Major facilitator superfamily (MFS) comprises a large family of fungal transporters. In this work four Penicillium digitatum MFS transporters named PdMFS2–5 were identified and functionally characterized through gene elimination and gene overexpression with aim of unveil the similarities and differences among members of the same family during pathogen-fruit interaction. Fungal mutants in which each of the MFS transporters were individually deleted, displayed a clear effect on their infective capacity during citrus fruit infection especially in two of them. In contrast, the observed effect on fungicide sensitivity limits PdMFS2 and PdMFS3 as transporters underlying fungicide resistance. Moreover, overexpression transformants confirmed P. digitatum MFS transporters function and PdMFS2 and PdMFS3 were able to confer fungicide resistance to P. digitatum strains originally fungicide sensitive. Gene transcription rate depended on each MFS transporter being PdMFS4 the one with higher gene expression. Transcriptional profiling was similar regardless the P. digitatum strain. The gene expression analysis showed an increase of PdMFSs transcription in all overexpression transformants, particularly in Pd27 strain. Expression analysis carried out during P. digitatum-citrus fruit interaction confirmed the contribution of all PdMFSs, excepting PdMFS5, in fungal virulence. These results indicate that MFS fungal transporters might be part of different processes and can replace other genes functions giving them a very high degree of versatility.

中号ajor ˚F acilitator小号uperfamily（MFS）包括一大家族真菌转运。在此工作相关的四个青霉名为MFS转运PdMFS2 - 5进行鉴定，并通过消除基因和基因过表达与病原体相互作用的水果在揭开同一个家庭的成员之间的相似性和差异的目标功能特点。其中每个MFS转运蛋白均被删除的真菌突变体，在柑橘类水果感染期间，尤其是其中两个，对其感染力表现出明显的影响。相反，观察到的对杀菌剂敏感性的影响限制了PdMFS2和PdMFS3作为抗真菌剂的转运蛋白。此外，过表达转化体证实了数指疟原虫MFS转运蛋白功能，PdMFS2和PdMFS3能够赋予最初对杀真菌剂敏感的数指疟原虫菌株抗药性。基因转录率取决于每个MFS转运蛋白是PdMFS4，即具有较高基因表达的转运蛋白。转录谱相似，无论指趾疟原虫菌株如何。基因表达分析表明，在所有过表达转化子中，特别是在Pd27菌株中，PdMFS转录均增加。在数指假单胞菌中进行表达分析-柑橘类水果的相互作用证实了除PdMFS5外的所有PdMFS对真菌毒力的贡献。这些结果表明，MFS真菌转运蛋白可能是不同过程的一部分，并且可以替代其他基因功能，从而使其具有高度的通用性。