Johnson et al. (2002) showed that long-tubed flowers in Zaluzianskya section Nycterinia, to which Z. microsiphon belongs, are mostly specialized for pollination by hawkmoths, predominantly Hippotion celerio. The hawkmoths carry large pollen loads (100–3 000 grains) on their proboscides and heads. In sharp contrast, Z. microsiphon with its unique floral traits (zygomorphy, diurnal flowering and the lack of scent), is adapted for pollination by long proboscid flies. Access to the nectar, which accumulates at the base of the long floral tubes, is restricted to these long-proboscid insects.

Even though Z. microsiphon and Z. natalensis have flowers of the same colour and about the same size, hawkmoths strongly prefer the latter, as its flowers are open and fragrant in the evening, and are presented vertically face up to the pollinator, as opposed to horizontally in Z microsiphon. Changing the orientation of Z. natalensis flowers to face horizontally decreased its attraction to hawkmoths (Campbell et al. 2015).

The study by Anderson et al. (2005) confirmed that Z. microsiphon is exclusively, or almost exclusively, pollinated by the long-proboscid fly Prosoeca ganglbaueri belonging to the family Nemestrinideae. Furthermore, flowers of Z. microsiphon in the Drakensberg region appear to be the most important sources of nectar for P. ganglbaueri during a large proportion of its flight season (Anderson et al. 2005).

Unmanipulated and self-pollinated flowers of Z. microsiphon do not produce seeds nor fruits, whereas fruits develop fully by cross-pollination. This shows that this species is self-incompatible, depending on pollinators for seed production. Johnson et al. (2002) speculate that the hairs around the mouth of the corolla tube may be responsible for the production of scent, since they are abundant in the scented night-flowering species of section Nycterinia, but rare in the unscented day-flowering Z. microsiphon. The gland-tipped hairs that cover most of the corolla may serve as a defence mechanism for the plant against herbivores that feed on the flowers, not involved in pollination (Johnson et al. 2002).