Abstract:As the direct receptor of abscisic acid (ABA), Pyrabactin resistance 1-like (PYL) protein plays an important role in plant growth, development and stress. In order to explore the function of PYL gene family in strawberry, this study identified PYL gene family members in woodland strawberry (Fragaria vesca) and pineapple strawberry (F. ananassa) using bioinformatic methods, and analyzed the expression of PYL gene under different simulated stress conditions by qPCR. The results showed that there were 16 FvPYL genes distributed on 7 chromosomes in woodland strawberry and 40 FaPYL genes distributed on 18 chromosomes in pineapple strawberry. The gene size span of FaPYL gene family was larger than that of FvPYL. Their protein structure was similar and mainly contained α-helix and random coil. In addition, except FvPYL15 and FaPYL40, the other proteins were hydrophilic and mainly expressed in chloroplast, cytoplasm and nucleus. Phylogenetic analysis showed that FaPYL and FvPYL genes were distributed in 5 subfamilies, and the gene structure and motif position of the same species in the same subtribe were basically the same. There were only 3 pairs of segmental duplications in FvPYL genes, but a large number of segmental duplications and some tandem repeats in FaPYL genes. Selection pressure analysis of genes found that strawberry PYL genes were biased toward purification selection. Cis-acting elements analysis found that PYL family genes mainly contained elements related to light, hormone, stress, growth and development. As detected by qPCR, the relative expression levels of strawberry PYL genes were significantly different under different treatments. Under ABA treatment, FaPYL and FvPYL were up-regulated except FvPYL8. Under NaCl treatment, most FaPYL genes were up-regulated to varying degrees, among which FaPYL21 and FaPYL40 were strongly induced by 200 μmol/L NaCl. FvPYL genes expression were overall upregulated, with the relative expression of FvPYL11 was 60 times higher than that of the control. Under PEG treatment, FvPYL genes were positively regulated, FvPYL1 and FvPYL7 were significantly up-regulated. Compared with woodland strawberry, the upregulation range of FaPYL genes were significantly different, among which the relative up-regulation levels of FaPYL2 and FaPYL7 were extremely significant. This study provides a theoretical basis for functional mining and application research of PYL gene family in strawberry.
郭丽丽, 卢世雄, 乃国洁, 马维峰, 毛娟. 草莓PYL基因家族的鉴定与非生物胁迫下的表达分析[J]. 农业生物技术学报, 2022, 30(12): 2315-2332.
GUO Li-Li, LU Shi-Xiong, NAI Guo-Jie, MA Wei-Feng, MAO Juan. Identification of PYL Gene Family and Expression Analysis Under Abiotic Stress in Strawberry. 农业生物技术学报, 2022, 30(12): 2315-2332.
[1] 丁冰杰, 孔祥强, 董合忠. 2020. 脱落酸受体PYLs的结构与功能研究进展[J]. 分子植物育种, 18(20): 6844-6852. (Ding B J, Kong X Q, Dong H Z.2020. Research progress on the structure and function of abscisic acid receptor PYLs[J]. Molecular Plant Breeding, 18(20): 6844-6852.) [2] 郭贵华, 刘海艳, 李刚华, 等. 2014. ABA缓解水稻孕穗期干旱胁迫生理特性的分析[J]. 中国农业科学, 47(22): 4380-4391. (Guo G H, Liu H Y, Li G H, et al.2014. Analysis of physiological characteristics about ABA alleviating rice booting stage drought stress[J]. Scientia Agricultura Sinica, 47(22): 4380-4391.) [3] 胡帅, 王芳展, 刘振宁, 等. 2012. PYR/PYL/RCAR蛋白介导植物ABA的信号转导[J]. 遗传, 34(05): 560-572. (Hu S, Wang F Z, Liu Z N, et al.2012. ABA signaling mediated by PYR/PYL/RCAR in plants[J]. Hereditas (Beijing), 34(5): 560-572.) [4] 黄思源, 呼天明, 杨培志. 2019. 蒺藜苜蓿PYL基因家族的全基因组鉴定、表达和功能分析[J]. 草业科学, 36(02): 422-433. (Huang S Y, Hu T M, Yang P Z.2019. Identification and function analysis of the PYL gene family in Medicago truncatula[J]. Pratacultural Science, 36(02): 422-433.) [5] 金龙飞, 尹欣幸, 曹红星. 2021. 油棕脱落酸受体PYL基因家族的全基因组鉴定及表达分析[J]. 南方农业学报, 52(06): 1545-1556. (Jin L F, Yin X X, Cao H X.2021. Genome-wide identification and expression analysis of abscisic acid receptor PYL gene family in oil palm[J]. Journal of Southern Agriculture, 52(06): 1545-1556.) [6] 李鸿杰, 王盈阁, 张夏伟, 等. 2015. 玉米脱落酸受体基因家族的生物信息学分析[J]. 核农学报, 29(09): 1657-1667. (Li H J, Wang Y G, Zhang X W, et al.2015. Bioinformatic analysis for abscisic acid aerceptor gene family in maize[J]. Journal of Nuclear Agricultural Sciences, 29(09): 1657-1667.) [7] 刘海洋, 陈玉珍. 2019. ABA受体结构及功能与ABA信号通路研究进展[J]. 中国农学通报, 35(21): 75-81. (Liu H Y, Chen Y Z.2019. Abscisic acid receptor structure and function and signaling pathway:Researchadvances[J]. Chinese Agricultural Science Bulletin, 35(21): 75-81.) [8] 刘丽. 2021. 葡萄抗霜霉病相关基因挖掘及VvPYL4功能分析[D]. 博士学位论文, 沈阳农业大学, 导师: 刘长远, pp. 92-97. (Liu Li.2021. Gene mining and functional analysis of resistance gene VvPYL4 from grapevine resistant to Plasmopara viticola[D]. Thises for ph.D., Agricultural University Of Shenyang, Supervisor: Liu C Y, pp. 92-97.) [9] 马宗桓, 陈佰鸿, 李文芳, 等. 2018. 葡萄PYL基因家族的鉴定与表达分析[J]. 果树学报, 35(03): 265-274. (Ma Z H, Chen B H, Li W F, et al.2018. ldentification and expression analysis of PYL gene families in grape[J]. Journal of Fruit Science, 35(03): 265-274.) [10] 彭晶晶, 张圣也, 阮美丽, 等. 2020. 高粱PYL基因家族进化及表达分析[C]. 中国作物学会, 第十九届中国作物学会学术年会论文摘要集. pp. 142. (Peng J J, Zhang S Y, Ruan M L, et al.2020. Evolution and expression analysis of sorghum PYL gene family[C]. Chinese Crop Society. Proceedings of the Nineteenth Annual Conference of the Chinese Crop Society. pp. 142.) [11] 史梦梦. 2020. 小麦TaPYL基因家族的分析及功能鉴定[D]. 山西大学, 导师: 郑军, 郑兴卫, pp. 13-15. (Shi M M.2020. Analysis and functional identification of TaPYL gene family in wheat[D]. Shanxi University, Suppervisor: Zheng J, Zheng X W, pp. 13-15.) [12] 王帅磊, 李子琪, 陈辉龙, 等. 2020. 谷子PYR/PYL/RCAR基因家族进化及表达分析[J]. 分子植物育种, 18(17): 5544-5554. (Wang S L, Li Z Q, Chen H L, et al.2020. Evolution and expression analysis of the PYR/PYL/RCAR gene family in Setariaitalica[J]. Molecular Plant Breeding, 18(17): 5544-5554.) [13] 王振宇, 杨光, 赛娜, 等. 2022. 野生二粒小麦PYL基因家族的鉴定及其在逆境胁迫下的表达特性分析[J]. 麦类作物学报, 42(01): 1-10. (Wang Z Y, Yang G, Sai N, et al.2022. ldentification and expression analysis of PYL gene family in wild emmer wheat (Triticum dicoccoides L.) under abiotic stress[J]. Journal of Triticeae Crops, 42(01): 1-10.) [14] 夏云, 颜渊. 2016. 正选择位点及其计算软件研究进展[J]. 长江大学学报(自科版), 13(27): 51-53. (Xia Y, Yan Y.2016. Advances in positive selection sites and their computational software[J]. Journal of Yangtze University (Natural Science Edition), 13(27): 51-53.) [15] 颜志明, 王全智, 冯英娜, 等. 2015. FaPYL9基因调控草莓果实成熟的分子机理[J]. 西北植物学报, 35(12): 2379-2384. (Yan Z M, Wang Q Z, Feng Y N, et al.2015 Molecular mechanism of FaPYL9 gene regulation in strawberry fruit development[J]. Acta Botanica Boreali-Occidentalia Sinica, 35(12): 2379-2384.) [16] 张建, 黄芸. 2021. 草莓FaPYL5基因的鉴定及功能分析[J]. 农业与技术, 41(03): 8-12. (Zhang J, Huang Y.2021. Identification and functional analysis of strawberry FaPYL5 gene[J]. Agriculture and Technology, 41(03): 8-12.) [17] 张欣馨, 王菲, 李浪, 等. 2016. 中国草莓生产中面临的主要问题及发展对策[J]. 中国林副特产, 02: 92-96. (Zhang X X, Wang F, Li L, et al.2016. Main problems and development countermeasures during the strawberry production in China[J]. Forest By-Product and Speciality in China, 02: 92-96.) [18] 张兴政, 赵豫川, 陈玥, 等. 2021. 平榛PYL基因家族全基因组鉴定及果实发育表达分析[J]. 江西农业大学学报, 43(02): 435-444. (Zhang X Z, Zhao Y C, Chen Y, et al.2021. Genome-wide identification and expression analysis of PYL gene family in Corylus heterophylla during fruit development[J]. Acta Agriculturae Universitatis Jiangxiensis, 43(02): 435-444.) [19] 张兆涵, 张天旭, 王万鹏, 等. 2020. 大豆脱落酸受体基因家族鉴定、系统发育进化及表达模式分析[J]. 南方农业学报, 51(08): 1904-1916. (Zhang Z H, Zhang T X, Wang W P, et al.2020. ldentification,phylogenetic evolution and expression analysis of abscisic acid receptors gene family in Glycine max L. Merr[J]. Journal of Southern Agriculture, 51(08): 1904-1916.) [20] Adie B A T, Pérez-Pérez J, Pérez-Pérez M M, et al.2007. ABA is an essential signal for plant resistance to pathogens affecting JA biosynthesis and the activation of defenses in Arabidopsis[J]. The Plant Cell, 19(5): 1665-1681. [21] Chen Y, Feng L, Wei N, et al.2017a. Overexpression of cotton PYL genes in Arabidopsis enhances the transgenic plant tolerance to drought stress[J]. Plant Physiology and Biochemistry, 115: 229-238. [22] Chen Z Q, Kong L, Zhou Y, et al.2017b. Endosperm-specific OsPYL8 and OsPYL9 act as positive regulators of the ABA signaling pathway in rice seed germination[J]. Functional Plant Biology: FPB, 44(6): 635-645. [23] Dalal M, Inupakutika M.2014. Transcriptional regulation of ABA core signaling component genes in sorghum (Sorghum bicolor L. Moench)[J]. Molecular Breeding, 34(3): 1517-1525. [24] Fujii H, Chinnusamy V, Rodrigues A, et al.2009. In vitro reconstitution of an abscisic acid signalling pathway[J]. Nature, 462(7273): 660-664. [25] González-Guzmán M, Rodríguez L, Lorenzo-Orts L, et al.2014. Tomato PYR/PYL/RCAR abscisic acid receptors show high expression in root, differential sensitivity to the abscisic acid agonist quinabactin, and the capability to enhance plant drought resistance[J]. Journal of Experimental Botany, 15: 4451-4464. [26] Guo D, Zhou Y, Li H L, et al.2017. Identification and characterization of the abscisic acid (ABA) receptor gene family and its expression in response to hormones in the rubber tree[J]. Scientific Reports, 7: 45157. [27] Hadiarto T, Ran L S P.2011. Progress studies of drought-responsive genes in rice[J]. Plant Cell Reports, 30(3): 297-310. [28] He Y, Hao Q, Li W Q, et al.2014. Identification and characterization of ABA receptors in Oryza sativa[J]. PLOS ONE, 9(4): e95246. [29] He Z, Zhong J, Sun X,et al.2018. The maize ABA receptors ZmPYL8, 9, and 12 facilitate plant drought resistance[J]. Frontiers in Plant Science, 9: 422. [30] Hu W, Yan Y, Shi H T, et al.2017. The core regulatory network of the abscisic acid pathway in banana: Genome-wide identification and expression analyses during development, ripening, and abiotic stress[J]. BMC Plant Biology, 17(1): 145. [31] Iyer L M, Koonin E V, Aravind L.2001. Adaptations of the helix-grip fold for ligand binding and catalysis in the START domain superfamily[J]. Proteins: Structure, Function, and Bioinformatics, 43(2): 134-144. [32] Lee S C, Luan S.2012. ABA signal transduction at the crossroad of biotic and abiotic stress responses[J]. Plant Cell Environment, 35(1): 53-60. [33] Kim H, Lee K, Hwang H,et al.2014. Overexpression of PYL5 in rice enhances drought tolerance, inhibits growth, and modulates gene expression[J]. Journal of Experimental Botany, 65(2): 453-464. [34] Ma Y, Izabela S, Arthur K, et al.2009. Regulators of PP2C phosphatase activity function as abscisic acid sensors[J]. Science, 324(5930): 1064-1068. [35] Mega R, Abe F, Kim J S, et al.2019. Tuning water-use efficiency and drought tolerance in wheat using abscisic acid receptors[J]. Nature Plants, 5(2): 153-159. [36] Park S Y, Pauline F, Noriyuki N, et al.2009. Abscisic acid inhibits type 2C protein phosphatases via the PYR/PYL family of START proteins[J]. Science, 324(5930): 1068-1071. [37] Santiago J, Rodrigues A, Saez A, et al.2009. Modulation of drought resistance by the abscisic acid receptor PYL5 through inhibition of clade APP2Cs[J]. Plant Journal, 60(4): 575-588. [38] Wang Y P, Wang Y, Kai W B, et al.2014. Transcriptional regulation of abscisic acid signal core components during cucumber seed germination and under Cu2+, Zn2+, NaCl and simulated acid rain stresses[J]. Plant Physiology and Biochemistry, 76: 67-76. [39] Xing L, Zhao Y, Gao J H, et al.2016. The ABA receptor PYL9 together with PYL8 plays an important role in regulating lateral root growth[J]. Scientific Reports, 6(1): 27177. [40] Yin P, Fan H, Hao Q, et al.2009. Structural insights into the mechanism of abscisic acid signaling by PYL proteins[J]. Nature Structural & Molecular Biology, 16(12): 1230-1236. [41] Yu J L, Ge H M, Wang X K, T, et al.2017. Overexpression of pyrabactin resistance-like abscisic acid receptors enhances drought, osmotic, and cold tolerance in transgenic poplars[J]. Frontiers in Plant Science, 8: 1752.