Neptunomonas sp. BPy-1 is an epiphytic bacterium isolated from in vitro culture of the red alga Pyropia yezoensis. It uses ethanol as a sole carbon source and promotes the growth of host alga. A related bacterium, Neptunomonas sp. BZm-1, was isolated from leaves of Zostera marina found in the Yatsushiro Sea (Japan). BZm-1 showed 99% 16S rRNA sequence identity with Neptunomonas sp. BPy-1. Similar to BPy-1, BZm-1 grew in artificial seawater (ASW) medium containing ethanol or butanol. When thalli were treated with a multi-enzyme cleaner, the growth of treated thalli was retarded, but the addition of BZm-1 to the medium promoted growth. To explore the benefits of epiphytic bacteria, indoleacetic acid (IAA) production by isolated bacteria was examined under conditions of limited nutrients. Salkowski assays and GC-MS analysis revealed that both BZm-1 and BPy-1 excreted IAA during growth in ASW medium containing glucose or ethanol in the presence of tryptophan. In ASW medium containing tryptophan but lacking a carbon source, neither isolate grow, but produced IAA. ASW medium includes nitrate as the sole nitrogen source. In the absence of carbon source, different nitrogen forms in the presence of tryptophan did not affect IAA production by the two isolates. These findings indicate that IAA production by the two isolates is strictly dependent on tryptophan but less affected by carbon and nitrogen sources. Based on the different origins of BPy-1 and BZm-1, this mode of IAA production seems to be conserved among relatives of BPy-1.

The established culture of gametophytes of the red alga Pyropia yezoensis (TU-1) is superficially colonized by epiphytic bacteria. By 16S rRNA sequencing, 6 bacterial species were identified, and a culturable bacterium, Neptunomonas sp. BPy-1, was isolated. The 16S rRNA sequences of BPy-1 showed 100% identity with that of Neptunomonas sp. 0536, a probiotic bacterium found in greenshell mussels in New Zealand. Physiological tests revealed that 22 characters were identical between BPy-1 and 0536, but that 4 characters differed. BPy-1 cannot grow in the artificial seawater used for the culture of gametophytes. BPy-1 can grow in the artificial seawater with ethanol or butanol but not in methanol or propanol. To determine the effect of BPy-1 on gametophyte growth, BPy-1 colonization was reduced by 80% using a multi-enzyme cleaner. Changing the cleaner concentration yielded two types of gametophytes, a compressed or callus-like form and a nearly normal form. BPy-1 promoted the growth of the treated gametophytes with relatively normal form, while it showed less effect on compressed gametophytes. These findings suggested that BPy-1 promotes the growth of damaged gametophytes but does not affect the development of normal gametophytes.