We exploited the advantages of genomic in situ hybridization (GISH) to monitor the introgression process at the chromosome level using a simple and robust molecular marker in the interspecific breeding of bulb onion (Allium cepa L.) that is resistant to downy mildew. Downy mildew (Peronospora destructor [Berk.] Casp.) is the most destructive fungal disease for bulb onions. With the application of genomic in situ hybridization (GISH) and previously developed DMR1 marker, homozygous introgression lines that are resistant to downy mildew were successfully produced in a rather short breeding time. Considering that the bulb onion is a biennial plant, it took seven years from the F1 hybrid production to the creation of S2BC2 homozygous lines that are resistant to downy mildew. Using GISH, it was shown that three progeny plants of S2BC2 possessed an A. roylei homozygous fragment in the distal region of the long arm of chromosomes 3 in an A. cepa genetic background. Previously, it was hypothesized that a lethal gene(s) was linked to the downy mildew resistance gene. With the molecular cytogenetic approach, we physically mapped more precisely the lethal gene(s) using the homozygous introgression lines that differed in the size of the A. roylei fragments on chromosome 3.

The data provided in this article were obtained from fresh and dry samples of green Basilic ( Ocimum Basillicum L. ), red Basilic ( Ocimum basilicum 'purpurascens' ), green Mizuna (Brassica rapa var. niposinica), and red Mizuna ( Brassica rapa var. pipposinica) microgreens grown in climatic chamber (phytotron) on jute mats for 15 days. Phytonutrients contents including chlorophylls, pheophytins, carotenoids pigments, total antioxidant capacity, total phenolic content, ascorbic acid, as well as organic acids contents varied between all cultivars. Spectrometry, electrophoresis, coulometric, and liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry (LC-ESI-QTOF/MS) were the principal employed methods. Data of antioxidants and phytonutrients contents contribute to the understanding of the benefits of microgreens as a newly emerging product. Data of pigments content compares the difference of accumulation of chlorophylls, pheophytins, and carotenoids between red and green cultivars of the studied microgreens, and the variability of their concentrations along with the contents of organic acids provide insights to plants physiology during the differentiation phase.

There is little information on the use of pollen in molecular research, despite the increased interest in genome editing by pollen-mediated transformation. This paper presents an essential toolbox of technical procedures and observations for molecular studies on onion (Allium cepa L.) pollen. PCR is a useful tool as an express method to evaluate editing results before pollination. A direct PCR protocol for pollen suspension has been adapted without needing DNA pre-extraction. We showed that the outer layer of lipids known as pollenkitt is a limiting factor for successful PCR on pollen. A simple pre-washing step of pollen suspension was able to eliminate the pollenkitt and enormously affect the PCR results. Additionally, our pollenkitt study helped us develop a simple and effective pollination method using wetted onion pollen grains. Classical manual pollination usually is conducted by intact pollen without wetting. Most existing methods of the editing system delivery into pollen are carried out in a wet medium with consequent drying before pollination, which adversely affects the viability of pollen. The optimal medium for wet pollination was 12% sucrose water solution. Our method of using wetted pollen grains for pollination might be very beneficial for pollen genetic manipulation.

There is little information on the use of pollen in molecular research, despite the increased interest to genome editing by pollen mediated transformation. PCR is useful tool as an express-method to evaluate editing results before pollination. A direct PCR protocol for pollen suspension has been adapted without the need for DNA pre-extraction. We showed that pollenkitt is a limiting factor for successful PCR on pollen. A simple pre-washing step of pollen suspension was able to eliminate the pollenkitt and enormously affect the PCR results. All currently existing methods of delivery of the editing system to pollen are carried out in a wet medium. Our pollenkitt study helped us develop a simple and effective pollination method using wetted onion pollen grains.