Christian R. Jensen

Soluble Invertase Expression Is an Early Target of Drought Stress during the Critical, Abortion-Sensitive Phase of Young Ovary Development in Maize

To distinguish their roles in early kernel development and stress, expression of soluble (Ivr2) and insoluble (Incw2) acid invertases was analyzed in young ovaries of maize (Zea mays) from 6 d before (−6 d) to 7 d after pollination (+7 d) and in response to perturbation by drought stress treatments. The Ivr2 soluble invertase mRNA was more abundant than the Incw2 mRNA throughout pre- and early post-pollination development (peaking at +3 d). In contrast,Incw2 mRNAs increased only after pollination. Drought repression of the Ivr2 soluble invertase also preceded changes in Incw2, with soluble activity responding before pollination (−4 d). Distinct profiles of Ivr2and Incw2 mRNAs correlated with respective enzyme activities and indicated separate roles for these invertases during ovary development and stress. In addition, the drought-induced decrease and developmental changes of ovary hexose to sucrose ratio correlated with activity of soluble but not insoluble invertase. Ovary abscisic acid levels were increased by severe drought only at −6 d and did not appear to directly affect Ivr2 expression. In situ analysis showed localized activity and Ivr2 mRNA for soluble invertase at sites of phloem-unloading and expanding maternal tissues (greatest in terminal vascular zones and nearby cells of pericarp, pedicel, and basal nucellus). This early pattern of maternal invertase localization is clearly distinct from the well-characterized association of insoluble invertase with the basal endosperm later in development. This localization, the shifts in endogenous hexose to sucrose environment, and the distinct timing of soluble and insoluble invertase expression during development and stress collectively indicate a key role and critical sensitivity of the Ivr2soluble invertase gene during the early, abortion-susceptible phase of development.

Drought stress effect on carbohydrate concentration in soybean leaves and pods during early reproductive development: its implication in altering pod set

Abstract To advance our understanding of the physiological mechanisms by which pod abortion is regulated in soybean ( Glycine max L. Merr.), leaf photosynthetic rates, carbohydrate concentrations and soluble invertase activities in leaves, flowers and pods were determined in plants subjected to drought stress during early reproductive development. Soybeans were grown in pots in an environmentally controlled glasshouse. Drought stress was imposed from −11 to 10 days after anthesis (DAA). Drought decreased photosynthetic rates and water potentials in leaves, flowers and pods. Drought decreased leaf sucrose and starch concentrations but increased hexose (glucose+fructose) concentrations. Drought did not affect the activity of soluble invertase in leaves. In flowers and pods, sucrose concentrations were higher under drought as compared with well-watered controls. Hexose and starch concentrations of flowers and pods were also higher under drought until 7 and 3 DAA, respectively; thereafter they were significantly lower than those of the well-watered controls. Soluble invertase activity was decreased by drought in pods 5–10 DAA. Although the concentrations of non-structural carbohydrate (sucrose+hexose+starch) were higher in droughted flowers and pods, the total amount of non-structural carbohydrate accumulated in pods during the sampling periods was substantially reduced by drought. Pod growth was decreased by drought 3–5 DAA, which coincided with a decrease in hexose to sucrose ratio, when a significant reduction of pod set was occurred. Collectively, the results indicate that both source and sink restrictions are involved in regulating pod set in drought-stressed soybeans. It is suggested that a low availability of current and reverse photosynthate in leaves coupled with an impaired ability to utilize the incoming sucrose by pods resulted in a decreased carbohydrate flux from leaves to pods, together with a decreased hexose to sucrose ratio in pods are potential factors contributing to pod abortion in drought-stressed soybeans.

Loss of pod set caused by drought stress is associated with water status and ABA content of reproductive structures in soybean

Drought stress occurring during flowering and early pod expansion decreases pod set in soybean (Glycine max L. Merr.). The failure of pod set may be associated with changes in water status and ABA content in soybean reproductive structures under drought stress. To test this, pot experiments in an environmentally-controlled greenhouse were conducted, in which soybeans were exposed to drought stress around anthesis. In a preliminary experiment (Expt. I), irrigation was withheld at -6 (D1), -4 (D2) and -2 (D3) to 11 days after anthesis (DAA), then the droughted plants were re-watered to control levels until physiological maturity. Pod set percentage, seed yield and yield components were recorded. In the main experiment (Expt. II), irrigation was withheld from -11 to 10DAA. During the drying cycle, parts of the droughted plants were re-watered at 0, 3, 5, 7 and 10 DAA and kept well-watered until physiological maturity. In Expt. II, water status, ABA contents in xylem sap, leaves, flowers and pods were measured at 0, 3, 5, 7 and 10 DAA. The water potential in the flowers and pods was always lower than the leaf water potential. Turgor was decreased in leaves by drought 3 DAA, but remained at control levels in flowers and pods. Compared with well-watered plants, in severely droughted plants (10 DAA), xylem [ABA] increased about 60-fold; leaf [ABA] increased 9-fold; pod [ABA] increased 6-fold. During soil drying, flower and pod [ABA] was linearly correlated with xylem [ABA] and leaf [ABA], indicating that root-originated ABA and/or leaf ABA were the likely sources of ABA accumulated in the flowers and pods. In Expt. I, pod set and seed number per pod was unaffected by drought stress, while seed yield and individual seed weight was significantly decreased by drought. In Expt. II, significant reductions in pod set and seed yield were observed when re-watering the droughted plants at 3-5 DAA, re-watering the droughted plants later than this stage resulted in a similar pod set. Collectively, these results suggest that drought-induced decrease in water potential and increase in ABA content in flowers and pods at critical developmental stage (3-5 DAA) contribute to pod abortion in soybean.

A review of drought adaptation in crop plants: changes in vegetative and reproductive physiology induced by ABA-based chemical signals

This review discusses the role of abscisic acid (ABA)-based drought stress chemical signalling in regulating crop vegetative and reproductive development and its contributions to crop drought adaptation. Increased concentrations of ABA in the root induced by soil drying may maintain root growth and increase root hydraulic conductivity; both lead to an increase in water uptake and thereby postpone the development of water deficit in the shoot. Root ABA is also transported in the xylem to the shoot and is perceived at the acting sites, where it causes stomatal closure and reduced leaf expansion, thereby preventing dehydration of leaf tissues and enhancing the chance for survival under prolonged drought. ABA-based chemical signalling can be amplified by several factors, particularly increased pH in the xylem/apoplast, which retains anionic ABA. Such an increase in xylem pH detected in field-grown maize might have been brought about by reduced nitrate uptake by plants during soil drying. In contrast, xylem sap alkalinisation was not found in soybeans, which depend on fixing nitrogen through their association with Rhizobium japonicum. Evidence has also shown that the xylem-borne ABA can be transported to plant reproductive structures and influence their development, presumably by regulating gene expression that controls cell division and carbohydrate metabolic enzyme activity under drought conditions. The possible involvement of ABA in the up- and down-regulation of acid invertase in crop source (adult leaves) v. sink (young ovaries) organs indicates a crucial role of the hormone in balancing source and sink relationship in plants according to the availability of water in the soil. A novel irrigation technique named partial root-zone drying (PRD), has been developed to allow exploitation of ABA-based drought stress signalling to improve water-use efficiency (WUE) based on its roles in regulating stomatal aperture and leaf expansion. However, little is known about how crop reproductive development is regulated when irrigated under PRD. We suggest that more attention should be paid to the latter aspect as it directly relates to crop yield and quality.

Hydraulic and chemical signals in the control of leaf expansion and stomatal conductance in soybean exposed to drought stress

Both hydraulic and chemical signals are probably important in regulating leaf growth and stomatal conductance of soybean (Glycine max L. Merr.) under drought stress. However, until now they have not been investigated concomitantly in this species. To explore this, a pot experiment in a temperature-regulated greenhouse was conducted, in which plants were subjected to progressive drought during early reproductive stages. Biophysical parameters, viz. relative leaf expansion rate, stomatal conductance, leaf turgor, leaf [ABA], xylem pH and xylem [ABA] were followed in control and stressed plants. Drought stress decreased relative leaf expansion rate, stomatal conductance and leaf turgor, whereas it increased leaf [ABA], xylem pH and xylem [ABA]. As soil dried, significant differences between water treatments for relative leaf expansion rate, stomatal conductance, leaf turgor, leaf [ABA], xylem pH and xylem [ABA] were observed at 14, 9, 14, 14, 14 and 9 d after imposition of stress, respectively. The relationships of relative values for relative leaf expansion rate, stomatal conductance, leaf turgor, leaf [ABA] and xylem pH to the fraction of transpirable soil water (FTSW) were well described by linear-plateau functions that allowed calculation of the soil-water thresholds at which processes in stressed plants began to diverge from well-watered controls. The soil-water threshold for stomatal conductance (0.64) was significantly higher than that for relative leaf expansion rate (0.29), xylem pH (0.28), leaf [ABA] (0.27) and leaf turgor (0.25). Relative xylem [ABA] increased, first linearly (when FTSW > 0.5) and then exponentially (when FTSW < 0.5) with decreasing FTSW. Relative stomatal conductance decreased exponentially with increasing relative xylem [ABA] (r2=0.98). Decreased stomatal conductance coincided with an increase in xylem [ABA] and occurred before any significant change of leaf turgor could be detected, indicating that chemical signals (seemingly root-originated ABA) control stomatal behaviour at moderate soil water deficits. Relative relative leaf expansion rate was linearly correlated with relative leaf turgor (r2=0.93), relative xylem pH (r2=0.97) and relative leaf [ABA] (r2=0.98), implying that both hydraulic and chemical signals were probably involved in regulation of leaf expansion at severe soil water deficits.

Improved plant nitrogen nutrition contributes to higher water use efficiency in tomatoes under alternate partial root-zone irrigation

Comparative effects of partial root-zone irrigation (PRI) and deficit irrigation (DI) on stomatal conductance (gs), nitrogen accumulation and distribution in tomato (Lycopersicon esculentum L.) plants were investigated in a split-root pot experiment. Results showed that both PRI and DI saved 25% water and led to 10.0% and 17.5% decreases in dry biomass, respectively, compared with the fully irrigated (FI) controls. Consequently, water use efficiency (WUE) was increased by 18.6% and 10.8% in the PRI and DI plants, respectively. The highest WUE in the PRI plants was associated with the highest carbon isotope composition (δ13C), indicating that the improvement of WUE might have been a result of long-term optimisation of stomatal control over gas exchange. The constantly higher xylem sap ABA concentration in PRI compared with DI plants was seemingly responsible for the greater control over stomatal conductance during the treatment. At the end of the experiment, N accumulation and 15N recovery was highest in FI, intermediate in PRI and lowest in DI. In addition, PRI plants consistently allocated more N into the upper and middle leaf layers than in the FI and DI treatments. The improved N nutrition and distribution in the canopy may indicate that PRI plants have a greater photosynthetic capacity than DI plants; this is confirmed by the observed positive linear relationship between specific leaf N content and δ13C. It is concluded that PRI improves N nutrition and optimises N distribution in the canopy, which might have been partly contributed to the high WUE in PRI tomato plants.

The effect of lupins as compared with peas and oats on the yield of the subsequent winter barley crop

Abstract New high yielding early maturing cultivars of lupins have been introduced in north-west Europe as grain protein crops in crop rotations. This paper reports on a comparative study of lupins with peas and oats, and of their effect on yield of subsequent winter barley crops. These crops were given five levels of N under irrigated and non-irrigated conditions on sand and loam. Under rain fed conditions the grain yield of pea, oat and lupin varied between 24–36, 34–53 and 18–37 hkg DM ha−1, respectively. Supplemental irrigation raised grain yield of oat to 50–60 hkg DM ha−1, while grain yield in pea was not affected and grain yield in lupin in most cases decreased due to gray mould attack and excessive vegetative growth in the indeterminate lupin variety. Under rain fed conditions, the grain nitrogen content of pea, oat and lupin varied between 137–172, 61–80 and 189–226 kg N ha−1, respectively, and was significantly higher in lupin as compared with pea. On sandy soil, similar low-root densities were found for pea, oat and lupin below 30 cm depth. On sand, at final harvest the residual soil-N of lupin and pea, as measured in a subsequent winter barley crop not supplied with N fertilizer, was 15 and 8–10 kg N ha−1 higher than in winter barley following oat, respectively. The nature of the probably more N-root residues of lupin is discussed. On loam, the residual N of lupin and pea was similar, 18–27 kg N ha−1. On sand, under rain fed conditions preceding lupin and pea as compared with oat, increased the barley grain yield at zero N-application 77 and 49%, respectively; the effect of lupin was significantly higher than that of pea until the highest N-level 120 kg N-application ha−1. On loam under rain fed conditions preceding lupin and pea increased the barley grain yield at zero N-application by 36 and 62%, respectively, as compared with oat; at N-application >60 kg N ha−1 the grain yield was similar after all three crops. For both soil types the same level of effect was found under irrigated conditions. Conclusions: Supplemental irrigation might result in lower grain yield in lupin due to gray mould attack and excessive growth if indeterminate lupin varieties are used. Grain nitrogen yield of lupin is significantly higher than that of pea. On sand, the effect of lupin on the subsequent winter barley grain yield is significantly higher than that of pea, probably due to greater N-root nitrogen residues. On loam, lupin and pea have similar effects on the subsequent winter barley crop.

Changes in carbohydrates, ABA and bark proteins during seasonal cold acclimation and deacclimation in Hydrangea species differing in cold hardiness

Cold injury is frequently seen in the commercially important shrub Hydrangea macrophylla but not in Hydrangea paniculata. Cold acclimation and deacclimation and associated physiological adaptations were investigated from late September 2006 to early May 2007 in stems of field-grown H. macrophylla ssp. macrophylla (Thunb.) Ser. cv. Blaumeise and H. paniculata Sieb. cv. Kyushu. Acclimation and deacclimation appeared approximately synchronized in the two species, but they differed significantly in levels of mid-winter cold hardiness, rates of acclimation and deacclimation and physiological traits conferring tolerance to freezing conditions. Accumulation patterns of sucrose and raffinose in stems paralleled fluctuations in cold hardiness in both species, but H. macrophylla additionally accumulated glucose and fructose during winter, indicating species-specific differences in carbohydrate metabolism. Protein profiles differed between H. macrophylla and H. paniculata, but distinct seasonal patterns associated with winter acclimation were observed in both species. In H. paniculata concurrent increases in xylem sap abscisic acid (ABA) concentrations ([ABA](xylem)) and freezing tolerance suggests an involvement of ABA in cold acclimation. In contrast, ABA from the root system was seemingly not involved in cold acclimation in H. macrophylla, suggesting that species-specific differences in cold hardiness may be related to differences in [ABA](xylem). In both species a significant increase in stem freezing tolerance appeared long after growth ceased, suggesting that cold acclimation is more regulated by temperature than by photoperiod.

Ovary abscisic acid concentration does not induce kernel abortion in field-grown maize subjected to drought

This study investigated the effects of drought of different duration and severity on ovary abscisic acid (ABA) concentration and yield components in field-grown maize (Zea mays L. cv. Loft). The study was conducted in a field lysimeter of the Royal Veterinary and Agricultural University (KVL) in Hojbakkegaard (55°40′N; 12°18′E; 28 masl), Denmark in 1997. Irrigation was withheld at four different dates to induce drought of different duration and severity at the reproductive stage of the plants. Plots were re-watered shortly after silking and kept at field capacity for the remainder of the season. Soil water status, plant height, and early morning leaf water potential were monitored during the treatment. Ovary ABA concentration was determined at four dates before and after fertilization. Final grain yield, total DM, harvest index (HI), mean kernel weight, kernel weight distribution, and kernel number per cob were determined at maturity. Plant height was significantly (P<0.05) reduced by 40 and 25%, respectively, in the two most severe drought treatments. In the two shorter drought treatments no effect of drought stress on plant height or biomass was observed. Leaf water potential decreased slowly as a function of relative available soil water content and resulted in −0.4 MPa at the end of the longest and −0.12 MPa at the end of the shortest stress period. Under fully watered conditions, plot yields averaged 1400 g m−2 for total dry matter (DM) and 700 g m−2 for grain yield, with a HI of about 0.5. Initiation of a drying cycle close to flowering did not change yields. Long drying cycles resulted in significant (P<0.05) yield reductions up to 70% of the fully watered controls. Kernel number per cob was reduced up to 60% under long drought conditions and not affected under short-term drought. Drought imposed about two weeks prior to fertilization resulted in 30% reduction in kernel number per cob, but this effect was balanced by an increase of 25% in mean kernel weight. Long and severe drought increased ovary ABA concentration prior to fertilization, whereas short-term drought did not. At fertilization no increase of ovary ABA as compared to fully watered controls was found in any treatment. It is concluded that drought induced grain yield losses in field grown maize cannot be attributed to kernel size reduction or kernel abortion due to ovary ABA concentrations as reported by some authors for studies on maize and wheat under controlled conditions, as ovary ABA concentrations peaked before zygote formation and endosperm development.

Comparative effects of deficit irrigation and alternate partial root-zone irrigation on xylem pH, ABA and ionic concentrations in tomatoes

Comparative effects of partial root-zone irrigation (PRI) and deficit irrigation (DI) on xylem pH, ABA, and ionic concentrations of tomato (Lycopersicon esculentum L.) plants were investigated in two split-root pot experiments. Results showed that PRI plants had similar or significantly higher xylem pH, which was increased by 0.2 units relative to DI plants. Nitrate and total ionic concentrations (cations+anions), and the proportion of cations influenced xylem pH such that xylem pH increases as nitrate and total ionic concentrations decrease, and the proportion of cations increases. In most cases, the xylem ABA concentration was similar for PRI and DI plants, and a clear association between increases in xylem pH with increasing xylem ABA concentration was only found when the soil water content was relatively low. The concentrations of anions, cations, and the sum of anions and cations in PRI were higher than in the DI treatment when soil water content was relatively high in the wetted soil compartment. However, when water content in both soil compartments of the PRI pots were very low before the next irrigation, the acquisition of nutrients by roots was reduced, resulting in lower concentrations of anions and cations in the PRI than in the DI treatment. It is therefore essential that the soil water content in the wet zone should be maintained relatively high while that in the drying soil zone should not be very low, both conditions are crucial to maintain high soil and plant water status while sustaining ABA signalling of the plants.