为探寻alg3、vps1301、rad51基因缺失对粟酒裂殖酵母(Schizosaccharomyces pombe)生长及接合生殖的影响,在25 ℃条件下,培养野生型和3种基因缺失的粟酒裂殖酵母,统计并分析其生长情况;将野生型和3种基因缺失菌株分别各自交配产孢,分析对比野生型和基因缺失菌株的接合生殖情况. 结果显示,生长实验中,25 ℃条件下,12 h内,野生型菌株、alg3Δ菌株、vps1301Δ菌株和rad51Δ菌株的平均生长速度分别为0.042 5 、0.009 8 、0.019 3 、0.036 9 OD595/h. 结果提示,粟酒裂殖酵母缺失alg3基因后,其生长速度受影响最大.在细胞接合生殖实验中,3种基因缺失菌株产孢数目都出现异常,其中alg3Δ菌株产孢数与野生型表现出显著差异.同时,alg3基因和vps1301基因缺失后孢子长度与野生型存在极显著差异.结果表明,alg3基因缺失对粟酒裂殖酵母的接合生殖影响最大.综上所述,缺失上述3种基因都对粟酒裂殖酵母的生长和接合生殖有不同程度的影响,其中alg3基因缺失对其影响最严重,其次是vps1301基因,rad51基因对其影响最小.
In order to study the effects of deletion of alg3,vps1301 and rad51 genes on the growth and conjugal reproduction of Schizosaccharomyce spombe,under the condition of 25 ℃,the wild-type and three kinds of gene deleted strains were cultured, and their growth was counted and analyzed; the wild-type and three kinds of gene deleted strains were mated to produce spores respectively,and the conjugal reproduction of wild-type and gene deleted strains were also analyzed and compared.The results showed that the average growth rates of wild-type strain, alg3Δ strain, vps1301Δ strain and rad51Δ strain were 0.042 5、- DOI:10.3969/j.issn.1000-1565.2021.06.012alg3、vps1301、rad51基因缺失对粟酒裂殖酵母接合生殖的影响白鑫1,丁祥2,李慧1,杨彤1,陈茜2,侯怡铃1(1.西华师范大学 生命科学学院,西南野生动植物资源保护教育部重点实验室,四川 南充 637009;2.西华师范大学 环境科学与工程学院,四川 南充 637009)摘 要:为探寻alg3、vps1301、rad51基因缺失对粟酒裂殖酵母(Schizosaccharomyces pombe)生长及接合生殖的影响,在25 ℃条件下,培养野生型和3种基因缺失的粟酒裂殖酵母,统计并分析其生长情况;将野生型和3种基因缺失菌株分别各自交配产孢,分析对比野生型和基因缺失菌株的接合生殖情况. 结果显示,生长实验中,25 ℃条件下,12 h内,野生型菌株、alg3Δ菌株、vps1301Δ菌株和rad51Δ菌株的平均生长速度分别为0.042 5 、0.009 8 、0.019 3 、0.036 9 OD595/h. 结果提示,粟酒裂殖酵母缺失alg3基因后,其生长速度受影响最大.在细胞接合生殖实验中,3种基因缺失菌株产孢数目都出现异常,其中alg3Δ菌株产孢数与野生型表现出显著差异.同时,alg3基因和vps1301基因缺失后孢子长度与野生型存在极显著差异.结果表明,alg3基因缺失对粟酒裂殖酵母的接合生殖影响最大.综上所述,缺失上述3种基因都对粟酒裂殖酵母的生长和接合生殖有不同程度的影响,其中alg3基因缺失对其影响最严重,其次是vps1301基因,rad51基因对其影响最小.关键词:粟酒裂殖酵母;alg3基因;vps1301基因;rad51基因;接合生殖中图分类号:Q932 文献标志码:A 文章编号:1000-1565(2021)06-0720-08Effects of the deletions of alg3, vps1301 and rad51 genes on the growth and conjugal reproduction of Schizosaccharomyces pombeBAI Xin1, DING Xiang2, LI Hui1, YANG Tong1, CHEN Xi2, HOU Yiling1(1. Key Laboratory of Southwest China Wildlife Resources Conservation( Ministry of Education), College of Life Sciences, China West Normal University, Nanchong 637009, China; 2. College of Environmental Science and Engineering, China West Normal University, Nanchong 637009, China)Abstract: In order to study the effects of deletion of alg3,vps1301 and rad51 genes on the growth and conjugal reproduction of Schizosaccharomyce spombe,under the condition of 25 ℃,the wild-type and three kinds of gene deleted strains were cultured, and their growth was counted and analyzed; the wild-type and three kinds of gene deleted strains were mated to produce spores respectively,and the conjugal reproduction of wild-type and gene deleted strains were also analyzed and compared.The results showed that the average growth rates of wild-type strain, alg3Δ strain, vps1301Δ strain and rad51Δ strain were 0.042 5、- 收稿日期:2021-03-23 基金项目:四川省科技厅应用基础重点项目(2018JY0087);四川省科技厅重点研发项目(2018NZ0055) 第一作者:白鑫(1996—),男,四川绵阳人,西华师范大学在读硕士研究生,主要从事细胞生物学研究.E-mail: 2432460273@qq.com 通信作者:侯怡铃(1983—),女,四川南充人,西华师范大学教授,博士,主要从事细胞生物学研究.E-mail: starthlh@126.com第6期白鑫等:alg3、vps1301、rad51基因缺失对粟酒裂殖酵母接合生殖的影响0.009 8、0.019 3、0.036 9 OD595/h at 25 ℃ for 12 h, respectively. The results showed that the growth rate of S. pombe was most affected by the deletion of alg3 gene. In the experiment of cell conjugal reproduction, the number of spores produced by the three gene deletion strains were abnormal, and the number of spores produced by the alg3Δ strain was significantly different from that of the wild type. At the same time, there was a significant difference in spore length between the wild type and the alg3Δ strain and vps1301Δ strain. which indicated that the deletion of alg3 gene had the greatest effect on the conjugal reproduction of S. pombe. In conclusion, deletion of the above three genes had different effects on the growth and conjugal reproduction of S. pombe, among which the deletion of alg3 gene had the most serious effect, followed by vps1301 gene deletion, and rad51 gene deletion had the least effect.
[1] 户银芝, 商巾杰. 粟酒裂殖酵母中指示线粒体形态的荧光标签的构建[J]. 基因组学与应用生物学, 2021, 40(3): 1115-1121. DOI:10.13417/j.gab.040.001115.
[2] PAGE S L, HAWLEY R S. Chromosome choreography: the meiotic ballet[J]. Science, 2003, 301(5634): 785-789. DOI:10.1126/science.1086605.
[3] LIU N N, HAN T X, DU L L, et al. A genome-wide screen for Schizosaccharomyces pombe deletion mutants that affect telomere length[J]. Cell Res, 2010, 20(8): 963-965. DOI:10.1038/cr.2010.107.
[4] DUDIN O, MERLINI L, BENDEZU ' F O, et al. A systematic screen for morphological abnormalities during fission yeast sexual reproduction identifies a mechanism of actin aster formation for cell fusion[J]. PLoS Genet, 2017, 13(4): e1006721. DOI:10.1371/journal.pgen.1006721.
[5] SHARMA C B, KNAUER R, LEHLE L. Biosynthesis of lipid-linked oligosaccharides in yeast: the ALG3 gene encodes the Dol-P-Man: Man5GlcNAc2-PP-Dol mannosyltransferase[J]. Biol Chem, 2001, 382(2): 321-328. DOI:10.1515/BC.2001.039.
[6] SAUVAGEAU S, STASIAK A Z, BANVILLE I, et al. Fission yeast rad51 and dmc1, two efficient DNA recombinases forming helical nucleoprotein filaments[J]. Mol Cell Biol, 2005, 25(11): 4377-4387. DOI:10.1128/mcb.25.11.4377-4387.2005.
[7] 钟志宏, 李启星, 唐小异, 等. 甘油保存菌种方法的改良[J]. 实用预防医学, 2005, 12(3): 670-671. DOI:10.3969/j.issn.1006-3110.2005.03.114.
[8] 王文超. 裂殖酵母表现遗传调控产孢及异染色质沉默的研究[D]. 上海: 复旦大学, 2014.
[9] DAVIDSON R C, NETT J H, RENFER E, et al. Functional analysis of the ALG3 gene encoding the Dol-P-Man: Man5GlcNAc2-PP-Dol mannosyltransferase enzyme of P. pastoris[J]. Glycobiology, 2004, 14(5): 399-407. DOI:10.1093/glycob/cwh023.
[10] LEES J A, REINISCH K M. Inter-organelle lipid transfer: a channel model for Vps13 and chorein-N motif proteins[J]. Curr Opin Cell Biol, 2020, 65: 66-71. DOI:10.1016/j.ceb.2020.02.008.
[11] KUMAR N, LEONZINO M, HANCOCK-CERUTTI W, et al. VPS13A and VPS13C are lipid transport proteins differentially localized at ER contact sites[J]. J Cell Biol, 2018, 217(10): 3625-3639. DOI:10.1083/jcb.201807019.
[12] LI P Q, LEES J A, LUSK C P, et al. Cryo-EM reconstruction of a VPS13 fragment reveals a long groove to channel lipids between membranes[J]. J Cell Biol, 2020, 219(5): e202001161. DOI:10.1083/jcb.202001161.
[13] PARK J S, THORSNESS M K, POLICASTRO R, et al. Yeast Vps13 promotes mitochondrial function and is localized at membrane contact sites[J]. Mol Biol Cell, 2016, 27(15): 2435-2449. DOI:10.1091/mbc.e16-02-0112.
[14] LANG A B, JOHN PETER A T, WALTER P, et al. ER-mitochondrial junctions can be bypassed by dominant mutations in the endosomal protein Vps13[J]. J Cell Biol, 2015, 210(6): 883-890. DOI:10.1083/jcb.201502105.
[15] JOHN PETER A T, HERRMANN B, ANTUNES D, et al. Vps13-Mcp1 interact at vacuole-mitochondria interfaces and bypass ER-mitochondria contact sites[J]. J Cell Biol, 2017, 216(10): 3219-3229. DOI:10.1083/jcb.201610055.
[16] PARK J S, NEIMAN A M. VPS13 regulates membrane morphogenesis during sporulation in Saccharomyces cerevisiae[J]. J Cell Sci, 2012, 125(pt 12): 3004-3011. DOI:10.1242/jcs.105114.
[17] ARGUNHAN B, SAKAKURA M, AFSHAR N, et al. Cooperative interactions facilitate stimulation of Rad51 by the Swi5-Sfr1 auxiliary factor complex [J]. Elife, 2020, 9: e52566. DOI: 10.7554/elife.52566 (
