Selenium (Se) is an essential microelement for human, and Se enriched products are important sources for Se intake in human. Tea plants (Camellia sinensis) have strong selenium enrichment ability. However, there is limited research on the molecular mechanism of phosphate transporters involved in Se absorption. In this study, CsPHT1;3 gene was cloned and its characteristics and responses to Se concentrations, valence, pH, time and expression in various Se-enriched tea resources were investigated. Gene characteristic analysis of CsPHT1;3 shows that CsPHT1;3 is grouped into phosphate transporter PHT1 subfamily and localized in the plasma membrane. The CsPHT1;3 protein contains the conserved domain GGDYPLSATIxSE, which belongs to the PHT1 protein. Expression pattern analysis of CsPHT1;3 in various tissues suggests that the expression levels of CsPHT1;3 in mature leaves and root tissues were significantly higher than those in other tissues. The induction results of different Se concentrations and valence states indicate that CsPHT1;3 was significantly induced by Se4+ at 1 d and 7 d after treatments. The expression of CsPHT1;3 in roots was obviously induced by Se6+ except for 3 d after treatment but largely unaffected by Se6+ concentration. The results of different pH and Se4+ treatments show that, at pH5, the highest expression of CsPHT1;3 in tea roots was observed at 24 h. While at pH3, the highest expression of CsPHT1;3 in tea roots was observed at 48 h. Moreover, at pH7, the highest expression of CsPHT1;3 in tea roots was observed at 72 h. The results of sodium selenate treatment on different Se-enriched tea resources indicate that the expression of CsPHT1;3 in the leaves and roots did not respond to sodium selenate treatment. However, the results of sodium selenite treatment on different Se-enriched tea resources suggest that CsPHT1;3 is significantly up-regulated in the leaves of Se-enriched tea resources. The above studies indicated that CsPHT1;3 may participate in the absorption and redistribution of selenite by roots in tea plants, which is important for the breeding of Se-enriched tea cultivars.
[1] 王雪儿, 王玲, 杨欣宇, 等. 不同化学形态硒的毒理学研究[J]. 湖北民族学院学报(自然科学版), 2022, 40(2): 150-156.
Wang X E, Wang L, Yang X Y, et al.Toxicological studies on different chemical forms of selenium[J]. Journal of Hubei Minzu University (Natural Science Edition), 2022, 40(2): 150-156.
[2] Guignardi Z, Schiavon M.Biochemistry of plant selenium uptake and metabolism[M]//Pilon-Smits E A H, Winkel L H E, Lin Z Q. Selenium in plants: molecular, physiological, ecological and evolutionary aspects. Cham: Springer, 2017: 21-34.
[3] Dinh Q T, Cui Z W, Huang J, et al.Selenium distribution in the Chinese environment and its relationship with human health: a review[J]. Environment International,2018, 112: 294-309.
[4] 高柱, 蔡荟梅, 彭传燚, 等. 富硒茶叶中硒的赋存形态研究[J]. 中国食物与营养,2014, 20(1): 31-33.
Gao Z, Cai H M, Peng C Y, et al.Distribution rule and combined forms of selenium in selenium-enriched tea[J]. Food and Nutrition in China, 2014, 20(1): 31-33.
[5] Zhang H J, Hao X Y, Zhang J J, et al.Genome-wide identification of SULTR genes in tea plant and analysis of their expression in response to sulfur and selenium[J]. Protoplasma,2022, 259: 127-140.
[6] 张晶晶, 钱文俊, 郝心愿, 等. 茶树硫酸盐转运蛋白基因CsSULTR3.1的克隆及其对硫和硒的响应分析[J]. 园艺学报,2018, 45(2): 321-332.
Zhang J J, Qian W J, Hao X Y, et al.Cloning and expression analysis ofCsSULTR3.1implicated in sulfate and selenate treatments in tea plant(Camellia sinensis)[J]. Acta Horticulturae Sinica, 2018, 45(2): 321-332.
[7] Shrift A, Ulrich J M.Transport of selenate and selenite into astragalus roots[J]. 1969, 44: 893-896.
[8] Broyer T C, Johnson C M, Huston R P.Selenium and nutrition of astragalus. II. ionic sorption interactions among selenium, phosphate, and macronutrient and micronutrient cations[J]. Plant and Soil, 1972, 36: 651-669.
[9] Zhang L H, Hu B, Li W, et al.OsPT2, a phosphate transporter, is involved in the active uptake of selenite in rice[J]. New Phytologist, 2014, 201(4): 1183-1191.
[10] Song Z, Shao H, Huang H, et al.Overexpression of the phosphate transporter geneOsPT8improves the Pi and selenium contents inNicotiana tabacum[J]. Environmental and Experimental Botany, 2017, 137: 158-165.
[11] Chang M X, Gu M, Xia Y W, et al.OsPHT1;3mediates uptake, translocation and remobilization of phosphate under extremely low phosphate regimes[J]. Plant physiology, 2019, 179(2): 656-670.
[12] Ren H Z, Li X M, Guo L N, et al.Integrative transcriptome and proteome analysis reveals the absorption and metabolism of selenium in tea plants [Camellia sinensis(L.) O. Kuntze][J]. Frontiers in Plant Science, 2022, 13: 848349. doi: 10.3389/fpls.2022.848349.
[13] Cao D, Liu Y, Ma L, et al.Genome-wide identification and characterization of phosphate transporter gene family members in tea plants [Camellia sinensis(L.) O. Kuntze] under different selenite levels[J]. Plant Physiology and Biochemistry, 2021, 166: 668-676.
[14] 辛华洪. 茶树磷转运蛋白基因CsPT4、CsPT1克隆和表达及CsPT4功能分析[D]. 南京: 南京农业大学, 2017.
Xin H H.Molecular cloning and expression analysis ofCsPT4andCsPT1gene from tea plant (Camellia Sinensis), subcellular localization and function analysis ofCsPT4[D]. Nanjing: Nanjing Agricultural University, 2017.
[15] 郭丽娜, 王璐, 郝心愿, 等. 茶树根系吸收硒的生理特性研究[J]. 园艺学报, 2022, 49(9): 1967-1976.
Guo L N, Wang L, Hao X Y, et al.Physiological characteristics of selenium uptake in tea plant root[J]. Acta Horticulturae Sinica, 2022, 49(9): 1967-1976.
[16] Wang X C, Feng H, Chang Y X, et al.Population sequencing enhances understanding of tea plant evolution[J]. Nature Communications, 2020, 11(1): 4447. doi: 10.1038/s41467-020-18228-8.
[17] Yoo S D, Cho Y H, Sheen J.Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis[J]. Nature Protocols, 2007, 2(7): 1565-1572.
[18] Hao X Y, Horvath D P, Chao W S, et al.Identification and evaluation of reliable reference genes for quantitative real-time PCR analysis in tea plant [Camellia Sinensis(L.) O. Kuntze][J]. International Journal of Molecular Sciences, 2014, 15(12): 22155-22172.
[19] Paszkowski U, Kroken S, Roux C, et al.Rice phosphate transporters include an evolutionarily divergent gene specifically activated in arbuscular mycorrhizal symbiosis[J]. PNAS, 2002, 99(20): 13324-13329.
[20] Lian X M.Comprehensive sequence and whole-life-cycle expression profile analysis of the phosphate transporter gene family in rice[J]. Molecular Plant, 2011, 4(6): 1105-1122.
[21] Zhou Q, Zhang S S, Chen F, et al.Genome-wide identification and characterization of the sbp-box gene family in petunia[J]. BioMed Central, 2018, 19(1): 193. doi: 10.1186/s12864-018-4537-9.
[22] 张豪杰, 郝心愿, 周超, 等. 富硒区茶树鲜叶中硒累积与土壤因子的相关性分析[J]. 茶叶科学, 2020, 40(4): 465-477.
Zhang H J, Hao X Y, Zhou C, et al.Correlation analysis between selenium accumulation in tea leaves and soil factors in selenium-rich areas[J]. Journal of Tea Science, 2020, 40(4): 465-477.
[23] Wang M K, Yang W X, Zhou F, et al.Effect of phosphate and silicate on selenite uptake and phloem-mediated transport in tomato (Solanum lycopersicumL.)[J]. Environmental Science and Pollution Research, 2019, 26(20): 20475-20484.
[24] Milne J.Haloselenate (IV) formation and selenous acid dissociation equilibria in hydrochloric and hydrofluoric acids[J]. Canadian Journal of Chemistry, 1987, 18(35): 8735. doi: 10.1002/chin.198735010.
[25] Cabannes E, Buchner P, Broadley M R, et al.A comparison of sulfate and selenium accumulation in relation to the expression of sulfate transporter genes in astragalus species[J]. Plant Physiology, 2011, 157: 2227-2239.
[26] Freeman J L, Tamaoki M, Stushnoff C, et al.Molecular mechanisms of selenium tolerance and hyperaccumulation inStanleya pinnata[J]. Plant Physiology, 2010, 153(4): 1630-1652.