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Impedance flow cytometry (IFC) is a versatile lab-on-chip technology which enables fast and label-free analysis of pollen grains in various plant species, promising new research possibilities in agriculture and plant breeding. Hazelnut is a monoecious, anemophilous species, exhibiting sporophytic self-incompatibility. Its pollen is dispersed by wind in midwinter when temperatures are still low and relative humidity is usually high. Previous research found that hazelnut can be characterized by high degrees of pollen sterility following a reciprocal chromosome translocation occurring in some cultivated genotypes. In this study, IFC was used for the first time to characterize hazelnut pollen biology. IFC was validated via dye exclusion in microscopy and employed to (i) follow pollen hydration over time to define the best pre-hydration treatment for pollen viability evaluation; (ii) test hazelnut pollen viability and sterility on 33 cultivars grown in a collection field located in central Italy, and two wild hazelnuts. The accessions were also characterized by their amount and distribution of catkins in the tree canopy. Pollen sterility rate greatly varied among hazelnut accessions, with one main group of highly sterile cultivars and a second group, comprising wild genotypes and the remaining cultivars, producing good quality pollen. The results support the hypothesis of recurring reciprocal translocation events in Corylus avellana cultivars, leading to the observed gametic semi-sterility. The measured hazelnut pollen viability was also strongly influenced by pollen hydration (Radj2 = 0.83, P ≤ 0.0001) and reached its maximum at around 6 h of pre-hydration in humid chambers. Viable and dead pollen were best discriminated at around the same time of pollen pre-hydration, suggesting that high humidity levels are required for hazelnut pollen to maintain its functionality. Altogether, our results detail the value of impedance flow cytometry for high throughput phenotyping of hazelnut pollen. Further research is required to clarify the causes of pollen sterility in hazelnut, to confirm the role of reciprocal chromosome translocations and to investigate its effects on plant productivity.

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Hazelnut cultivation is rapidly expanding in regions outside its native range. New hazelnut plantations in South Africa are facing adverse environmental conditions which threaten the pollination process and hamper nut yield. Artificial pollination can increase fruit yield and fill pollination gaps in some fruit crops, but its application on hazelnut is still not well explored. This study investigated biological factors and technical procedures in the first artificial pollination experiment on hazelnut in South Africa. A suspension media for artificial application composed of 10% sucrose, 0,1% agar and 0,02% boric acid was used. In addition, alternative low-cost suspension media containing other forms of sugar were also evaluated. Moreover, a novel and practical approach to assess pollen conditions in liquid solutions was designed. Pollen viability was tested in the different suspension media. This varied greatly among the examined cultivars. Wild hazelnut produced the pollen with the highest viability, while Tonda Gentile delle Langhe (TGL) and Barcelona the lowest. Sterile grains were very abundant, especially in cultivated varieties (up to 65% in TGL). Preliminary data on nut yield were also collected. Altogether, this study indicated that artificial pollination is a promising approach for hazelnut cultivation in increasing the final yield. Further research is needed to develop integrated pollination strategies, especially in areas where the environmental conditions are adverse to the pollination process.