Synthesis of male sterile,triazine-resistant Brassica napus by somatic hybridization between cytoplasmic male sterile B. oleracea and atrazine-resistant B. campestris

Summary Fusion of leaf protoplasts from an inbred line of Brassica oleracea ssp. botrytis (cauliflower, n=9) carrying the Ogura (R1) male sterile cytoplasm with hypocotyl protoplasts of B. campestris ssp. oleifera (cv Candle, n=10) carrying an atrazine-resistant (ATR) cytoplasm resulted in the production of synthetic B. napus (n=19). Thirty-four somatic hybrids were produced; they were characterized for morphology, phosphoglucose isomerase isoenzymes, ribosomal DNA hybridization patterns, chromosome numbers, and organelle composition. All somatic hybrids carried atrazine-resistant chloroplasts derived from B. campestris. The mitochondrial genomes in 19 hybrids were examined by restriction endonuclease and Southern blot analyses. Twelve of the 19 hybrids contained mitochondria showing novel DNA restriction patterns; of these 12 hybrids, 5 were male sterile and 7 were male fertile. The remaining hybrids contained mitochondrial DNA that was identical to that of the ATR parent and all were male fertile.     

Inheritance of resistance to bacterial blight in common bean

Summary Inheritance of resistance to common bacterial blight in the trifoliate leaf, plant canopy, and pods was controlled by a single major gene. Additive followed by dominance effects were more important than epistatic interactions. Narrow-sense heritability values ranged from 0.18 to 0.87 for trifoliate leaf, from 0.26 to 0.76 for canopy, and from 0.11 to 0.36 for pods. Observed gains from selection for resistance were higher than expected gains. Implications of these results in breeding for resistance are discussed.     

Water storage and osmotic pressure influences on the water relations of a dicotyledonous desert succulent

Abstract Water storage and nocturnal increases in osmotic pressure affect the water relations of the desert succulent Ferocactus acanthodes, which was studied using an electrical circuit analog based on the anatomy and morphology of a representative individual. Transpiration rates and osmotic pressures over a 24-h period were used as input variables. The model predicted water potential, turgor pressure and water flow for various tissues. Plant capacitances, storage resistances and nocturnal increases in osmotic pressure were varied to determine their role in the water relations of this dicotyledonous succulent. Water coming from storage tissues contributed about one-third of the water transpired at night: the majority of this water came from the nonphotosynthetic, water storage parenchyma of the stem. Time lags of 4 h were predicted between maximum transpiration and maximum water uptake from the soil. Varying the capacitance of the plant caused proportional changes in osmotically driven water movement but changes in storage resistance had only minor effects. Turgor pressure in the chlorenchyma depended on osmotic pressure, but was fairly insensitive to doubling or halving of the capacitance or storage resistance of the plant. Water uptake from the soil was only slightly affected by osmotic pressure changes in the chlorenchyma. For this stem succulent, the movement of water from the chlorenchyma to the xylem and the internal redistribution of water among stem tissues were dominated by nocturnal changes in chlorenchyma osmotic pressure, not by transpiration.     

Protein and cDNA sequences of Bowman-Birk protease inhibitors from the cowpea (Vigna unguiculata Walp.)

The protein and gene sequences of the cowpea Bowman-Birk type trypsin inhibitor which confers enhanced insect resistance to transgenic tobacco plants, and of cowpea trypsin/chymotrypsin inhibitors are presented. There are regions of high conservation and high divergence within the 5 leader, mature protein and 3 non-coding regions of the Bowman-Birk inhibitors and in the genes which encode them in different members of this family within the Leguminosae. The practical implications of this finding for studies on the evolution of plants and the utilization of these genes for enhancing insect resistance is discussed.     

Kinetic measurement of bacterial adherence to eukaryotic cells

We developed a kinetic assay using a monolayer of differentiated respiratory epithelium in culture to assess bacterial adherence. Mean residence time of bacteria in the tissue culture chamber was estimated from a model-independent (moment) analysis of the rate of bacterial washout from perfused Rose chambers. Results with this method compared favorably with visual assessment of adherence and double radiolabel method with H. influenzae. Adherence was assessed with low inoculae of H. influenzae, P. cepacia and P. aeruginosa avoiding cytotoxic effects seen when large inoculae are added to eukaryotic cells. This method will provide a means of assessing adherence of pathogenic respiratory bacteria to their cellular target at low inoculae.     

Heritability estimates of phenthoate resistance in the diamond-back moth

Realized heritabilities were estimated for the character of phenthoate resistance in two local strains of the diamond-back moth, Plutella xylostella L. (Lepidoptera: Yponomeutidae), by performing artificial laboratory selection for resistance and susceptibility to phenthoate. Heritability estimates indicated that such traits are moderately heritable ( ²=0.42 and 0.41 in the resistant selection and ²=0.31 and 0.21 in the susceptible selection), and give an experimental basis accounting for rapid evolutionary changes of phenthoate resistance observed in field populations of this insect.The observed changes in variances of phenthoate susceptibility are discussed in relation to the additive genetic variance eliminated by directional selection. The explanation stresses the importance on the underlying genetic system.

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Résumé L'héritabilité de la résistance au phenthoate obtenue chez deux souches locales de P. xylostella L. (Lep.; Yponomeutidae) a été calculée en procédant au laboratoire à des sélections artificielles pour la résistance et pour la sensibilité à cet insecticide. Les calculs de l'héritabilité ont montré que de tels caractères sont moyennement héritables ( ²=0,42 et 0,41 lors de la sélection pour la résistance, et ²=0,31 et 0,21 lors de la sélection pour la sensibilité), et ont fourni la base expérimentale rendant compte des changements évolutifs rapides observés pour la résistance au phenthoate chez des populations naturelles de cet insecte.Les changements observés de la variance de la sensibilité au phenthoate sont discutés en fonction de la variance génétique additive éliminée par la sélection orientée. L'explication insiste sur l'importance du système génétique souligné.

     

Glandular trichome exudate is the critical factor in geranium resistance to foxglove aphid

Resistance to the foxglove-aphid (Acyrthosiphon solani Kaltenbach) has been demonstrated in some inbred geranium lines (Pelargonium Xhortorum Bailey). To establish more definitively the cause/effect relationship between tall glandular trichome exudate and resistance, an intact plant bioassay was performed comparing a resistant plant line, a resistant plant line from which the tall glandular trichome exudate had been removed using a basic buffer solution, a susceptible line and a susceptible line treated with the buffer wash. After 5 days of isolation on the respective surfaces, the number of surviving adult aphids as well as the number of nymphs produced and remaining alive were determined. Aphids on the buffer washed, resistant line exhibited mortality and fecundity which was not significantly different from that produced by the susceptible line. In contrast, the untreated resistance line was clearly resistant with lower adult survival and fewer living nymphs. The tall glandular trichome exudate must therefore be a critical factor in geranium resistance to the foxglove aphid.

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Zusammenfassung Widerstandsfähigkeit dem Fingerhut-Blattlaus (Acyrthosiphon solani Kaltenbach) gegenüber wurde in einigen durch Inzucht erzeugten Pelargonie-Linien (Pelargonium Xhortorum Bailey) gezeigt. Um das Verhältnis von Ursache und Wirkung zwischen dem hochgewachsenen glandulären Trichom-Exudat und Widerstandsfähigkeit genauer zu bestimmen, wurde eine Bio-Untersuchung an intakten Pflanzen unternommen. Dabei wurden eine widerstandsfähige Pflanzenlinie, eine widerstandsfähige Pflanzenlinie, von der das hochgewachsene glanduläre Trichom-Exudat durch eine basische Pufferlösung entfernt worden war, eine anfällige Linie und eine mit Pufferlösung behandelte Linie verglichen. Zwei erwachsene weibliche Blattläuse wurden fünf Tage durch ein engmaschiges Netz auf den zu untersuchenden dritten und vierten Knotenblättern eingesperrt. Bei jeder Pflanze wurde die Untersuchung an einem nichtbehandelten Blatt und an einem Blat, von dem das Exudat durch Waschen mit der Pufferlösung entfernt worden war, durchgeführt. Für jede Linie wurden fünf Pflanzen gebraucht, und der ganze Versuch wurde sechsmal wiederholt. Nach einer fünftägigen Isolierung auf den jeweiligen Oberflächen wurden die Blätter von der Pflanze entfernt, und sowohl die Zahl der überlebenden erwachsenen Blattläuse wie auch die der produzierten und noch am Leben gebliebenen Nymphen festgestellt. Mit einer niedrigeren Überlebensrate der Erwachsenen und weniger noch lebenden Nymphen war die nichtbehandelte widerstandsfähige Linie deutlich widerstandsfähig. Im Gegensatz dazu zeigten die Blattläuse auf der mit Puffer gewaschenen widerstandsfähigen Linie eine Sterblichkeit und Fruchtbarkeit, die nicht erheblich höher waren, als die auf der anfälligen Linie, was beweist, daß das Waschen mit der Pufferlösung den Widerstandsfaktor entfernt hatte. Das hochgewachsene glanduläre Trichom-Exudat muß deshalb ein kritischer Faktor in der Widerstandsfähigkeit gegen Pelargonie-Schädlinge sein.

     

Stylet penetration by two aphid species on susceptible and resistant lettuce

The probing behavior of two aphid species, Myzus persicae (Sulz.) and Nasonovia ribisnigri (Mosley), was electronically monitored on susceptible and resistant lettuce lines using a DC amplifier. A waveform pattern associated with extracellular stylet pathway activities, pattern C, occurred for longer periods when either aphid species probed resistant plants. This pattern is usually regularly interrupted by drops in electrical potential lasting a few seconds, reflecting cell membrane punctures followed by rapid withdrawal of the stylet tips. For M. persicae on resistant lettuce a large increase in pattern C without these potential drops accounted for the increased duration of this pattern. For N. ribisnigri the increase in pathway activity on resistant plants was due to an increase in the more typical pattern C with potential drops, as well as to an increased duration of pattern F, associated with a curious type of stylet penetration within cell walls. Both aphids made more but shorter probes on resistant than on susceptible plants, and these probes led less frequently to periods of sieve element contact and ingestion. The effects of resistance appear to involve both mesophyll and phloem factors. The underlying mechanisms, however, remain unclear. The results indicate which stylet penetration activities or waveform patterns are of interest for further investigation of resistance mechanisms.

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Résumé Le comportement de sondage de variétés sensibles et résistantes de laitues par deux espèces de pucerons, Myzus persicae (Sulz.) et Nasonovia ribisnigri (Mosley), a été enregistré électroniquement en utilisant un amplificateur DC. Les deux espèces de pucerons ont présenté une amplification de l'onde de type C associée au cheminement extracellulaire des stylets lors du sondage des variétés résistantes. Cette onde est normalement interrompue par des chutes de quelques secondes du potentiel électrique, traduisant des ponctions de la membrane cellulaire suivies par un rapide retrait de l'extrémité des stylets. La forte augmentation de l'onde C mais sans chutes de potentiel rend compte de la plus longue durée de cette onde chez M. persicae sur laitue résistante. L'accroissement du cheminement des stylets chez N. ribisnigri sur plantes résistantes est dû à une augmentation de l'onde C typique avec chutes de potentiel, ainsi qu'à une prolongation de l'onde F liée à la pénétration des stylets dans les parois cellulaires. Les deux espèces font des sondages plus brefs et plus nombreux sur variétés résistantes, et ces sondages entraînent des contacts moins fréquents avec les éléments criblés et débouchent moins souvent sur de l'ingestion. Les effects de la résistance semblent impliquer des facteurs liés à la fois au mésophylle et au phloème. Les mécanismes sous-jacents, cependant, ne sont pas encore clairs. Ces résultats Montrent que l'examen des ondes liées à la pénétration des stylets est important pour des études ultérieures sur les mécanismes de résistance.

     

Host plant resistance in Brachiaria grasses to the spittlebug Zulia colombiana

Twelve forage grass accessions including 11 accessions of Brachiaria Griseb, were evaluated in a glasshouse for host plant resistance to nymphs and tolerance to feeding damage caused by adults of Zulia colombiana (Lallemand) (Homoptera: Cercopidae). Resistance to nymphs was evaluated with a technique that provided uniform environmental conditions and abundant feeding sites. B. brizantha Stapf (cv. Marandú) was the most resistant of the accessions tested based on nymphal mortality, duration of nymphal stadia, and weight of adult females. Andropogon gayanus Kunth, resistant to spittlebug attack in the field, was susceptible under the conditions of this study. While growth habit and rooting characteristics may contribute to field resistance, other resistance factors are present within the genus Brachiaria, particularly in the case of B. brizantha cv. Marandú. The number of insect-days causing severe damage in the most tolerant species (B. dictyoneura Stapf and B. humidicola Schweick) was approximately six times greater than that necessary to cause the same level of damage to the most susceptible species (B. ruziziensis Germain & Evrard and B. decumbens Stapf). No difference was found in regrowth capacity between infested and noninfested plants within accessions. There was a significant positive correlation between number of insect-days causing severe damage (tolerance) and regrowth of infested plants.

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Résumé Brachiaria est une graminée fourragère prometteuse pour les sols tropicaux acides, saturés d'aluminium. Z. colombiana est un Cercopidae très répandu, limitant l'utilisation de Brachiaria en Amérique Latine. La résistance (antibiose et tolérance) à Z. colombiana, de Brachiaria d'origines diverses a été examinée. B. brizantha cv. Marandù s'est révélé le plus résistant d'après la forte mortalité larvaire, la prolongation du développement larvaire, et le poids réduit des femelles adultes de Z. colombiana. Andropogon gayanus, résistant dans la nature, s'est révélé sensible. Ces résultats suggèrent que cette résistance de A. gayanus dans la nature pourrait être due à la structure du végétal et à son mode de croissance. Dans le cas de B. brizantha cv. Marandù, des facteurs supplémentaires de résistance, mis en évidence à partir de différents modes de croissance, ont été éliminés, de façon à identifier les mécanismes de l'antibiose présents chez Brachiaria. Une grande gamme de résistance aux attaques alimentaires a été observée chez Brachiaria. Les plus résistants ont besoin de 6 fois plus de jours d'attaque par Z. colombiana pour provoquer les dégâts observés sur individus sensibles.

     

Increase in hepatic microsomal lipid peroxidation mediated by α-adrenergic system under cold stress and noradrenaline treatment

A membrane protein recognized by monoclonal antibody SQM1 was identified in human squamous carcinomas, including those originating in the head and neck (SqCHN), lung and cervix. Cell lines derived from SqCHN of previously untreated patients expressed high amounts of this protein. In contrast, many cell lines established from SqCHN of patients previously treated with chemotherapy and/or radiation showed diminished amounts of this SQM1 protein. The expression of SQM1 antigen was determined in several SgCHN cell lines made resistant by exposure to methotrexate (MTX) in vitro. The parent cell lines all exhibited strong binding to SQM1 antibody. The MTX-resistant sublines showed much lower membrane binding of SQM1. The lowest SQM1 reactivity was found in cell lines with high resistance to MTX and with diminished rate of MTX transport. Some highly MTX-resistant cell lines which had high levels of dihydrofolate reductase, but which retained a high rate of MTX transport, also retained high levels of SQM1 binding. Reduced SQM1 protein was also found in SgCHN cells which developed resistance to the alkylating drug cis-platinum (CDDP) and which showed reduced membrane transport of CDDP. Cell growth kinetics and non-specific antigenic shifts were not responsible for the differences in SQM1 binding between the parent cell lines and their drug-resistant sublines. The finding of a novel protein which is reduced in cells resistant to MTX and CDDP could contribute to our understanding of the basic mechanisms of drug resistance. By detecting SQM1 protein in clinical specimens, it may be possible to monitor the development of drug resistance in tumors.Abbreviations SqCHN Squamous Carcinoma of the Head and Neck - MTX Methotrexate - CDDP Cis-Platinum - DHFR Dihydrofolate Reductase