Somatic hybridization has been frequently used to overcome sexual incompatibility between potato and its secondary germplasm. The primary objective of this study was to produce and evaluate somatic hybrids of Solanum tuberosum (Stub) and S. bulbocastanum (Sblb) for breeding purposes. In 2007, 23 somatic hybrids were produced using an electrofusion of mesophyll protoplasts of diploid (2n = 2x = 24) potato line StubDH165 and S. bulbocastanum PI24351 (Sblb66). Phenotype of somatic hybrids in field conditions were evaluated, together with constitution and stability of 30 nuclear (ncSSR) and 27 cytoplasmic (cpSSR) microsatellite markers and content of main glycoalkaloids. All somatic hybrids had very high field resistance against late blight, but the plants were infertile: the viability of pollen grains insignificantly varied between 0.58 and 8.97%. A significant somaclonal variation was observed in terms of the morphology of plants, the date of emergence, the quantity of harvested tubers, the content of glycoalkaloids in foliage, and nuclear microsatellite markers (ncSSR). The analysis of ncSSR identified five distinct genotypes of hybrids partly associated with phenotype variations. The process of somatic hybridization with regeneration of shoots was identified as the most likely source of somaclonal variation because the ncSSR genotypes of hybrids, which were maintained in vitro, remained stable for more than 10 years. The infertile somatic hybrids have no practical breeding potential, but they are considered very suitable for advanced studies of the differential expression of genes in the pathways linked to dormancy of tubers and synthesis of glycoalkaloids.

» Articles | Agricultural Journals

» Articles | Agricultural Journals

The identification of sex in larvae of insects is usually challenging or even impossible, while in adults the sexual dimorphism is usually evident. Here, we used copy number analysis to develop a method of sex detection in Colorado potato beetle (Leptinotarsa decemlineata), which has an X0 sex determination system. The X linked geneLdVsscand autosomal geneLdUBE3Bwere identified as appropriate target and reference loci, respectively. The copy numbers (CNV) ofLdVsscin males and females were estimated using standard droplet digital PCR (ddPCR) and real-time PCR (qPCR). With both methods, CNVs were bimodally distributed (BA_ddPCR = 0.709 and BA_qPCR = 0.683) with 100% ability to distinguish females from males. The use of qPCR-based sex detection in a broad collection of 448 random CPB adults showed a perfect association (Phi = 1.0,p < 0.05) with the true sexes of adults, with mean CNV in females of 2.032 (SD = 0.227) and 0.989 in males (SD = 0.147). In the collection of 50 random 4th instar larvae, 27 females and 23 males were identified, consistent with the expected 1:1 sex ratio (p = 0.689). The method is suitable for sexing in all stages of ontogenesis. The optimal cost-effective application of the method in large populations requires the DNA extraction using CTAB, the qPCR assay in one biological replicate and three technical replicates of each marker, and the use of one randomly chosen male per run to calibrate calculation of CNV.

To evaluate the frequency and stability of the occurrence of P. infestans races and mating types in the Czech Republic, 338 monosporic isolates were collected from 31 sites in different potato-growing areas from 2012 to 2014 and in 2016. In total, 142 isolates were evaluated for virulence and race structure using the detached leaflet assay on Black’s differential set, supplemented with cultivar Sarpo Mira and somatic hybrid REG 46F. With the exception of virulence for resistance genes R9 and Rpi-blb-1, all virulence genes were detected among isolates, with a predominance of genes R1, R3, R7, R10, and R11. Most isolates were virulent to five or more R-genes, with a mean virulence complexity of 7.1. Among the 38 races detected, the most commonly occurring races were 1.2.3.4.6.7.10.11 and 1.2.3.4.7.10.11. Of the 338 isolates tested by the pairing test and the cleaved amplified polymorphism sequence (CAPS) marker, 40% were of the A1 mating type and 60% were of the A2 mating type, with an A1 : A2 isolate ratio demonstrating the predominance of the A2 mating type each year of the survey.

This study describes a newly developed index for predicting and forecasting the first (and potentially subsequent) timing of fungicide application against late blight in potato crops based on weather variables measured close to the crop. Inputs for index calculation were the following: daily minimum temperature, mean relative air humidity and daily precipitation. The decisive moment in the process of forecasting is the sum of daily index values for the previous 5 days. The index was tested in various localities of the Czech and the Slovak Republics for several years with a relatively high success rate exceeding the accuracy of previously applied strategies-NoBlight and negative prognosis. In comparison to the mentioned methods, the calculated index corresponded very well to long-term wet periods and indicated the first application date correctly. In years with no wet periods (in this case, 2015 and 2017), it allowed postponing the first application and reducing the number of required sprays during the growing season. The method does not depend on determining the emergence date, so it can be presented on the internet without cooperation with specific growers in a given locality, and thus supply information for a wider range of users. With knowledge about crop development and the degree of resistance to late blight of grown varieties, users can subsequently choose a specific fungicide and its application date.