Screening of Heat Tolerance Genes in Tomato (Solanum lycopersicum) Based on BSA-seq and Plant Hormone Combined Analysis
JIANG Yue-Chang1, DUAN Jin-Hu1, LI Yi-Feng1, MAO Xiu-Jie1,2,3,4,*, WANG Shuai1,2,4,*, ZHANG Ning1,2
1 College of Horticulture Science and Technology, Hebei Normal University of Science and Technology, Qinhuangdao 066004, China; 2 Key Laboratory of Specialty Horticulture Germplasm Mining and Innovative Utilization of Hebei Province, Qinhuangdao 066004, China; 3 Yanshan Agricultural Characteristic Industry Technology Research Institute of Hebei Province, Qinhuangdao 066004, China; 4 Hebei Higher Institute Application Technology Research and Development Center of Horticultural Plant Biological Breeding, Hebei Normal University of Science & Technology, Qinhuangdao 066004, China
Abstract：High temperatures affect the physiology and hormone metabolism of tomatoes (Solanum lycopersicum). Localization of genes involved in heat tolerance in tomato can provide theoretical and technical support for subsequent gene cloning and functional analysis. In this study, electrical conductivity was used as the identification index, and 2 tomato strains at seedling stage were identified by high temperature and heat resistance, and the F2 generation isolated population formed by heat-resistant strain CH and heat-sensitive strain DH and their F1 generation were used as materials to detect plant hormones in parents CH and DH. The F2 generation isolated population was resequenced by bulked sergeant analysis (BSA), and candidate genes related to tomato heat resistance were screened through the combined analysis. The results showed that 4 metabolic pathways related to abiotic stress were obtained by phytohormone differential metabolite screening and KEGG annotation, namely glycine, serine and threonine metabolic pathways, amino acid biosynthetic pathway, tryptophan metabolic pathway and aminoacyl-tRNA biosynthetic pathway; 6.32 Mb size candidate interval was obtained on chromosome 3 by BSA-seq sequencing. which included 752 candidate genes. Combined with the differential metabolic pathways, 5 candidate genes associated with heat tolerance in tomato were finally screened within the candidate regional metabolic pathways, namely: D-glycerate 3-kinase (DG3K), Hydroxypyruvate reductase (HYDR), prolyl-tRNA synthetase (PARS2) and ribulose phosphate 3-epimerase (RPE), aldehyde dehydrogenase (ALDH). The expression of the candidate genes was analysed by qPCR, the expression of candidate genes DG3K, PARS2, and RPE increased extremely significant after 3 hours of high temperature stress (P<0.01), it was hypothesized that these 3 candidate genes might play an important role in the regulation of heat tolerance in tomato. This study has important implications for molecular assisted selection of heat-resistant tomato lines.
[1] 曹云英. 2009. 高温对水稻产量和品质的影响及其生理机制[D]. 博士学位论文, 扬州大学, 导师: 杨建昌, pp. 73-82. (Cao Y Y.2009. Effect of high temperature on yield and quality of rice and its physiological mechanism[D]. Thesis for Ph.D., Yangzhou University, Suppervisor: Yang J C, pp. 73-82.) [2] 陈雷. 2021. 水稻花期耐热性与耐热QTL定位及候选基因分析[D]. 博士学位论文, 广西大学, 导师: 江立庚, pp. 45-53. (Chen L.Mapping and candidate gene analysis of QTL for heat tolerance and heat resistance in flowering rice[D]. Thesis for Ph.D., Guangxi University, Suppervisor: Jiang L G, pp. 45-53.) [3] 高丽红, 李式军. 1998. 高温胁迫对蔬菜作物的影响[J]. 园艺学进展, (2): 344-349. (Gao L H, Li S J. 1998. Effects of high temperature stress on vegetable crops[J]. Advances in Horticulture, (2): 344-349.) [4] 李锡香, 杜永臣. 2006. 番茄种质资源描述规范和数据标准[M]. 中国农业出版社, 北京, pp. 52-54. (Li X X, Du Y C.2006. Description and Data Standard of Tomato Germplasm Resources[M]. China Agriculture Press, Beijing, pp. 52-54.) [5] 刘梦雨, 刘小丰, 边东, 等. 2019. 利用重测序-BSA分析鉴定金柑油胞的发育相关基因[J]. 园艺学报, 46(5): 841-854. (Liu M Y, Liu X F, Bian D.2019. The genes related to oil cell development of Kumquat were identified by resequencing and BSA analysis[J]. Journal of Horticulture, 46(5): 841-854.) [6] 毛胜利, 杜永臣, 王孝宣. 2001. 番茄耐热育种研究进展[J]. 园艺学报, 28(增刊): 655-660. (Mao S L, Du Y C, Wang X X.2001. Advances in heat tolerance breeding of tomato[J]. Chinese Journal of Horticulture, 28(Supplement): 655-660.) [7] 潘光辉, 王文强, 尹贵贤, 等. 2001. 电导法鉴定番茄耐热性与田间结果比较[J]. 西南园艺, 29(2): 26-27. (Pan G H, Wang W Q, Yin G X, et al.2001. Determination of tomato heat resistance by conductivity method and comparison of field results[J]. South West Horticulture, 29(2): 26-27.) [8] 沈火林, 刘燕燕, 余进安. 2006. 不结球白菜耐高温胁迫与内源激素含量的关系[J]. 江苏农业科学, (6): 207-210. (Shen H L, Liu Y Y, Yu J A. 2006. Relationship between high temperature tolerance and endogenous hormone content in non-heading Chinese cabbage[J]. Jiangsu Agricultural Sciences, (6): 207-210.) [9] 王冬梅, 许向阳, 李景富. 2003. 番茄耐热性研究进展[J]. 中国蔬菜, (2): 58-60. (Wang D M, Xu X Y, Li J F. 2003. Research progress on heat resistance of tomato[J]. Chinese Vegetables, (2): 58-60.) [10] 王学奎. 2006. 植物生理生化实验原理和技术[M]. 高等教育出版社, 北京, pp. 120-122. (Wang X K.2006. Principles and techniques of plant physiological and biochemical experiments[M]. Higher Education Pressr, Beijing, pp. 120-122.) [11] 张洁, 李天来, 徐晶. 2004. 不同时期昼间亚高温处理对日光温室番茄生长发育的影响[J]. 华中农业大学学报, (Z2): 230-235. (Zhang J, Li T L, Xu J. 2004. Effects of diurnal sub-high temperature treatment at different periods on the growth and development of tomato in solar greenhouse[J]. Journal of Huazhong Agricultural University, (Z2): 230-235.) [12] 张娜, 2020. 番茄封顶花序数遗传规律及BSA分析[D]. 硕士学位论文, 河北科技师范学院, 导师: 毛秀杰, pp. 33-43. (Zhang N.2020. Genetic pattern of capped inflorescence number in tomato and BSA analysis[D]. Thesis for M.S., Hebei Science and Technology Teachers College, Suppervisor: Mao X J, pp. 33-43.) [13] 赵海燕. 2017. 基于BSA技术的水稻(Oryza sativa L.)粒长基因初步定位及候选基因分析[D]. 硕士学位论文, 华南农业大学,导师: 王兰, pp. 22-36. (Zhao H Y.2017. Preliminary mapping and candidate gene analysis of grain length genes in rice (Oryza sativa L.) based on BSA technology[D]. Thesis for M.S., South China Agricultural University, Suppervisor: Wang L, pp. 22-36.) [14] Colebrook E H, Thomas S G, Phillips A L, et al.2014. The role of gibberellin signaling in plant responses to abiotic stress[J]. Journal of Experimental Biology, 217(1): 67-75. [15] Gray W M, Ostin A, Sandberg G, et al.1998. High temperature promotes auxin-mediated hypocotyl elongation in Arabidopsis[J]. Proceedings of the National Academy of Sciences of the USA, 95(12): 7197-7202. [16] Han Y, Fan S, Zhang Q, et al.2013. Effect of heat stress on the MDA, proline and soluble sugar content in leaf lettuce seedlings[J]. Agricultural Sciences, 4(5): 112-115. [17] Hill J T, Demarest B L, Bisgrove B W, et al.2013. MMAPPR: Mutation mapping analysis pipeline for pooled RNA-seq[J]. Genome Research, 23(4): 687-697. [18] Kanehisa M, Goto S.2000. KEGG: Kyoto encyclopedia of genes and genomes[J]. Nucleic Acids Research, 28(1):27-30. [19] Michelmore R W, Paran I, Kesseli R V.1991. Identification of markers linked to disease-resistance genes by bulked segregant analysis: A rapid method to detect markers in specific genomic regions by using segregating populations[J]. Proceedings of the National Academy of Sciences of the USA, 88(21): 9828-9832. [20] Paliyath G, Fletcher R A.1995. Paclobutrazol treatment alters peroxidase and catalase activities in heat-stressed maize coleoptiles[J]. Physiology and Molecular Biology of Plants, 1(1): 171-178. [21] Popko J, Hansch R, MendelL R R, et al.2010. The role of abscisic acid and auxin in the response of poplar to abiotic stress[J]. Plant Biology, 12(2): 242-258. [22] Skubacz A, Daszkowska G A, Szarejko I.2016. The role and regulation of ABI5 (ABA-Insensitive 5) in plant development, abiotic stress responses and phytohormone crosstalk[J]. Frontiers in Plant Science, 7(16): 1884-1901. [23] Stainforth D A, Aina T, Christensen C, et al.2005. Uncertainty in predictions of the climate response to rising levels of greenhouse gases.[J]. Nature, 433(27): 403-406. [24] Tian J Y, Ma Y, Chen Y B, et,al.2022. Plant hormone response to low-temperature stress in cold-tolerant and cold-sensitive varieties of Zanthoxylum bungeanum Maxim[J]. Frontiers in Plant Science, 13(4): 1-18. [25] Wit M D, Lorrain S, Fankhauser C.2014. Auxin-mediated plant architectural changes in response to shade and high temperature[J]. Physiol Plantarum, 151(1): 13-24. [26] Zhao C, Liu B, Piao S, et al.2017. Temperature increase reduces global yields of major crops in four inde-pendent estimates[J]. Proceedings of the National Academy of Sciences of USA, 114(35): 9326-9331. [27] Zhou Y G, Liu W C, Li X W, et,al.2020. Integration of sRNA, degradome,transcriptome analysis and functional investigation reveals gma-miR398c negatively regulates drought tolerance via GmCSDs and GmCCS in transgenic Arabidopsis and soybean[J]. BMC Plant Biology, 20(1): 190. [28] Zou C, Wang P X, Xu Y B.2016. Bulked sample analysis in genetics,genomics and crop improvement[J]. Plant Biotechnol, 14(1): 1-15.
