Simulation and Prediction of Plant Phenology for Five Crops Based on PhotoperiodxTemperature Interaction

This paper presents a plant phenological model based on genotypextemperaturexphotoperiodinteraction (GPTmodel). In the model, rate of development towardsa specified stage (e.g. flowering) for a given genotype is composedof three components: the genotype's maximum rate of development;any delay due to a non-optimal temperature; and any delay dueto a photoperiod response. It is assumed that development tothe specified stage is an autonomous process established bymost, if not all, genes other than the vernalization genes andthe photoperiod genes; and that this autonomous process is delayedby any activity of the photoperiod genes. Since all physiologicalprocesses are modulated by temperature, any photoperiod responseis inevitably a photoperiodxtemperature interaction. This interactionis simulated by assuming that the photoperiod gene activityoccurs only beyond a critical photoperiod (P_c) and is enlargedby temperature above a base temperature (T_bp) that allows thephotoperiod gene activity. The model is written asR=1/D_b-S_t(T-T_opt)²-S_p(T-T_bp)|P-P_c|, whereRis the expected rate of development to the specifiedstage under any combination of temperature (T) and photoperiod(P). The other model parameters are:S_p, the sensitivity to adelaying photoperiod;T_opt, the optimum temperature for developmentin the absence of the photoperiod response;S_t, the sensitivityto a non-optimum temperature; andD_b, the basic duration to thespecified stage (or intrinsic earliness), the inverse of whichis the maximum rate of development.D_bis observable only ifT=T_optandsimultaneouslyP     

影响叶螨磷酸酯酶活性的四因子数学模型

应用二次回归通用旋转组合设计,组建了影响叶螨磷酸酯酶（酸性和碱性）活性的四因子（缓冲液Ph值X₁、温浴时间X₂、反应温度X₃、底物浓度X₄）数学模型: Y_酸性=0.456380+0.107889X₂+0.069027X₃-0.026836X₁²-0.030794X₃², F=24.98,P<0.01;Y_碱性=0.267286-0.200736X₁+0.049541X₂+0.030930X₃-.049063X₁X₂+0.053585X₁²-0.049665X₂^2, F=57.68,P<0.01。结果表明,温浴时间是影响叶螨酸性磷酸酯酶活性的关键因子,在缓冲液pH 4.4、底物浓度8.5×10^-3 mol/L、42℃温浴40 min测得该酶活性最强。影响碱性磷酸酯酶活性的关键因子则是缓冲液pH值,pH 9.0、37℃恒温30 min、底物7.5×10^-3 mol/L的条件下,光密度值最大。两种酶的最大吸收峰波长为405 nm。     

Dependence of Yields of Wheat Varieties on their Leaf Area Durations

In a field experiment three wheat varieties: autumn-sown CappelleDesprez, Prestige, both autumn-sown and spring-sown, and spring-sownJufy I, each supplied with 0.5 or 1.0 cwt nitrogen/acre (6₃or 126 kg/ha), had grain yields nearly proportional to theirLeaf Area Durations (D) during grain development. Squarehead'sMaster had a smaller grain yield relative to its total D, butwith 0.5 cwt N/acre the ratio of its grain yield to D of partsabove the flag leaf node (D_F) was similar to the other varieties.The difference in this ratio between varieties was less whenD_Fwas calculated from anthesis than from ear emergence to ripening.Squarehead’s Master with 1.0 cwt N/acre had a smallerratio of grain yield to D_F than other treatments, implying lessgrain per ear relative to D_Y per shoot, perhaps because of lodging,or because factors limiting size of ears restricted their abilityto accept all the assimilate the shoots could produce with thisamount of N.     

环境因子对角倍蚜秋迁蚜生殖和雌性蚜发育的影响

研究了环境因子对角倍蚜Schlechtendalia chinensis (Bell) 秋迁蚜生殖和雌性蚜发育的影响。温、湿度单因子试验表明,秋迁蚜在26℃和80%RH条件下有最大生殖量;温、湿度对秋迁蚜生殖量的影响均符合开口向下的二次抛物线变化趋势,极端温、湿度会导致生殖量的下降。采用三元一次正交组合设计,研究了环境温度（X₁）、湿度（X₂）和光照强度（X₃）三因子不同水平组合对雌性蚜发育的影响,表明温度是影响发育历期的主要因子,其次是光照强度,最后是湿度。因此,适当高温、强光照条件可以加快雌性蚜发育;而适当高湿条件可以降低雌性蚜的发育速率而延长其发育历期。在人工培育角倍蚜生产中,创造有利于秋迁蚜生殖的温、湿度条件可以使秋迁蚜产下较多的越冬侨蚜;在适当降低温度、增加湿度的阴暗条件下贮留雌性蚜可以适当延长其发育,以使角倍蚜与盐肤木在物候上达到最佳吻合。     

Positive Association Among Three Binary Variables and Cross-Product Ratios

We show that, when the three-way association level among thethree binary variables, X, U₁ and U₂ is fixed, D_P = pr(X = 1¦U₁= 1) – pr(X = 1¦U₁ = 0) increases as the cross-productratio of U₁ and U₂ increases under the assumption that X ispositively associated with U₁ and U₂. We then discuss some implicationsof this property.     

Dynamics of Competition in Populations of Carrot (Daucus carota)

Populations of carrot (Daucus carota) were raised over a widerange of densities (79–5763 plants m^-2) to examine thedynamics of competition in terms of yield–density relationshipsand size variability, and to investigate the effects of nutrientsupply on competition. While the relationship between shootyield and density was asymptotic, the relationship between rootand total yield and density tended to be parabolic. For a giventime and density series the relationship between yield per unitarea and density could best be described by the model: y=w_mD_(1+aD)^b wherey is the yield per unit area,D is density,w_m, a andb arefitted parameters. The parametersw_m anda increased over timebut nutrient availability affected onlyw_m. An extension of thebasic yield-density model is proposed to describe the dynamicsof the yield–density relationship over time: y=kD_{[1+cexp(-rt)]{1+}     

Reproductive Allometry in Soybean, Maize and Sunflower

We compared the relationship between grain yield per plant (Y_P)and shoot biomass per plant (S_P) in three annual crops withcontrasting reproductive strategies: sunflower, a determinatespecies with a single inflorescence; maize, a determinate specieswith a limited capacity to adjust the number of ears in responseto resource availability; and indeterminate soybean, a specieswith a large capacity to adjust the number of inflorescences.Our working hypotheses were: H₁—the relationship betweenY_PandS_P is linear; H₂—the intercept of the model is zero,i.e. there is not a threshold plant mass for reproduction. Awide range of Y_Pand S_Pwas generated by manipulation of plantdensity;S_Pvaried between 0.3 and 196 g per plant in soybean,between 6 and 873 g per plant in sunflower and between 23 and697 g per plant in maize. Within these broad ranges of plantsize, both hypotheses were rejected in five out of six experiments,i.e. the relationship between Y_Pand S_Pdeparted from linearityand there was a threshold for S_Pbelow which no grain set occurred.TheS_P threshold for grain set varied widely among species; itwas close to 2 g per plant for soybean, 27 g per plant for sunflowerand 43–71 g per plant for maize. Because of this sizethreshold and non-linearity, harvest index (HI = Y_PS_P^-1) wasstable for mid-size plants, diminished slightly for large plants,and diminished sharply for smaller plants in all three crops.Harvest index stability was highest in soybean, intermediatein sunflower and lowest in maize. Differential stability ofreproductive partitioning partially derived from contrastingpatterns of meristem allocation. Copyright 2000 Annals of BotanyCompany Helianthus annuus L., Zea mays L., Glycine max(L.) Merrill, grain yield, harvest index, plant density, reproductive allocation, meristem allocation, plasticity     

Expression, regulation, and function of P2X4 purinergic receptor in human cervical epithelial cells

Micromolarconcentrations of ATP stimulate biphasic change in transepithelialconductance across CaSki cultures on filters, an acute transientincrease (phase I response; triggered by P2Y₂ receptor and mediated by calcium mobilization-dependent cell volume decrease) followed by a slower decrease in permeability (phase II response). Phase II response is mediated byaugmented calcium influx and protein kinase C-dependent increase intight junctional resistance. The objective of the study was todetermine the role of P2X₄ receptor as a mediator ofphase II response. Human cervical epithelial cells expressP2X₄ receptor mRNA (1.4-, 2.2-, and 4.4-kb isoforms byNorthern blot analysis) and P2X₄ protein. Depletion ofvitamin A reversibly downregulated P2X₄ receptor mRNA andprotein and ATP-induced calcium influx. Depletion of vitamin Aabrogated phase II response, and the effect could bepartially reversed only with retinoic acid receptor (RAR)-selectiveretinoids but not retinoid X receptor (RXR) agonists. Depletion ofvitamin A also abrogated protein kinase C increase in tight junctionalresistance, and the effect could not be reversed with retinoids.Depletion of vitamin A also abrogated phase I increase inpermeability and reversibly downregulated P2Y₂ receptormRNA and ATP-induced calcium mobilization. However, in contrast tophase II response, both RAR and RXR agonists could fullyreverse those effects. These results suggest that phase IIresponse is mediated by a P2X₄ receptor mechanism.

     

Resource competition between two rotifer species (Brachionus rubens and B.calyciflorus): an experimental test of a mechanistic model

A mechanistic model of competition on a single resource wastested experimentally with two freshwater rotifers, Brachionusrubens and B.calyciflorus, both grown on the alga Chlorellapyrenoidosa. Using open culture systems for each species wemeasured: (i) the resource-saturated exponential growth rate,µ_max, and (ii) the relationship between specific growthrate, µ, at steady-state and the residual algal concentrationover a range of system turnover rates, or dilution rates, D.The µ_max of B.calyciflorus was {small tilde}60% higherthan B. rubens. These results were then used to construct agraphical model for predicting the victor in interspecific competitionbetween the two rotifers. Since the two resource-dependent growthrates crossed, one species, B.calyciflorus, was predicted tobe the victor at a high D while B. rubens was predicted as thevictor at low D. Finally, the outcome of competition was determinedfor two turnover rates. As predicted by the graphical competitionmodels, B.calyciflorus was the dominant species at rapid D (0.029h^–1) and B.rubens was dominant at slow D (0.0044 h^–1).These studies support recent conclusions that mechanistic competitionmodels may be applied to predict dominant species from a prioriinformation on growth potential and resource levels, which isnot possible with traditional Lotka-Volterra models.     

Isotonic contractile and fatigue properties of developing rat diaphragm muscle

Postnatal transitions in myosin heavy chain (MHC) isoformexpression were found to be associated with changes in both isometric and isotonic contractile properties of rat diaphragm muscle(Dia_m). Expression of MHC_neo predominated inneonatal Dia_m fibers but was usually coexpressed withMHC_slow or MHC_2A isoforms. Expression ofMHC_neo disappeared by day 28. Expression ofMHC_2X and MHC_2B emerged at day 14 andincreased thereafter. Associated with these MHC transitions in theDia_m, maximum isometric tetanic force (P_o), maximum shortening velocity, and maximum power output progressively increased during early postnatal development. Maximum power output ofthe Dia_m occurred at ~40% P_o at days0 and 7 and at ~30% P_o in older animals.Susceptibility to isometric and isotonic fatigue, defined as a declinein force and power output during repetitive activation, respectively,increased with maturation. Isotonic endurance time, defined as the timefor maximum power output to decline to zero, progressively decreasedwith maturation. In contrast, isometric endurance time, defined as thetime for force to decline to 30-40% P_o, remained>300 s until after day 28. We speculate that with thepostnatal transition to MHC_2X and MHC_2Bexpression energy requirements for contraction increase, especiallyduring isotonic shortening, leading to a greater imbalance betweenenergy supply and demand.

     

Common canonical variates

Canonical correlation analysis measures the linear relationshipbetween two random vectors X₁ and X₂ as the maximum correlationbetween linear combinations of X₁ and linear combinations ofX₂. Several generalisations of canonical correlation analysisto k2 random vectors X₁ ..., X_k have been proposed in the literature(Kettenring, 1971, 1985), based on the principle of maximisingsome generalised measure of correlation. In this paper we proposean alternative generalisation, called common canonical variates,based on the assumption that the canonical variates have thesame coefficients in all k sets of variables. This generalisationis applicable in situations where all X_i have the same dimension.We present normal theory maximum likelihood estimation of commoncanonical variates, and illustrate their use on a morphometricdata set.     

Photosynthesis in Panicum milioides, a Species with Reduced Photorespiration

The capacity for C₄ photosynthesis in Panicum milioides, a specieshaving reduced levels of photorespiration, was investigatedby examining the activity of certain key enzymes of the C₄ pathwayand by pulse-chase experiments with ¹⁴CO₂. The ATP$P₁ dependentactivity of pyruvate,P₁ dikinase in the species was extremelylow (0.14–0.18 µmol mg chlorophyll^–1 min^–1).Low activity of the enzyme was also found in Panicum decipiensand Panicum hians (related species with reduced photorespiration)and in Panicum laxum (a C₃ species). The antibody to pyruvate,P₁dikinase caused about 70% inhibition of the ATP$P₁ dependentactivity of the enzyme in P. milioides. The activity of NAD-malicenzyme and NADP-malic enzyme in ^P. ^milioides was equally low(approximately 0.1–0.2 µmol mg chlorophyll^–1min^–1) and similar to the activity in P. decipiens, P.hians and P. laxum. Photosynthetic pulse-chase experiments underatmospheric conditions showed a typical C₃-like pattern of carbonassimilation including the labelling of glycine and serine asexpected during photorespiration. During the pulse with ¹⁴CO₂only about 1% of the labelled products appeared in malate and2–3% in aspartate. During a chase in atmospheric levelsof CO₂ for up to 6 min there was a slight increase in labellingin the C₄ acids. The amount of label in carbon 4 of aspartatedid not change during the chase, indicating little or no turnoverof the C₄ acid via decarboxylation. The results indicate thatunder atmospheric conditions P. milioides assimilates carbondirectly through the C₃ pathway. Photorespiration as indicatedby the CO₂ compensation point may be repressed in the speciesby a more efficient recycling of photorespired CO₂. (Received June 8, 1982; Accepted July 22, 1982)     

Time-dependent stimulation by aldosterone of blocker-sensitive ENaCs in A6 epithelia

To study and define the early time-dependent response (6 h) ofblocker-sensitive epithelial Na⁺channels (ENaCs) to stimulation ofNa⁺ transport by aldosterone, weused a new modified method of blocker-induced noise analysis todetermine the changes of single-channel current (i_Na) channel open probability(P_o), andchannel density(N_T) undertransient conditions of transport as measured by macroscopic short-circuit currents(I_sc). In threegroups of experiments in which spontaneous baseline rates of transportaveraged 1.06, 5.40, and 15.14 µA/cm², stimulation of transportoccurred due to increase of blocker-sensitive channels.N_T variedlinearly over a 70-fold range of transport (0.5-35µA/cm²). Relatively small andslow time-dependent but aldosterone-independent decreases ofP_o occurredduring control (10-20% over 2 h) and aldosterone experimentalperiods (10-30% over 6 h). When theP_o of control andaldosterone-treated tissues was examined over the 70-fold extendedrange of Na⁺ transport,P_o was observedto vary inversely withI_sc, falling from~0.5 to ~0.15 at the highest rates ofNa⁺ transport or ~25% per3-fold increase of transport. Because decreases ofP_o from anysource cannot explain stimulation of transport by aldosterone, it isconcluded that the early time-dependent stimulation ofNa⁺ transport in A6 epithelia isdue exclusively to increase of apical membraneN_T.

     

Differential Effects of Day and Night Temperature on Development to Flowering in Rice

There are conflicting reports with regard to difference in effectsof day temperature (T_D) and night temperatures (T_N) on plantdevelopment. The objective of this study is to determine whetherthere are different effects ofT_DandT_Non development from sowingto flowering in rice (Oryza sativaL.). Plants of 24 rice cultivars were grown in naturally-lightedgrowth chambers at five diurnally constant (22, 24, 26, 28 and32 °C) and four diurnally fluctuating temperatures (26 /22,30 /22, 22 /26 and 22 /30 °C forT_D/T_Nwith 12hd^-1each) witha constant photoperiod of 12hd^-1. The treatments were selectedto enable the separation of effects ofT_DandT_Non developmentrate (DR). The response of DR to constant temperatures was typically nonlinear.This nonlinearity could not explain the difference in floweringdates between fluctuating temperatures with the same mean dailyvalue but oppositeT_D/T_Ndifferences. Differential effects ofT_DandT_NonDR to flowering were detected in all but one cultivar. In mostcases,T_Dexerted a greater influence thanT_N, in contrast withmany previous reports based on the assumption of a linearitybetween DR and temperature. The data were further analysed bya nonlinear model which separated effects ofT_DandT_N. The estimatedvalue for the optimumT_Nwas generally 25 –29 °C, about2 –4 °C lower than the estimated optimumT_Din mostcultivars. The effects ofT_DandT_Non DR were found to be interactivein some cultivars. These results form a new basis for modellingflowering dates in rice. Oryza sativa; rice; flowering; development; day and night temperature; thermoperiodicity     

利用人工合成性信息素诱捕器诱捕印度谷螟的影响因素分析

采用自行设计式及仿制圆筒式诱捕器,以人工合成性信息素(Z,E)-9,12-tetradecadienyl acetate (简称TDA) 为引诱源在实验室条件下对影响诱捕器诱捕印度谷螟Plodia interpunctella效果的几个因素进行分析测定。多元线性回归分析结果表明：日平均温度在18.5～26.2℃,人工合成性信息素TDA散发日数9～37天,温度(X₁)、TDA散发日数(X₂)、当日释放蛾(雄)量(X₃)、累计2日释放蛾量(X₄)、累计3日释放蛾量(X₅)等5个因素与每日诱捕蛾量间存在着相关关系。对5个因素进行逐步回归分析和筛选,得出线性回归方程：Y=-27.31+1.37X₁+0.28X₃,回归系数R（0.90）>R_0.01(n-2,0.63)。统计分析结果表明：日平均温度(X₁)、当日释放蛾量(X₃)与诱蛾量(Y)之间呈显著线性相关关系。卡方测验表明预测值与实测值之间差异不显著。     

Semiparametric regression estimation in the presence of dependent censoring

We propose a semiparametric estimation procedure for estimatingthe regression of an outcome Y, measured at the end of a fixedfollow-up period, on baseline explanatory variables X, measuredprior to start of follow-up, in the presence of dependent censoringgiven X. The proposed estimators are consistent when the dataare ‘missing at random’ but not ‘missing completelyat random’ (Rubin, 1976), and do not require full specificationof the complete data likelihood. Specifically, we assume thatthe probability of censoring at time t is independent of theoutcome Y conditional on the recorded history up to t of a vectorof time-dependent covariates that are correlated with Y. Ourestimators can be used to adjust for dependent censoring andnonrandom noncompliance in randomised trials studying the effectof a treatment on the mean of a response variable of interest.Even with independent censoring, our methods allow the investigatorto increase efficiency by exploiting the correlation of theoutcome with a vector of time-dependent covariates.     

洛氏脊漠甲抗冻蛋白基因的克隆、表达和功能检测

为了研究抗冻蛋白基因是否在新疆荒漠昆虫洛氏脊漠甲 Pterocoma loczyi 中存在,利用 RT-PCR 技术克隆获得了洛氏脊漠甲抗冻蛋白的 cDNA 片段,命名为 Plafp743。测序结果表明洛氏脊漠甲 Plafp743 所编码的蛋白质由 94 个氨基酸组成,蛋白序列呈现规则结构 CTX₁X₂X₃X₄CX₅X₆X₇X₈X ₉。利用原核表达载体构建重组质粒 pGEX-4T-1-Plafp743,转化 Escherichia coli BL21 进行融合蛋白表达,SDS-PAGE 结果表明抗冻蛋白PLAFP基因以可溶性融合蛋白形式表达,相对分子质量 36 kD。用 Glutathione Sepharose 4B 亲和柱对表达蛋白进行纯化后,通过赤翅甲 Dendroides canadensis 抗冻蛋白的小鼠抗血清进行 Western blot 分析,结果表明纯化的 GST-PLAFP 融合蛋白与赤翅甲抗血清能够发生特异性的免疫反应。大肠杆菌的低温抗冻保护实验结果证明,30 μg/mL 的 GST-PLAFP 在-6℃ 对细菌具有显著的保护作用,且随着抗冻蛋白浓度的增加,抗冻保护作用的效 果也随之增加。本研究首次报道了从荒漠昆虫洛氏脊漠甲扩增得到抗冻蛋白基因,提示抗冻蛋白可能是荒漠昆虫普遍采取的过冬生存策略,为进一步开发利用昆虫抗冻蛋白奠定了基础。     

上海地区三带喙库蚊春季首次出现、季节分布及其与气象条件的关系

根据1982～1992年和2001～2002年上海奉贤地区每年3月～6月灯诱捕蚊的现场调查数据,采取相关对比分析、统计分析、天气图分析法,探讨三带喙库蚊春季首次出现、季节分布及其与气象条件的关系。结果表明,三带喙库蚊季节分布按蚊虫密度变化划分为首次出现期、季节增多期和混合发生期三个时期。三带喙库蚊春季首次出现的温度条件是：首现日当天和前1天的日平均气温均≥11.0℃,其2天的日平均气温累加值≥25.5℃,或首现日当天、前2天和后1天任意连续3天的日平均气温均≥11.0℃,其3天的日平均气温累加值≥34.0℃,结果三带喙库蚊春季首现日和首现日温度日期的差异在1～2天内(91.7%)。三带喙库蚊季节增多期其逐候蚊虫密度（Y₁）与平均气温（X₁）呈显著的正相关,用一元回归方程表示为：Y₁=-9.3534+0.7217X₁;至候平均气温≥18℃（平均始日为5月3日）进入三带喙库蚊混合发生期。总体上三带喙库蚊蚊虫密度季节分布与平均气温、“S”系风向变化存在密切关系。首次出现期至季节增多期三带喙库蚊突增日与锋面天气活动存在着较大的关联,候平均气温≥18℃是三带喙库蚊大量发生和进入峰期重要的温度指标。上述结果提示上海地区三带喙库蚊春季和初夏随气流北迁降落的可能性相当大。     

Nucleotide regulation of the voltage-dependent nonselective cation conductance in murine colonic myocytes

ATP is proposed to be a major inhibitory neurotransmitter in the gastrointestinal (GI) tract, causing hyperpolarization and smooth muscle relaxation. ATP activates small-conductance Ca²⁺-activated K⁺ channels that are involved in setting the resting membrane potential and causing inhibitory junction potentials. No reports are available examining the effects of ATP on voltage-dependent inward currents in GI smooth muscle cells. We previously reported two types of voltage-dependent inward currents in murine proximal colonic myocytes: a low-threshold voltage-activated, nonselective cation current (I_VNSCC) and a relatively high-threshold voltage-activated (L-type) Ca²⁺ current (I_L). Here we have investigated the effects of ATP on these currents. External application of ATP (1 mM) did not affect I_VNSCC or I_L in dialyzed cells. ATP (1 mM) increased I_VNSCC and decreased I_L in the perforated whole-cell configuration. UTP and UDP (1 mM) were more potent than ATP on I_VNSCC. ADP decreased I_L but had no effect on I_VNSCC. The order of effectiveness was UTP = UDP > ATP > ADP. These effects were not blocked by pyridoxal phosphate-6-azo(benzene-2,4-disulfonic acid) (PPADS), but the phospholipase C inhibitor U-73122 reversed the effects of ATP on I_VNSCC. ATP stimulation of I_VNSCC was also reversed by protein kinase C (PKC) inhibitors chelerythrine chloride or bisindolylmaleimide I. Phorbol 12,13-dibutyrate mimicked the effects of ATP. RT-PCR showed that P2Y₄ is expressed by murine colonic myocytes, and this receptor is relatively insensitive to PPADS. Our data suggest that ATP activates I_VNSCC and depresses I_L via binding of P2Y₄ receptors and stimulation of the phospholipase C/PKC pathway. inhibitory junction potentials; smooth muscle; enteric nervous system     

In situ Effects of Elevated Atmospheric CO2on Leaf Freezing Resistance and Carbohydrates in a Native Temperate Grassland

The objectives of this study were to quantify changes in leaffreezing resistance and carbohydrate concentrations caused bylong-term (6 years) exposure to elevated CO₂(ambient: 360 µll^-1, elevated: 600 µl l^-1) in five dominant plant speciesgrowing in situ in a native temperate grassland. Across allfive species tested from three functional groups, the mean temperatureat which all leaves were damaged (T₁₀₀) significantly (P = 0.016)increased from -9.6 to -8.5 °C under elevated CO₂ , anda similar marginally significant (P = 0.079) reduction was observedfor the mean temperature that caused 50% leaf damage (T₅₀),from -6.7 to -6.0 °C. The mean temperature at which initialleaf damage was observed (T₀) was not significantly influencedby elevated CO₂ . Although concentrations of soluble sugars(+25%,P = 0.042), starch (+53%, P < 0.001), and total non-structuralcarbohydrates (TNC, +40%, P < 0.001) were significantly higherunder elevated CO₂ , leaf freezing resistance actually decreasedunder elevated CO₂ . Concentrations of soluble sugars were positivelycorrelated with freezing resistance when viewed across all fivecommunity dominants, but within any individual species, no suchrelationships were found. We also found no evidence for ouroriginal hypothesis that increased concentrations of solublesugars increase freezing resistance. Thus, future atmosphericCO₂levels may instead increase the risk of late spring freezingdamage. Furthermore, the strong differences in freezing resistanceobserved among the species, along with decreased freezing resistance,may increase the risk of losing species that have inherentlyweak freezing resistances from the plant community. Copyright2001 Annals of Botany Company CO₂enrichment, frost hardiness, sugar, starch, total non-structural carbohydrates (TNC)