Comparative Studies of Net Photosynthesis and Transpiration of Some Citrus Species and Relatives

Comparisqns were made between ‘Campbell Valencia’ orange (Citrus sinensis (L.) Osbeck), and ‘Gabon Cherry-orange’ (Citropsis gabunensis (Engl.) Swing.), ‘Frost Lisbon’ lemon (Citrus limon (L.) Burm. f.) and ‘Eremolemon’ (Eremocitrus glauca (Lindl.) Swing. ×Citrus limon (L.) Burm. f.) and between diploid and autotetraploid ‘Lisbon’ lemons with respect to the influences of temperature and humidity on net photosynthesis and transpiration. Net photosynthesis, leaf conductance to water vapor and water-use-efficiency of Citropsis gabunensis were lower than with Citrus sinensis.‘Eremolemon’ had higher net photosynthesis and higher water-use-efficiency than ‘Lisbon’ lemon, but only small differences were observed between the two species in leaf conductance to water vapor. Small, nonsignificant, differences were observed between diploid and tetraploid ‘Lisbon’ lemons in responses of net photosynthesis and leaf conductance to temperature and humidity. Temperatures above 30°C and increases in vapor pressure difference caused declines in net photosynthesis and increases in vapor pressure difference resulted in decreases in leaf conductance to water vapor by all of the species used in these studies.     

Cryopreservation and storage of embryogenic callus cultures of several Citrus species and cultivars

Nucellus-derived embryogenic callus cultures of Salustiana sweet orange were subjected to cryoconservation assays. Cryoprotection with 10%(vol/vol) dimethylsulfoxide, freezing by slow cooling and thawing by fast warming was suitable to recover viable growing cultures and whole plants through embryogenesis. Evaluation of liquid phase R ₁ and solid phase R ₂ cooling rates using a programmable freezing unit indicated that 100% of embryogenic cultures survived when frozen using a range of cooling rates (R ₁ not above 0.5°C min^–1 and R ₂ not above 1°C min ¹) and thawed by fast warming. Storage up to 2 years in liquid nitrogen did not affect the growth of the cryopreserved cultures and the recovery of whole plants. Cultures of four cultivars of sweet orange (C. sinensis Osb.), three cultivars of grapefruit (C. paradisi Macf.), and one cultivar each of lemon [C. limon (L.) Burm. f.], Cleopatra mandarin (C. reshni Hort. ex Tan.), sour orange (C. aurantium L.) and Mexican lime [C. aurantifolia (Christm.) Swing.] have been successfully cryopreserved. Problems using a viability assessment using fluorescein diacetate staining are discussed. Received: 15 April 1996 / Revision received: 22 July 1996 / Accepted: 6 August 1996     

The introduction of cultivated citrus to Europe via Northern Africa and the Iberian Peninsula

The circumstances concerning the diffusion of the main cultivated citrus from their places of origin in Asia are studied here, showing that the citron (’Citrus medica L.) was the only one knew in Ancient times in Europe, while the lemon (C. limon [L.] Osbeck), lime (C. aurantiifolia [Christm.] Swingle), pomelo (C. maxima [Burm.] Merr.) and sour orange (C. x aurantium L.) were introduced to Europe by the Muslims via the Iberian Peninsula and Sicily, and that the grapefruit (C. paradisi Macfad.), mandarin (C. reticulata Blanco) and sweet orange (C. x aurantium L.) arrived to the West between the fifteenth and nineteenth centuries as a result of the trade with the British and Portuguese colonies.     

Relationships of leaf Fatty acids to cold hardening of citrus seedlings

Three cultivars of citrus with different sensitivities to freezing temperatures (citron, Citrus medica L.; rough lemon, C. limon Burm. F; sour orange, C. aurantium L.) were cold hardened for 4 weeks. Lipids from leaves of hardened and control seedlings were fractionated and analyzed for fatty acids. The absolute amount of triglycerides and phospholipids increased in the leaves upon hardening. With hardening, total linoleic acid also increased 141% in citron, 210% in rough lemon, and 233% in sour orange. Specific increases in linoleic acid were found in triglycerides, in the four phospholipids (phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylglycerol), and in neutral lipids more polar than triglycerides. Trans-3-hexadecenoic acid was found only in phosphatidylglycerol.     

A Low-Moisture-Content Limit to Logarithmic Relations Between Seed Moisture Content and Longevity

Seeds of quinoa (Chenopodium quinoa Willd.), sunflower (Helianthusannuus L.) and linseed (Linum usitatissimum L.) showed negativelogarithmic relations between longevity and moisture contentsbetween 4.4 and 15.4, 3.2 and 13.0, and 3.2 and 15.5%, respectively,in hermetic storage at 65 °C. However, between 1.8 and 3.1,1.1 and 1.9, and 1.1 and 2.1%, respectively, longevity did notvary. The critical moisture content, below which further reductionin moisture content no longer increased longevity in hermeticstorage at 65 °C, for each species was 4.1, 2.04 and 2.7%,respectively. Quinoa, Chenopodium quinoa Willd., sunflower, Helianthus annuus L., linseed, Linum usitatissimum L., seed storage, improved viability equation, seed longevity, seed moisture content     

Morphogenesis and tissue cultures of three citrus species

Studies were performed to define tissue culture techniques and culture conditions for morphogenesis, callus culture and plantlet culture of sweet orange (Citrus sinensis (L.) Osb.), citron (C. medica L.) and lime (C. aurantifolia) (Christm. Swing). The optimal concentrations of NAA to induce root formation on stem segments were 10 mg l^-1 for sweet orange and lime, and 3 mg l^-1 for citron. The optimal BA concentration for shoot and bud proliferation was 3 mg l^-1 for sweet orange and citron, and 1 mg l^-1 for lime. Callus initiation was accomplished in a culture medium containing 10 mg l^-1 NAA and 0.25 mg l^-1 BA. Callus was maintained by periodical subculture into the same medium supplemented with 10% (v:v) organge juice. In vitro plantlets of the three species were obtained by rooting of shoots developed from bud cultures, and of citron and lime by development of shoots from root cultures. The plants were successfully established on soil.     

Temperature and Seed Storage Longevity

Seed survival data for eight diverse species, namely the cerealbarley (Hordeum vulgare L.), the grain legumes chickpea (Cicerarietinum L.), cowpea [Vigna unguiculata (L.) Walp.] and soyabean [Glycine max (L.) Merr.], the timber trees elm (Ulmus carpinifoliaGleditsch.), mahogany (Swietenia humilis Zucc.), and terb (Terminaliabrassii Exell.), and the leaf vegetable lettuce (Lactuca sativaL.) were compared over a wide range of storage environments(temperatures from –13 °C to 90 °C, seed moisturecontents from 1.8 to 25% f. wt) using a viability equation developedpreviously. In accordance with that equation, the effect oftemperature on seed longevity was dependent upon the temperaturerange. The temperature coefficients of the viability equationdid not differ significantly (P > 0.05) among the eight speciesdespite their contrasting taxonomy. Thus the quantitative relationbetween seed longevity and temperature does not vary among diversespecies. The same conclusion was obtained for the coefficientsof a proposed alternative model of the relation between seedlongevity and temperature. The implications of the two temperaturemodels in the viability equation for extrapolations to low andvery low temperatures are discussed. Seed storage, seed longevity, seed moisture, temperature, viability equation, genetic resources conservation, Cicer arietinum L., Glycine max (L.) Merr., Hordeum vulgare L., Lactuca sativa L., Swietenia humilis Zucc., Terminalia brassii Exell., Ulmus carpinifolia Gleditsch., Vigna unguiculata (L.) Walp     

Water status indicators of lemon trees in response to flooding and recovery

Potted 2-year-old lemon trees [Citrus limon (L.) Burm. fil, cv. Verna] grafted on sour orange (C. aurantium L.) rootstock were subjected to flooding for 3 d. Control plants were irrigated daily to field capacity. Continuously (sap flow, trunk diameter fluctuations) and discretely (predawn and midday leaf water potential, leaf conductance) measured plant-based water status indicators were compared. The sensitivity of the maximum daily trunk shrinkage signal intensity to flooding and its behaviour during the recovery period demonstrated that this indicator is more feasible than the others for use in automatic irrigation. The responses to flooding of continuously and discretely measured plant-based water status indicators were very similar to those observed in response to drought stress indicating that it necessary to use soil water measurement automatic sensors to detect the cause of the stress. The results underlined the robustness of the compensation heat-pulse technique for estimating instantaneous and daily transpiration rates on flooding stress and recovery.     

Effects of Volatiles Evolved from Excised Plant Tissues and Lemon Oil on the Growth of Mung Bean Seedlings

Volatiles evolved from excised segments of leaves of oleander(Nerium oleander) and cedar (Cedrus deodara), peels of lemon(Citrus limon), orange (Citrus sinensis) and lime (Citrus aurantifolia),and roots of wasabi (Wasabia japonica) inhibited growth of mungbean seedlings. The volatiles evolved from lemon oil also markedlyinhibited elongation of mung bean seedlings. The active constituent(s)in the volatiles were distinct from ethylene. The mode of inhibitionby the volatiles from lemon oil was different from that of ethylene.The volatiles from lemon oil alleviated the inhibitory effectof ethylene on the growth of seedlings. The volatiles from lemonoil supressed the formation of lamellar structure in chloroplasts,thus resulting in etiolation of seedlings. (Received October 25, 1982; Accepted December 29, 1982)     

Influence of virus and virus-like agents on the development of citrus buds cultured in vitro

Tissue culture in vitro was used to determine the effect of six major citrus virus and virus-like agents. Nodal stem segments from inoculated Pineapple sweet orange (Citrus sinensis (L.) Osb.), Mexican lime (C. aurantifolia (Christm.) Swing.) and Arizona Etrog citron 861-Sl (C. medica L.) were cultured in vitro to induce shoots. Some virus and virus-like agents had a marked effect on bud development and further recovery of plantlets. The number and size of the shoots that developed from each bud were affected as a result of infection. The effect depended on the specific virus, the isolate and the host-disease combination. The possible implications of these results are discussed.     

Effect of Evaporative Demand on Water Relations of Citrus limon

Conductance, transpirational flux and xylem pressure potentialwere measured in leaves of well-watered 5-year-old lemon trees(Citrus limon (L.) Burm. f.) subjected to different levels ofevaporative demand. Increased leaf-to-air absolute humiditydifference generally decreased stomatal conductance and increasedxylem pressure potential, with a good correlation between thelast two parameters; but this trend was reversed on days withvery high evaporative demand, when stomata opened in spite ofthe low humidity. Citrus limon (L.) Burm. f., lemon, water stress, stomatal conductance, leaf water potential, transpiration, air humidity     

The Influence of Genotype, Temperature and Moisture on Seed Longevity in Chickpea, Cowpea and Soya bean

Seed of three chickpea (Cicer arietinum L.), three cowpea [Vignaunguiculata (L.) Walp.] and four soya bean [Glycine max (L.)Merr.] cultivars were hermetically stored for up to 2 yearsin various constant environments which included temperaturesfrom —20 to 70 °C and moisture contents (fresh weightbasis) from 5 to 25 per cent. In all cases the survival curvescould be described by negative cumulative normal distributions.The longevity of the various seed lots differed but the valueof the standard deviation (the reciprocal of which gives theslope of the survival curve when percentage germination is transformedto probit) was the same for all cultivars within a species whenstored under similar conditions. Within each species the relativeeffects of moisture and temperature on longevity did not differsignificantly between cultivars. In all three species therewas a negative logarithmic relationship between seed moisturecontent and longevity, but the relative effect of moisture contentdiffered between the species: differences in the longevity ofsoya bean seed as a function of moisture content were less thanfor either cowpea or chickpea. The relative effect of temperatureon seed longevity did not differ between the three species,and the seed of all three species showed increasing temperaturecoefficients for the change in rate of loss of viability withincrease in temperature. The complete pattern of loss in viabilityin all three species can be described by a single equation whichwas developed for barley and has also been shown to apply toonion seed. The constants applicable to the three grain legumeshave been calculated so that it is now possible to predict percentageviability of any seed lot of these species after any storageperiod under a very wide range of storage conditions. Cicer arietinum L., chickpea, Glycine max (L.) Merr., soya bean, Vigna unguiculata (L.) Walp., cowpea, seed longevity, seed storage, moisture content, temperature     

Antioxidant properties of essential oils from lemon,grapefruit, coriander,clove, and their mixtures

Antioxidant properties of individual essential oils from lemon (Citrus limon L.), pink grapefruit (Citrus paradisi L.), coriander (Coriandrum sativum L.), and clove (Caryophyllus aromaticus L.) buds and their mixtures were studied by capillary gas-liquid chromatography. Antioxidant activity was assessed by oxidation of the aliphatic aldehyde hexanal to the carboxylic acid. The lowest and highest antioxidant activities were exhibited by grapefruit and clove bud essential oils, respectively. Mixtures containing clove bud essential oil also strongly inhibited oxidation of hexanal. Changes in the composition of essential oils and their mixtures in the course of long-term storage in the light were studied. The stability of components of lemon and coriander essential oils in mixtures increased compared to individual essential oils.     

Carbon Economy of Sour Orange in Relation to Mycorrhizal Colonization and Phosphorus Status

The effects of plant phosphorus (P) status and the mycorrhizal(M) fungus, Glomus intraradices Schenck & Smith, on thecarbon (C) economy of sour orange (Citrus aurantium L.) weredetermined during and following active M colonization. Therewere four treatments: M seedlings grown at standard-strength(1 mM) P (M1) and nonmycorrhizal (NM) plants grown at one, twoand five times standard-strength P (NM1, NM2 and NM5). Mycorrhizalcolonization, tissue dry mass, P content, root length and leafarea were determined in five harvests from 6 to 15 weeks ofage. Rate of C assimilation (A) was determined at 7, 8 and 12weeks by gas exchange. Partitioning of ¹⁴ C was determined from7 to 15 weeks using a 10-min pulse followed by a 24-h chaseperiod. For a given attribute, M1 plants were compared to thecurve defining the NM response as a function of tissue P concentration.In contrast to the large effects of P nutrition on C economyof sour orange, M effects were generally subtle. Mycorrhizaeincreased the root biomass fraction, the root length/leaf arearatio and the percentage of ¹⁴C recovered from below-groundcomponents. A higher percentage of below-ground ¹⁴ C was inthe respiration and soil fractions in M than NM plants of equivalentP status. Mycorrhizal plants tended to enhance A only for abrief period. Mycorrhizal plants had lower relative growth ratesthan NM plants of equivalent P status, suggesting that the temporarilyenhance A of M plants did not fully compensate for their greaterbelow-ground carbon expenditure. Problems of interpreting thedynamic effects of mycorrhizae on C economy that are independentof P nutrition are discussed.Copyright 1993, 1999 Academic Press Citrus aurantium L., sour orange, carbon economy, ¹⁴carbon, CO₂ assimilation, vesicular-arbuscular mycorrhizae, phosphorus fertilization, phosphorus nutrition     

Comparison of continuously recorded plant-based water stress indicators for young lemon trees

Continuously recorded plant-based water stress indicators (sap flow and parameters derived from trunk diameter fluctuations) were compared in potted young lemon trees (Citrus limon (L.) Burm. fil, cv. Verna) grafted on sour orange (C. aurantium L.) rootstock submitted to deficit irrigation. Daily maximum (MXTD) and minimum (MNTD) trunk diameters and daily trunk diameter shrinkage (MDS) were directly influenced by the water supply to the trees from the soil. The continuously recorded plant-based water stress indicators presented different degrees of sensitivity when used to estimate the water status of the plants. Sap flow (SF) and MDS were more immediate and sensitive than MXTD and MNTD. However, the higher signal intensity: noise for SF and the fact that its signal intensities remained clearly above unity during the stress period, indicating that the soil water was depleted, point to the greater reliability of this indicator. Also, the possibility of developing further baseline relationships between SF and air vapour pressure deficit in fully irrigated trees in field conditions increases the feasibility of using this parameter in automatic irrigation systems.     

Soil temperature and flooding effects on two species of citrus

Summary Rough lemon (Citrus jambhiri Lush.) and sour orange (C. aurantium L.) seedlings were grown at constant soil temperatures of 16, 24, and 33 C for 3 months. Shoot and root growth of rough lemon was greatest at 33 C while growth of sour orange was greatest at 24 C. There were no significant effects of soil temperature on shoot: root ratio, leaf water potential or stomatal conductance. The hydraulic conductivity of intact root systems of both species was highest when seedlings were grown at 16 C. Thus, acclimation through greater root conductivity at low soil temperature may have compensated for decreased root growth at 16 C and negated effects of soil temperature on plant water relations. Half the plants growing at each soil temperature were subsequently flooded. Within 1 week, the soil redox potential (Eh) dropped below zero mV, reaching a minimum Eh of –250mV after 3 weeks of flooded conditions. Flooded plants exhibited lower root conductivity, a cessation of shoot growth, lower leaf water potentials, lower stomatal conductances, and visual sloughing of fibrous roots. Decreases in root conductivity in response to flooding were large enough to account for the observed decreases in stomatal conductance.Florida Agricultural Experiment Stations Journal Series No. 4080.     

Citrus somatic hybridization with potential for improved blight and CTV resistance

Summary The production of five new somatic hybrids with potential for improved disease resistance is reported herein. Protoplast isolation, fusion, and plant regeneration was achieved from Caipira sweet orange (Citrus sinensis L. Osbeck) as an embryogenic parental source and Volkamer lemon (C. volkameriana Pasquale), Cleopatra mandarin (C. reticulata Blanco), and Rough lemon (C. jambhiri Lushington) as non-embryogenic parental sources. Fusion involving Cleopatra mandarin and Rangpur lime (C. limonia L. Osbeck) as embryogenic parental sources with Sour orange (C. aurantium L.) also resulted in somatic hybrid plants. Somatic hybridization was confirmed by leaf morphology evaluation, chromosome counting, and randomly amplified polymorphic DNA (RAPD) analyses. Somatic hybrids may combine complementary characteristics from both parental sources and have potential for tolerance to blight and citrus tristeza virus (CTV).     

Leaf Characteristics and Net Gas Exchange of Diploid and Autotetraploid Citrus

The effect of tetraploidy on leaf characteristics and net gasexchange was studied in diploid (2x ) and autotetraploid (4x) ‘Valencia’ sweet orange (Citrus sinensis (L.)Osb.) and ‘Femminello’ lemon (Citrus limon (L.)Burm. f.) leaves. Comparisons between ploidy levels were madeunder high irradiance (I) in a growth chamber or low total Iin a glasshouse. Tetraploids of both species had thicker leaves,larger mesophyll cell volume and lower light transmittance thandiploids regardless of growth I. Mesophyll surface area perunit leaf area of 2x leaves was 5–15% greater than on4x leaves. Leaf thickness and mesophyll cell volume were greaterin high I leaves than low I leaves. In high I, average leafarea was similar for 2x and 4x leaves, whereas in low I it was30% greater in 4x than in 2x leaves. Nitrogen and chlorophyllconcentration per cell increased with ploidy level in both growthconditions. The ratio of chlorophyll a:b was 25% greater in2x than in 4x leaves. When net CO₂assimilation rate (A_CO2) wasbased on leaf area, 4x orange leaves had 24–35% lowerA_CO2than their diploids. There were no significant differencesin A_CO2between 2x and 4x orange or lemon leaves when expressedon a per cell basis. Overall, lower A_CO2per unit leaf area oftetraploids was related to increase in leaf thickness, largermesophyll cell volume, the decrease in mesophyll area exposedto internal air spaces, and the lower ratio between cell surfaceto cell volume. Such changes probably increased the resistanceto CO₂diffusion to the site of carboyxlation in the chloroplasts. Cell volume; chlorophyll; irradiance; leaf thickness; nitrogen; photosynthesis; ploidy; Citrus limon ; C. sinensis ; ‘Valencia’ sweet orange; ‘Femminello’ lemon     

Growth and root hydraulic conductivity of several citrus rootstocks under salt and polyethylene glycol stresses

The water relations responses to salt of several important citrus rootstocks such as Swingle citrumelo, sour orange, and Milam lemon have not been studied in detail before. Studies were set up to compare growth and root hydraulic properties of these rootstocks to other citrus rootstocks by exposing them to NaCl and polyethylene glycol (PEG) stresses. Seedlings of 7 citrus rootstocks were irrigated for 5 months with nutrient solutions containing NaCl or PEG that had been adjusted to osmotic potentials of -0.10, -0.20 or -0.35 MPa. The 7 rootstocks studied were sour orange (Citrus aurantium), Cleopatra mandarin (Citrus reticulata Blanco), Swingle citrumelo (C. paradisi x P. trifoliata), Carrizo citrange (C. sinensis x P. trifoliata), rough lemon (Citrus jambhiri Lush), Milam lemon (C. jambhiri hybrid), and trifoliate orange (Poncirus trifoliata [L.] Raf.). In both shoot and root growth, Cleopatra mandarin and sour orange were the least sensitive to salt, Milam and trifoliate orange were the most sensitive, and rough lemon, Swingle, and Carrizo were intermediate in sensitivity. Even though the roots were exposed to solutions of equal osmotic potentials, plant growth and root conductivity were reduced more by the PEG treatments than the corresponding NaCl treatments. At -0.10 and -0.20 MPa, shoot and root dry weights were reduced 16 to 55% by NaCl and 24 to 68% by PEG. Shoot root ratio was lowered at the higher concentrations, particularly by PEG. There was a major decrease in root conductivity caused by NaCl at -0.10 MPa (19 to 30% in sour orange and Cleopatra mandarin and 78 to 85% in trifoliate orange and Milam). Conductivity decreased more at -0.20 and -0.35 MPa, but not proportionally as much as at -0.10 MPa. Root weight per unit length increased at the higher salt levels, particularly in trifoliate orange. Water flow rate through root systems followed the same trend as root conductivity; salt affected sour orange and Cleopatra mandarin the least and trifoliate orange and Milam the most. However, reductions in fibrous root length by salt treatment differed. Root lengths of Swingle and Carrizo were least affected by salt while sour orange. Milam, and rough lemon were the most affected. Hence, even though sour orange and Cleopatra mandarin were more tolerant than the other rootstocks in terms of water flow rate or root conductivity, these 2 rootstocks showed a proportionally greater decrease in root length than Carrizo, Swingle, or trifoliate orange.     

Logarithmic Relationship between Moisture Content and Longevity in Sesame Seeds

The relationship between seed moisture content and seed longevityin sesame (Sesamum indicum L.) in hermetic storage at 50 °Cis logarithmic. The logarithmic relationship is maintained from15 per cent down to 2 per cent – the lowest moisture contenttested — but above 15 per cent this ‘air-dry’relationship no longer holds since further increase in seedmoisture content does not reduce longevity. Tentative estimatesof constant values for the improved seed viability equationare provided, and implications for long-term storage are discussed. Sesame, Sesamum indicum L., seed storage, improved viability equation, seed moisture content, seed longevity prediction