Structure-function relationships of α and β elicitins,signal proteins involved in the plant-Phytophthora interaction

Elicitins form a family of 10-kDa holoproteins secreted by various Phytophthora species. The large-scale purification of parasiticein, a novel elicitin secreted by P. parasitica, led to the determination of its sequence. We have compared the necrotic activities and the primary and secondary structures (determined through circular dichroism) of four elicitins. On tobacco plants, they could be classified into two classes: a, comprising capsicein and parasiticein (less necrotic), and , comprising cryptogein and cinnamomin (very toxic with a necrosis threshold of 0.1 g per leaf). The features of elicitin structure which might be involved in the interaction of elicitins with the leaf target cells and that could explain the different necrosis-inducing properties of the two proteins are investigated. About 75% sequence identity was observed between the four elicitins: only two short terminal regions are heterologous, while the central core is mainly conserved. The circular-dichroism spectra showed that the secondary structure of the elicitins was largely conserved. All of them consisted of approx. 50% -helix with little or no -structure. Comparisons of the complete sequences, amino-acid compositions, isoelectric points, hydropathy indices and the secondary-structure predictions correlated with the necrotic classification. Alpha elicitins corresponded to acidic molecules with a valine residue at position 13, while elicitins were basic with a lysine at this position, which appeared to be a putative active site responsible for necrosis induction.Abbreviations CD circular dichroism - RPLC reversed-phase liquid chromatography - SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis The authors are indebted to Dr. A. Van Dorrselaer (Laboratoire de Chimie Organique des Substances Naturelles, Strasbourg, France) for mass-spectrometry measurements. They are grateful to their staff in Versailles, more particularly to Marc Sallantin for electrophoreses, to Françoise Beauvais for biological-activity determinations and to Monique Mansion and Christian Ouali for their skilful technical assistance.     

Amino acid sequence of cinnamomin, a new member of the elicitin family, and its comparison to cryptogein and capsicein

The phytopathogenic fungi Phytophthora cinnamomi cause systemic leaf necrosis on its non-host tobacco; in culture, it secretes a protein, called cinnamomin, which elicits leaf necrosis and protects tobacco against the pathogen Phytophthora nicotianoe, in a way similar to cryptogein and different from capsicein, elicitins of known amino acid sequences. The cinnamomin sequence has been determined and compared to other elicitins. The differences in the 3 elicitin sequences were correlated to the biological activities: 2 lysines were ascribed as the key amino acids involved in the differential control of protection with respect to necrosis.     

Relationship between Active Oxygen Species,Lipid Peroxidation,Necrosis, and Phytoalexin Production Induced by Elicitins in Nicotiana

Excised leaves of Nicotiana tabacum var Xanthi and Nicotiana rustica were treated with cryptogein and capsicein, basic and acidic elicitins, respectively. Both compounds induced leaf necrosis, the intensity of which depended on concentration and duration of treatment. N. tabacum var Xanthi was the most sensitive species and cryptogein was the most active elicitin. Lipid peroxidation in elicitin-treated Nicotiana leaves was closely correlated with the appearance of necrosis. Elicitin treatments induced a rapid and transient burst of active oxygen species (AOS) in cell cultures of both Nicotiana species, with the production by Xanthi cells being 6-fold greater than that by N. rustica. Similar maximum AOS production levels were observed with both elicitins, but capsicein required 10-fold higher concentrations than those of cryptogein. Phytoalexin production was lower in response to both elicitins in N. tabacum var Xanthi cells than in N. rustica cells, and capsicein was the most efficient elicitor of this response. In cryptogein-treated cell suspensions, phytoalexin synthesis was unaffected by diphenyleneiodonium, which inhibited AOS generation, nor was it affected by tiron or catalase, which suppressed AOS accumulation in the extracellular medium. These results suggest that AOS production, lipid peroxidation, and necrosis are directly related, whereas phytoalexin production depends on neither the presence nor the intensity of these responses.     

Biosynthesis and Secretion of Cryptogein, a Protein Elicitor Secreted by Phytophthora cryptogea

The phytopathogenic fungi Phytophthora subspecies elicit hypersensitive-like necroses on their nonhost tobacco (Nicotiana tabacum), with the exception of the tobacco pathogen Phytophthora nicotianæ. In culture, these fungi—except P. nicotianæ—secrete proteins, called elicitins, that cause these remote leaf necroses and are responsible for the incompatible reaction. These proteins protect tobacco against invasion by the agent of the tobacco black shank, P. nicotianæ, which is unable to produce such an elicitor. Cryptogein, secreted by Phytophthora cryptogea, has been purified, sequenced, and characterized as an elicitin, a novel family of 10 kilodalton holoproteins. In the present paper, we examined the secretion and biosynthesis of this protein elicitor from P. cryptogea culture. Results showed that the secretion of cryptogein began later than its synthesis and stopped earlier, simultaneously with mycelium growth, when the nitrogen source in the culture medium was nearly exhausted. Electrophoretic patterns of total protein from mycelium extracts and N-terminal sequence analysis showed that cryptogein accumulated in the mycelium in its mature form. The comparison of the immunoselected in vitro translation products with ³⁵S in vivo-labeled cryptogein showed that cryptogein was synthesized as a preprotein with a signal peptide removed cotranslationally before the secretion into the culture medium. Immunoselected in vitro-synthesized products were subjected to radiosequencing to clearly determine the N-terminal position and the size (20 amino acids) of the signal peptide. Cryptogein did not undergo any other posttranslational modification.     

Investigations on the action of Phytophthora quercina,P. citricola and P. gonapodyides toxins on tobacco plants

Phytophthora quercina, P. citricola and P. gonapodyides isolated from declining oak roots and from soil rhizosphere in the field, released proteins into their culture medium. The proteins of P. quercina and P. gonapodyides caused severe chlorosis and necrosis on tobacco leaves. Only few symptoms were seen for the P. citricola protein. Surprisingly leaf chlorosis and necrosis were only visible in the light, whereas wilt symptoms were light-independent. The proteins were characterized on SDS gels as small peptides with basic and acidic isoelectric points. All proteins were heat stable. Even boiling for 15 min did not affect their activity. However, pronase treatment totally destroyed their activity. Transmission electron microscopy studies clearly showed that membrane structures especially of chloroplasts were damaged. The proteins of P. quercina and P. gonapodyides strongly crossreacted with the antibody raised against the P. cryptogea protein cryptogein. Therefore, these proteins might belong to the family of Phytophthora leaf necrotic proteins called elicitins.     

Production de deux glycopeptides par le Phialophora cinerescens (Wr.) van beyma cultive in vitro

Résumé Le Phialophora cinerescens alcalinise nettement le milieu liquide sur lequel il est cultivé durant 7 semaines; Le saccharose fourni est rapidement hydrolysé en glucose et fructose. La production des deux glycopeptides neutre et anodique — qui affectent la croissance des jeunes plantes d'Oeillet — a été appréciée. Le glycopeptite neutre est synthétisé dès le début de la culture, le glycopeptide anodique plus tardivement; les variations de la croissance du P. cinerescens cultivé in vitro suggèrent que les deux glycopeptides ne sont pas des produits de l'autolyse du mycélium.

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In vitro growth of the fungus Phialophora cinerescens was studied for 7 weeks. During this period the pH values were shown to increase in the culture filtrates; the added sucrose was rapidly hydrolyzed into glucose and fructose. The production of both neutral and anodic glycopeptides — which are know to affect the growth of carnation seedlings — was periodically monitored. The neutral glycopeptide was synthesized at the beginning of the culture period, the anodic one was synthesized later. The P. cinerescens growth curves indicated these two metabolites were probably not autolytic products of fungal cells.

     

Population differences in larval hostplant use in the checkerspot butterfly, Euphydryas chalcedona

Larvae from two populations of Euphydryas chalcedona Doubleday & Hewitson (Nymphalidae) were reared on their own hostplant and that of the other population, in both pre-diapause and post-diapause instars. One population, Chico, uses Penstemon breviflorus Lindl. (Scrophulariaceae), and the other, Echo Lake, uses P. newberryi Gray. Growth rate and survival were determined for pre-diapause and post-diapause larvae from both populations on both plant species; and digestive efficiencies were calculated during the prediapause instars. The results showed that larvae from the two populations differed in their responses to the two plant species. Pre-diapause larvae from Chico performed equally well on both plant species—survival and digestive indices were not significantly different for two Penstemon species. In contrast, pre-diapause larvae from Echo Lake performed significantly worse on the non-hostplant—growth and survival were significantly lower on the non-host, P. breviflorus. In addition, comparison of digestive efficiencies for the two plants showed that larvae from Echo Lake digested P. breviflorus better than P. newberryi, but were significantly less able to convert P. breviflorus to body mass. In the post-diapause instars, larvae from Chico grew faster on the host than on the non-host. Larvae from Echo Lake grew quite slowly on both plant species and significantly more of the Echo Lake larvae returned to diapause instead of completing development.

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Résumé Des chenilles de deux populations d'E. chalcedona ont été élevées sur leur propre plante-hôte et sur celle de l'autre population, aux stades avant et après diapause. Les deux populations s'alimentent sur différentes espèces de Penstemon (Scrophulariaceae), et une population—Echo Lake—est monophage sur P. newberry, tandis que l'autre—Chico—utilise d'abord P. breviflorus, mais les chenilles après diapause sont trouvées sur au moins deux autres espèces de plantes. Les taux de croissance et de survie ont été déterminés pour des chenilles avant et après diapause pour les deux populations sur les deux plantes; les efficacités digestives ont été calculées sur les chenilles avant diapause.Les résultats ont montré que les chenilles des deux populations différaient par leur degré de spécialisation digestive sur leur plante hôte normale: les chenilles de Chico ont utilisé aussi bien les deux plantes, tandis que celles d'Echo Lake le faisaient significativement moins bien sur la plante non-hôte, par suite de l'inaptitude à la digérer. Ainsi la population oligophage est alimentairement moins spécialisée et plus capable de se débrouiller avec une plante non-hôte. Après diapause, les chenilles de Chico s'alimentaient significativement mieux sur plante hôte que non-hôte, ce qui était le cas aussi pour la population monophage. Dans l'ensemble, les chenilles de la population monophage semblaient moins capables de se débrouiller dans des conditions défavorables ou moins avantageuses.

     

Influence of method of presentation on results of plant-host preference tests with two species of grasshopper

Leaf material of two Piper spp. (Piperaceae) was presented to two different generalist herbivores (Orthoptera: Acrididae) to determine whether method of leaf presentation in preference trials affects choice. For the grasshopper Leptomerinthoprora brevipennis, the method of presentation had no effect on preference ranking: more of Piper sancti-felicis was eaten than of P. arieianum, regardless of whether leaf material was presented as leaf disks, whole leaves supplemented with water, or whole leaves without water. In contrast, preference of the grasshopper Abracis flavolineata depended on presentation method. In that case, whole leaves of P. scancti-felicis supplemented with water were preferred over the same of P. arieianum, while dry weight consumption of leaf disks of P. arieianum was greater than that for disks of P. sancti-felicis. When presented with leaves not supplemented with water, A. flavolineata showed no significant preference for one plant species over another.

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Résumé Des feuilles provenant de deux espèces de Piper (Piperacae) ont été proposées à deux Orthoptères, Acrididae, herbivores généralistes, pour déterminer si la méthode de présentation des feuilles modifiait le choix. Pour Leptomerinthoprora brevipennis, la méthode de présentation est sans influence sur l'ordre de préférence; la consommation de Piper sancti-felicis est toujours supérieure à celle de P. arieianum, quelque soit le mode de présentation: disques de feuilles, feuilles entières humidifiées, ou feuilles entières sans addition d'eau. A l'opposé, l'ordre de préférence de Abracis flavolineata dépend du type de présentation; les feuilles entières humidifiées de P. sancti-felicis sont préférées à celles de P. arieianum proposées de la même façon, tandis que la consommation de disques secs de P. arieianum est plus importante que celle des disques secs de P. sancti-felicis. En présence de feuilles entières humidifiées, A. flavolineata ne manifeste aucune préférence.

     

Cytoplasmic acidification as an early phosphorylation-dependent response of tobacco cells to elicitors

Elicitor-induced cytoplasmic pH changes of tobacco (Nicotiana tabacum L. cv. Xanthi) cells grown in suspension cultures were explored under a variety of conditions by using a flexible technique based on the distribution of [¹⁴C] benzoic acid between the intracellular and extracellular compartments. Comparison of data obtained by this technique and by ³¹P-nuclear magnetic resonance spectrometry qualifies the benzoic acid distribution method as a convenient and reliable way to probe cytoplasmic pH variations. Various elicitors shown to induce several defense-related responses in tobacco cells, namely oligogalacturonides of degree of polymerization 7–20, pectolyase from Aspergillus japonicus, Phytophthora megasperma crude elicitors and purified cryptogein, triggered cytoplasmic acidifications differing in intensity and kinetics according to the signal molecule. In contrast, no changes in cytoplasmic protons and external pH were observed in cells treated with short galacturonide oligomers, or with soybean-specific hepta -glucoside from P. megasperma, which are devoid of elicitor activity in tobacco cells. The oligogalacturonide-induced cytoplasmic acidification was inhibited by two structurally unrelated protein kinase inhibitors, staurosporine and 6-dimethylaminopurine, which both reduced the external alkalinization response to the elicitor. The protein phosphatase inhibitor calyculin A alone behaved as an elicitormimicking molecule in triggering cytoplasmic acidification, again associated with extracellular alkalinization. These results indicate that the increase in the cytoplasmic concentration of protons may be considered as a common early intracellular response of tobacco cells to elicitors, associated with the extracellular alkalinization response and controlled by protein phosphorylation.Abbreviations BA(H) benzoic acid (protonated form) - 6-DMAP 6-dimethylaminopurine - DP degree of polymerization - Mes 2-(N-morpholino)ethanesulfonic acid - OG oligogalacturonide - pHc cytoplasmic pH - ³¹P-NMR nuclear magnetic resonance spectroscopy of ³¹P atoms The authors thank P. Albersheim (CCRC, Athens, Georgia, USA) for providing the purified oligogalacturonides and the hepta -glucoside and P. Ricci (INRA, Antibes, France) for providing the purified cryptogein.     

Feeding by adult sweet potato weevils, Cylas formicarius elegantulus, on sweet potato leaves

A bioassay was developed to quantify the feeding of adult sweet potato weevils, Cylas formicarius elegantulus (Summers) (Coleoptera; Curculionidae) on the foliage of four cultivars (Centennial, Jewel, Resisto and Regal) of sweet potato (Ipomoea batatas (L.) Lam.) (Convolvulaceae). Weevils fed along the leaf veins, preferring the lower to the upper leaf surface. Males and females had similar levels of feeding. Different levels of feeding by female weevils were observed between cultivars in dual-choice bioassays with Centennial, a susceptible cultivar in field-plot experiments, being most preferred and Resisto least preferred. However, these feeding differences were not observed in no-choice bioassays. Little difference was observed in the leaf surface chemistry of the four cultivars.

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Résumé L'étude a porté sur la consommation pendant 12 h, en boîtes de Pétri de diamètre 9 cm, de rondelles de 2 cm de diamètre de feuilles de 4 cultivars,—Centennial, Jewel, Resisto, Regal—, d'I. batatas par C. formicarius elegantulus. Les charançons ont consommé le long des nervures, préférant la face inférieure à la face supérieure des feuilles. Un index de consommation a été calculé en fonction de la longueur de nervure ayant servi à la consommation, rapportée à la longueur totale des nervures de la surface exposée.Les niveaux de consommation des mâles et des femelles étaient semblables. En présence de choix binaires, le cultivar Centennial a été le plus consommé, et le cultivar Resisto, le moins, par les femelles. De telles préférences n'ont pas été observées en l'absence de choix.La composition chimique de la surface de la feuille a été analysée par chromatographie en phase gazeuse. 8 pics principaux identiques ont été observés chez les 4 cultivars, mais ils avaient différentes hauteurs.

     

Rumex subgen.Rumex sect.Axillares (Polygonaceae) in South America

Rumex sect.Axillares first described by the author in 1937 is known to be represented in N. America by 20 endemic species. From S. America only five species, all endemic, were previously known; this number is raised in the present revision to 15. Outside America there exist small groups of endemic species in S. Africa and in the islands of Hawaii as well as in Australia and scattered single species in C. Africa, on the remote Atlantic islands of Tristan de Cunha and Gough Island, and in eastern Asia.—In Europe the section is completely absent, except for three American species which occur as rare aliens. TheAxillares are not only morphologically well-characterized by their growth—no leaf rosettes, flowering period practically unlimited by formation of later flowering axillary shoots, flowers often monoecious—but also by their genetic structure expressed by a different behaviour in hybridization. Hybrids are frequent between species of sect.Rumex (syn.Simplices) and are easy to recognize by their sterility which results in a certain peculiarity in habit. In contrast, no spontaneous hybrids are known with certainty among species of sect.Axillares. Most artificial hybrids are fairly fertile. So far only three hybrids between species of the two sections have been known. Three more have been found by the author in Chile, all being highly sterile and showing the growth characteristics resulting from sterility as occur in hybrids between species of sect.Rumex.

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Resumen Rumex secciónAxillares fue descrito por primera vez por el autor en 1937. Se conoce en América del Norte con 20 especies endémicas. De América del Sur se conocían sólo 5 especies, todas ellas endemicas; este número asciende a 15 en la presente revision. Fuera de las Américas se encuentran pequeños grupos de especies endémicas en Sudáfrica, en las Islas de Hawaii y en Australia; especies aisladas se encuentran en Africa Central, en las lejanas Islas Atlánticas de Tristan de Cunha y Gough Island y en Asia Oriental. La sección no tiene representantes en Europa, con excepción de tres especies americanas introducidas, poco frecuentes. La secciónAxillares se caracteriza bien, tanto morfológicamente por su forma de creciemiento—sin rosetas foliares, período floral prácticamente ilimitado por la formación de inflorescencias axilares parciales, flores a menudo monoicas—como por su estructura genética que se manifiesta en un comportamiento diferente de hibridación. Híbridos son frecuentes en las especies de la secciónRumex (syn.Simplices) y reconocibles por su esterilidad que se expresa en cierta particularidad del habito. Al contrario, no hay seguridad de híbridos espontáneos en la secciónAxillares. La mayor parte de los híbridos artificiales son totalmente fértiles. Hasta ahora se conocían solo tres híbridos entre las especies de ambas secciones. Otros tres han sido encontrados por el autor en Chile. Todos completamente estériles y con las características de hábito típicas resultantes la esterilidad, como ocurre en los híbridos de la secciónRumex.

     

Pollen morphology ofSonchus and related genera,and a general discussion

The pollen morphology of the taxa belonging to the generaAetheorhiza Cass.,Launaea Cass.,Reichardia Roth andSonchus L. in the Iberian Peninsula has been studied with light and electron microscopy. The pollen is 3(-4)-zonocolporate and echinolophate (without polar lacunae, but in general with prelacunae), with equatorial ridges and 15–20 lacunae: 3–4 poral, 6–8 abporal and 6–8 paraporal. Small to medium size, P × E = 19–36 × 23–42 µm; sometimes two different sizes have been found. Exine 3–9 µm thick and ornamentation microreticulate and echinate. The results clearly show the relationships between genera. Moreno-Socías, E., Mejías, J. A., Díez, M. J., 1994: Morfología polínica deLactuceae (Asteraceae) en la Península Ibérica, I.Lactuca y géneros relacionados. — Acta Bot. Malacitana.19: 103–113.     

Responses of Microplitis croceipes to host and nonhost plants of Heliothis virescens in a wind tunnel

The relative attractiveness of velvet leaf (Abutilon theophrasti Medicus), cotton (Gossypium hirsutum L.) (host plants) and groundcherry (Physalis angulata L.) (nonhost plant), and cotton plants with or without nectaries and with or without glands to Microplitis croceipes Cresson (Hymenoptera: Braconidae) was determined in a wind tunnel. Female parasitoids flew significantly more to glandless than to glanded cotton; response to nectaried and nectariless cotton was similar. Velvet leaf and cotton were favored significantly over groundcherry; parasitoids being equally responsive to both host plant species. Addition of larval frass alone or in combination with host larvae significantly improved the attractivity of the nonhost plant (groundcherry) to the parasitoids. There was no difference in attractiveness of groundcherry terminals with or without host larvae. Parasitoid search time was significantly increased with addition of larval frass. In the presence of cotton, however, kairomone-treated groundcherry remained unattractive.

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Résumé L'influence des nectaires et des glandes du coton, et l'attractivité des feuilles veloutées du coton (plante hôte) et de Physalis angulata sur Microplitis croceipes, ont été déterminées dans des expériences avec tunnel à vent. Les résultats ont montré que les parasitoïdes femelles sont significativement plus attirés par les cotons sans glandes que par les cotons glanduleux, tandis que la présence ou l'absence de nectaires ne modifie pas l'attractivité du coton. Elles réagissent de la même façon aux feuilles veloutées. P. angulata traité avec des crottes de chenilles, présenté seul ou en combinaison avec du coton, attire plus de femelles que des pousses de P. angulata non traitées. L'addition de chenilles hôtes seules n'améliore pas significativement l'attractivité de la plante. P. angulata traité avec des kairomones est demeuré inattractif quand il était proposé en même temps que du coton.

     

Effects of diet on longevity and fecundity of the spined stilt bug, Jalysus wickhami

We evaluated the suitability of selected food items for the adult spined stilt bug, Jalysus wickhami Van Duzee (Hemiptera: Berytidae), by providing one of seven diet treatments: (1) prepupae of the parasitoid, Cotesia congregata (Say) (Hymenoptera: Braconidae), on a leaf of tobacco (Nicotiana tabacum L., NC 2326) (Solanaceae); (2) C. congregata prepupae alone (i.e., no tobacco leaf); (3) eggs of the parasitoid's host, Manduca sexta (L.) (Lepidoptera: Sphingidae), on a tobacco leaf; (4) M. sexta eggs alone; (5) tobacco aphids, Myzus nicotianae Blackman (Homoptera: Aphididae), on a tobacco leaf; (6) a tobacco leaf alone; and (7) no prey and no tobacco. A tobacco leaf was necessary for the long-term survival of stilt bugs, but prey source did not affect longevity. Regardless of the prey type, stilt bugs with access to a tobacco leaf lived 21–23 days, while stilt bugs without access to tobacco lived less than 12 days. Animal prey provided a protein source required for egg deposition in J. wickhami, and prey species differed in their relative nutritive values. Female stilt bugs that fed on M. sexta eggs or on C. congregata prepupae were significantly more fecund (102 and 106 nymphs per female, respectively) than females that fed on tobacco aphids (24 nymphs per female). Results suggest that stilt bugs may feed on tobacco aphids and C. congregata prepupae when other prey are unavailable.     

Metabolism of amino acids, organic acids and sugars extracted from the xylem fluid of four host plants by adult Homalodisca coagulata

The efficiency of amino acid, organic acid and sugar metabolism was quantified for adult Homalodisca coagulata (Say) (Homoptera: Cicadellidae) by comparing chemical profiles of xylem fluid (food source) and insect exudate. Leafhoppers were confined in Parafilm^® sachets to stems of 4 host plants: [Baccharis halimifolia (L.), Lagerstroemia indica (L.), Prunus salicina (Lindl.), Prunus persica (L.), Batsch]. Insect feeding rates (0.09–0.27 ml h^–1), exudate osmolarity (7.8–12.8 mM) and exudate composition (mainly inorganic entities) were characteristic of a xylem feeder. Total organic solute concentration in the xylem fluid of B. halimifolia, L. indica, P. salicina and P. persica was ca. 9.4, 13.8, 5.5 and 1.8 mM, respectively. Nineteen protein amino acids, 7 organic acids and 3 or 4 sugars were identifid in the xylem fluid. Total amino acids, organic acids and sugars were metabolized with ca. 99% efficiency. Glutamine, asparagine, arginine and citric, malic and succinic acids, the predominant organic compounds in the xylem fluid of all four plant species, were metabolized with greater than 99% efficiency. Cysteine (51%), methionine (74%) and oxalic acid (77%) were metabolized with the lowest efficiency. The primary nitrogenous waste was NH _inf4 ^sup+ ; uric acid or urea were not detected. Nitrogen retention was generally less than 60% of dietary nitrogen. High feeding rates, ammonotelism and an extremely high metabolic efficiency of organic compounds permit H. coagulata to subsist on the dilute and skewed chemical profile of xylem fluid.

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Résumé L'efficacité du métabolisme des amino-acides, des acides organiques et des sucres a été quantifiée chez des H. coagulata Say (Hom. Cicadellidae) adultes, en comparant la composition chimique de la sève du xylème et du miellat des insectes. Les cicadelles ont été maintenues dans des sachets de parafilm avec des tiges de 4 plantes hôtes: Baccharis halimifolia L., Lagerstroemia indica L., Prunus salicina Lindl. et Prunus persica Batsch. Le taux de consommation (0,09 à 0,27 ml hr^–1), l'osmolarité du miellat (7,8 à 12,8 mM) et la composition du miellat (principalement des éléments inorganiques) sont caractéristiques des consommateurs de xylème. Les concentrations organiques totales en soluté de la sève du xylème étaient respectivement: 9,4; 13,8; 5,5; et 1,8 mM chez B. halimifolia, L. indica, P. salicina et P. persica. 19 amino acides protéiques, 7 acides organiques et 3 ou 4 sucres ont été identifiés dans la sève du xylème. Les acides aminés, les acides organiques et les sucres ont été métabolisés dans leur ensemble avec une efficacité de 99%. La glutamine, l'asparagine, l'arginine et les acides citrique, malique et succinique,-les principaux composés organiques de la sève du xylème de ces 4 plantes,-ont été métabolisés avec plus de 99% d'efficacité. La cystéine (51%), la méthionine (74%) et l'acide oxalique (77%) ont été métabolisés avec une plus faible efficacité. Le déchet azoté primarie était NH _inf4 ^sup+ ; l'acide urique et lurée n'ont pas été décelés. La fixation d'azote a été généralement inférieure à 60% de l'azote consommé. Des taux de consommation élevés, l'ammonotélisme et une efficacité extrêmement élevée du métabolisme des composés organiques permettent à H. coagulata de survivre malgré la composition chimique biaisée et la dilution de la sève du xylème.

     

Transgenic mimicry of pathogen attack stimulates growth and secondary metabolite accumulation

Plant secondary metabolites, including pharmaceuticals, flavorings and aromas, are often produced in response to stress. We used chemical inducers of the pathogen defense response (jasmonic acid, salicylate, killed fungi, oligosaccharides and the fungal elicitor protein, cryptogein) to increase metabolite and biomass production in transformed root cultures of the medicinal plant, Withania somnifera, and the weed, Convolvulus sepium. In an effort to genetically mimic the observed effects of cryptogein, we employed Agrobacterium rhizogenes to insert a synthetic gene encoding cryptogein into the roots of C. sepium, W. somnifera and Tylophora tanakae. This genetic transformation was associated with stimulation in both secondary metabolite production and growth in the first two species, and in growth in the third. In whole plants of Convolvulus arvensis and Arabidopsis thaliana, transformation with the cryptogein gene led, respectively, to increases in the calystegines and certain flavonoids. A similar transgenic mimicry of pathogen attack was previously employed to stimulate resistance to the pathogen and abiotic stress. In the present study of biochemical phenotype, we show that transgenic mimicry is correlated with increased secondary metabolite production in transformed root cultures and whole plants. We propose that natural transformation with genes encoding the production of microbial elicitors could influence interactions between plants and other organisms.     

Fruit dimorphism inPhaseolus sublobatus (Fabaceae) and its evolutionary significance

Populations of the annualPhaseolus sublobatus from different ecogeographical zones are genetically differentiated. In twelve populations from the western ghats (Maharashtra range) chasmogamous flowers are arranged in peduncled capitate racemes borne in leaf axils of higher nodes. One population (Poona Race S₄) has additional inflorescences, also with chasmogamous flowers, on the main axis between the cotyledonary node and the ground. When the pods of these flowers ripen, the inflorescence gradually coils and, eventually, gets bury the fruits in the soil. This phenomenon is not known in any other plant.—The seed-coat patterns as revealed by SEM, and germination behaviour of both aerial and subterranean seeds are similar, and both types of seeds lack dormancy.—Heavy seed predation selection pressure is probably the principal cause for the evolution of fruit dimorphism inP. sublobatus. The origin of geocarpy in relation to fruit dimorphism and seed predation is discussed.