectively, at 84 DAP for no-
P plants than for high-P plants. Comparing the shape and relative areas encompassed by the polygonal diagrams for VAM plants with those for NM plants demonstrates the advantage of the VAM symbiosis to plant growth and development under P-limiting conditions (Fig. 3). By 84 DAP, VAM plants grown without supplemental P had a dry weight equivalent to that of NM plants grown with low P (0.5 mM), and no-P VAM plants had a 24% higher total leaf area than low-P NM plants. Furthermore, low-P VAM plants were comparable to high-P NM plants in terms of overall growth and development. For highP plants, however, the influence of the VAM symbiosis on leaf number, stem dry weight, and leaf dry weight was almost negligible, although total leaf area, branch number, and root dry weight were marginally higher for VAM plants than for NM plants. Significant differences among species in their ability to influence production of various yield components (Table I) were consistent with those for plant dry mass; however, for the sake of brevity, separate presentation of these data has been omitted. Relative to roots, dry matter accumulation in leaves and stems was highly responsive to P nutrition. For each 1 mM [P] increase in the nutrient solution supplied to NM plants during growth, dry weights of stems and leaves increased by 7.1 and 6.6 g plant-', respectively, at 84 DAP ( r = 0.99). In contrast, the root response to increasing P was only 0.7 g plant-' ( r = 0.92). These differential responses resulted in a lower allocation of dry matter to shoot growth (Sw/W) with low P nutrition and, conversely, a higher proportion in roots (Rw/W) (Table 11). Tuber dry mass made a relatively minor contribution to total plant dry mass (Tw/W) under our growth conditions, and, although tuber growth was enhanced by increasing abiotic I' supply, VAM plants were less responsive than NM plants in this regard (VAM X Plinear,P < 0.05). Tuberization occurred earlier in NM plants (before 56 DAP) than in VAM plants (after 56 DAP), and possibly for this reason, tuber dry weight averaged over abiotic P levels was 52% lower at 84 DAI' for VAM plants than for NM plants.
This altered partitioning of dry matter by the VAM symbiosis favored a greater allocation of photoassimilates to shoot growth. Plant growth analysis demonstrated that P nutrition influenced both the net assimilatory capacity of leaf biomass (NAR)and plant dry matter partitioning (Table 111). The NAR, an index of the productive efficiency of leaves, declined linearly (r = -0.91) with decreasing abiotic P leve1 for NM plants. Similarly, the production of photosynthetic surface area relative to plant dry m a s (LAR) for NM plants was lower for no-P or low-P plants than for high-P plants. Reductions in both the photosynthetic surface area per unit
Table II. Relative dry matter allocation of NM and VAM potato plants grown with various levels ofabiotic P supply Plants were inoculated with NM or VAM (C. dimorphicum, C . intraradices, or C. mosseae) clover pot cultures and fertilized with a complete nutrient solution containing 0.0, 0.5, or 2.5 mM P. The data presented are for 84 DAP only, and VAM data are the average
for three species.
,
Treatment
5WlW
[PI
0.5 2.5
TwlW
%
illM
0.0
RwIW
VAM
-
+ + +
VAMa P VAM X PL VAM X Pn
79.7 85.2 80.9 83.7 82.5 85.7 0.01b L
ns ns
17.6 14.7 15.0 13.9 9.9 11.3
ns L 0.10
ns
2.8 0.6 4.1 2.5 7.7 3.O 0.01 L 0.05
ns
Sources of variation (L, linear trend; D, deviations). Significance levels or significant trend (L, linear with P c 0.05) for indicated sources of variation (ns, not significant). a
McArthur and Knowles
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Table 111. Crowth analysis of NM and VAM potato plants grown with various levels of abiotic P supply Plants were inoculated with NM or V A M (C. dimorphicum, C.
intraradices, or C. mosseae) clover pot cultures and fertilized with a complete nutrient solution containing 0.0, 0.5, or 2.5 mM P. Data were pooled from plants harvested at 28, 56, and 84 DAP, and the V A M data are the average for three species. Crowth indices were calculated from the data presented in Figures 2 and 3. Treatment rpi mM
0.0
0.5 2.5
NAR
LAR
SLA
mg cm-2 d-'
cm2 mg-'
cm2 mg-'
0.51 0.50 0.55 0.49 0.58 0.51
0.11 0.13 0.11 0.14 0.12 0.14
0.23 0.26 0.24
0.01b L 0.10
0.01
0.01 D ns
LwIW
RCR
VAM
-
+ + +
VAMa P V A M X PL V A M X PD
ns
L, D
ns ns
0.27
0.25 0.26
ns
g g-' d-'
0.36
0.38 0.36 0.40 0.39 0.40 0.01 L
0.10 ns
0.059 0.065 0.063 0.068 0.070
Plant Physiol. Vol. 102, 1993
and low-P NM plants had accumulated 76 and 55% les!; I', respectively, than high-P NM plants. The rate at which roots absorbed P (mg of P g-' root dry weight d-') was lineiirly related to the abiotic P supply (SAR = 0.13[P] + 0.44,r = 0.99). The SAR declined with time for no-P and low-P plants; however, for high-P plants, a decrease in SAR was evidlent only at the last harvest date (Fig. 4, insets). For a pot study, such results likely reflect increasing overlap of root absorptive surface area with time and indicate depletion of soil P for no-P and low-P plants (Sanders et al., 1977). Differences between NM and VAM plants for P accumulation and SAR were initially negligible (Fig. 4), reflecting relatively high soil P levels and low levels of VAM infection early in the study. However, differences in P status between NM and VAM plants were evident by 56 DAP at all levels of abiotic P. At 134 DAP, VAM plants grown with no P, low
0.071 0.01 L, D 0.01
ns
Sources of variation (L, linear trend; D, deviations). Significance levels or significant trend (L, linear; D, deviations with P < 0.05) for indicated sources of variation (ns, not significant). a
120
+ 80i . -2 c
c
40
O
leaf dry mass (SLA) and the relative allocation of plant dry mass to leaf production (Lw/W) with decreasing abiotic P level showed that the relative leaf area produced by NM plants was restricted by both diminished leaf expansion and allocation of dry mass to leaf growth. Because the RGR of plants is a product of NAR and LAR and because LAR = SLA X Lw/W, these results characterize the significant negative influence of inadequate P nutrition on the RGR of potato plants (Table 111). As expected, VAM plants maintained a higher average RGR than NM plants (Table 111), although a highly significant VAM X Plinea,interaction from ANOVA indicated that differences between NM and VAM plants were diminished by increasing abiotic P supply. At 2.5 m P, NM plants had an RGR equivalent to that of VAM plants. A major factor in the VAM-stimulated increase in RGR under no-P or low-P conditions was an average 18% increase in LAR for VAM plants, resulting from a combination of enhanced leaf expansion (SLA) and relatively greater allocation of plant dry matter to leaf growth (Lw/W). Conversely, the NAR for VAM plants was lower than that for NM plants, thus indicating that the photosynthetic surface area of VAM plants was somehow less efficient in producing dry weight gain than that of NM plants. An increase in root respiration of VAM plants compared to NM plants has been observed (Baas et al., 1989), and this could at least partly account for the significantly lower NAR for VAM plants than for comparable NM plants. '
P Acquisition The trend in whole-plant P accumulation depended on P nutrition and the VAM symbiosis (Fig. 4). By 84 DAP, no-P
1
/ / /
40 6 0 80
"'"20
-
4 '50.020
O
40
60 80
%
40 60 D A SAFER P M N G
20
80
Figure 4. Time course of P accumulation for NM (O) plants and V A M plants (C. dimorphicum, A; C. intraradices, O; C. mosseae, O ) grown with 0.0, 0.5, or 2.5 mM P for 84 d . F values for the main effects of VAM, C. dimorphicum versus (average of C. intrara,dices
and C. mosseae), C. mosseae versus C. intraradices, f'l,,,,,, Timequadra,,=, and the interaction of V A M x Timequadratlc were significant at the 0.01 level. Insets, Time course of SAR for P for NM plants and the average of V A M (O) plants. F values for Timellnear, VAM, P,I and the interaction of V A M X Timed,.v,at,on Timedevlatlon, were significant at the 0.01 level. dwt, Dry weight.
777
Potato, P Deficiency, and Mycorrhizal Fungi
P, or high P had accumulated an average of 115, 71, and 34% more P, respectively, than comparably fertilized NM plants. It is of interest that, although the relative contribution of the VAM fungi was decreased by high P, the actual increment in milligrams of P from VAM fungi marginally increased with abiotic P supply (P < 0.01). The basis for higher P accumulation by VAM plants was a 63% higher SAR for VAM roots than for NM roots, when averaged over P levels for the last two harvest dates (Fig. 4, insets). Species differed significantly with respect to their contribution to plant P accumulation; G. intraradices increased P uptake the most and G. dimorphicum the least (P < 0.01). In spite of the relatively small amount of total P accumulated by no-P NM plants, they had a ratio of P accumulated/ P available at 84 DAI' that was 25% greater than that of high-P NM plants (Fig. 5 ) . Moreover, comparison of the P accumulated/P available ratio for no-P and low-P plants indicated that VAM roots were twice as effective with respect to absorbing available P as were NM roots (Fig. 5). This benefit of the VAM symbiosis to root efficiency of P acquisition was inversely related to increasing abiotic P supply but was still evident for high-P plants.
2.5 mM P
0.80
-
20
40 60 DAYSAFTER PLANnNG
80
Figure 5. Time course of P accumulated/P available for N M (O) plants and the average of VAM plants (O) grown with 0.0, 0.5, or 2.5 mM P for 84 d. F values for the main effects of P,,I VAM, Time,,,, Timedevlatlon, and the interaction of VAM X P,I X Timedevlatlon were significant at the 0.01 level.
Shoot Mineral Physiology
Although plants accounted for an increasing proportion of available P over time, P uptake could not match dry matter accumulation, as was evident by the declining concentration of P in shoots (Fig. 6). P concentration declined most rapidly between O and 28 DAP, reflecting rapid early growth in spite of limited P resources (seedpiece reserves were excised from sprouts at the time of transplanting) and a relatively undeveloped root system. For each 1 m~ [P] increase, shoot P increased by 0.16 mg g-' dry weight during the 84-d growth period ([shoot P] = 0.16 [P] 3.35, r = 0.99) (Fig. 6, inset). At 28 DAP, the VAM symbiosis marginally increased shoot P concentration ( L S D ~= . ~0.13 ~ mg g-' dry weight); however, this increase was more substantive at 56 (31%)and 84 DAP (22%). Whereas shoot P concentration was highest at the transplant stage, K, Mg, and N (mg g-' dry weight) increased from transplanting to about 56 DAP, indicating rapid mineral nutrient uptake in relation to dry matter accumulation (Fig. 6). These levels then declined during the period of greatest dry mass accumulation, which is in agreement with previous reports concerning the mineral nutrition of potato (Harris, 1978). In contrast to shoot P concentration; K, Mg, and N levels did not show any significant response to abiotic P supply (data not shown). However, a highly significant main interaction from effect of VAM and a VAM X Timesuadratic ANOVA demonstrated that VAM plants had higher shoot concentrations of K, Mg, and N than did NM plants on two of three harvests dates after inoculation and, for N and Mg, as early as 28 DAP (Fig. 6). Table IV shows that, although the level of shoot N was not altered by P nutrition, concentrations of soluble protein N, free amino N, and nitrate N were higher in P-deficient plants. Higher levels of such soluble N compounds in shoots reflect the slow growth and nitrate assimilation relative to N accumulation by P-deficient plants (McArthur and Knowles, 1993). Insoluble N in shoots increased linearly (P < 0.01) with increasing abiotic P supply, possibly indicating increased incorporation of soluble N into nucleic acids, imino acids, and structural N compounds, which were not detected by our assays. Most of the difference in the concentration of total shoot N between VAM and NM plants was attributable to an enhanced level of insoluble N in VAM plants. Because an enhanced level of insoluble N in shoots was also associated with high P nutrition, the VAM effect on insoluble N was probably mediated indirectly through the improved plant P status afforded by the VAM symbiosis. Shoot Fe concentration (pg g-' dry weight) declined with time and was not altered by VAM or abiotic P supply (Fig. 7). Although such a trend may indicate a developing deficiency, comparison of these shoot Fe levels with published values for potato (Harris, 1978) indicated that Fe was not likely limiting to plant growth in our study. The effects of P nutrition and VAM infection on Zn concentrations of shoots were in contrast to those on shoot P. Shoot Zn concentration decreased 21% as abiotic P supply was increased to 2.5 mM (Fig. 7, inset) and was 18% less for VAM plants, compared with NM plants, when averaged over the study interval (Fig. 7). Shoot Zn concentration decreased by 3.5 pg g-' dry weight
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Plant Physiol. Vol. 102, 1993
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lnteractions between the effects of VAM fungi and time on shoot P, K, Mg, and N concentrations of potato plants harvested at O, 28, 56, and 84 DAP. Data are for NM (O)plants and the average of VAM (O)plants. For P, F values and the interaction of VAM X Timequadratic were significant at the 0.01 level for the main effects of VAM, TimequadraIlc, ( L S D ~= . ~ 0.1 ~ 3 mg g-' dry weight). Inset, Main effect of abiotic P supply on shoot P concentration. F value for the linear were significant at the trend was significant at the 0.01 level. For K, F values for the main effects of VAM and Timequadratlc 0.01 level ( L S D ~ = , ~ 2~. 3 mg g-' dry weight). For Mg, F values for the main effects of VAM, Timequadratlc, and the interaction were significant at the 0.01 level ( L S D ~ .=~ ~0.6 mg g-' dry weight). For N, F values for the main of VAM X TimequadraIlc effects of VAM, TimequadraIlc, and the interaction of VAM X Timequadratic were significant at the 0.01 level ( L S D ~ = . ~ 2~. 3 mg g-' dry weight). Figure 6.
+
with each 1 m~ [E'] increase ([shoot Zn] = -3.5[P] 60.1, Y = 0.99) when averaged over the harvest interval. Increasing the abiotic P supply possibly produced interactions between Zn and Fe oxides in the soil, rendering Zn less available (Loneragan et al., 1979). Although P nutrition had no effect on the concentrations of N, K, Mg, or Fe in shoots, it strongly influenced shoot mineral accumulation, as depicted in the polygonal diagrams of Figure 8. Each polygonal diagram summarizes the content of N, P, K, Mg, Fe, and Zn in shoots of potato plants at 28 (inner polygon), 56, and 84 DAP for NM and VAM plants grown with a particular abiotic P level. Treatment effects for each element as characterized by ANOVA are summarized in Table V. Comparison of polygonal diagrams for no-P NM plants with low-P or high-P NM plants indicated that mineral nutrient accumulation was restricted by P stress and maximally so between 56 and 84 DAP. Shoot content of P, K, Mg, N, Fe, and Zn at 84 DAP was 74, 68, 66, 64, 56, and 45% lower, respectively, for NM plants grown with no supplemental P than for plants grown with 2.5 mM P. Comparison of the polygonal figures for VAM plants with those for NM plants shows that even partia1 alleviation of P deficiency by the VAM symbiosis markedly improved mineral nutrient accumulation in shoots (Fig. 8). Similar to the yield variables shown in the polygons of Figure 3, at 84 DAP,
no-P VAM plants matched or even exceeded the mineral nutrient accumiilation of low-P NM plants. Uptake of nutrients was not limited by P stress for plants grown with 2.5 mM P; however, high-P VAM plants demonstrated enhanced ~ mg accumulation of not only P but also K ( L S D ~=~ 156 shoot-') in shoots. Although the VAM symbiosis enhanced shoot Zn content for P-deficient plants, high-P VAM plants paradoxically had a lower shoot Zn content (VAM X Plmea,, P < 0.01) than high-P NM plants. Significant differences among species with respect to their ability to influence mineral accumulation (Table V) were similar to those for plant dry mass; however, for the sake of brevity, separate presentation of these data has been omitted. DISCUSSION
Growth and development of potato plants were dernonstrated to be responsive to P nutrition. Moreover, our results characterized the progressive development of P deficiency between 28 and 84 DAP, a period in which the leve1 of available soil P had clearly become inadequate to maintain the initial RGR of plants. No-P or low-P treatment of NM plants resulted in substantially less growth and photosynthetic surface area, lower tissue-P concentrations, and an altered plant morphology when compared with high-P treat-
Potato,
P Deficiency, and Mycorrhizal Fungi
Table IV. Effect of VAM fungi and P nutrition on N pools in shoots oípotato plants Plants were inoculated with NM or VAM (C. dimorphicum, C. intraradices, or C. mosseae) clover pot cultures and fertilized with a complete nutrient solution with 0.0, 0.5, or 2.5 mM P. Data were pooled for plants harvested at 28, 56, and 84 DAP, and VAM data are the average for three species. Soluble NO3 N Amino
Treatment [Pl
VAM
N
Soluble
Protein
lnsoluble
0.0 mM P
Reduced Na
(soluble ~
f,
~ - ,
N
-
0.0
9.7 10.0 9.6
+ + +
0.5 2.5
10.2 9.0 8.4
nsc
VAM* P VAM X PL VAM X Po
L
ns ns
2.5 2.9 2.3 2.8 2.4 2.4
0.01 L 0.10 ns
9.7 9.2 9.0
9.0 8.6 9.0
ns L 0.05
ns
17.6 19.3 17.9 19.0 18.3 20.9
i'"
0.56 0.54 0.55 0.54 0.56 0.58
0.5 mM P
ns L ns ns
0.01 L
ns ns
@ 14.8
PhosphoNs
Phosphorus
Ni
"(Soluble amirio N + protein N)/(total soluble N) ratio for shoots. Sources of variation. Significance levels or trend (L, linear; D, deviations with P < 0.05) for indicated sources oí variation (ns, not significant).
2.5
lron (mg)
Non-mycorrhizal
$ 160
@m f4.E
E
mg g-' shoot dry wt
mM
779
lron (mg)
VA-mycorrhizal
e
O
BE
Figure 8. Polygonal diagrams describing total mineral nutrient ac120
80
I
I 75
cumulation in shoots of NM and VAM potato plants grown with 0.0, 0.5, or 2.5 m M P and harvested at 28 (inner polygon), 56, and 84 DAP (outer polygon). Data are pooled for three VAM species. Axes are defined on the 2.5 mM P polygonal diagrams. A summary of statistical analysis for each mineral nutrient is presented in Table V. For N, L S D O . = ~ ~108 mg shoot-'; for P, L S D ~=. ~4.5 ~ mg shoot-'; for K, L S D ~ = . ~ 156 ~ mg shoot-'; and for Mg, L S D ~ = . ~ 19.5 ~ mg shoot-'.
5 t.
O
9 W
3 55
35 O
20 40 60 D A S AFfER P W N G
80
Figure 7. Change in shoot Fe and Zn concentrations oí NM (O)and (a)potato plants harvested at O, 28, 56, and 84 DAP. Data are pooled for plants grown with 0.0, 0.5, or 2.5 mM P, and the
VAM
VAM data are an average of three speciei. For Fe, F value for the main effect oí Timequadratlc was significant at the 0.01 level. For Zn, and the interaction Fvalues for t h e main effects of VAM, Timequadratlc, of VAM X Timequadratlc were significant at the 0.01 level. Inset, Main effect of abiotic P supply on shoot Zn concentration, averaged for both NM and VAM plants. F value for the linear trend was significant at the 0.01 level.
ment of NM plants. Furthermore, these changes to the growth and P status of plants due to P deficiency were accompanied by a concomitant decrease in the accumulation of mineral nutrients other than P (Fig. 8), probably because of a reduced capacity to intercept and absorb ions. In this regard, an average 27% lower root dry mass for no-P NM plants than for high-P NM plants between 28 and 84 DAP (Fig. 3) indicated a reduction in root surface absorptive area by P stress (Newman and Andrews, 1973). Changes in the ion selectivity of roots and a diminished metabolic capacity for ion uptake have also been implicated in lower mineral nutrient accumulation by P-deficient plants (Lee, 1982; Baas et al., 1989; Rufty et al., 1990; McArthur and Knowles, 1993). Although shoot N concentration was not altered by P nutrition, N assimilation appears to be less efficient in P-deficient plants (McArthur and Knowles, 1993; Table IV). These responses of potato plants to low-P nutrition are similar to
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Table V. Partia/ summary of ANOVA for mineral elements accumulated in shoots of NM and VAM potato plants grown with various levels of abiotic P supply
Plants were inoculated with NM or VAM (C. dimorphicum [ C d ] ,C. inl‘raradices [Gil, or C. mosseae [ C m ] )clover pot cultures and fertilized with a complete nutrient solution containing 0.0, 0.5, or 2.5 mM P. Plants were harvested 28, 56, and 84 DAP. Data are presented in Figure 7. Source of Variation
VAM P
VAM X PL VAM X TimeQ PL X TimeQ VAM X PL X TimeQ Ci and Cm > Cd Ci > Cm
N
P
K
Mg
Fe
0.01” L, D 0.01 0.01 0.01 0.01
0.01 L, D
0.01
0.01 L 0.05
0.01
0.01
0.01 L, D 0.01 0.01 0.01 0.01 0.01
0.01
0.01
0.01
ns 0.01 0.01
ns
Zn
0.01 0.01 0.01
o1.01
0.10 0.05
ns ns 0.01 0.05 ns 0.01 ns
ns
ns
ns
L, D 0.01 0.05
ns
Total Plant P
0.01 L, D ns
0.01 0.01
ns 0.01 0.01
Significance levels or significant trends (L, linear; Q, quadratic; D, deviations with P < 0.05) for indicated sources of variation Ins. not sianificant). a
those reported for other plants grown with inadequate P supply (Lambert et al., 1979; Fredeen et al., 1989) or exposed to short-tem P deprivation (Cogliatti and Clarkson, 1983; Goldstein et al., 1988; Rufty et al., 1990). Such responses are generally attributed to P-deficiency stress. Comparison of root and shoot growth of plants grown with different levels of P nutrition demonstrated that roots were less affected by P deficiency than were shoots (Fig. 3, Table 11). Because P tends to be immobile in soils, continued root elongation at the expense of shoot growth appears to be a primary plant strategy for maintaining P acquisition under deficiency conditions (Clarkson, 1985). Roots of P-deficient plants accounted for a higher proportion of total available P than those of high-P plants (Fig. 5 ) , a finding that may relate to observations of enhanced activities of acid phosphatases and ATPases in roots of low-P plants compared to those of high-P plants (McArthur and Knowles, 1993). These results are consistent with the view that P starvation enhances the affinity and transport capacity of potato roots for P (Lee, 1982; Cogliatti and Clarkson, 1983). Unlike the rapid induction of a high-affinity system (within 3 d) by complete P deprivation as observed by Cogliatti and Clarkson (1983), depletion of available soil P (28-56 DAI’), as with no-P plants, would have been required before such physiological changes could be anticipated in our study. Collectively, these responses to P deficiency appear to constitute part of a ‘phosphate starvation rescue system” in higher plants (Goldstein et al., 1988). Growth and development of P-deficient plants were markedly improved by inoculation with any one of the three species of VAM fungi, although the plant response to G. dimorphicum was generally less than that to G. intraradices or G. mosseae. Such differences among VAM species appear to relate to their ability to colonize roots (Abbott and Robson, 1982; Fig. 1) and to the production of externa1 hyphae required for P acquisition (Sanders et al., 1977). VAM roots were more efficient at P uptake than NM roots (Fig. 5), as observed by others (Sanders et al., 1977; Cress et al., 1979; Baas et al., 1989; McArthur and Knowles, 1993).
Although the VAM symbiosis produced marginally h igher shoot P concentrations in plants by 28 DAP, significant changes to plant P accumulation occurred only after 28 d (Figs. 4 and 5). It is of interest that, in spite of the inhitlitory influence that high abiotic P supply to plants had on fungal growth (Fig. 1, inset), the fungal symbionts were clearly successful in maintaining the exchange of P and carbohydrates with the host under non-tuber-inducing conditions, as evidenced by the high level of root infection and by the enhanced P content of high-P VAM plants at 84 DAP (Fig. 4). Under tuber-inducing conditions in which sink-source relations favor dry matter partitioning to tuber growth, VAM fungi appear to be less effective in maintaining root colonization and nutrient exchange with the host (McArthur and Knowles, 1993’). Comparison of the polygonal diagrams in Figure 21 and Figure 8 shows that improvements to growth and development of P-deficient plants by the VAM symbiosis were concomitant with an enhanced mineral nutrient accuxnulation in shoots. One of the most immediate benefits of the VAM symbiosis to plants was a stimulation of leaf gsowth and expansion, which was evident by 56 DAP and associated with a significantly greater total accumulation of P, N, K, and Mg for plants grown with each level of abiotic P supply. Even preceding these effects, however, the concentrations of P, N, and Mg in shoots were already higher by 28 D24Pin VAM plants than in NM plants. By enhancement of the concentration of these nutrients in shoots, the VAM symbiont appeared to play a role in stimulating the development of plant photosynthetic surface area (Woolley and Wareing, 1972; Radin and Boyer, 1982). Conversely, tuber growth, which might compete with the VAM symbiont for photoassimilates, was negatively influenced by VAM (Table I]), opposite to what was found under tuber-inducing conditions (McArthur and Knowles, 1993). This could be specifically ascribed to enhanced N accumulation and subsequently higher tissue N levels for VAM plants (Figs. 6 and 8), 3which are known to delay tuber initiation and growth (Krauss, 1985).
Potato, P Deficiency, and Mycorrhizal Fungi A rather unusual response by potato plants to the VAM
symbiosis was the decrease in shoot Zn concentrations (Fig. 7B). Whereas the shoot Zn concentration response to increasing abiotic P supply was consistent with that previously reported (Lambert et al., 1979; Loneragan et al., 1979), our result of a significantly lower shoot Zn concentration in VAMinfected plants is contrary to most reports (Lambert et al., 1979; Marschner, 1986; Kucey and Janzen, 1987). Because concentrations of shoot Zn in our study were high relative to other studies in which Zn levels were noticeably deficient, transfer of Zn by VAM fungi to plants in our study was probably not of significant benefit relative to the transfer of P. Therefore, it seems possible that the greater accumulation of Zn under P-deficient conditions in VAM plants than in NM plants (Fig. 8) reflects improved uptake of Zn by roots after alleviation of P-deficiency stress, similar to that for N, K, Mg, and Fe. Finally, the lower shoot Zn concentrations of VAM plants than of NM plants may be attributable to chemical interactions between P and Zn involving intraradical hyphae and root cells, which have yet to be characterized (Robson and Pitman, 1983). In summary, our results provide the first detailed characterization of the influence and interactions of P-deficiency stress and VAM symbiosis on mineral physiology and early growth of potato under non-tuber-inducing conditions. The effect of plant P status, as achieved by either abiotic P supply or VAM symbiosis, on the N, K, Mg, and Zn status of plants demonstrated that plant nutrition does not involve elements in a singular fashion but is certainly one of numerous interactions. In this regard, the prohibitive influence of P-deficiency stress on mineral nutrient acquisition of potato underscores the importance of P nutrition to maintaining optimal nutrient-utilization efficiency and plant growth in general. The development and nutrition of potato were demonstrated by this and our previous reports (McArthur and Knowles, 1992, 1993) to have a major impact on the growth and establishment of VAM fungi. Conversely, the VAM symbiosis appeared capable of influencing plant development, e.g. leaf expansion, apparently through changes to the mineral status of shoots. Species of VAM fungi differed with respect to their ability to benefit plant P nutrition under our growth conditions; however, further investigation is recommended to determine why this was so and whether similar species differences exist under field conditions. Finally, the significant contribution of the VAM symbiosis to potato P nutrition under non-tuber-inducing conditions and relative plant P sufficiency suggests that VAM fungi may benefit the yield of potato crops by enhancing P accumulation during early- to mid-season growth. Received October 8, 1992; accepted March 19, 1993. Copyright Clearance Center: 0032-0889/93/102/0771/12. LITERATURE ClTED
Abbott LK, Robson AD (1982) The role of vesicular-arbuscular mycorrhizal fungi in agriculture and the selection of fungi for inoculation. Aust J Agric Res 3 3 389-408 Anderson AJ (1992) The influence of the plant root on mycorrhizal formation. In MF Allen, ed, Mycorrhizal Functioning. Chapman and Hall, New York, pp 37-64
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Baas R, van der Werf A, Lambers H (1989) Root respiration and growth in Plantago major as affected by vesicular-arbuscular mycorrhizal infection. Plant Physiol91: 227-232 Bensadoun A, Weinstein D (1976) Assay of proteins in the presence of interfering materials. Anal Biochem 7 0 241-250 Boyetchko SM, Tewari JP (1986) A new species of Glomus (Endogonaceae, Zygomycotina),mycorrhizal with barley in Alberta. Can J Bot 6 4 90-95 Cataldo DA, Haroon M, Schrader LE, Youngs VL (1975) Rapid colorimetric determination of nitrate in plant tissue by nitration of salicylic acid. Commun Soil Sci Plant Anal 6: 71-80 Clarkson DT (1985) Factors affecting mineral nutrient acquisition by plants. Annu Rev Plant Physiol36 77-115 Cogliatti DH, Clarkson DT (1983) Physiological changes in potato plants during development of, and recovery from, phosphate stress. Physiol Plant 58: 287-294 Cress WA, Throneberry GO, Lindsey DL (1979) Kinetics of phosphorus absorption by mycorrhizal and nonmycorrhizal tomato roots. Plant Physiol64 484-487 Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Colorimetric method for determination of sugars and related substances. Anal Chem 2 8 350- 356 Duff SMG, Moorhead GBG, Lefebvre DD, Plaxton WC (1989) Phosphate starvation inducible ‘bypasses’ of adenylate and phosphate dependent glycolytic enzymes in Brassica nigra suspension cells. Plant Physiol90 1275-1278 Dwelle RB (1985) Photosynthesis and assimilate partitioning. In PH Li, ed, Potato Physiology. Academic Press, Toronto, Canada, pp 36-58 Fawcett JK, Scott JE (1960) A rapid and precise method for the determination of urea. J Clin Patholl3: 156-159 Fredeen AL, Rao IM, Terry N (1989) Influence of phosphorus nutrition on growth and carbon partitioning in Glycine max. Plant Physiol89 225-230 Giovanetti M, Mosse B (1980) An evaluation of techniques for measuring vesicular-arbuscular mycorrhizal infection in roots. New Phytol84 489-500 Goldstein AH, Baertlein DA, McDaniel RG (1988) Phosphate starvation inducible metabolism in Lycopersicon esculentum. Plant Physiol87: 711-714 Harley JL, Smith SE (1983) Mycorrhizal Symbiosis.Academic Press, London Harris PM (1978) Mineral nutrition. In PM Harris, ed, The Potato Crop. Chapman and Hall, London, pp 195-243 Hunt R (1982) Plant Growth Curves. Edward Amold, London Krauss A (1985) Interaction of nitrogen nutrition, phytohormones, and tuberization. In PH Li, ed, Potato Physiology. Academic Press, Toronto, Canada, pp 209-230 Kucey RMN, Janzen HH (1987) Effects of VAM and reduced nutrient availability on growth and phosphorus and micronutrient uptake of wheat and field beans under greenhouse conditions. Plant SoillO4 71-78 Lambert DH, Baker DE, Cole H (1979) The role of mycorrhizae in the interaction of phosphorus with zinc, copper, and other elements. Soil Sci SOCAm J 4 3 976-980 Lee RB (1982) Selectivityand kinetics of ion uptake by barley plants following nutrient deficiency. Ann Bot 5 0 429-449 Lefebvre DD, Duff SMG, Fife CA, Julien-Inalsingh C, Plaxton WC (1990) Response to phosphate deprivation in Brassica nigra suspension cells. Plant Physiol93 504-511 Loneragan JF, Grove TS, Robson AD, Snowball K (1979) Phosphorus toxicity as a factor in zinc-phosphorus interactions in plants. Soil Sci SOCAm J 4 3 966-972 MacKay DC, Carefoot JM, Entz T (1988) Detection and correction of midseason P deficiency in irrigated potatoes. Can J Plant Sci 6 8 523-534 Marschner H (1986) Mineral Nutrition of Higher Plants. Academic Press, London McArthur DAJ, Knowles NR (1992) Resistance responses of potato to vesicular-arbuscular mycorrhizal fungi under varying abiotic phosphorus levels. Plant Physiol 100 341-351
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Plant Physiol. Vol. 102, 1993
limiting leaf expansion of phosphorus-deficient cotton plants. Plant Physiol7!j: 372-377 Robson AD, Pitnnan MG (1983) Interactions between nutrients in higher plants. In A Lauchli, RL Bieleski, eds, Inorganic 1Plant Nutrition. Springer-Verlag,New York, pp 147-179 Rosen H (1956)A modified ninhydrin colorimetric analysis for amino acids. Arch Biochem Biophys 67: 10-15 Rufty TW, MacKown CT, Israel DW (1990) Phosphorus stress effects on assimilation of nitrate. Plant Physiol94 328-333 Sanders FE, Tinker PB, Black RLB, Palmerley SM (1977) The development of' endomycorrhizal root systems. I. Spread of infection and growth-promoting effects with four species of vesiculararbuscular endophyte. New Phytol78 257-268 Serrano R, Kanner BI, Racker E (1976) Purification and properties of the proton-translocating adenosine triphosphatase complex of bovine heart mitochondria. J Biol Chem 251: 2453-2461 Skinner PW, Matthews MA (1990) A nove1 interaction of magnesium translocation with the supply of phosphorus to roots of grapevine (Vitir; vinifera L.). Plant Cell Environ 1 3 821-826 Smith SE, Gianinazzi-Pearson V (1988) Physiological interactions between symbionts in vesicular-arbuscular mycorrhizal plants. Annu Rev Plant Physiol Plant Mo1 Biol 3 9 221-244 Somogyi M (1952) Notes on sugar determination. J Biol Chem 1 9 5 19-23 Woolley DJ, Wareing PF (1972) The interaction between growth promoters in apical dominance. 11. Environmental effects 011 endogenous cytoltinin and gibberellin levels in Solanum andigena. New Phytol71: 1015-1025
