Low temperature effect on selective fertilization by pollen mixtures of wild and cultivated tomato species

Summary In vitro pollen germination of cultivated tomato, Lycopersicon esculentum Mill., is inhibited by an ambient temperature of 5°C, more so than pollen from a Peruvian ecotype of Lycopersicon hirsutum Humb. & Bonpl. originating from an altitude of 3200 m. The frequency of L. hirsutum gametes contributing to hybrid zygote formation is more than doubled when controlled fertilizations with pollen mixtures of the two species occurs at 12/6°C as compared to crosses with the same mixtures at 24/19°C. The results suggest that differential selection at the gametophytic level occurs in response to low temperature regimes. To our knowledge this is the first time in higher plants that alteration of an environmental factor has been demonstrated to change selection values of male gametophytes in a fashion predicted by the ecology of the parental sporophytes.     

Pollen selection for low temperature adaptation in tomato

Summary Pollen selection experiments were conducted in tomato to determine the effects of low temperature conditions during pollination on the rate of root elongation of the progeny. Pollen was harvested from an F₁ interspecific hybrid between a high altitude Lycopersicon hirsutum accession and the cultivated tomato L. esculentum. The pollen was applied to stigmas of malesterile L. esculentum plants maintained in growth chambers set at either 12°C/7°C or 24°C/18°C. BC₁ seeds from the low and normal temperature crosses were germinated and root elongation rate was measured at either 9°C or 24°C. At 9°C, the rate of root elongation for progeny of the low temperature crosses was higher than for progeny of crosses at normal temperatures; at 24°C the rate of root elongation was similar for the two crossing treatments. To compare the temperature responses of the two backcross populations we also calculated the relative inhibitory effect of low temperature on the rate of root elongation: the ratio between the rate of root elongation at 9°C to that at 24°C. Root elongation of seedlings from the low temperature crosses was less inhibited by the cold than root elongation for progeny of the normal temperature crosses. These results suggest a relationship between pollen selection at low temperatures and the expression of a sporophytic trait under the same environmental stress.     

Effects of chilling on the biochemical and functional properties of thylakoid membranes

The mechanism of chilling resistance was investigated in 4-week-old plants of the chilling-sensitive cultivated tomato, Lycopersicon esculentum Mill. cv H722, and rooted cuttings of its chilling-resistant wild relative, L. hirsutum Humb. and Bonpl., which were chilled for 3 days at 2°C with a 14-hour photoperiod and light intensity of 250 micromoles per square meter per second. This chilling stress reduced the chlorophyll fluorescence ratio, stomatal conductance, and dry matter accumulation more in the sensitive L. esculentum than in the resistant L. hirsutum. Photosynthetic CO₂ uptake at the end of the chilling treatment was reduced more in the resistant L. hirsutum than in L. esculentum, but recovered at a faster rate when the plants were returned to 25°C. The reduction of the spin trap, Tiron, by isolated thylakoids at 750 micromoles per square meter per second light intensity was taken as a relative indication of the tendency for the thylakoids to produce activated oxygen. Thylakoids isolated from the resistant L. hirsutum with or without chilling treatment were essentially similar, whereas those from chilled leaves of L. esculentum reduced more Tiron than the nonchilled controls. Whole chain photosynthetic electron transport was measured on thylakoids isolated from chilled and control leaves of the two species at a range of assay temperatures from 5 to 25°C. In both species, electron transport of the thylakoids from chilled leaves was lower than the controls when measured at 25°C, and electron transport declined as the assay temperature was reduced. However, the temperature sensitivity of thylakoids from chilled L. esculentum was altered such that at all temperatures below 20°C, the rate of electron transport exceeded the control values. In contrast, the thylakoids from chilled L. hirsutum maintained their temperature sensitivity, and the electron transport rates were proportionately reduced at all temperatures. This sublethal chilling stress caused no significant changes in thylakoid galactolipid, phospholipid, or protein levels in either species. Nonchilled thylakoid membranes from L. hirsutum had fourfold higher levels of the fatty acid 16:1, than those from L. esculentum. Chilling caused retailoring of the acyl chains in L. hirsutum but not in L. esculentum. The chilling resistance of L. hirsutum may be related to an ability to reduce the potential for free radical production by close regulation of electron transport within the chloroplast.     

Sensitivity of Altitudinal Ecotypes of the Wild Tomato Lycopersicon hirsutum to Chilling Injury

The transition temperature of the leaf polar lipids and the critical temperature for chill-induced inhibition of photosynthesis was determined for three altitudinal ecotypes of the wild tomato Lycopersicon hirsutum. Photosynthesis was measured as CO₂-dependent O₂ evolution at 25°C after leaf slices were exposed to chilling temperatures for 2 hours at a moderate photon flux density of 450 micromoles per square meter per second. The transition temperature of the leaf polar lipids was detected from the change in the temperature coefficient of the fluorescence intensity of trans-parinaric acid. Chill-induced photoinhibition was evident in the three tomato ecotypes when they were chilled below a critical temperature of 10°, 11°, and 13°C, respectively, for the high (LA1777), mid (LA1625), and low (LA1361) altitudinal ecotypes. The temperature differential, below the critical temperature, required to produce a 50% inhibition was also similar for the three ecotypes. A transition was detected in the leaf polar lipids of these plants at a temperature similar to that of the critical temperature for photoinhibition. The results show that the three tomato ecotypes are similar with respect to their critical temperature for chilling-induced photoinhibition and the rate of their response to the chilling stress. They are, thus, similarly sensitive to chilling.     

Tomato pollen development: stages sensitive to chilling and a natural environment for the selection of resistant genotypes

Abstract The time during which pollen development is most sensitive to chilling was investigated. Five cultivars of tomato (Lycopersicon esculentum Mill.) bearing flower buds at different stages of development were kept at 7°C for 1 week under 12-h light periods, during which time growth stopped. After returning the plants to minimum temperatures of 18°C, the presence of chromatin in the pollen was assessed daily as the flowers reached anthesis. The results suggested that there are two stages of acute sensitivity to cold during pollen development, each of which results in cold-stressed plants having pollen empty of chromatin. The first and most sensitive stage is about 11.2 d (SE = 0.3 d) before anthesis, and this is followed by a second stage of sensitivity about 5.6±0.2 d before anthesis. Flowers that had wholly developed under simulated natural temperatures that decreased diurnally from a maximum of 18°C to a minimum of 7°C also had defective pollen, but pollen of normal appearance was regained within 14°d on return to higher temperatures. Plants of L. esculentum, and a form (LA 1363) of the wild species L. hirsutum from high altitudes in the Andes, as well as F1 and F3 generations of their hybrid, were grown to the flowering stage at an altitude of 600 m in Hawaii and then grown for a further 30°d at 2000 m, where night temperature was below 10°C. The high altitude environment severely affected the quality of pollen produced and its release from the stamen in L. esculentum, but not in L. hirsutum LA 1363. The results with the hybrids suggested that such tropical mountain environments can be used as a natural phytotron in the selection of chilling resistance that is only expressed in the mature plant.     

Assay of chilling injury in wild and domestic tomatoes based on photosystem activity of the chilled leaves

Tomato leaves were detached and stored at 0 C for various periods of time. Chloroplasts were isolated from the leaves and their photoreductive activities were determined. Comparisons were made between two altitudinal forms of the wild tomato Lycopersicon hirsutum Humb. and Bonpl. (a tropical lowlands form and a highlands form adapted to growth at 3,100 meters), and two cultivars of the domestic tomato L. esculentum Mill. In each case the capacity of the isolated chloroplasts to photoreduce ferricyanide declined linearly with time of storage of the leaves at 0 C, but not at 10 C. This injury developed more slowly in the high altitudinal form of the wild tomato compared with the low altitudinal form and the two domestic cultivars indicating an enhanced resistance toward chilling injury in the tomato from 3,100 meters. Chloroplast activity declined in green tomato fruit held at 0 C, at about the same rate as in the chilled leaves.     

Effect of Chilling Temperatures on the Protoplasmic Streaming of Plants from Different Climates

A microscope mount was designed so that specimen temperaturescould be monitored and controlled without impairing phase contrastoptics and used to measure rates of protoplasmic streaming between0 and 25 ?C in trichome cells of Lycopersicon esculentum, Lycopersiconhirsutum, Citrullus vulgaris, Tradescantia albiflora, Digitalispurpurea, and Veronica persica. Between 10 and 20 ?C the rates of streaming varied from 2–6µm s^–1 depending on the temperature, and differencesbetween the species were small. The temperature coefficientof streaming rates was found to increase as the temperaturewas lowered so that the plot of log rate against temperaturehad a steeper slope at the lower temperatures. The largest temperature cofficients were for the warmth-requiringL. esculentum (tomato) and C. vulgaris (water melon), and thesmallest for the temperate-zone plants V. persica (speedwell)and D. purpurea (foxglove). The changes in rate always occurredover a range of temperature; no ‘critical temperature’wasobserved below which streaming abruptly stopped and above whichit was active, although the amount of streaming as well as therate decreased as the lowest temperatures were approached. The temperatures experienced by the specimens during the experimentdid not affect the recovery of normal streaming rates betweenabout 10 and 20 ?C. In a population of a wild tomato, Lycopersicon hirsutum Humb.and Bonpl., collected from different altitudes in Peru and Ecuador,i.e. from locations of different environmental temperature,the rate of protoplasmic streaming at 5 ?C was greatest in thevarieties collected from the highest altitudes. The resultssuggest that streaming rates correlate with genetic adaptationto low temperature in the species examined.     

Variation among Species in the Temperature Dependence of the Reappearance of Variable Fluorescence following Illumination

The relationship between the thermal dependence of the reappearance of chlorophyll variable fluorescence following illumination and temperature dependence of the apparent Michaelis constant (K_m) of NADH hydroxypyruvate reductase for NADH was investigated in cool and warm season plant species. Brancker SF-20 and SF-30 fluorometers were used to evaluate induced fluorescence transients from detached leaves of wheat (Triticum aestivum L. cv TAM-101), cotton (Gossypium hirsutum L. cv Paymaster 145), tomato (Lycopersicon esculentum cv Del Oro), bell pepper (Capsicum annuum L. cv California Wonder), and petunia (Petunia hybrida cv. Red Sail). Following an illumination period at 25°C, the reappearance of variable fluorescence during a dark incubation was determined at 5°C intervals from 15°C to 45°C. Variable fluorescence recovery was normally distributed with the maximum recovery observed at 20°C in wheat, 30°C in cotton, 20°C to 25°C in tomato, 30 to 35°C in bell pepper and 25°C in petunia. Comparison of the thermal response of fluorescence recovery with the temperature sensitivity of the apparent K_m of hydroxypyruvate reductase for NADH showed that the range of temperatures providing fluorescence recovery corresponded with those temperatures providing the minimum apparent K_m values (viz. the thermal kinetic window).     

Temperature-induced leakage from chilling-sensitive and chilling-resistant plants

Leakage rates were determined from leaf cells loaded with rubidium and [³H]leucine. There was a differential response between leucine and rubidium leakage depending upon the species used. The rate of leucine leakage shows a small decline below 5 C for two altitudinal variants of Lycopersicon hirsutum Humb. and Bonpl., whereas Lycopersicon esculentum L. showed a marked increase below 5 C. Rubidium showed a marked increase in leakage rate below 10 C with the altitudinal variants, with only a slight increase for the L. esculentum species. A rough relationship existed between rubidium leakage rate at 1 C and the altitude of origin of the L. hirsutum race, the low altitudinal forms having higher leakage rates than the higher altitudinal variants. The L. esculentum lines show a rubidium leakage response similar to that of the high altitude L. hirsutum variants. Higher leakage rates were obtained if the calcium concentration in the medium was less than 1 millimolar and upon addition of metabolic poisons and detergents.     

Competence of the pollen of wild species and the cultivar of tomato to affect fertilization at low temperature

Summary The competitive ability of the pollen of wild tomato species (Solanum pennellii, Lycopersicum hirsutum and L. minutum) and a cultivar of tomato to affect fertilization at low temperature was investigated. Pollen grains of wild species were mixed with those of the cultivar and pollinated onto the stigma of the cultivar. The number of seedlings resembling the wild species, excluding L. minutum, were significantly higher when the plants were maintained at a low temperature (8°–12° C) than when they were maintained at a normal temperature (22°–28° C). This indicates that the pollen of S. pennellii and L. hirsutum wild species can compete better with pollen of the cultivar at a low temperature, while there was no change in the competitive ability of L. minutum pollen under these conditions.     

Re-Aeration following Hypoxia or Anoxia Leads to Activation of the Antioxidative Defense System in Roots of Wheat Seedlings

We previously reported that short exposure of tomato (Lycopersicon esculentum L.) fruits to high temperature protects them from chilling injury. To study the involvement of heat-shock proteins (HSPs) in the acquisition of low-temperature tolerance, we cloned two heat-shock-induced genes that are also expressed at low temperatures. The cloned cDNAs belong to the small HSP group. Sequence analyses of the clones showed perfect homology to the tomato-ripening gene tom66 and to the tomato chloroplastic HSP21 gene tom111. The expression of both genes was induced by high temperature in fruits, flowers, leaves, and stems, but not by low or ambient temperatures or by other stresses such as drought and anaerobic conditions. When the heated fruits were transferred to low temperature, tom66 and tom111 mRNA levels first decreased but were then reinduced. Induction was not observed in nonheated fruits at low temperature. Immunodetection of tom111-encoded protein indicated that this protein is present at low temperatures in the heated fruits. The results of this study show that the expression of tom66 and tom111 is correlated with protection against some, but not all, symptoms of chilling injury.     

Identification of a short rDNA spacer sequence highly specific of a tomato line containing Tm-1 gene introgressed from Lycopersicon hirsutum

Summary We studied rDNA restriction fragment length polymorphism between two tomato lines used for F₁ hybrid seed production: line A, containing the Tm-1 gene responsible for tobacco mosaic virus tolerance introgressed from the wild species Lycopersicon hirsutum, and line B, a tobacco mosaic virus sensitive line. Hybridization patterns led to distinct rDNA maps with two size classes, 10.4 and 10.7 kb, in line A and a single, 8.9-kb class in line B. Size differences were located in the intergenie sequence (IGS). A highly specific 54-bp TaqI fragment was cloned from the line A IGS and used in dot blot experiments to probe total DNA from line A, line B, and their F₁ hybrid. It proved capable of discriminating B from A and the hybrid. This probe could thus serve to screen inbreds in commercial seed lots where line A is used as male. This fragment showed 80–90% sequence homology with the 53-bp subrepeats previously characterized in a region of the tomato IGS close to the 25S rRNA gene. Preliminary comparison of rDNA in line A and several wild related species indicated that the L. hirsutum H₂ genotype was the closest to line A. rDNA variations between line A and this wild genotype could be explained by recombination during the introgression process involving numerous backcrosses or by an important intraspecific polymorphism. Our results strongly suggest that Tm-1 and the rDNA were introgressed together into tomato from L. hirsutum through linkage drag.     

Water relations under root chilling in a sensitive and tolerant tomato species

The shoots of cultivated tomato (Lycopersicon esculentum cv. T5) wilt if their roots are exposed to chilling temperatures of around 5 °C. Under the same treatment, a chilling‐tolerant congener (Lycopersicon hirsutum LA 1778) maintains shoot turgor. To determine the physiological basis of this differential response, the effect of chilling on both excised roots and roots of intact plants in pressure chambers were investigated. In excised roots and intact plants, root hydraulic conductance declined with temperature to nearly twice the extent expected from the temperature dependence of the viscosity of water, but the response was similar in both species. The species differed markedly, however, in stomatal behaviour: in L. hirsutum, stomatal conductance declined as root temperatures were lowered, whereas the stomata of L. esculentum remained open until the roots reached 5 °C, and the plants became flaccid and suffered damage. Grafted plants with the shoots of one genotype and roots of another indicated that the differential stomatal behaviour during root chilling has distinct shoot and root components.     

Distribution of acid invertase in the tomato plant

Acid invertase activity in Lycopersicon esculentum was highest in the locular wall of ripe fruit and lowest in roots. Activity was greater in leaf laminae than in petiole tissue and increased with leaf age, whereas there was more invertase in the upper part of the stem compared with the older portion. Activity in whole fruit increased with increasing ripeness and was greatest in overripe fruit. Of various tissues from a number of wild tomato species examined, the fruit of L. pimpinellifolium were particularly rich in the enzyme, in contrast to the fruit of L. hirsutum, L. hirsutum, var. glabratum and L. peruvianum which had low activity.     

PCR-generated molecular markers for the invertase gene and sucrose accumulation in tomato

The green-fruited tomato species, Lycopersicon hirsutum, unlike the domesticated red-fruited species, L. esculentum, accumulates sucrose during the final stages of fruit development, concomitant with the loss of soluble acid invertase activity. In order to study the genetic linkage of sucrose accumulation to the invertase gene, part of the invertase gene from L. hirsutum was cloned, sequenced and the sequence compared with the invertase sequence of the red-fruited L. esculentum. Several base changes were found in the coding region of the two invertase genes. Based on these base -pair differences, we developed a species-specific PCR assay capable of determining, in a single PCR reaction, the origin of the invertase gene in segregating seedlings of an interspecific cross. Our results indicate that the invertase gene is genetically linked to sucrose accumulation in the green-fruited L. hirsutum.     

Analysis of Low-temperature Tolerance of a Tomato (Lycopersicon esculentum) Cybrid with Chloroplasts from a more Chilling-tolerant L. hirsutum Accession

Growth and photosynthesis of an alloplasmic tomato (cybrid),i.e. line AH47, containing the nuclear genome of the chilling-sensitivecytoplasmic albino mutant of L. esculentum Mill. ‘LargeRed Cherry’ (LRC) and the plastome of a more chilling-toleranthigh-altitude accession of the related wild species L. hirsutumHumb. & Bonpl. LA 1777, were investigated at an optimal(25/20°C) and suboptimal (16/14°C) day/night temperatureregime and their performance compared with that of both euplasmicparents. The cybrid shoot had a similar biomass and developmentrate to the nuclear tomato (L. esculentum) parent at both temperatureregimes. Compared with the biomass production of shoots grownat optimal temperature, the reduction in shoot biomass at suboptimaltemperature was smaller for L. hirsutum than for L. esculentumand the cybrid. This difference was related to a stronger inhibitionof leaf area expansion in L. esculentum and the cybrid in thesuboptimal temperature regime than in L. hirsutum. Irrespectiveof the temperature regime under which the plants were grown,photosynthetic performance and leaf pigment, carbohydrate andsoluble-protein contents of the cybrid resembled those of thenuclear parent. No advantages of the alien L. hirsutum chloroplastwith respect to growth and photosynthesis-related characteristicswere observed in the cybrid in the suboptimal temperature regime,indicating that the temperature sensitivity of the photosyntheticapparatus is regulated by nuclear genes. An adverse consequenceof interspecific chloroplast transfer was the increased susceptibilityto chill-induced photoinhibition of the cybrid. It is concludedthat cybridization is not a useful tool for improving low-temperaturetolerance of tomato. Copyright 2000 Annals of Botany Company Alloplasmic tomato, chloroplast, cybrid(ization), growth, low-temperature tolerance, Lycopersicon esculentum, L. hirsutum, photosynthesis, plastome, tomato     

A major QTL introgressed from wild Lycopersicon hirsutum confers chilling tolerance to cultivated tomato (Lycopersicon esculentum)

Many plants of tropical or subtropical origin, such as tomato, suffer damage under chilling temperatures (under 10°C but above 0°C). An earlier study identified several quantitative trait loci (QTLs) for shoot turgor maintenance (stm) under root chilling in an interspecific backcross population derived from crossing chilling-susceptible cultivated tomato (Lycopersicon esculentum) and chilling-tolerant wild L. hirsutum. The QTL with the greatest phenotypic effect on stm was located in a 28 cM region on chromosome 9 (designated stm9), and enhanced chilling-tolerance was conferred by the presence of the Lycopersicon hirsutum allele at this QTL. Here, near-isogenic lines (NILs) were used to verify the effect of stm9, and recombinant sub-NILs were used to fine map its position. Replicated experiments were performed with NILs and sub-NILs in a refrigerated hydroponic tank in the greenhouse. Sub-NIL data was analyzed using least square means separations, marker-genotype mean t-tests, and composite interval mapping. A dominant QTL controlling shoot turgor maintenance under root chilling was confirmed on chromosome 9 using both NILs and sub-NILs. Furthermore, sub-NILs permitted localization of stm9 to a 2.7 cM interval within the original 28 cM QTL region. If the presence of the L. hirsutum allele at stm9 also confers chilling-tolerance in L. esculentum plants grown under field conditions, it has the potential to expand the geographic areas in which cultivated tomato can be grown for commercial production.     

Influence of temperature on the in vitro pollen germination and pollen tube growth of various native Iranian almonds (Prunus L. spp.) species

Pollen germination and pollen tube growth was quantified among various native Iranian wild almonds (P. dulcis (Mill.) D. A. Webb, P. eleaegnifolia Mill., P. orientalis Mill., P. lycioides Spach, P. reuteri Bioss. et Bushe, P. arabica Olivier, P. glauca Browick and P. scoparia Spach in order to identify differences in the tolerance of pollen to temperature variations. Pollen germination and pollen tube growth were observed after incubation in darkness in a germination medium for 24?h at 10?C50°C at 5°C intervals. Maximum pollen germination of the wild almond species and specify that 60% was obtained for P. orientalis pollen and 98% for P. scoparia. Pollen tube length ranged from 860???m was obtained in P. lycioides and 1490???m in P. scoparia. A modified bilinear model best described the response to temperature of pollen germination and pollen tube length. Almond species variation was found for cardinal temperatures (T _min, T _opt and T _max) of pollen germination percentage and pollen tube growth. Mean cardinal temperatures averaged over eight almond species were 14.7, 24.2, and 43.7°C for maximum percentage pollen germination and 14.48, 25.3, and 44.4 °C for maximum pollen tube length. The principal component analysis (PCA) identified maximum percentage pollen germination and pollen tube length of the species, and T _max for the two processes as the most important pollen parameters in describing a species tolerance to high temperature. PCA also classified Prunus L. spp. into four groups according to the tolerance of pollen to temperature variations. The T _min and T _opt for pollen germination and tube growth, rate of pollen tube growth were less predictive in discriminating species for high temperature tolerance.     

Anther response to high-temperature stress during development and pollen thermotolerance heterosis as revealed by pollen tube growth and in vitro pollen vigor analysis in upland cotton

Main conclusion

Pollen tube growth in styles was strongly inhibited by temperature above 35 °C, and the yield of cotton decreased because of the adverse effect of high temperatures during square development. High-temperature stress during flowering influences the square development of upland cotton (Gossypium hirsutum L.) and cotton yield. Although it is well known that square development is sensitive to high temperature, high-temperature sensitive stages of square development and the effects of high temperature on pollen tube growth in the styles are unknown. The effect of high temperature on anther development corresponding to pollen vigor is unknown during anther development. The objectives of this study were to identify the stages of square development that are sensitive to high temperatures (37/30 and 40/34 °C), to determine whether the abnormal development of squares influenced by high temperature is responsible for the variation in the in vitro germination percent of pollen grains at anthesis, to identify the effect of high temperature on pollen germination in the styles, and to determine pollen thermotolerance heterosis. Our results show that the stages from the sporogenous cell to tetrad stage (square length <6.0 mm) were the most sensitive to high temperature, and the corresponding pollen viability at anthesis was consistent with the changes in the square development stage. Pollen tube growth in the styles was strongly inhibited by temperature above 35 °C, and the yield of cotton decreased because of the effect of high temperature during square development. The thermotolerance of hybrid F₁ pollen showed heterosis, and pollen viability could be used as a criterion for screening for high-temperature tolerance cultivars. These results can be used in breeding to develop new cotton cultivars that can withstand high-temperature conditions, particularly in a future warmer climate.

     

High temperature limits in vivo pollen tube growth rates by altering diurnal carbohydrate balance in field-grown Gossypium hirsutum pistils

It has recently been reported that high temperature slows in vivo pollen tube growth rates in Gossypium hirsutum pistils under field conditions. Although numerous physical and biochemical pollen-pistil interactions are necessary for in vivo pollen tube growth to occur, studies investigating the influence of heat-induced changes in pistil biochemistry on in vivo pollen tube growth rates are lacking. We hypothesized that high temperature would alter diurnal pistil biochemistry and that pollen tube growth rates would be dependent upon the soluble carbohydrate content of the pistil during pollen tube growth. G. hirsutum seeds were sown on different dates to obtain flowers exposed to contrasting ambient temperatures but at the same developmental stage. Diurnal pistil measurements included carbohydrate balance, glutathione reductase (GR; EC 1.8.1.7), soluble protein, superoxide dismutase (SOD; EC 1.15.1.1), NADPH oxidase (NOX; EC 1.6.3.1), adenosine triphosphate (ATP), and water-soluble calcium. Soluble carbohydrate levels in cotton pistils were as much as 67.5% lower under high temperature conditions (34.6 °C maximum air temperature; August 4, 2009) than under cooler conditions (29.9 °C maximum air temperature; August 14, 2009). Regression analysis revealed that pollen tube growth rates were highly correlated with the soluble carbohydrate content of the pistil during pollen tube growth (r² = 0.932). Higher ambient temperature conditions on August 4 increased GR activity in the pistil only during periods not associated with in vivo pollen tube growth; pistil protein content declined earlier in the day under high temperatures; SOD and NOX were unaffected by either sample date or time of day; pistil ATP and water soluble calcium were unaffected by the warmer temperatures. We conclude that moderate heat stress significantly alters diurnal carbohydrate balance in the pistil and suggest that pollen tube growth rate through the style may be limited by soluble carbohydrate supply in the pistil.