Low root‐to‐root transmission of a tobamovirus,yellow tailflower mild mottle virus,and resilience of its virions

Tobamoviruses are serious pathogens because they have extremely stable virions, they are transmitted by contact, and they often induce severe disease in crops. Knowledge of the routes of transmission and resilience of tobamovirus virions is essential in understanding the epidemiology of this group of viruses. Here, an isolate of the tobamovirus yellow tailflower mild mottle virus (YTMMV) was used to examine root‐to‐root transmission in soil and in a hydroponic growth environment. Root‐to‐root transmission occurred rarely, and when it occurred plants did not exhibit systemic movement of the virus from the roots to the shoots over a 30‐day period. The resilience of YTMMV virions was tested in dried leaf tissue over time periods from one hour to one year under temperatures ranging from ?80 to 160 °C. Infectivity was maintained for at least a year when incubated at ?80 or 22 °C, or at fluctuating ambient temperatures of 0.8 to 44.4 °C, but incubation under dry conditions at 160 °C for >4 days eliminated infectivity. Exposure of virions to 0.1 m sodium hydroxide or 20% w/v skimmed milk solution for 30 min, treatments recommended for tobamovirus inactivation, did not abolish infectivity of YTMMV.     

Advanced lentil lines screened for resistance to <Emphasis Type="BoldItalic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="BoldItalic">lentis</Emphasis> under greenhouse and field conditions

Fusarium oxysporum f. sp. lentis is the most important pathogen of lentil plants, and most areas under lentil cultivation are reported to have a fusarium wilt disease background. The plants are infected in the seedling stage and later stages of their development. Fusarium wilt disease, which has appeared at high incidence rates during recent years, has caused sharp drops in the yield, especially in Moghan, in the northwest of Iran. Forty-five isolates of the pathogen were collected from different regions of the country with two isolates from ICARDA in the summer of 2008 and identified using Nelson’s key. The pathogenicity of the collected isolates was studied on a sensitive line (ILL 4605) under greenhouse conditions and significant differences in pathogenicity were found among them. The most pathogenic isolates from three provinces, East Azerbaijan (EA 30), Ardebil (Ar 3) and Khorasan (Kh 45), were selected and used in screening of 55 developed lines under greenhouse and field conditions. In the greenhouse, test plants were inoculated by immersing root tips in spore suspension and sowing seeds in pre-infested pot soil. Field tests were carried out in a naturally highly infested farm. At all stages, the plant response to the disease was based on the percentage of dead plants. Cluster analyses of the greenhouse and field data led to the selection of three lines (81S15, FLIP2007-42 L and FLIP2009-18 L) that were resistant under greenhouse and field conditions.     

Three genes of miraculin-like proteins from Nicotiana benthamiana with dissimilar putative structures show highly similar patterns of induction following bacterial and fungal infections

Expression of three Nicotiana benthamiana miraculin-like protein genes, NbMLP1, NbMLP2 and NbMLP3, showed almost identical responses to wounding, an incompatible interaction with Pseudomonas syringae pv. tabaci and compatible interactions with P. syringae pv. tabaci, Colletotrichum destructivum or Colletotrichum orbiculare. However, only NbMLP1 expression responded to exogenous methyl jasmonate or ethylene. None exhibited expression in healthy leaves and stems, and all showed highest expression in seeds, except for NbMLP1, which had highest expression in roots. NbMLP1, NbMLP2 and NbMLP3 were in different subfamilies of miraculin-like protein sequences of N. benthamiana and Nicotiana tabacum. Subfamilies correlated well with predicted features of the reactive-site loop potentially affecting the bond that could react with serine proteinases. Despite considerable predicted structural diversity that might affect biological activity, the apparently coordinated expression of these genes to pathogen attack may reflect the need to produce diverse proteinase inhibitors to act against a potentially broad range of secreted microbial proteinases during basal resistance to pathogens.     

Seasonal changes in germination response of buried seeds of Orobanche crenata Forsk.

Orobanche crenata seeds, collected in Syria, Egypt and Spain, were buried in the field in Syria (all three seed lots) and Spain (only Spanish seeds) and at regular intervals exhumed and tested for germination, to investigate whether the seeds exhibit an annual dormancy/non-dor- mancy cycle. When exposed directly to the synthetic germination stimulant GR24 for 7 days at 20°C, seeds only germinated in autumn after the first rains and to a limited extent in winter. When the seeds were conditioned for 11 days at 20°C prior to exposure to GR24, germination occurred during summer and autumn, but seeds were dormant in winter and early spring. The observed seasonal pattern in germinability, in relation to rainfall and soil temperature, was largely consistent with the results of an in vitro experiment by Van Hezewijk et al. (1993), investigating the effect of conditioning temperature and conditioning period on germination capacity and the development of secondary dormancy. Moisture and temperature can therefore be considered the major factors regulating induction and alleviation of dormancy in buried O. crenata seeds. There were no basic differences in response owing to site of collection of O. crenata seeds, nor to the location where they were buried. Variations saisonnières des exigences de germination de graines enfouies d'Orobanche crenata Forsk. Des graines d'Orobanche crenata récoltées en Syrie, en Égypte et en Espagne ont été enfouies au champ en Syrie (les 3 lots) et en Espagne (seules les graines d'Espagne) puis ont été exhumées a intervalles régulier pour que leur aptitude à la germination soil évaluée. Le but était de déterminer si les graines possédaient un cycle annuel dormance/non dormance. Quand elles étaient directement exposées au stimulant de germination synthétique GR24 pendant 7 jours à 20°C, les graines ne germaient qu'à l'automne après les premières pluies et peu en hiver. Quand les graines restaient pendant 11 jours à 20°C avant leur exposition au GR24, la germination seproduisait en été et à l'automne mais les graines restaient dormantes en hiver et au début du prin-temps. Les variations saisonnières d'aptitude à la germination, liées aux précipitations et à la temperature du sol, étaient en accord avec les résultats d'une expérience in vitro de Van Hezewijk et al. (1993) concernant l'effet de la température et de la durée pendant laquelle elle est appliquée, sur l'aptitude à la germination et le développement de la dormance secondaire. L'humidité du sol et sa température peuvent ainsi être considérées comme les principaux facteurs qui induisent et lèvent la dormance de graines de O. crenata enfouies. On n'observait pas de différences importantes dues au lieu de récolte ou à l'endroit oü elles étaient enfouies. Jahreszeitliche Änderungen der Keimung von vergrabenen Samen von Orobanche crenata Forsk. Proben von in Syrien, Ägypten und Spanien gesammelten Orobanche-crenata-Samen wurden in Syrien und Proben nur spanischer Herkunft in Spanien im Freiland im Boden ausgelegt und in regelmäßigen Zeitabständen ausgegraben und auf ihre Keimfähigkeit getestet, um zu untersuchen, ob die Samen einen jährlichen Dormanz-Zyklus haben. Beim direktem Auslegen in dem synthetischen Keimungsmittel GR24 öber 7 d bei 20°C keimten die Samen nur im Herbst nach den ersten Regenfällen und in beschränktem Umfang im Winter. Wenn die Samen för 11 d bei 20°C vor dem Auslegen in GR24 vorbehandelt worden waren, keimten sie im Sommer und Herbst, aber im Winter und fröhen Fröhjahr waren sie dormant. Das jahreszeitliche Verhalten der Keimfähigkeit in Abhängigkeit von Niederschlag und Bodentemperatur stimmte weitgehend mit den Ergebnissen eines In-vitro-Versuches von Van Hezewijk et al. (1993) öber die Wirkung einer Wärmevorbehandlung und Vorbehandlungszeit auf die Keimfähigkeit und die Ausprägung sekundärer Dormanz öberein. Bodenfeuchte und -temperatur können deshalb als die wichtigsten Faktoren för die Induktion und Aufhebung der Dormanz von Orobanche-crenata-Samen im Boden angesehen werden. Herkunft und Versuchsort hatten keinen erheblichen Einfluß auf die Ergebnisse.     

Population studies of the citrus psylla,trioza erytreae: factors influencing dispersal

More adults of the citrus psylla,Trioza erytreae (Del Guercio) (Hemiptera: Triozidae), dispersed when the number of eggs, nymphs or adults on the citrus increased (positive correlations) and also when new leaves that were present decreased (negative correlation). More psylla were trapped close to high obstacles than in an open area, even though the obstacles were at a distance from the psylla-infested orchard.     

An enrichment microsphere immunoassay for the detection of <Emphasis Type="Italic">Pectobacterium atrosepticum</Emphasis> and <Emphasis Type="Italic">Dickeya dianthicola</Emphasis> in potato tuber extracts

An enrichment microsphere immunoassay (MIA) was developed, based on the Luminex xMAP® technology, for the simultaneous (duplex) detection of Pectobacterium atrosepticum (former name Erwinia carotovora subsp. atroseptica) (Pca) and Dickeya dianthicola (former name Erwinia chrysanthemi) (Dcd) in potato plant extracts. Target bacteria in the extracts were enriched for 48 h in a semi-selective broth containing polypectate under low oxygen conditions. Samples were subsequently incubated with antibody-coated colour-coded microspheres (beads) and with secondary antibodies conjugated with Alexa Fluor® 532, a reporter dye. Samples were analyzed with the Luminex analyzer, in which one laser identified each microsphere and another laser the reporter dye conjugated to the secondary antibodies. The assay required minimal sample preparation, could be completed in 1 h, was performed in 96 wells microtitreplates and required no wash steps. The limit of detection for the duplex enrichment MIA was 100–1000 cfu ml^?1, which was a hundred times lower than of an enrichment-ELISA. Without enrichment, the sensitivity of MIA and ELISA was largely similar and ranged between 10⁶ and 10⁷ cells ml^?1. No difference in sensitivity was found between a MIA in a single or duplex format. In a comparative test with non-infected potato plant extracts and extracts from plants infected with Pca or Dcd, results of the enrichment MIA correlated well with those of the enrichment ELISA and enrichment PCR. These results indicate that MIA can be reliably used for multiplex detection of soft rot Enterbacteriaceae in crude potato plant extracts. The technology is an attractive and cost-effective alternative to other detection methods, including ELISA.     

Evaluating diverse systems of tobacco genetic resistance to Phytophthora nicotianae in Yunnan,China

Black shank, caused by the soilborne pathogen Phytophthora nicotianae, is one of the most devastating diseases affecting tobacco production in China. The most effective strategy for reducing economic loss from this pathogen is development and use of resistant tobacco varieties. Multiple sources and systems of resistance have been developed in the Western Hemisphere; however, populations of P. nicotianae are variable around the world, including the predominance of different races. Different P. nicotianae isolates may react differently on tobacco plants with different systems of resistance, a possibility that could complicate the breeding of cultivars with resistance that is effective in different tobacco production regions worldwide. The objective of this research was to evaluate an array of tobacco germplasm possessing different systems of genetic resistance to black shank disease in tobacco-growing regions of Yunnan, China. Resistance types included simply inherited resistance mechanisms introgressed from wild Nicotiana relatives and polygenic partial resistance systems of N. tabacum origin. The loci of Wz exhibited high level resistance to black shank in the five diverse disease environments in Yunnan, China. K326 Php/−Wz/− genotype and Beinhart 1000 exhibited the greatest levels of resistance in both 2015 and 2016. Field observed results for 13 tobacco genotypes were highly correlated with those tested in growth chamber evaluation. These findings suggest that both Wz− and Beinhart 1000-mediated resistance have important commercial value in flue-cured tobacco breeding programmes in China. Cultivars developed for black shank resistance in China may also have utility in other tobacco-growing areas.     

Detection of pineapple closterovirus in pineapple plants and mealybugs using monoclonal antibodies

Monoclonal antibodies (MAbs) were produced to the pineapple closterovirus (PCV) in Hawaii. These antibodies were shown to be specific for PCV by decoration of the virus particles in immunosorbent electron microscopy (ISEM) and indirect enzyme-linked immunosorbent assay (ELISA). Several methods of ELISA were compared. An indirect DAS ELISA using a polyclonal antibody to trap virus particles followed by reaction with monoclonal antibody was shown to be the method of choice for detecting PCV in pineapple plants. Pineapple root tissue was found to be most suitable for detecting PCV in crude samples by indirect ELISA. PCV was detected in symptomatic and asymptomatic pineapple plants collected from Oahu and Maui, and pineapple collections in the USDA/ARS National Clonal Germplasm Repository, but was not detected from pineapple plants grown from seed. At least two serotypes of PCV were detected. In addition. PCV was detected from mealybugs collected from wilted pineapple plants, but not from mealybugs of the same species collected from a colony reared on squash. The role of PCV in mealybug wilt of pineapple is being investigated.     

Infection of Tomicus piniperda (Col., Scolytidae) with Canningia tomici sp. n. (Microsporidia, Unikaryonidae)

Canningia tomici sp. n. (Microsporidia, Unikaryonidae) infects the midgut epithelium, the gut muscules, Malpighian tubules, connective tissues, adipose tissues and the gonads of the pine shoot beetle, Tomicus piniperda (L.) (Coleoptera, Scolytidae). The infection is present in populations of Tomicus piniperda in Europe and in the United States. Uninucleate oval single spores occur in two sizes: 2.8±0.4× 1.4±0.4m and 3.8±0.3×2.0±0.2m. The polar filament of this microsporidium is fixed subapically in a flat anchoring disc. The thick posterior lamellae of the binary polaroplast are asymmetric due to the lateral fixation of the polar filament.     

Control of Fusarium Wilt of Radish by Combining Pseudomonas putida Strains that have Different Disease-Suppressive Mechanisms

ABSTRACT Biological control of soilborne plant pathogens in the field has given variable results. By combining specific strains of microorganisms, multiple traits antagonizing the pathogen can be combined and this may result in a higher level of protection. Pseudomonas putida WCS358 suppresses Fusarium wilt of radish by effectively competing for iron through the production of its pseudobactin siderophore. However, in some bioassays pseudobactin-negative mutants of WCS358 also suppressed disease to the same extent as WCS358, suggesting that an, as yet unknown, additional mechanism may be operative in this strain. P. putida strain RE8 induced systemic resistance against fusarium wilt. When WCS358 and RE8 were mixed through soil together, disease suppression was significantly enhanced to approximately 50% as compared to the 30% reduction for the single strain treatments. Moreover, when one strain failed to suppress disease in the single application, the combination still resulted in disease control. The enhanced disease suppression by the combination of P. putida strains WCS358 and RE8 is most likely the result of the combination of their different disease-suppressive mechanisms. These results demonstrate that combining biocontrol strains can lead to more effective, or at least, more reliable biocontrol of fusarium wilt of radish.