Zinc is critically required for pollen function and fertilisation in lentil

The role of zinc (Zn) in reproduction of lentil (Lens culinaris Medik. cv. DPL 15) and the extent to which the Zn requirement for reproduction can be met through supplementation of Zn at the time of initiation of the reproductive phase have been investigated.Low supply (0.1 μmol/L) of Zn reduced the size of anthers, the pollen producing capacity and the size and viability of the pollen grains. Scanning electron microscopy (SEM) of pollen grains of Zn deficient plants showed enhanced thickening of exine and wide and raised muri. In vitro germination of pollen grains was reduced by >50% and growth of pollen tubes was retarded. Unlike Zn sufficient plants, the cuticle around the stigmatic papillae of Zn deficient plants remained intact, preventing the interaction between pollen grains and stigmatic exudates that provides the polarity for the growth of pollen tubes through the stylar tract. Zn deficiency increased the activity of acid phosphatase and peroxidase in extracts of pollen grains. Histochemical localisation on the stigmatic surface and native PAGE of the enzyme extracts of pollen grain and stigma exudates showed enhanced expression of acid phosphatase and peroxidase and suppressed expression of esterase in response to Zn deficiency. Zn deficiency reduced the setting of seeds and also their viability. The effect on seed setting was more marked than on in vitro germination of pollen grains, suggesting that the latter was not the exclusive cause of inhibition of fertility. Possibly, loss of fertility was also caused by impairment in pollen–pistil interaction conducive to pollen tube growth and fertilisation. Impairment in pollen structure and function and seed setting was observed even when plants were deprived of Zn at the time of flowering, but to a lesser extent than in plants maintained with low Zn supply from the beginning. Increasing the Zn supply from deficient to sufficient at the initiation of flowering decreased the severity of Zn deficiency effects on pollen and stigma morphology, pollen fertility and seed yield.In conclusion, structural and functional changes induced in pollen grains and stigma of Zn deficient plants and associated decrease in seed setting of lentil indicate a critical requirement of Zn for pollen function and fertilisation that can be partially met by supplementing Zn at the onset of the reproductive phase.