In the present study, we identified and characterized the apricot ( Prunus armeniaca L.) homologs of three dormancy-related genes, namely the ParCBF1 ( C-repeat binding factor ), ParDAM5 ( dormancy-associated MADS-BOX ) and ParDAM6 genes. All highly conserved structural motifs and the 3D model of the DNA-binding domain indicate an unimpaired DNA-binding ability of Par CBF1. A phylogenetic analysis showed that ParCBF1 was most likely homologous to Prunus mume and Prunus dulcis CBF1 . Par DAM5 also contained all characteristic domains of the type II (MIKC C ) subfamily of MADS-box transcription factors. The homology modeling of protein domains and a phylogenetic analysis of ParDAM5 suggest its functional integrity. The amino acid positions or small motifs that are diagnostic characteristics of DAM5 and DAM6 were determined. For ParDAM6 , only a small part of the cDNA was sequenced, which was sufficient for the quantification of gene expression. The expression of ParCBF1 showed close association with decreasing ambient temperatures in autumn and winter. The expression levels of ParDAM5 and ParDAM6 changed according to CBF1 expression rates and the fulfillment of cultivar chilling requirements (CR). The concomitant decrease of gene expression with endodormancy release is consistent with a role of ParDAM5 and ParDAM6 genes in dormancy induction and maintenance. Cultivars with higher CR and delayed flowering time showed higher expression levels of ParDAM5 and ParDAM6 toward the end of endodormancy. Differences in the timing of anther developmental stages between early- and late-flowering cultivars and two dormant seasons confirmed the genetically and environmentally controlled mechanisms of dormancy release in apricot generative buds. These results support that the newly identified apricot gene homologs have a crucial role in dormancy-associated physiological mechanisms.
The walnut husk fly (Rhagoletis completa) is one of the main pests affecting common walnut in both Europe and America. This work examines the effects of abamectin on the development of walnut husk fly larvae by injecting the product Vertimec 1.8 EC (Syngenta) into the trunks of walnut trees in Hungary. In the case of properly injected trees, the infection rate was negligible and the abamectin content in the husk samples ranged between 1.54 and 3.00 ng/g; controls show a very high (> 90%) infestation rate and an abamectin content under the detection limit. Abamectin residue measured in walnut kernel did not exceed the maximum residue limit; moreover, the active ingredient content was below the detection limit (0.0003 mg/kg) in all the collected kernel samples. Our results confirm that trunk injection is a viable method for walnut pest control.
This study examined the larvicidal effect of trunk-injected abamectin and emamectin benzoate against the walnut husk fly (Rhagoletis completa Cresson, 1929). Walnut trees in two locations in two years were injected with the pesticides at different concentrations. For the toxicokinetic studies, the active ingredient content was measured in the leaves, flowers, husks, and kernels, using a UHPLC-MS/MS analytical method. The walnut husk fly infestation rates were between 3 and 70% and 10 and 34% for abamectin and emamectin benzoate, respectively, and were much lower compared to those measured for the control. The active ingredient content in the walnut husk showed a positive correlation with the larvicidal effect. The injections had a measurable but unsatisfactory insecticidal effect in the second year, when the economic threshold was exceeded. Trace amounts of the active ingredients were detected in the flowers. The residue analysis showed a declining concentration trend in the leaves over time. The largest quantities were detected in the leaves (≤439 ng/g of abamectin; ≤19,079 ng/g of emamectin benzoate), with concentrations in the husks of orders of magnitude lower (≤5.86 ng/g; ≤50.19 ng/g). The measurements showed no active ingredient residue above the MRLs in either fresh or dried kernels. The results indicate that trunk injections of abamectin, as well as trunk injections of emamectin benzoate, have the potential to suppress walnut husk fly populations.
Blossom blight caused by Monilia laxa (Ehr.) is the most important fungal disease in Hungarian apricot orchards. The cultivars traditionally grown in the country are susceptible to Monilia laxa (Ehr.) to various extents. In this study the shoots of one tree each of the varieties Zard and Korai Zamatos and 48 hybrids from their cross were artificially infected in vivo with Monilia laxa (Ehr.). The results indicated that when artificial infections are made to evaluate pathogen resistance, this should be carried out on one-year-old shoots, since this is the natural infection point of Monilia. It also appears that, due to the great variability in the size of destroyed tissues, the microscopic analysis of the infections could provide a more reliable evaluation of progeny resistance than comparing the sizes of destroyed shoot areas.
Genes encoding for proteins with nucleotide-binding site and leucine-rich repeat motifs (NBS-LRR) have been suggested to play a general role in plant defence mechanism. In Prunus species, many TIR (Toll / Interleukin-1 Receptor), and only very few non-TIR sequences were identified, which was explained either by the unequal distribution of TIR/non-TIR sequences in the Prunus genome or by the incapability of primers in the amplification of non-TIR RGAs. The objective of this work was to check whether a new semi-nested PCR strategy can be developed for the targeted isolation of non-TIR-NBS-LRR Resistance Gene Analog (RGA) sequences from apricot. Three primers (CUB-P-loop F, CUB-Kin2 F and CUB-HD R) were designed, from which CUB-Kin2 F and CUB-HD R were constructed to anneal selectively to the non-TIR sequences. A colony Polymerase Chain Reaction (PCR) indicated that out of the 96 clones tested 28 showed amplification using the newly developed primers, while no amplification occurred when using the formerly described primers. Half of the 28 positive clones were sequenced and they turned out to represent 11 different non-TIR RGA sequences. A phylogenetic analysis was carried out based on an alignment containing 293 Rosaceae and 21 non-Rosaceaa sequences. A significantly higher ratio (91%) of non-TIR sequences were arranged in multi-genera clades than that of (57%) the TIR groups confirming that non-TIR sequences might be of more ancient origin than TIR sequences.
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