Veluchamy Selvakumar, Rollins Jeffrey A
Department of Plant Pathology, University of Florida, 1453 Fifield Hall, Gainesville, FL 32611-0680, USA.
Fungal Genet Biol. 2008 Sep;45(9):1265-76. doi: 10.1016/j.fgb.2008.06.004. Epub 2008 Jun 27.
Apothecial development is the multicellular, sexual reproduction phase in the developmental life cycle of Sclerotinia sclerotiorum. This development begins within the sclerotium, a compact aggregation of vegetative hyphae contained within a melanized rind layer. Upon germination from the sclerotium, the apothecial stipe requires exposure to UV-A wavelengths of light to develop a fertile disc. We have identified a gene, cry1 from S. sclerotiorum that is most closely related to photolyase/cryptochrome proteins in the CRY-DASH family. We characterized this CRY-DASH ortholog from S. sclerotiorum and observed significant transcript accumulation only after exposure to UV-A and not in response to other wavelengths of light. Tissue-specific expression studies revealed that cry1 transcripts accumulate to low levels in vegetative mycelia and to higher levels in all light-exposed stages of apothecia development. Maximal cry1 transcript accumulation occurs in stipes between 2 and 6h of continuous UV-A exposure. Mutant strains carrying a deletion of cry1 exhibited a decrease in sclerotial mass and displayed greater numbers of pigmented hyphal projections on apothecial stipes under UV-A treatment but are otherwise developmentally normal. Tissue level localization of Cry1-GFP protein accumulation expressed from the native cry1 promoter was consistent with transcript localization. This study suggests that cry1 may have a function during UV exposure but is not essential for completing the developmental life cycle under laboratory conditions.
子囊盘发育是核盘菌发育生命周期中的多细胞有性繁殖阶段。这种发育始于菌核,菌核是由黑化外皮包裹的营养菌丝紧密聚集而成。从菌核萌发后,子囊盘柄需要暴露于UV-A波长的光下才能发育出可育盘。我们从核盘菌中鉴定出一个基因cry1,它与CRY-DASH家族中的光解酶/隐花色素蛋白关系最为密切。我们对核盘菌的这个CRY-DASH直系同源物进行了表征,发现只有在暴露于UV-A后才会有显著的转录本积累,而对其他波长的光没有反应。组织特异性表达研究表明,cry1转录本在营养菌丝中积累水平较低,而在子囊盘发育的所有光照阶段积累水平较高。在持续UV-A照射2至6小时之间,子囊盘柄中cry1转录本积累达到最大值。携带cry1缺失的突变菌株在UV-A处理下菌核质量下降,子囊盘柄上出现更多色素化的菌丝突起,但在其他方面发育正常。从天然cry1启动子表达的Cry1-GFP蛋白积累的组织水平定位与转录本定位一致。这项研究表明,cry1可能在UV暴露期间发挥作用,但在实验室条件下对完成发育生命周期不是必需的。