Jean-Marie Kinet

The use of the electrolyte leakage method for assessing cell membrane stability as a water stress tolerance test in durum wheat

This work was carried out to adapt the electrolyte leakage technique todurum wheat and then to evaluate its relevance in the assessment of the cellmembrane stability as a mechanism of water stress tolerance in this species.Themethod currently used is based on in vitro desiccation ofleaf tissues by a solution of polyethylene glycol (PEG) and a subsequentmeasurement of electrolyte leakage into deionised water. It consists of threesuccessive steps: (1) a washing treatment to remove solutes from both leafsurfaces and cells damaged by cutting; (2) a stress period during which theleaftissues are plunged in a PEG-solution and (3) a rehydration period during whichafter-effects of the stress are evaluated. During the washing period, the majorpart of electrolytes was removed within 15 min. Varying the stressconditions influenced both the percent and the kinetics of electrolyte leakageduring rehydration. Electrolyte leakage exhibited a characteristic patternreflecting the condition of cellular membranes (repair and hardening). Inpractice, we recommend a 15-minute washing time, a10-hour stress period and 4 h of rehydration. Theextent of the cell membrane damage not only correlated well with the growthresponses of wheat seedlings belonging to various cultivars to withholdingwaterbut also with the recognised field performances of these cultivars. Therelativeproportion of endogenous ions lost in the effusate during the rehydration stepmay vary strongly according to the element analysed and the precise nutritionalstatus of the plant should therefore be considered. However, an increase ininorganic ion leakage does not fully explain the recorded PEG-induced increasein electrical conductivity (EC) during the subsequent rehydration step andorganic ions are probably also involved in such an increase.

Water deficit effects on solute contribution to osmotic adjustment as a function of leaf ageing in three durum wheat (Triticum durum Desf.) cultivars performing differently in arid conditions.

A greenhouse study was carried out using three durum wheat (Triticum durum Desf.) cultivars differing in their field performances under arid conditions (Kabir 1, poor yield stability; Omrabi 5, high yield stability and Haurani, landrace well adapted to drought). Water stress was imposed by withholding water at the seedling stage. Water potential (Psi(w)), relative water content (RWC), stomatal resistance (SR), and changes in solute concentrations were quantified: (1) as a function of leaf development during the stress period; and (2) in young expanded and growing leaves harvested at the end of the stress treatment. Psi(w), RWC and SR were almost unaffected by leaf age in controls. In contrast, solute concentrations appeared to vary in the course of leaf development. During the stress treatment, Psi(w) and RWC decreased and SR increased in all cultivars; the changes were most often largest in Omrabi 5, lowest in Haurani and intermediate in Kabir 1. Water stress also increased sugar and proline concentrations and decreased nitrate levels. Young expanded and growing leaves differed in terms of Psi(w), RWC and osmotic adjustment (OA). The capacity of OA was greater in growing than in expanded leaves, especially in the two cultivars best adapted to aridity, and allowed turgor maintenance in these genotypes. Sugars were the main solutes that contributed to OA particularly in growing leaves followed by proline and then quaternary ammonium compounds. The contributions of these organic solutes to OA tended to be higher in Omrabi 5 and in Haurani than in Kabir 1. Inorganic solutes, however, did not seem to play an important role in OA despite their high proportion in total solutes.

Salt stress effects on roots and leaves of Atriplex halimus L. and their corresponding callus cultures

Salt stress effects on growth, osmotic adjustment, mineral and organic contents and soluble peroxidase activities were determined in roots and leaves of Atriplex halimus and their corresponding callus cultures. Low NaCl doses (150 mM) promoted shoot growth, corroborating the halophilic nature of this species; in these stress conditions, Na+ concentration markedly increased in the leaves indicating that salinity resistance was not associated with the ability of the plants to restrict sodium accumulation in the aerial part. Whole organs and their corresponding calli were able to cope with high NaCl doses but there was no clear correspondence between the physiological behaviour of cell culture and whole plant. For several physiological parameters (osmotic potential (Psi s), mineral content, proline accumulation), roots were less affected by NaCl than leaves while both root and leaf calli behaved in the same way in response to salinity. NaCl-induced modifications of the recorded parameters are discussed in relation to the mechanisms of salinity resistance in this species. Evidence indicated the existence of a cellular basis for salinity resistance in A. halimus, but the expression of this cellular property at organ level appeared to be masked by the physiological complexity of the intact plant and the nature of the whole organ response was apparently determined primarily by regulation mechanisms assigned by the differentiated tissue organisation

Cosuppression of a Plasma Membrane H + -ATPase Isoform Impairs Sucrose Translocation, Stomatal Opening, Plant Growth, and Male Fertility

The plasma membrane H+-ATPase builds up a pH and potential gradient across the plasma membrane, thus activating a series of secondary ion and metabolite transporters. pma4 (for plasma membrane H+-ATPase 4), the most widely expressed H+-ATPase isogene in Nicotiana plumbaginifolia, was overexpressed in tobacco. Plants that overexpressed PMA4 showed no major changes in plant growth under normal conditions. However, two transformants were identified by their stunted growth, slow leaf initiation, delayed stem bolting and flowering, and male sterility. Protein gel blot analysis showed that expression of the endogenous and transgenic pma4 was cosuppressed. Cosuppression was developmentally regulated because PMA4 was still present in developing leaves but was not detected in mature leaves. The glucose and fructose content increased threefold, whereas the sucrose content remained unchanged. The rate of sucrose exudation from mature leaves was reduced threefold and the sugar content of apical buds was reduced twofold, suggesting failure of sucrose loading and translocation to the sink tissues. Cosuppression of PMA4 also affected the guard cells, stomatal opening, and photosynthesis in mature leaves. These results show that a single H+-ATPase isoform plays a major role in several transport-dependent physiological processes.

Effect of light conditions on the development of the inflorescence in tomato

Abstract By growing tomato plants ( Lycopersicon esculentum Mill.) in 4 different light regimes (2 photoperiods — 8 and 16 h — combined with 2 light intensities — 9,000 and 18,000 ergs cm −2 s −1 , it was shown that increasing light integrals hastened flower initiation, greatly promoted the development of the inflorescence and increased the rates of leaf production and the growth of the stem. In similar light integrals, flower initiation was earlier and inflorescence development far better in short photoperiods than in long ones; the rates of leaf production were almost the same and stem growth was greater in long days. Transfer experiments from favourable to insufficient light conditions and inverse transfers at different times during the life of the plant indicated that light conditions were critical at the time of, and after, the macroscopic appearance of the inflorescence. At that stage, a transfer to low light conditions for 10 days induced complete abortion of the truss in our growth conditions. The effect of a transfer from insufficient to favourable light conditions was slower since at least 15 days in these latter conditions were required in order to achieve the development of the inflorescence.

Ethylene production by leaves of rice (Oryza sativa L.) in relation to salinity tolerance and exogenous putrescine application

The effects of salt stress on the production of ethylene in leaves of five rice cultivars differing in salinity tolerance were investigated after 15 and 30 days of stress exposure in plants sprayed every 2 days with deionised water or putrescine 100 mu M. NaCl induced an increase in ethylene synthesis which was higher in salt-tolerant than in salt-sensitive cultivars (cvs.), Salt-stress also induced an increase in 1-aminocyclopropane-1-carboxylic acid (ACC) content which was larger in salt-tolerant cvs, whereas a decrease in ACC conversion to ethylene suggested a reduction in ACC oxidase activity. Putrescine increased the growth and the leaf tissue viability (as indicated by the reduction of 2,3,5-triphenyltetrazolium chloride (TTC) to formazan) of salt-treated plants in all cvs. This positive effect was associated with an increase in ethylene biosynthesis through an increase in ACC content and a suppression of NaCl-induced inhibition of ACC conversion to ethylene. These data suggest that putrescine could be involved in salinity tolerance in rice.

Sucrose increase during floral induction in the phloem sap collected at the apical part of the shoot of the long-day plant Sinapis alba L.

Sinapis alba L., a long-day plant, has been induced to flower either by a single 22-h-long photoperiod or by an 8-h short photoperiod displaced by 10 h in a 24 h cycle. The ehtylenediametetraacetate method previously used for leaf exudation was modified to collect phloem sap at the apical part of the shoot. Carbohydrates in the phloem sap have been analysed comparatively in vegetative and induced plants, using high-performance liquid chromatography and refractometry. Sucrose was the major sugar detected. A dramatic increase of its flux in the apical sap occurred early and transiently during the floral transition in plants induced by both long days and displaced short days. These results indicate a message-like role for sucrose since they fit nicely with previous observations indicating that an early event in the floral transition in S. alba is the accumulation of sucrose in the meristem.

Morphology of Nectaries and Biology of Nectar Production in the Distylous Species Fagopyrum esculentum

BACKGROUND AND AIMS The mechanisms of floral nectar production in buckwheat (Fagopyrum esculentum, Polygonaceae), a distylous pseudo-cereal, have received relatively little attention, prompting an investigation of the factors that regulate this process. The aim was to perform a refined study of the structures that secrete nectar and of the internal and external parameters influencing nectar volumes and sugar concentrations. METHODS In order to control environmental parameters, plants were cultivated in growth rooms under controlled conditions. The structure of nectaries was studied based on histological sections from flowers and flower buds. Nectar was extracted using glass micropipettes and the sugar concentration was measured with a hand refractometer. Sugar concentration in the phloem sap was measured using the anthrone method. To test the influence of photosynthesis on nectar production, different light and defoliation treatments were applied. KEY RESULTS Unicellular trichomes were located in the epidermis at the ventral part of eight nectary glands situated on the flower receptacle alternately with stamens. Vascular bundles consisting of both phloem and xylem were identified at the boundary between a multilayered nectary parenchyma and a sub-nectary parenchyma with chloroplasts. A higher volume of nectar in thrum morphs was observed. No other difference was found in morphology or in sugar supply to inflorescences between morphs. Nectar secretion was strongly influenced by plant age and inflorescence position. Nectar volumes were higher in the upper inflorescences and during the flowering peak. Light had a dual role, (1) acting directly on reproductive structures to trigger flower opening, which conditions nectar secretion, and (2) stimulating photosynthetic activity, which regulates nectar accumulation in open flowers. CONCLUSIONS In buckwheat, nectar is secreted by trichomes and probably proceeds, at least in part, from phloem sap. Nectar secretion is strongly influenced by floral morph type, plant age, inflorescence position and light.

Physiological changes after exposure to and recovery from polyethylene glycol-induced water deficit in callus cultures issued from durum wheat (Triticum durum Desf.) cultivars differing in drought resistance

Calli obtained from three durum wheat (Triticum durum Desf.) cultivars with different drought resistance levels were exposed for 30 days to a polyethylene glycol-induced water deficit followed by 30 days of recovery. Relative growth rate, percent dry weight, osmotic potential and the changes in inorganic and organic solutes were determined at the end of both the stress and the recovery periods. After the stress period, calli derived from the drought resistant cultivars, particularly Omrabi 5 and to some extent Haurani, showed a less reduced relative growth rate, lower osmotic potential and higher proline and other amino acid accumulations as compared with the sensitive cultivar Kabir I. Drought resistance could also be associated with a better protection of enzymes involved in nitrogen metabolism. At the end of the recovery period, most of the quantified parameters, except osmotic potential, recovered completely, indicating the reversibility of the changes induced by water deficit at the plant cell level in durum wheat. Our data show that a correlation exists between performances of the cultivars under drought and the responses of callus cultures to PEG-induced water deficit; this suggests that: in durum wheat the degree of resistance to drought at the plant level depends, at least in part, on the existence of mechanisms operating at the cellular level.

Effect of prolonged cold storage on the production capacity of strawberry plants

Abstract After cold storage for various periods (42, 99, 155, 189, 227 and 259 days) strawberry waiting-bed plants cultivar ‘Elsanta’ were planted on six dates between January and August. Plant performance was evaluated simultaneously in a growth chamber under controlled environmental conditions and in a greenhouse. Starch content in the crown tissue was clearly reduced during the cold storage period. In the greenhouse trials a variable decline in fruit number and yield per plant was found in relation to the length of cold storage and the temperature conditions during the growth and harvest period. Also in the growth chamber, under a constant temperature regime, a significant decline in the number of flowers per plant developing up to anthesis was observed after increasing periods of storage. This indicates that it is not only stressing climatical conditions after planting which are influencing the performance of cold stored plants. Also, the length of cold storage strongly affects plant vigour and productivity. This is related to the carbohydrate status of the plants after cold storage.