Ingenious　design　and　fabrication　of　advanced　carbon-based　sulfur　cathodes　are　extremely　important　to　the　development　of　high-energy　lithium-sulfur　batteries,　which　hold　promise　as　the　next-generation　power　source.　Herein,　for　the　first　time,　we　report　a　novel　versatile　hyphae-mediated　biological　assembly　technology　to　achieve　scale　production　of　hyphae　carbon　fibers　(HCFs)　derivatives,　in　which　different　components　including　carbon,　metal　compounds,　and　semiconductors　can　be　homogeneously　assembled　with　HCFs　to　form　composite　networks.　The　mechanism　of　biological　adsorption　assembly　is　also　proposed.　As　a　representative,　reduced　graphene　oxides　(rGOs)　decorated　with　hollow　carbon　spheres　(HCSs)　successfully　co-assemble　with　HCFs　to　form　HCSs@rGOs/HCFs　hosts　for　sulfur　cathodes.　In　this　unique　architecture,　not　only　large　accommodation　space　for　sulfur　but　also　restrained　volume　expansion　and　fast　charge　transport　paths　are　realized.　Meanwhile,　multiscale　physical　barriers　plus　chemisorption　sites　are　simultaneously　established　to　anchor　soluble　lithium　polysulfides.　Accordingly,　the　designed　HCSs@rGOs/HCFs-S　cathodes　deliver　a　high　capacity　(1189　mA　h　g−1　at　0.1　C)　and　good　high-rate　capability　(686　mA　h　g−1　at　5　C).　Our　work　provides　a　new　approach　for　the　preparation　of　high-performance　carbon-based　electrodes　for　energy　storage　devices.　©　2024　The　Authors.　Carbon　Energy　published　by　Wenzhou　University　and　John　Wiley　&　Sons　Australia,　Ltd.