Characteristic Changes in Relaxation Times of Water Protons in Vigna radiata Seedlings Exposed to Temperature Stress

Water protons in hypocotyl tissues from etiolated seedlingsof Vigna radiata that were exposed to temperature stress showedcharacteristic relaxation behaviors for ¹H-NMR. Cold stresstreatment (0C) caused gradual prolongation of NMR relaxationtimes (T₁). After exposure of tissues to cold stress for 24h, T₁ returned to the initial value as a result of subsequentincubation at normal temperature (20C). By contrast, heat stresstreatment (40C) induced a time-dependent decrease in T₁, whichdid not return completely to the initial value upon subsequentincubation at 20C after exposure to heat stress for 4 h. Weexamined changes in various physical factors that influencethe response of T₁ to temperature stress, namely, water contentand the concentrations of protein, diamagnetic (K⁺, Na⁺, Ca²⁺and Mg²⁺) and paramagnetic (Mn²⁺ and Fe²⁺) ions in the tissues.From the relationships between T₁ and these factors in vitro,we could not interpret the responses of T₁ to the temperaturestress only in terms of a change in water content. A synergisticeffect of an Mn²⁺ -protein complex and pH might be essentialfor the mechanism of changes in T₁ that are due to cold stress.The influence of heat stress on structural water in tissuesis discussed in terms of water-protein interactions. (Received December 28, 1992; Accepted May 6, 1993)     

Estimation of Chilling Sensitivity and Injury in Gloxinia Leaves by the Thermal Hysteresis of NMR Relaxation Times of Water Protons

Thermal hysteresis of longitudinal relaxation times (T₁) ofwater protons in leaf tissues of gloxinia was investigated bypulse nuclear magnetic resonance (NMR) spectroscopy. The profilesof T₁ hysteresis during a slow cool-warm cycle (20 to 0?C) variedwith the degree of chilling injury. General trends of T₁ changesupon warming were as follows: (1) a small increase of T₁ wasobserved when no chilling injury occurred or the degree of injurywas minor, (2) a large increase of T₁ occurred when injury increased,(3) a large decrease of T₁ occurred from the earlier stage ofwarming when serious injury occured. The sum of T₁ ratio (theratio obtained from the difference between the T₁ value in thecooling process and that of the wanning process for eight stepsof cool-warm cycle, divided by the former) reflecting T₁ increaseor decrease during the warming process was related to varietaldifference and seasonal changes of chilling sensitivity. Therefore,T₁ hysteresis can be used as a diagnostic tool in detectingthe chilling sensitivity and the degree of chilling injury. (Received August 27, 1986; Accepted February 24, 1987)     

Monitoring Stress Sensitivity by Water Proton NMR Relaxation Times in Leaves of Azaleas That Originated in Different Ecological Habitats

Responses of the leaves of five species of azalea to environmentalstresses, such as freezing, dehydration, high temperature andsalt spray, were measured in terms of water proton NMR relaxationtimes (T₁), supercooling ability, and water content. Three subtropicalspecies (R. scabrum cv. Shounoshin, R. eriocarpum and R. tashiroivar. lasiophyllum) and two northern species (R. indicum cv.Kumano-satsuki and R. yedoense f. poukhanense), which originatedin different ecological habitats, showed characteristic behaviorsin terms of T₁ relaxation times. In general, a species witha large change in T₁ is more stress-sensitive than a speciesshowing the opposite tendency. The relative sensitivity to variousstresses of each species appears to be related to the severityof conditions in its natural habitat. It seems possible thatthose species of azalea with higher sensitivity to a particularsingle stress may also exhibit higher sensitivity to severalor even most stresses, and vice versa. (Received August 27, 1992; Accepted February 26, 1993)     

Role of aminolevulinic acid in improving biomass production in Vigna catjung, V. mungo, and V. radiata

A precursor in tetrapyrrole biosynthesis, 5-aminolevulinic acid (ALA), was applied via presowing soaking in Vigna catjung, V. mungo, and V. radiata. ALA increased plant growth and influenced dry matter accumulation in leaves, stems, and pods through increased chlorophyll content and photosynthetic CO₂ absorption. At harvest, ALA treated plants had increased number of pods per plant, seeds per pod, 100 seed dry matter, biological yield, and the harvest index. Therefore, pretreatment of seeds with optimal concentration of ALA is recommended for improving the growth and productivity of tropical legumes. This revised version was published online in July 2006 with corrections to the Cover Date.     

Statistical Studies Using AIC Method to Decide the Question of "Break" or "Straight" in Arrhenius Plots of Water Proton NMR Relaxation Times in Chilling-Sensitive Vigna and Insensitive Pisum Seedlings

The question of "break" or "straight" in Arrhenius plots forthe temperature dependency of NMR relaxation times (T₁) of waterprotons in etiolated intact seedlings for chilling-sensitivetwo Vigna species and chilling-insensitive Pisum was statisticallystudied using the Akaike's Information Criterion (AIC), a versatileprocedure for statistical model identification. Among sevenmodels, the most appropriate was based on the following assumptions:individual k (preparation numbers) two-half lines connectingat the break point (TCB) and imposing no restrictions for thegradient and the break point (Model 7). The worst two modelswere one straight line or one TCB (Model 1 or 4). Thus, thedata obtained from replicated preparations should not be treatedas a whole but as individual in each sample case. Break pointsdetermined with Model 7 ranged around 11-5°C for V. radiataand 17-10°C for V. mungo. The implication of the occurrenceof "break" for Pisum clearly differed from Vigna judging bythe model fitness based on the AIC values. The question of "break"or "straight" in Arrhenius plots is therefore fairly dependenton the validity of the model selection in the statistical analysis,and the AIC method is a useful procedure for the resolutionof the problem of the "use" or "misuse" of Arrhenius plots inplant physiology. (Received November 2, 1988; Accepted January 17, 1989)     

Anthocyanin Influence on Water Proton NMR Relaxation Times and Water Contents in Leaves of Evergreen Woody Plants During the Winter

An investigation was made to determine the effect of anthocyaninin red-colored evergreen leaves during the mid-winter on waterproton NMR relaxation times (T₁). Water contents, anthocyanincontentsand histologic localization of red-coloration in mesophyllswere determined by using both red-colored and green leaves fromthe same branches of Rhododendron, Viburnum and Mahonia, respectively.Although the decrease of water contents in the red-colored leavesin Mahonia was insignificant, decreases in the former two specieswere clearly observable. T₁ differences between red-coloredand green leaves for the three species were insignificant. Increasesof anthocyanin contents and histologic localization of red colorationin mesophylls for the red-colored leaves were more pronouncedin Rhododendron and Viburnum than in Mahonia. These observationssuggest that the pronounced increases of histologic localizationof red-colored mesophyll cells and anthocyanin contents in red-coloredleaves for the former two species contribute to maintenanceof T₁ relaxation times in spite of the marked decrease of watercontents in leaves. It is assumed that the increase of localizationareas of red-colored parenchymatous cells in mesophylls is moreeffective than the total contents of anthocyanins in leavestowards the maintenance of the T₁ level in red-colored leaves,and this appears to be dependent on the vacuolar compartmentationof anthocyanin in mesophyll cells. (Received August 3, 1991; Accepted December 12, 1991)     

甜瓜幼苗叶片内源一氧化氮和蔗糖代谢对低温胁迫的响应

以低温敏感型甜瓜品种‘XL-1’和耐低温型品种‘红优’为试材,采用6℃低温处理0、1、3、6、12、24h及3d、5d和7d,研究低温胁迫下甜瓜幼苗叶片中NO合成和蔗糖代谢的变化特征。结果表明:(1)低温胁迫能提高甜瓜幼苗叶片中硝酸还原酶(NR)活性,诱导促进NO生成,其中耐低温型甜瓜‘红优’中NO对低温的响应时间更早,变化幅度更大。(2)与对照相比,低温胁迫处理提高了2种甜瓜幼苗叶片中蔗糖、果糖和葡萄糖含量,增加了蔗糖磷酸合成酶(SPS)、蔗糖合成酶(SS)、酸性转化酶(AI)和中性转化酶(NI)活性,降低了淀粉含量。(3)低温胁迫处理使2种甜瓜叶片中渗透调节物质可溶性糖、可溶性蛋白、脯氨酸的含量上升。研究发现,低温胁迫通过增加甜瓜体内NO合成酶的活性刺激体内NO合成,通过促进蔗糖代谢相关酶的活性,提高蔗糖、果糖和葡萄糖的含量,从而响应低温胁迫,且低温胁迫诱导的糖分物质积累时间晚于NO的产生时间。     

Response of Vigna radiata L. (Mung Bean) to Ozone Phytotoxicity Using Ethylenediurea and Magnesium Nitrate

Journal of Plant Growth Regulation - The response of Vigna radiata L. (mung bean) to tropospheric ozone (O3) phytotoxicity using Ethylenediurea (EDU) and magnesium nitrate (Mg(NO3)2,...     

The wild ancestors of urid and mung beans (Vigna mungo (L.) Hepper and V. radiata (L.) Wilczek)

The crop plants Vigna mungo (urid, urd or black gram) and V. radiata (mung bean or green gram: and related taxa have been studied by seed protein electrophoresis, leaf phenolics chromatography, vegetative morphology, and seed testa patterns. The results disprove the theory that these species are very closely related and have evolved from a single wild taxon. The present evidence shows that V. mungo var. mungo and I. radiata var. radiata have independent lineages and were domesticated from two very distinct taxa. namely V. mungo var. silvestris and V. radoata var. sublobata respectively.     

Irreversible Effects of Water Stress on Growth and Stomatal Development in Cotyledons of Etiolated Squash Seedlings

Growth of etiolated squash cotyledons and hypocotyls was suppressedwhen the seedlings were subjected to 60 mM polyethylene glycol(PEG) in 1/5 strength of Hoagland solution. The fresh weightof the hypocotyl completely recovered when the water stresswas relieved after one day of PEG treatment. The fresh weightof the cotyledons, however, did not completely recover eventhree days after the relief of water stress. The transpiration rate of the cotyledons was substantially reducedby the water stress, and it also did not completely recoverafter the water stress was relieved. Microscopic observationof stomata of the cotyledons by a replica method revealed thatthe water stress reduced the increase in both stomatal widthand density, and this reduction did not completely recover afterthe water stress was relieved. After one day of water stress, the endogenous ABA content ofthe cotyledon was increased from 68 to 114 ng/g fr wt. Afterthe water stress was relieved, the increased ABA content decreasedlinearly over two days to 20 ng/sg fr wt, the same value asin unstressed cotyledons. These results indicate that the effect of water stress on thehypocotyl growth was reversible but that the effect on the cotyledonswas irreversible. The irreversible effect of water stress onthe growth of the cotyledons probably resulted from the inhibitionof stomatal development, but endogenous ABA did not appear toinhibit the development. (Received March 24, 1986; Accepted June 25, 1986)     

Mimosine Mitigates Oxidative Stress in Selenium Deficient Seedlings of Vigna radiata. Part II: Mitochondrial Uptake of 75Selenium and Mimosine

During the growth of selenium (Se)-deficient seedlings of Vigna radiata, exposure to mimosine [2-amino-3-(3-hydroxy-4-oxo-1H-pyridin-1-yl)-propanoic acid], a nonprotein plant amino acid, effectively mitigated stress at 0.1 mM, as reflected in enhancement of growth and efficiency of mitochondrial functions. Since the changes in the seedlings elicited by exposure to mimosine were similar to those effected by Se at an optimal exposure level of 0.75 ppm (Sreekala et al., Biol Trace Elem Res 70:193–207, 1999), the uptake of Se and that of mimosine itself was individually studied in the respiring mitochondria of Se-deficient seedlings (−Se-stressed group) in comparison with those exposed to mimosine during growth at 0.1 mM (Mim 0.1 group). In both groups, the mitochondrial uptake of ⁷⁵Se at 10 μM added increased linearly up to 2 min, attaining steady-state levels thereafter. Uptake levels were 2.3-fold higher in the Mim 0.1 group than in the −Se-stressed group. Double-reciprocal plots of mitochondrial ⁷⁵Se uptake against 2–20 μM in the medium were nonlinear and negative cooperative effects during the uptake were confirmed by Scatchard plots, whereas Hill coefficients were 0.8 and 0.85 for the two groups. Mitochondrial uptake of mimosine, at added levels of 25 or 50 μM, increased linearly up to 1 min and decelerated thereafter. Initial uptake levels of mimosine at 1 min were higher by 6.5-fold at 25 μM and 4-fold at 50 μM in the Mim 0.1 group than those in the −Se-stressed group. Initial uptake levels with added mimosine up to 50 or 100 μM yielded nonlinear double-reciprocal plots; and kinetic analyses at 5 to 50 μM revealed the prevalence of positive cooperativity in the −Se-stressed group and negative cooperativity in the Mim 0.1 group. Involvement of active thiol groups in the uptake of both Se and mimosine were indicated by inhibition studies. Evidence presented for mimosine mediated increase in mitochondrial Se uptake and cooperative interactions thereof underscores the metabolic significance of mimosine.     

Impairment of Tonoplast H-ATPase as an Initial Physiological Response of Cells to Chilling in Mung Bean (Vigna radiata [L.] Wilczek)

Biochemical alterations of cellular membranes in chilling-sensitive mung bean (Vigna radiata [L.] Wilczek) hypocotyls were investigated with reference to chilling injury. Reversible decreases in activities of tonoplast H⁺-ATPase and in vivo respiration became manifest within 24 hours of chilling when tissues suffered no permanent injury as assessed by electrolyte leakage and regrowth capacity. These changes were found to be the earliest cellular responses to chilling. A density-shift on a sucrose density gradient was observed in Golgi membranes early in the chilling treatment, suggesting that Golgi function and/or membrane biogenesis via the Golgi may have been altered upon chilling. After chilling more than 2 days, irreversible changes were generally produced in cellular membranes including the plasma membrane, endoplasmic reticulum, and mitochondria. Respiratory functions remained intact in mitochondria isolated from tissues prechilled for 24 hours, but were impaired after prechilling for 3 days. Given the important role of the tonoplast H⁺-ATPase in the active transport of ions and metabolites, the early decline in the tonoplast H⁺-ATPase activity may give rise to an alteration of the cytoplasmic environment and, consequently, trigger a series of degenerative reactions in the cells.     

Enhancement in Growth and Yield by Pre-sowing Chilling Treatment to Seeds of Phaseolus mungo cv. T 9

Pre-sowing chill treatments to seeds of Phaseolus mungo induceenhancement in growth, branching, nodulation and seed yieldper plant. The concentrations of certain biochemical constituentsare also higher in the seeds harvested from treated plants. Phaseolus mungo cv. T 9, pre-sowing chill treatment, yield enhancement     

Root Regeneration and Plant Water Status of Pinus radiata D. Don Seedlings Transplanted to Different Soil Temperatures

The regeneration and configuration of new roots on transplanted8 month old Pinus radiata D. Don seedlings were measured inrelation to soil temperature and time after transplanting. Theeffects of root wrenching and nutrition on root regenerationwere compared. Low soil temperature adversely affected the initiationand elongation of new roots. The branching and morphology ofthe new roots were also influenced by soil temperature. Theprocesses of initiation and elongation are greatly retardedbelow a critical root temperature between 11 and 14 ?C. Wrenchingand fertilization stimulated new root growth at low temperatures.There is a close interrelationship between the constituent partsof the network which forms the new root system. The first-orderlaterals on transplants are critical as a basic framework forextension and production of second-order laterals which formthe major part of the regenerating root system. Needle water potential was closely related to the new root growth.The water stress experienced by the transplants for severalweeks after planting was due primarily to the suppressive effectof low soil temperature on root regeneration. Wrenched plantswere less water-stressed than unwrenched plants. Field measurements confirmed the finding from controlled experimentsthat, in southern Australia, the low soil temperature in theplanting season imposes a major restriction on early root regenerationwhich in turn inflicts water stress in transplants. This needsto be considered during the planning of planting and fertilization.     

The Basal Level Ethylene Response is Important to the Wall and Endomembrane Structure in the Hypocotyl Cells of Etiolated Arabidopsis Seedlings

The sub-cellular events that occur during the ethylene-modulated cell elongation were characterized by examining the ultra-structure of etiolated Arabidopsis seedling hypocotyl cells. Preventing the basal level ethylene response facilitated cell elongation, and the cells exhibited wall loosening and separation phenotype. Nearby the wall separation sites were frequently associated with an increase in the cortical rough endoplasmic reticulum (rER) membranes, the presence of paramural bodies, and the circular Golgi formation. The cortical rER proliferation and circular Golgi phenotype were reverted by the protein biosynthesis inhibitor cycloheximide. The cortical rER membranes were longer when the ethylene response was prevented and shortened with elevated ethylene responses. Proteomic changes between wild type and the ethylene-insensitive mutant ethylene insensitive2 (ein2) seedling hypocotyls indicated that distinct subsets of proteins involving endomembrane trafficking, remodeling, and wall modifications were differentially expressed. FM4-64 staining supported the proteomic changes, which indicated reduced endocytosis activity with alleviation of the ethylene response. The basal level ethylene response has an important role in endomembrane trafficking, biological materials transport and maintenance of the endomembrane organization. It is possible that endomembrane alterations may partly associate with the wall modifications, though the biological significance of the alterations should be addressed in future studies.     

Distribution and Redistribution of Molybdenum in Black Gram (Vigna mungo L. Hepper) in Relation to Molybdenum Supply

The effect of seven rates of molybdenum (Mo) supply on the distributionand redistribution of Mo in Vigna mungo (black gram) cv. Reguron a Mo-deficient sandy loam was examined from flower bud appearanceto pod set in one experiment and during pod filling to maturityin another. At the three lowest Mo supply rates, N deficiency symptoms typicalof Mo deficiency appeared, and shoot dry matter and shoot nitrogencontent were depressed. Increasing Mo supply increased Mo concentrationsin all plant parts but the response varied with Mo supply andwith plant part. In leaf blades and petioles, Mo concentrationsincreased slightly when the Mo supply increased from severelydeficient to deficient levels but further increases in Mo supplymarkedly increased the Mo concentrations, particularly in immatureand recently matured leaves. In petioles, Mo concentrationsgenerally exceeded those in the blades which they supportedat all levels of Mo supply. At Mo rates greater than that requiredfor maximum growth, Mo concentrations in basal stem segmentsexceeded those in petioles. Molybdenum concentrations in nodulesexceeded those in above ground plant parts except at the highestlevel of Mo supply where the concentrations in basal stem segmentsexceeded those in nodules. In Mo-adequate plants, Mo contents in the trifoliolate leavesdecreased with time suggesting that Mo was readily remobilized.By contrast, in stem segments at all levels of Mo supply, andin trifoliolate leaves in Mo-deficient plants, Mo contents remainedconstant or increased with time suggesting that Mo was not remobilizedin all plant parts or at all levels of Mo supply. Thus, theresults suggest that in black gram Mo was variably mobile, beingphloem immobile at low Mo supply, but phloem-mobile in all plantparts with the possible exception of stem segments at adequateMo supply. The relevance of these results for the developmentof plant tests for Mo deficiency diagnosis is discussed.Copyright1994, 1999 Academic Press Molybdenum, phloem-mobility, redistribution, black gram, Vigna mungo L. Hepper