Initial Study on the Response of Potato (Solanum tuberosum) StCYS6 Gene to Salt Stress and Phytophthora infestans Infection
ZHAO Ding1,2, WANG Lu-Yao1,2,4, SHU Hai-Dong1,2, LI Zi-Hao1,2,3,*, DONG Suo-Meng1,2,*
1 College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China; 2 The Key Laboratory of Integrated Management of Crop Diseases and Pests (Ministry of Education), Nanjing Agricultural University, Nanjing 210095, China; 3 School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China; 4 Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518120, China
Abstract：Breeding salt tolerance and late blight-resistant potatoes (Solanum tuberosum) is an important requirement for the development of the potato industry. In this study, key genes responding to salt stress and the invasion of the late blight pathogen Phytophthora infestans were screened by transcriptome sequencing. The results showed that the infection area of P. infestans on the leaves significantly increased after being treated with 100 mmol/L NaCl for 1 week, namely, salt stress significantly weakened the resistance of tissue-cultured potato to P. infestans. Transcriptome sequencing revealed that the expression of 3 780 potato genes changed significantly under salt stress, in which the cystatin 6 (CYS6) gene significantly downregulated. qRT-PCR results further confirmed that StCYS6 gene expression was downregulated in potato leaves after salt stress, but upregulated after P. infestans invasion. Transient expression of the candidate gene StCYS6 in Nicotiana benthamiana leaves significantly enhanced resistance to P. infestans. Inactivation mutations showed that StCYS6 protein promoted plant resistance to P. infestans independent of its cysteine protease inhibitor activity. The analysis of salt tolerance and pathogen resistance of StCYS6 transgenic potato plants showed that the degree of wilting of potato leaves was significantly reduced, indicating that StCYS6 could significantly improve the salt tolerance of potatoes; However, StCYS6 transgenic potato plants did not significantly improve their resistance to P. infestans. This study obtained a candidate gene StCYS6 that could enhance the salt tolerance of potatoes, providing basic data for cultivating new varieties of stress resistant potatoes.
赵鼎, 王路遥, 舒海东, 李子豪, 董莎萌. 马铃薯StCYS6基因响应盐胁迫和致病疫霉侵染的初步研究[J]. 农业生物技术学报, 2024, 32(6): 1236-1249.
ZHAO Ding, WANG Lu-Yao, SHU Hai-Dong, LI Zi-Hao, DONG Suo-Meng. Initial Study on the Response of Potato (Solanum tuberosum) StCYS6 Gene to Salt Stress and Phytophthora infestans Infection. 农业生物技术学报, 2024, 32(6): 1236-1249.
[1] 黄晶, 孔亚丽, 徐青, 等. 2022. 盐渍土壤特征及改良措施研究进展[J]. 土壤, 54(01): 18-23. (Huang J, Kong Y L, Xu Q, et al.2022. Progresses for characteristics and amelioration measures of saline soil[J]. Soils, 54(01): 18-23.) [2] 李建国, 濮励杰, 韩明芳, 等. 2014. 中国土壤盐渍化研究现状及未来研究热点[J]. 地理学报, 24(05): 943-960. (Li J G, Pu L J, Han M F, et al.2014. The present situation and hot issues in the salt-affected soil research[J]. Acta Geographica Sinica, 24(05): 943-960.) [3] 李青, 秦玉芝, 胡新喜, 等. 2017. 马铃薯耐盐性研究进展[J]. 园艺学报, 44(12): 2408-2424. (Li Q, Qing Y Z, Hu X X, et al.2017. Advances in the research on salt tolerance of potato[J]. Acta Horticulturae Sinica, 44(12): 2408-2424.) [4] 卢珍红, 武欢, 焦元辰, 等. 2022. 非洲菊CAT2基因的克隆及表达[J]. 福建农林大学学报(自然科学版), 51(06): 774-781. (Lu Z H, Wu H, Jiao Y C, et al.2022. Cloning and expression of CAT2 gene in Gerbera jamesonii[J]. Journal of Fujian Agriculture and Forestry University (Nutural Science Edition), 51(06): 774-781.) [5] 徐进, 朱杰华, 杨艳丽, 等. 2019. 中国马铃薯病虫害发生情况与农药使用现状[J]. 中国农业科学, 52(16): 2800-2808. (Xu J, Zhu J H, Yang Y L, et al.2019. Status of major diseases and insect pests of potato and pesticide usage in China[J]. Scientia Agricultura Sinica, 52(16): 2800-2808.) [6] Ahmad R, Zuily-Fodil Y, Passaquet C, et al.2015. Molecular cloning, characterization and differential expression of novel phytocystatin gene during tropospheric ozone stress in maize (Zea mays) leaves[J]. Comptes Rendus Biologies, 338(3): 141-148. [7] Basu R, Dutta S, Pal A, et al.2021. Calmodulin7: Recent insights into emerging roles in plant development and stress[J]. Plant Molecular Biology, 107(1-2): 1-20. [8] Benchabane M, Schlüter U, Vorster J, et al.2014. Plant cystatins[J]. Biochimie, 92(11): 1657-1666. [9] Cheng M L, Tzen J T, Shyu D J, et al.2014. Functional characterization of the N-terminal and C-terminal domains of a sesame group II phytocystatin[J]. Botanical Studies, 55(1): 1-10. [10] Christova P K, Christov N K, Imai R J P.2006. A cold inducible multidomain cystatin from winter wheat inhibits growth of the snow mold fungus, Microdochium nivale[J]. Planta, 223(6): 1207-1218. [11] Cramer G R, Ergül A, Grimplet J, et al.2007. Water and salinity stress in grapevines: Early and late changes in transcript and metabolite profiles[J]. Functional & Integrative Genomics, 7(2): 111-134. [12] Gaddour K, Vicente-Carbajosa J, Lara P, et al.2001. A constitutive cystatin-encoding gene from barley (Icy) responds differentially to abiotic stimuli[J]. Plant Molecular Biology, 45(5): 599-608. [13] Guo K, Bu Y, Takano T, et al.2013. Arabidopsis cysteine proteinase inhibitor AtCYSb interacts with a Ca2+-dependent nuclease, AtCaN2[J]. FEBS Letters, 587(21): 3417-3421. [14] Grajek H, Rydzyński D, Piotrowicz-Cieślak A, et al.2020. Cadmium ion-chlorophyll interaction-examination of spectral properties and structure of the cadmium-chlorophyll complex and their relevance to photosynthesis inhibition[J]. Chemosphere, 261: 127434. [15] Haas B J, Kamoun S, Zody M C, et al.2009. Genome sequence and analysis of the Irish potato famine pathogen Phytophthora infestans[J]. Nature, 461(7262): 393-398. [16] Hwang J E, Hong J K, Je J H, et al.2009. Regulation of seed germination and seedling growth by an Arabidopsis phytocystatin isoform, AtCYS6[J]. Plant Cell Reports, 28(11): 1623-1632. [17] Imbo M C, Budambula N, Mweu C M, et al.2016. Genetic transformation of sweet potato for improved tolerance to stress: A review[J]. Advances in Life Science and Technology, 49: 67-76. [18] Judelson H S, Randall T A.1998. Families of repeated DNA in the oomycete Phytophthora infestans and their distribution within the genus[J]. Genome, 41(4): 605-615. [19] Kamle M, Borah R, Bora H, et al.2020. Systemic acquired resistance (SAR) and induced systemic resistance (ISR): Role and mechanism of action against phytopathogens. In: Hesham A L, Upadhyay R, Sharma G, Manoharachary C, Gupta V, eds. Fungal Biotechnology and Bioengineering. Fungal Biology[M]. Springer Nature, Switzerland, pp. 457-470. [20] Litalien A, Zeeb B.2020. Curing the earth: A review of anthropogenic soil salinization and plant-based strategies for sustainable mitigation[J]. Science of the Total Environment, 698: 134235. [21] Liu L, Wang B.2021. Protection of halophytes and their uses for cultivation of saline-alkali soil in China[J]. Biology, 10(5): 353. [22] Martínez M, Abraham Z, Carbonero P, et al.2005. Comparative phylogenetic analysis of cystatin gene families from Arabidopsis, rice and barley[J]. Molecular Genetics and Genomics, 273(5): 423-432. [23] Munns R, Tester M.2008. Mechanisms of salinity tolerance[J]. Annual Review of Plant Biology, 59: 651-681. [24] Nguyen X C, Kim S H, Lee K, et al.2012. Identification of a C2H2-type zinc finger transcription factor (ZAT10) from Arabidopsis as a substrate of MAP kinase[J]. Plant Cell Reports, 31(4): 737-745. [25] Pinheiro H A, Damatta F M, Chaves A R, et al.2005. Drought tolerance is associated with rooting depth and stomatal control of water use in clones of Coffea canephora[J]. Annals of Botany, 96(1): 101-108. [26] Popovic M, Andjelkovic U, Burazer L, et al.2013. Biochemical and immunological characterization of a recombinantly-produced antifungal cysteine proteinase inhibitor from green kiwifruit (Actinidia deliciosa)[J]. Phytochemistry, 94: 53-59. [27] Quain M D, Makgopa M E, Cooper J W, et al.2015. Ectopic phytocystatin expression increases nodule numbers and influences the responses of soybean (Glycine max) to nitrogen deficiency[J]. Phytochemistry, 112: 179-187. [28] Quan R, Lin H, Mendoza I, et al.2007. SCABP8/CBL10, a putative calcium sensor, interacts with the protein kinase SOS2 to protect Arabidopsis shoots from salt stress[J]. The Plant Cell, 19(4): 1415-1431. [29] Sun X, Yang S, Sun M, et al.2014. A novel Glycine soja cysteine proteinase inhibitor GsCPI14, interacting with the calcium/calmodulin-binding receptor-like kinase GsCBRLK, regulated plant tolerance to alkali stress[J]. Plant Molecular Biology, 85(1-2): 33-48. [30] Suzuki N, Rivero R M, Shulaev V, et al.2014. Abiotic and biotic stress combinations[J]. New Phytologist, 203(1): 32-43. [31] Van der Hoorn R A L.2008. Plant proteases: From phenotypes to molecular mechanisms[J]. Annual Review of Plant Biology, 59: 191-223. [32] Wang Y, Zhan Y, Wu C, et al.2012. Cloning of a cystatin gene from sugar beet M14 that can enhance plant salt tolerance[J]. Plant Science, 191: 93-99. [33] Zhang X X, Liu S K, Takano T.2008. Two cysteine proteinase inhibitors from Arabidopsis thaliana, AtCYSa and AtCYSb, increasing the salt, drought, oxidation and cold tolerance[J]. Plant Molecular Biology, 68(1-2): 131-143.
