Development of a co-dominant SCAR marker linked to the Ph-3 gene for Phytophthora infestans resistance in tomato (Solanum lycopersicum)

Random amplified polymorphism DNA (RAPD) and bulk segregant analysis (BSA) approaches were used to characterize the molecular marker linked to the Phytophthora infestans resistance gene Ph-3 in tomato. A total of 800 RAPD primers were screened. One RAPD marker UBC#602 was identified to be tightly linked to the Ph-3 gene. The marker was successfully converted into a co-dominant sequence characterized amplified region (SCAR) marker. The SCAR marker SCU602 was used to analyze 96 F₂ progenies and fitted the expected 1:2:1 Mendelian segregation ratio. Forty one tomato inbred lines were screened using the SCAR marker in comparison with a reference marker linked to the Ph-3 gene and both markers gave the same results. SCU602 was further validated for association to resistance and its potential in MAS in 72 tomato lines and cultivars. The marker identified three genotypes harbouring the resistance allele. This SCAR marker can be used in breeding programs for the selection of the Ph-3 gene for Phytophthora infestans resistance.     

Plant Disease Control

Abstract

Experiments have been carried out in the Ivory Coast to assess the performance of a prototype rapid release system to apply larvicides for the control of S. damnosum, the vector of onchocerciasis. The equipment was fitted to a Pilatus Porter aircraft and was used to apply Abate larvicide to a large river during conditions of low water level. It was shown that the equipment could accurately deliver volumes of larvicide between one and 50 l and that these quantities could be placed exactly along chosen points in the river, provided the aircraft was flown along the direction of the watercourse. Satisfactory control of S. damnosum larvae was achieved at three sites using a dosage rate of 0.05 ppm/10 min of waterflow. However, it was recommended that a dosage of 0.1 ppm/10 min be adopted in future work because some of the more extensive sites treated were underdosed. It was not possible to assess the biological effectiveness of the equipment for treating small rivers, but physical tests were carried out which suggest that the system should be suitable for applying larvicide to rivers as narrow as five metres width. Recommendations were made to improve the reliability of the equipment for use in an operational onchocerciasis programme.     

Large subclonal variation in Phytophthora infestans populations associated with Ecuadorian potato landraces

The population of Phytophthora infestans on potato landraces in three provinces (Carchi, Chimborazo and Loja) of Ecuador was analysed. All isolates (n = 66) were of the A1 mating type. Simple sequence repeats (SSR) were used to assess the genetic diversity of the isolates. The P. infestans isolates from the potato landraces grouped in a single clade together with reference isolates belonging to the clonal lineage EC‐1. In the 66 SSR profiles obtained, 31 multilocus genotypes were identified. The 66 isolates constituted 49 different races according to the Solanum demissum differential set ( R1 to R11). The P. infestans population was complex and virulent on 4 to 11 R genes. Analysis showed that the subclonal variation in the Ecuadorian EC‐1 clone is increasing over time and is much larger than clonal variation in lineages in the Netherlands and Nicaragua, suggesting high mutation rates and little or no selection in Ecuador.     

An antibiotic fusaricidin: a cyclic depsipeptide from Paenibacillus polymyxa E681 induces systemic resistance against Phytophthora blight of red-pepper

In this study fusaricidin, a cyclic depsipeptide isolated from Paenibacillus polymyxa E681 (E681), was demonstrated to control Phytophthora blight infection caused by Phytophthora capsici in red-pepper. The minimal inhibitory concentration (MIC) of fusaricidin was found to be 16 ppm against P. capsici. The disease severity of P. capsici was 40% at 0.1 ppm of fusaricidin when compared with water-treated control (81.7%) on post-treatment, whereas the disease severities on pre-treatment were 45% and 83.3% in fusaricidin (0.1 ppm) and water-treated control, respectively, in red-pepper plants. Significant (P?<?0.05) disease suppression was observed on treatment with fusaricidin (0.1 ppm) by foliar spray and soil drench. The disease severity was drastically reduced to 3.3% by soil drench of fusaricidin (1.0 ppm), whereas in water-treated control, the disease severity was 83.3% in the first experiment. Fusaricidin at 0.1 ppm reduced disease severity of P. capsici to 27.5% when compared with positive control (43.1%) and water-treated control (66.2%) in the second experiment. Soft rot disease in tobacco was suppressed upon treatment with fusaricidin at 1.0 ppm by leaf infiltration. RT-PCR analyses of Arabidopsis thaliana revealed that there was an up-regulation of pathogenesis-related (PR) gene expression in wild type A. thaliana (Col-0), while there was no accumulation of PR genes, which implies that the mechanism of protection might be based on a salicylic acid-dependent pathway. This is the first report that fusaricidin exhibits protection against plant pathogens in addition to activity as an antibiotic agent. Hence, E681 can play a role in plant protection by secretion of ISR elicitors including fusaricidin.     

First molecular genotyping of voltage gated sodium channel alleles in Culex quinquefasciatus populations in Malaysia

A nationwide investigation was performed to detect the presence of 1014 mutation(s) in voltage gated sodium channel (kdr) gene of Culex quinquefasciatus from 14 residential areas across 13 states and a federal territory in Malaysia. Molecular genotyping of kdr mutation was performed via a modified three tubes allele-specific-polymerase chain reaction (AS-PCR) and direct sequencing of kdr gene. Based on the results of AS-PCR, homozygous susceptible (SS) genotype was found in nine out of 14 populations with 38 individuals from a total sample size of 140. Heterozygous (RS) genotype was most predominant (99 individuals) and distributed across all study sites. Homozygous resistance (RR) genotype was detected in Perak (one individual) and Selangor (two individuals). The resistance kdr allele frequencies ranged from 0.1 to 0.55, with the highest being detected in Cx. quinquefasciatus population from Selangor. This study has documented the first field-evolved instance of 1014F mutation in Malaysian mosquitoes and the findings of this study could be utilized in the implementation of strategic measures in vector control programs in Malaysia.     

Climate and interrelated tree regeneration drivers in mixed temperate–boreal forests

Forest compositional shifts in response to climate change are likely to be initially detectable in the understory tree regeneration layer near species range limits. Because many factors in addition to climate, such as seedbed and soil characteristics, overstory composition, and interactions with other understory biota, drive tree regeneration trends, a thorough understanding of the relative importance of all variables as well as their interrelationships is needed. The range limits of several widespread temperate and boreal tree species overlap in the upper Great Lakes region, USA, thus facilitating an observational study over relatively short regional climate gradients. We used redundancy analysis and variation partitioning to quantify the unique, shared, and total explanatory power of four sets of explanatory variables. The results showed that all four variable sets (climate 9.5 %, understory environment 13.7 %, overstory composition 26.3 %, and understory biota 13.8 %) were significantly associated with tree regeneration compositional variation in mixed temperate–boreal forests. Partitioning also revealed high confounded or shared explanatory power, but also that each set contributed significant unique explanatory power not shared with other sets. Spatial patterning in regeneration composition was strongly related to broad scale environmental patterns, while the large majority of unexplained variation did not have a detectable spatial structure, suggesting factors with local scale variability. Future forest shifts across the landscape will depend not only on the rate and direction of climate change but also on how the strengths and interrelationships among other explanatory variables, such as overstory composition and understory biota, shift with a changing climate.     

Assessing the risk of recurrence of porcine epidemic diarrhea virus in affected farms on Jeju Island,South Korea

BackgroundPorcine epidemic diarrhea virus (PEDV) is a swine enteropathogenic coronavirus that has devastated the swine industry in South Korea over the last 30 years. The lack of an effective method to control the endemics has led to a surge in PEDV recurrences in affected farms throughout the country.ObjectivesIn the first step toward establishing systematic monitoring of and active control measures over the swine populations, we constructed an assessment model that evaluates the status of (1) biosecurity, (2) herd immunity, and (3) virus circulation in each of the PEDV-infected farms.MethodsA total of 13 farrow-to-finish pig farms with a history of acute PEDV infection on Jeju Island were chosen for this study. The potential risk of the recurrence in these farms was estimated through on-site data collection and laboratory examination.ResultsOverall, the data indicated that a considerable number of the PEDV-infected farms had lax biosecurity, achieved incomplete protective immunity in the sows despite multi-dose vaccination, and served as incubators of the circulating virus; thus, they face an increased risk of recurrent outbreaks. Intriguingly, our results suggest that after an outbreak, a farm requires proactive tasks, including reinforcing biosecurity, conducting serological and virus monitoring to check the sows’ immunity and to identify the animals exposed to PEDV, and improving the vaccination scheme and disinfection practices if needed.ConclusionsThe present study highlights the significance of coordinated PEDV management in infected farms to reduce the risk of recurrence and further contribute towards the national eradication of PEDV.     

Factors Affecting the Storage Behaviour of Papaya Fruit

Fruits of the papaya cultivar Solo 63/2 were damaged when stored at or below 45 °F (7 °C). Wrapping the fruit in polyethylene bags reduced weight loss during storage but increased the development of saprophytic fungi. Perforated polyethylene bags similarly reduced weight loss but did not increase fungal development.

Careful harvesting significantly reduced fungal infection, thus giving the fruits better flavour and longer storage life. The optimum stage of maturity for satisfactory storage and subsequent ripening was when the yellow colour was just beginning to develop in the funicles. Fruits from hermaphrodite trees were superior in flavour to those from female trees but behaved similarly during storage.     

Development of new microsatellite markers for molecular diversity analysis of Citrus species

Summary

Thirty microsatellite loci for further genetic analysis of Citrus species were developed by constructing a microsatellite-enriched library using capture with streptavidin-coated magnetic beads and used to assess genetic diversity in 40 Citrus accessions. In total, 150 alleles were detected, with an average of five alleles per locus. The average gene diversity and polymorphism information content values were 0.58 and 0.52, and ranged from 0.35 – 0.74 and from 0.32 – 0.70, respectively. Values for the observed and expected heterozygosities ranged from 0.13 – 1.00 and from 0.36 – 0.75, respectively. Fifteen loci deviated from the Hardy-Weinberg equilibrium (P < 0.05). The mean similarity coefficient among accessions was 0.5906. Based on the UPGMA algorithm, two main groups were successfully identified. These new microsatellite markers can be used to further investigate the genetics of, and phylogenic relationships in Citrus spp.