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[Objective] To explore genes related to the regulation of yeast spore germination, we previously screened for mutants defective in maintaining the quiescent state of spores. [Methods] Using the temperature-sensitive yeast mutant J33 with defective spore quiescence maintenance as the research material, heterozygous diploid strains were constructed through tetrad dissection and haploid fusion, and genetic segregation analysis was performed to clarify the genetic regulatory pattern of mutant phenotypes. The yeast genomic library complementation screening was applied to identify candidate mutant genes, and gene cloning and sequence alignment were conducted to confirm the mutation site. Furthermore, recombinant plasmids carrying wild-type and mutant TIP20 were constructed for functional complementation assays, and site-directed mutagenesis was used to verify the correlation between TIP20 mutation and strain phenotypes. [Results] Functional complementation experiments demonstrated that introduction of the wild-type TIP20 restored the growth at 37 ℃ and the normal spore germination phenotype of J33, while the mutant TIP20 and empty vector had no such effect. Site-directed mutagenesis analysis further confirmed that this single-base mutation was T1219G, a direct cause of the TS phenotype and defective spore germination of J33. [Conclusion] TIP20 is involved in the regulation of yeast spore germination. TIP20 encodes a subunit of the tethering complex involved in endoplasmic reticulum-Golgi retrograde transport. However, TIP20 is likely a multifunctional protein, and its mammalian homologue RINT1 has been reported to regulate the cell cycle checkpoint. Thus, TIP20 may also be involved in regulating the quiescent state of yeast cells.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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