Localization of pectins in the pollen tube wall of Ornithogalum virens L. Does the pattern of pectin distribution depend on the growth rate of the pollen tube?

Monoclonal antibodies that recognize pectins were used for the localization of esterified (JIM7) and acidic, unesterified (JIM5) forms of pectin in pollen tube walls of Ornithogalum virens L. (x = n = 3). The results indicated that the distribution of the two forms of pectin in the pollen tube wall depended on the medium (liquid or solid) used for pollen germination. In pollen tubes grown in the liquid medium, the localization of JIM7 was limited to the very tip of the pollen tube, whereas the localization of JIM5 indicated a uniform distribution of unesterified pectins in the very tip of the tube and along the subapical parts of the tube wall. In tubes germinated on the medium stabilized with agar (1–2%) the localization of JIM7 and JIM5 indicated the presence of both forms of pectin in the tube tip and along the whole length of the pollen tube wall in a ring-like pattern. Thus, the localization of esterified pectins in the sub-apical part of the pollen tube wall, below the apex of the tube, is described for the first time. Measurements of the growth rates of pollen tubes growing on the two types of medium indicated that oscillations in tube growth rate occur but these do not coincide with the pattern of pectin distribution in the tube wall. Our results complement the previous data obtained for the localization of JIM5 and JIM7 in pollen tube walls of other plant species. (Y.-Q. Li et al. 1994, Sex Plant Reprod 7: 145–150) and provide new insight into an understanding of the construction of the pollen tube wall and the physiology of pollen grain germination. Received: 25 January 1999 / Accepted: 23 June 1999     

Xyloglucan−pectin linkages are formed intra-protoplasmically,contribute to wall-assembly,and remain stable in the cell wall

We tested two hypotheses for the mechanism by which xyloglucan–pectin covalent bonds are formed in Arabidopsis cell cultures. Hypothesis 1 proposed hetero-transglycosylation, with xyloglucan as donor substrate and a rhamnogalacturonan-I (RG-I) side-chain as acceptor. We looked for enzyme activities that catalyse this reaction using α-(1→5)-l-[³H]arabino- or β-(1→4)-d-[³H]galacto-oligosaccharides as model acceptor substrates. The ³H-oligosaccharides were supplied (with or without added xyloglucans) to living Arabidopsis cell-cultures, permeabilised cells, cell-free extracts, or four authentic XTHs. No hetero-transglycosylation occurred. Therefore, we cannot support hypothesis 1. Hypothesis 2 proposed that some xyloglucan is manufactured de novo as a side-chain on RG-I. To test this, we pulse-labelled Arabidopsis cell-cultures with [³H]arabinose and monitored the radiolabelling of anionic (pectin-bonded) xyloglucan, which was resolved from free xyloglucan by ion-exchange chromatography. [³H]Xyloglucan–pectin complexes were detectable <4 min after [³H]arabinose feeding, which is shorter than the transit-time for polysaccharide secretion, indicating that xyloglucan–pectin bonds were formed intra-protoplasmically. Thereafter, the proportion of the wall-bound [³H]xyloglucan that was anionic remained almost constant at ∼50% for ≥6 days, showing that the xyloglucan–pectin bond was stable in vivo. Some [³H]xyloglucan was rapidly sloughed into the medium instead of becoming wall-bound. Only ∼30% of the sloughed [³H]xyloglucan was anionic, indicating that bonding to pectin promoted the integration of xyloglucan into the wall. We conclude that ∼50% of xyloglucan in cultured Arabidopsis cells is synthesised on a pectic primer, then secreted into the apoplast, where the xyloglucan–pectin bonds are stable and the pectic moiety aids wall-assembly.     

A suberized perimedullary reaction zone in Populus balsamifera novel for compartmentalization in trees

 Following artificial inoculation of nonhost Populus balsamifera with Ophiostoma ulmi, structural defensive tissues were formed in the xylem. Among these tissues there was a perimedullary sheath of cells, located adjacent to the invaded xylem, that originated from the dedifferentiation of perimedullary and xylem parenchyma cells. Histochemical tests revealed that this sheath was intensively suberized. A band of lignified cells was frequently detected on both sides of this suberized tissue. The formation of such a tissue at the pith margin represents a new type of anatomical barrier in relation to compartmentalization processes described for trees. Ultrastructural examination showed that the wall of cells forming this zone was generally composed of a compound middle lamella, a suberized secondary wall and a tertiary wall layer. Using colloidal gold conjugated to monoclonal antibodies against pectin and to an exoglucanase for cellulose, only limited labelling was obtained for pectin whereas labelling for cellulose was abundant in the compound middle lamella and the tertiary wall layer. In a few fibres close to this suberized zone, the latter probe also made it possible to distinguish the occasional presence of several alternating wall layers mainly composed of either suberin or cellulose. In Salix sp., another tree species belonging to the Salicaceae, this type of suberized reaction zone was also observed. The new reaction zone is similar in structure and location to a suberized barrier formed in nonwoody carnation (Dianthus caryophyllus) plants in the defense against vascular fungi. Received: 30 July 1996 / Accepted: 7 January 1997     

Foliage consumption and larval development of parasitized and unparasitized soybean looper,Pseudoplusia includens [Lep.: Noctuidae], reared on a resistant soybean genotype and effects on an associated parasitoid,Compidosoma truncatellum [Hym.: Encyrtidae]

Field grown foliage from the resistant soybean [Glycine max (L.) Merrill] breeding line GAT “81–327” and the susceptible cultivar “Ransom” was used to rear unparasitized larvae of the soybean looper (SBL),Pseudoplusia includens (Walker), and larvae parasitized byCopidosoma truncatellum (Dalman). SBL larvae, whether parasitized or not, consumed more foliage when fed “Ransom”. Unparasitized larvae reared on “81–327” had longer developmental times and suffered greater mortality than unparasitized larvae reared on “Ransom”. Parasitization of SBL larvae byC. truncatellum increased total foliage consumption of both soybean lines. Parasitized larvae reared on the resistant “81–327” weighed less and yielded fewer parasitoid adults.

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Résumé Des larves dePseudoplusia includens (Walker) parasitées ou non parCopidosoma truncatellum (Dalman) ont été nourries des feuilles de deux lignées de soja [Glycine max (L.) Merrill] l'une, GAT “81–327” résistante et l'autre, “Ransom” sensible. Les larves deP. includens qu'elles soient parasitées ou non consommaient plus de feuillage lorsqu'elles étaient nourries de la lignée “Ransom”. Les larves non parasitées élevées sur “81–327” avaient un cycle de développement beaucoup plus long et un taux de mortalité beaucoup plus élevé que les larves non parasitées élevées comparativement sur feuilles de “Ransom”. Par contre, les larves parasitées manifestaient une consommation accrue du feuillage des deux lignées de soja. Les larves parasitées élevées sur les feuilles de la variété résistante GAT “81–327” pesaient moins et produisaient moins également de parasites adultes.

     

Pectin immunolocalization and calcium visualization in differentiating derivatives from poplar cambium

Summary Calcium distribution and pectin esterification patterns in the cambial zone of poplar branches were studied with ionic microscopy and immunological tools respectively. Dynamic changes correlating with cell growth and cell differentiation were observed both on the xylem and on the phloem sides. In expanding cell walls of xylem derivatives, unesterified pectins were restricted to cell junctions and middle lamellae, occasionally accompanied by calcium ions. In contrast, in differentiating and mature phloem cells, acidic pectins and Ca²⁺ were present all over the walls leading to early stiffening of the polysaccharide network. Significant labelling was detected with JIM5 antibodies in some dictyosomes suggesting exocytosis of low methylated polymers towards the cell walls. At cell junctions, unesterified pectins might originate from the activity of pectinmethylesterases localized in these areas. Thus un- and deesterified pectins might be located in different cell wall domains whose distribution, varying with cell type, will confer specific extensibility to the wall matrix.Abbreviations BSA bovine serum albumin - DM degree of methylation - FITC fluorescein isothiocyanate - HM highly methylated pectins - LM low methylated pectins - PME pectin methylesterase - SIMS secondary ion mass spectrometry - TBS tris-buffered saline     

Effects ofBacillus thuringiensis var.San diego on predation effectiveness,development and mortality ofColeomegilla maculata lengi (Col.: Coccinellidae) larvae

Laboratory experiments were conducted to determine the effects of M-One^™ (Bacillus thuringiensis var.san diego) on larval instars ofColeomegilla maculata lengi Timberlake. Coccinellid larval development (from egg hatch to adult), completed on pollen treated with suspensions of M-One^™ at 20 ml/litre (5.6×10⁸ CPBIU/litre) and 200 ml/litre, took respectively 29.3 and 38.5 days compared with 21.9 days for the control (water). M-One^™ did not cause larval mortality.C. maculata third instars did not show any preference between eggs ofLeptinotarsa decemlineata (Say) treated with water or with M-One^™ at 20 ml/litre. However, at 200 ml M-One^™/litre, the number of eggs attacked was 34.7% lower than the eggs treated with water only, 48 h after the beginning of the test. These results indicate that the use of M-One^™, at the manufacturer's recommended field rate of 20 ml/litre, does not cause a major threat to larvalC. maculata populations.

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Résumé Des bioessais en laboratoire ont été effectués afin de déterminer si les larves de la coccinelle maculée,Coleomegilla maculata lengi Timberlake (Col.: Coccinellidae), sont affectées par M-One^™, un insecticide biologique préparé à partir de la bactérieBacillus thuringiensis var.san diego Berliner et utilisé dans la lutte contre le doryphore de la pomme de terre,Leptinotarsa decemlineata (Say) (Col.: Chrysomelidae). Le développement larvaire, effectué sur du pollen traité avec des concentrations de 20 ml M-One^™/litre (5,6×10⁸ unités internationales de doryphore/litre) et 200 ml M-One^™/litre, a nécessité 29,3 et 38,5 jours respectivement, comparativement à 21,9 jours pour le témoin (eau). M-One^™ n'a pas causé de mortalité chez les larves. Au cours d'un test de 48 h, les larves de stade III n'ont montré aucune préférence entre des œufs traités avec 20 ml M-One^™/litre et des œufs traités avec de l'eau. Par contre avec 200 ml M-One^™/litre, le nombre d'œufs attaqués a diminué significativement de 34,7% par rapport au témoin, 48 h après le début du test. Ces résultats indiquent que l'utilisation de M-One^™ à la concentration recommandée de 20 ml/litre ne représente pas une menace pour les populations larvaires de la coccinelle maculée.

     

Predation byIridomyrmex humilis [Hym.: Formicidae] on eggs ofChrysoperla carnea [Neu.: Chrysopidae] released for inundative control ofIllinoia liriodendri [Hom.: Aphididae] infestingLiriodendron tulipifera

In an effort to suppress the tuliptree aphidIllinoia liriodendri (Monell), approximately 2,000 eggs ofChrysoperla carnea (Stephens) from a commercial insectary were released 4 times on each of 8 tuliptreesLiriodendron tulipifera L. in Berkeley, California, during the spring of 1984. On trees foraged by the Argentine antIridomyrmex humilis (Mayr), 98% of the eggs ofC. carnea were removed from the egg release tapes by the ants. A total of about 1,250 larvae per tree eclosed from the 8,000 eggs released on each tree without ants. Fifty percent of the larvae that did eclose died due to cannibalism or entrapment in the sticky egg release tapes and approximately 625 first instar larvae on each tree were free to forage for aphids. Inundative lacewing releases ofC. carnea did not suppress populations ofI. liriodendri due to ant predation, the low viability of commercial eggs (0–73% eclosion),

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Résumé Dans le but de limiter les populations du puceron du tulipierIllinoia liriodendri (Monell), 4 lachers d'environ 2.000 œufs de provenance commerciale deChrysoperla carnea (Stephens) ont été réalisés au cours du printemps 1984 sur 8 tulipiersLiriodendron tulipifera L., à Berkeley en Californie. cannibalism by emerged larvae, and inadequate release technology. Sur les arbres visités par la fourmi d'ArgentineIridomyrmex humilis (Mayr), 98% des œufs deC. carnea ont été enlevés du support artificiel par les fourmis. Sur les arbres exempts de fourmis, la mortalité de 50% des larves est due au cannibalisme ou à leur engluement sur le support de lacher. A partir de 8.000 œufs déposés sur chaque arbre sans fourmi, on aboutit à environ 625 larves de 1^er stade susceptibles de rechercher des pucerons. Les lachers inondatifs deC. carnea n'ont pas limité les populations d'I. liriodendri. Les raisons en sont: la consommation par les fourmis, une faible viabilité de la plus grande partie des œufs commercialisés (0–73% d'éclosions), une technique inadaptée pour le lacher des œufs et le cannibalisme par les larves elles-mêmes deC. carnea.

     

Acetyl- and methyl-esterification of pectins of friable and compact sugar-beet calli: consequences for intercellular adhesion

Monoclonal antibodies (2F4), specific for a conformational epitope of homopolygalacturonic acid induced by calcium ions, were used to compare the nature and the distribution of the pectic polysaccharides in cell walls of compact and friable sugar-beet (Beta vulgaris L. var. altissima) calli, at the electron-microscope level. Labelings performed before or after de-esterification pretreatments of callus sections enabled three major types of pectic polysaccharides to be distinguished within compact calli: (i) acidic pectins, probably with few acetyl ester groups, detected without any de-esterification treatment in expanded areas of cell separation but never on middle lamellae between tightly associated cells; (ii) highly methyl-esterified pectins with an expected low acetyl ester content, recognized by the 2F4 antibodies after pectin methylesterase de-esterification, and mostly located on intercellular junctions and on middle lamellae in the central zones of the calli; (iii) highly methyl-esterified and largely acetylated pectins, only localized after alkaline de-esterification, in all primary walls of the compact calli. By contrast, all pectins of friable calli were highly methyland acetyl-esterified. This was consistent with an average degree of methyl-esterification of about 60% measured in both calli, and a higher average degree of acetylation for the friable callus line (85%) compared to the compact one (60%). Accordingly, the pectic fraction (acid-soluble) predominant in both calli was acetyl-esterified to 85% in friable callus and to 22% in compact callus cell walls. Friability of sugar-beet callus is thus correlated with an increase in acetylation of its pectin. Labelings of the Golgi apparatus indicate that the pectic polymers of both callus types are synthesized in dictyosomes in a highly methyl-esterified form and are probably subsequently acetyl-esterified.Abbreviations AIR alcohol-insoluble residue - DA degree of acetylation - DM degree of methyl-esterification - MAbs monoclonal antibodies - PME pectin methylesterase Many thanks are due to Mrs. Ch. Devignon (Unité interfacultaire de microscopie électronique, FUNDP, Namur, Belgium) for her technical assistance. F.L. gratefully acknowledges Dr. J.-F. Thibault (Laboratoire de Biochimie et Technologie des glucides, INRA, Nantes, France) for allowing her to stay in his laboratory and Dr. C. Renard for her help with biochemical analyses and her comments on the results. Appreciation is also expressed to P. Vandersmissen (Unité Cell, I.C.P., Bruxelles, Belgium) and to P. Cambier and C. Vinals (FUNDP) for their contribution. The work reported here was supported in part by grants from IRSIA and CGRI, Belgium, to F.L.     

Biological activity of five strains ofMetarhizium anisopliae,upon the coffee berry borerHypothenemus hampei (Col.: Scolytidae)

Five strains of the entomopathogenic fungusMetarhizium anisopliae (Metschnikoff) Sorokin were bioassayed against the coffee berry borerHypothenemus hampei (Ferrari) (Coleoptera: Scolytidae) under laboratory conditions (T=27±2°C; R.H.=90±5% and 12∶12 L∶D light cycle). The LC₅₀ values for the three most virulent strains, Ma4, Ma5 and Ma3, were 4.2, 5.9 and 6.7×10⁶ conidia/ml of suspension respectively and median lethal times (LT₅₀) observed were between 9.7 and 13.8 days. The maximum percentage sporulation was obtained with strains Ma3 and Ma4 both with 90% sporulation and Ma10 with 60.7%. These results suggest that the fungusM. anisopliae could be incorporated into an integrated control programme for the biological control ofH. hampei.

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Résumé Cinq souches du champignon entomopathogèneMetarhizium anisopliae (Metschnikoff) Sorokin, ont été évaluées dans les conditions du laboratoire, pour mesurer leur activité vis-à-vis du scolyte des fruits du caféierHypothenemus hampei (Ferrari) (Coleoptera: Scolytidae). Les résultats montrent que les souches Ma4, Ma5 et Ma3 sont les plus virulentes contreH. hampei, avec des valeurs de concentration létale moyenne (CL₅₀, valeurs minimales et maximales, ± limite de confiance à 95%) de 0,0226 (0,0195–0,00260), 0,0328 (0,0267–0,0390) et 0,0354 (0,0297–0,0417) équivalentes à 4,2, 5,9 et 6,7×10⁶ conidies/ml de suspension et un temps létal moyen (TL₅₀) entre 9,7 et 13,8 jours. Les pourcentages maximaux de sporulation ont été obtenus avec les souches Ma3 et Ma4 avec 90%, la souche Ma10 présentant le pourcentage le plus bas de sporulation (67%). Ces résultats démontrent queM. anisopliae peut être introduit dans un programme de lutte intégrée contreH. hampei.

     

Lutte biologique et intégrée dans les vergers de citrus en zone méditerranéenne

Les problèmes posés par les ravageurs des Citrus ont pris une importance considérable au cours de ces 3 dernières décennies et cela pour plusieurs raisons:

Predation byCheilomenes vicina [Coleoptera: Coccinellidae] on the cowpea aphid,Aphis craccivora [Homoptera: Aphididae]: Effect of prey stage and density

Consumption of larvae and females ofAphis craccivora Koch by 1st and 4th larvae and adults ofCheilomenes vicina (Muls.) was studied under fluctuating temperature (24–30°C). The early aphid instars were consumed in significantly greater numbers than later instars and females. The feeding rates ofC. vicina were significantly positively correlated with the population density of prey. The number of prey consumed daily by each predator stage tested, increased more steeply at lower than at higher prey densities, exhibiting thus the type 2 functional response

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Résumé Consommation des larves et des ♀ d'Aphis craccivora par les11^e stades, les 4^e stades et les adultes deCheilomenes vicina a été étudiée à une température variant de 24 à 30°C. Les jeunes stades du puceron sont consommés en nombre plus grand significativement que les derniers stades et les ♂. Les taux d'alimentation deC. vicina manifestent une corrélation hautement significative avec la densité de population de la proie. Le nombre de proies consommées quotidiennement par chaque stade considéré du prédateur augmentait plus brusquement aux faibles qu'aux fortes densités de proies, manifestant ainsi chez le prédateur le type 2 de réponse fonctionnelle.

     

Localization of cell wall polysaccharides in nonarticulated laticifers ofAsclepias speciosa Torr.

Summary Asclepias speciosa Torr, has latex-containing cells known as nonarticulated laticifers. In stem sections of this species, we have analyzed the cell walls of nonarticulated laticifers and surrounding cells with various stains, lectins, and monoclonal antibodies. These analyses revealed that laticifer walls are rich in (1→4) β-D-glucans and pectin polymers. Immunolocalization of pectic epitopes with the antihomogalacturonan antibodies JIM5 and JIM7 produced distinct labeling patterns. JIM7 labeled all cells including laticifers, while JIM5 only labeled mature epidermal cells and xylem elements. Two antibodies, LM5 and LM6, which recognize rhamnogalacturonan I epitopes distinctly labeled laticifer walls. LM6, which binds to a (l→5) α-arabinan epitope, labeled laticifer walls more intensely than walls of other cells. LM5, which recognizes a (1→4) β-D-galac-tan epitope, did not label laticifer segments at the shoot apex but labeled more mature portions of laticifers. Also the LM5 antibody did not label cells at the shoot apical meristem, but as cells grew and matured the LM5 epitope was expressed in all cells. LM2, a monoclonal antibody that binds to β-D-glucuronic acid residues in arabinogalactan proteins, did not label laticifers but specifically labeled sieve tubes. Sieve tubes were also specifically labeled byRicinus communis agglutinin, a lectin that binds to terminal β-D-galactosyl residues. Taken together, the analyses conducted showed that laticifer walls have distinctive cytochemical properties and that these properties change along the length of laticifers. In addition, this study revealed differences in the expression of pectin and arabinogalactan protein epitopes during shoot development or among different cell types.     

Ultrastructural immunolocalization of periodic pectin depositions in the cell wall ofNicotiana tabacum pollen tubes

Summary The monoclonal antibody (MAb) JIM5, marking acidic pectins, was used to localize ultrastructurally pectin molecules in the pollen tube wall ofNicotiana tabacum. Longitudinal sections of LR-White embedded pollen tubes were exposed to antibody treatment; accumulations of pectins were identified by counting the density of the gold particles representing the pectin epitopes along the pollen tube wall. Significant accumulations of gold grains were marked and the distances between them were measured. In many pollen tubes a more or less regular distribution of the accumulations was observed along the tube indicating a periodical deposition of pectin. The distances between the accumulations were 4–6 m. Most of the label was found in the inner part of the outer layer of the bilayered cell wall. These findings correspond to and confirm the earlier observation by our group reporting ring-shaped periodical deposits in pollen tubes after immunofluorescence labelling with the MAb JIM5 under the confocal laser scanning microscope.Abbreviations Ab antibody - MAb monoclonal antibody     

ON THE ULTRASTRUCTURE OF CAMBIUM AND ITS VASCULAR DERIVATIVES. III. THE SECONDARY WALLS OF THE SIEVE ELEMENTS OF PINUS STROBUS

The sieve elements of Pinus strobus have thick, lamellate secondary walls, which are composed predominantly of cellulose and lesser amounts of polyuronides and pectins. Eight to 10 lamellae may be present in walls 2-3 μ in thickness. Each lamella represents a plane of high cellulose density which results from intersection of two parallel sets of fibrils. Polyuronides and pectins are more or less evenly distributed in the wall, possibly with a greater concentration near the middle lamella and the inner surface. Resemblance of these walls to the so-called nacré walls is indicated, and it is possible that the two represent the same structure.     

Development of genotype-independent regeneration system for transformation of rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> ssp. <Emphasis Type="Italic">indica</Emphasis>)

Rice (Oryza sativa ssp. indica) is an important economic crop in many countries. Although a variety of conventional methods have been developed to improve this plant, manipulation by genetic engineering is still complicated. We have established a system of multiple shoot regeneration from rice shoot apical meristem. By use of MS medium containing 4 mg L⁻¹ thidiazuron (TDZ) multiple shoots were successfully developed directly from the meristem without an intervening callus stage. All rice cultivars tested responded well on the medium and regenerated to plantlets that were readily transferred to soil within 5–8 weeks. The tissue culture system was suitable for Agrobacterium-mediated transformation and different factors affecting transformation efficiency were investigated. Agrobacterium strain EHA105 containing the plasmid pCAMBIA1301 was used. The lowest concentration of hygromycin B in combined with either 250 mg L⁻¹ carbenicillin or 250 mg L⁻¹ cefotaxime to kill the rice shoot apical meristem was 50 mg L⁻¹ and carbenicillin was more effective than cefotaxime. Two-hundred micromolar acetosyringone had no effect on the efficiency of transient expression. Sonication of rice shoot apical meristem for 10 s during bacterial immersion increased transient GUS expression in three-day co-cultivated seedlings. The gus gene was found to be integrated into the genome of the T₀ transformant plantlets.     

Identification par immunofluorescence des cellules corticotropes et mélanotropes de l'hypophyse des Amphibiens

Résumé Les antisérums préparés à l'égard de trois hormones de synthèse [β(1–24) corticotropine, α et β MSH] nous ont servi pour localiser par immunofluorescence dans l'adénohypophyse des Amphibiens les sites d'élaboration de l'ACTH, de l'α et de la β MSH. Les anticorps anti-α et anti-β MSH se fixent exclusivement sur les cellules de lapars intermedia tandis que l'anti β-(1–24) corticotropine révèle simultanément une catégorie de cellules de lapars distalis et des cellules de lapars intermedia. D'après le contr?le cytologique effectué par la technique au bleu Alcian PAS, l'ensemble de ces cellules sont PAS positives et bleu alcian négatives. Elles réagissent également à la paraldéhyde fuchsine en l'absence d'oxydation permanganique. La signification de ces résultats est discutée.

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Identification by immunofluorescence of the corticotropin and melanotropin cells in Amphibian pituitary

Summary Antisera prepared against synthetic β(1–24)corticotropin (synacthen) α and β MSH were used to localize, by immunofluorescent procedure in the adenohypophysis of Amphibians, the site of production of ACTH, α and β MSH. Anti-α and anti-β MSH antibodies appeared to be bound only to thepars intermedia cells whereas anti-β(1–24) corticotropin reacted simultaneously with specific cells of thepars distalis and cells of thepars intermedia. Cytological control by alcian blue (pH 3) PAS technique proved that all these cells are PAS positive and alcian blue negative. They react also with paraldehyde fuchsine without oxidation. The possible significance of these results is discussed.

     

Biology of the spiderChiracanthium mildei [Arachnida: Clubionidae]

The biology ofChiracanthium mildei L. Koch was studied under standardized laboratory conditions (24±1°C, 55–60% RH). Specimens were reared from eggs, with 1- to 6-day-oldSpodoptera larvae serving as prey throughout their life cycle. Biological studies included an investigation of the rate of development, life cycle, reproductive potential and behavior, as well as the influence of certain environmental factors on these parameters. Under standard laboratory conditions,C. mildei showed 87% survival from egg to functional fertility. Males required a mean of 182 (137–207) days after hatching to reach maturity, became adults following 7–8 molts, and lived for an average of 73 days as adults. Females required a mean of 231 (191–286) days after hatching to mature, reached adulthood after 9–10 molts, and lived for an average of 240 days as adults. Females were found to mate only once and to oviposit from 1 to 5 times (average 1.8), at 30-day intervals. They produced a mean of 35 eggs in the 1st hatch and 31 eggs in the 2nd.

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Résumé La biologie de l'araignéeChiracanthium mildei L. Koch a été étudiée au laboratoire (24±1°C. 55–60% H.R.). Les élevages ont été conduits à partir des œufs, avec des larves deSpodoptera de 1 à 6 j fournies comme proies pendant toute la durée du cycle évolutif. Les études biologiques ont porté sur la vitesse de développement, le cycle évolutif, le potentiel et le comportement de reproduction, ainsi que sur l'influence de certains facteurs du milieu sur ces paramètres. Dans les conditions du laboratoire, le taux de survie deC. mildei fut de 87% de l'œuf à la maturité sexuelle. Chez les males la maturité est atteinte en moyenne 182 j (137–207) après l'éclosion de l'œuf. Ils deviennent adultes après 7 ou 8 mues et vivent en moyenne 73 j. Chez les femelles on compte une moyenne de 231 j (191–386) de l'éclosion de l'œuf à la maturité. Elles deviennent adultes après 9 ou 10 mues et vivent en moyenne 240 j. Les femelles s'accouplent 1 fois mais pondent 1 à 5 fois (moyenne 1,8) en 30 j. Leur fécondité moyenne est de 35 œufs lors de la 1^re ponte et de 31 œufs pour la 2^e.

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Contribution from the Agricultural Research Organization, Bet Dagan, Israel. No. 203-E, 1979 series.     

Cytochemical studies of pectin digestion in epidermis with specific cell separation

Summary This investigation concerns a unique type of epidermal cells in the anther ofStrelitzia reginae. At dehiscence these cells are released and form multicellular threads. The radial and tangential middle lamella regions of their cell walls disintegrate by the formation of numerous growing and fusing cavities. The possibility that this process could be due to digestion by pectinase was elucidated by use of cytochemical methods. In immature ordinary and thread-forming cells staining for pectin with hydroxylamine-ferric chloride yielded reaction products mainly in the middle lamella region and the subcuticular layer. After the appearance of cavities reaction took place around but not inside these formations. Treatment with fungal pectinase caused degradation of cell walls in ordinary epidermal tissue. Mature cell walls appeared more resistant to the lytic action than immature ones. In thread-forming tissue, independent of the stage of maturation, digestion of the pectin rich regions was induced. However, the fungal enzyme was not able to produce cavities. No pectin reaction with hydroxylamine-ferric chloride was obtained after pectinase treatment.     

Etudein situ de l'influence de l'humidité et de la teneur en nitrate d'un sol dunaire sur l'accumulation et la réduction du nitrate chez l'oyat (Ammophila arenaria L.)

Résumé L'étude du fonctionnement du cycle de l'azote dans les milieux dunaires des c?tes méditerranéennes fran?aises a conduit à analyser le comportement de l'oyat,Ammophila arenaria, au cours de son développement en réponse aux variations de l'humidité et de la teneur en nitrate du sol. Les mesures d'activité nitrate réductase et de la teneur en nitrate dans les feuilles ont été effectuées simultanément avec les mesures des teneurs en eau et en nitrate du sol. Cette plante a des activités nitrate réductase ne dépassant pas 0,27 μmoles h⁻¹ g⁻¹ matière fra?che, qui n'ont pu être mesuréesin vivo qu'en présence de nitrate exogène dans le milieu d'incubation. Il appara?t que la plante réduit ou accumule préférentiellement le nitrate selon que l'on se situe en période humide ou sèche. Pour des valeurs voisines ou inférieures à 2% d'humidité dans les vingt premiers centimètres de sol, l'activité nitrate réductase décro?t et des quantités importantes de nitrate s'accumulent (169 μg N.NO₃.g⁻¹ matière sèche). Dans ces conditions, le nitrate dans la solution du sol peut atteindre une concentration élevée (6 meq.l⁻¹). Inversement, au-delà de 2% d'humidité le nitrate endogène s'épuise et l'activité nitrate réductase augmente. La régulation de l'assimilation et de l'accumulation du nitrate par l'oyat dépend de son alimentation hydrique et done des quantités de nitrate absorbées à partir de la solution du sol et véhiculées dans la sève brute.

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In situ study of the influence of dune soil humidity and nitrate contents on the nitrate accumulation and reduction in marram (Ammophila arenaria L.)

Summary The search for a better understanding of the nitrogen cycle in the sand dunes of french mediterranean beaches have driven us to study the habits ofAmmophila arenaria. In this way, we have followed its responses to different soil water potentials and nitrate contents of the soil during its development. Nitrate reductase activities and nitrate contents of leaves have been measured simultaneously with water and nitrate contents of soil. This plant has a slight nitrate reduction activity, not more than 0,27 μmoles h⁻¹ g⁻¹ fresh matter, which could be measuredin vivo only by the addition of nitrate to the incubation media. It seems that the plant reduces or else it stores, in a preferential way the nitrate whether it is in a humid or dry period. If humidity values are near or below 2%, the marram diminishes its assimilation activity and stores important amounts of nitrate (169 μg N−NO₃ ⁻.g⁻¹ dry matter). In these conditions the nitrate can be highly concentrated in the soil solutions (6 meq.l⁻¹). Opposite, above 2% of humidity, the nitrate reductase activity decreases and the endogenous nitrate is consumed. The regulation of the assimilation and storage of nitrate by the marram is mainly a function of the soil humidity and so, of the amount of nitrate taken from the soil solution and carried into the xylem.

     

Incorporation of D-[U-14C]Glucose in the Cell Wall of Linum Plantlets during the First Steps of Growth

Flax plantlets, cultivated from day 3 in liquid medium and undercontinuous light showed linear growth. Electron microscopy observationsshowed that treatment of the cell walls with cdta-Na₂ clearedout the middle lamella and the cell junctions, whereas boilingwater extracted pectic polysaccharides from the primary cellwall in each tissue (epidermis, cortical parenchyma and phloem). Pulse-chase experiments showed that there was during the growthof flax plantlets a continuous redistribution of radioactivityin all parts of the cell walls: 1) from pectins to hemicellulosesand even to the cellulosic residues. 2) from oligomers to polymers.3) from neutral to acidic polymers in the core of the middlelamella. 4) from acidic to neutral pectins in the primary cellwalls. The elongation zone which was restricted to a small zoneback from the tip, involved strong synthesis of neutral pectinsin all the cell walls. Conversely, the redistribution of radioactivitywas related mainly to the plant cell maturation, and especiallyto the acidification of the cell wall. Demethylation of someneutral pectins occurred in the middle lamella whereas stronglyacidic pectins were synthetized in the primary cell wall. (Received October 1, 1990; Accepted April 9, 1991)