Abstract:Plants absorb and transport phosphorus through phosphorus transporters (PHO), and phosphorus is one of the essential nutrients for plant growth and development, which has an important effect on crop yield, quality and stress resistance. High temperature is an important environmental factor affecting potato yield and quality. In order to explore the influence of phosphorus transporter on potato (Solanum tuberosum) heat resistance, this study constructed a transgenic potato strain with overexpression of phosphorus transporter gene StPHO1.3 mediated by Agrobacterium tumefaciens. By comparing the growth and development of transgenic strain and wild-type strain at high temperature (35 ℃), it was found that overexpressing StPHO1.3 could enhance the heat resistance and promote the growth and development of potato. In addition, the results of subcellular localization showed that StPHO1.3 was expressed on the cell membrane. Therefore, in this study, the interacting proteins of StPHO1.3 were screened in the membrane system library plasmid by yeast two-hybridization and further verified by bimolecular fluorescence complementation (BiFC). The results showed that, StPHO1.3 interacted with both the photosystem Ⅱ protein subunit and calcium transport-associated proteins. In conclusion, overexpression of StPHO1.3 could enhance the resistance to high temperature stress and promote the growth and development of potato by affecting photosynthesis and signal transduction. These results provide theoretical basis and reference for further understanding of the function of phosphorus transporters and promote the breeding of new potato varieties.
李万, 李成, 程敏, 吴芳. 磷转运蛋白StPHO1.3对马铃薯耐热性的影响及其互作蛋白的筛选[J]. 农业生物技术学报, 2024, 32(2): 273-282.
LI Wan, LI Cheng, CHENG Min, WU Fang. Effects of Phosphorus Transporter StPHO1.3 on Heat Resistance of Potato (Solanum tuberosum) and Screening of Its Interacting Proteins. 农业生物技术学报, 2024, 32(2): 273-282.
[1] 窦海鸥. 2014. AtCBF3提高转基因马铃薯耐热性的研究[D]. 硕士学位论文, 山东农业大学, 导师: 杨兴洪. pp. 5-15. (Dou H O.2014. Arabidopsis thaliana CBF3 enhances the tolerance of potato to high temperature[D]. Thesis for M.S., Shandong Agricultural University, Supervisor: Yang X H, pp. 5-15.) [2] 李万, 杨明明, 高翔, 等. 2017. 西农538 LMW-GS基因的克隆, 原核表达及功能鉴定[J]. 麦类作物学报, 37(4): 445-451. (Li W, Yang M, Gao X, et al.2017. Isolation, prokaryotic expression and functional analysis of LMW-GS from Xinong 538 (Triticum aestivum L.)[J]. Journal of Triticeae Crops, 37(4): 445-451.) [3] 陆孙杰. 2012. 水稻OsMADS15基因功能研究及用酵母双杂筛选互作蛋白[D]. 博士学位论文, 浙江大学, 导师: 凃巨民, pp. 74-92. (Lu S.2012. Functional characterization of OsMADS15 and screening its interactors by yeast two hybrid system[D]. Thesis for Ph.D., Zhejiang University, Supervisor: Tu J M, pp. 74-92.) [4] 于人杰. 2019. 响应低磷干旱胁迫磷转运蛋白基因在大豆组织中的表达及功能分析[D]. 博士学位论文, 吉林农业大学, 导师: 杨美英, pp. 1-4. (Yu R J.2019. Expression and functional analysis of phosphorus transporter genes responding to low phosphorus and drought stress in soybean tissues[D]. Thesis for Ph.D., Jilin Agricultural University, Supervisor: Yang M Y, pp. 1-4.) [5] 叶明辉, 赵朋, 牛洋, 等. 2021. 马铃薯同源异形框基因家族的鉴定和表达分析[J]. 农业生物技术学报, 29(2): 224-239. (Ye M H, Zhao P, Niu Y, et al.2021. Identification and expression analysis of homeobox gene family in potato (Solanum tuberosum)[J]. Journal of Agricultural Biotechnology, 29(2): 224-239.) [6] 张超. 2021. 茉莉酸调控基因GH3家族的鉴定及在马铃薯中抗病及损伤分析[D]. 博士学位论文, 西北农林科技大学, 导师: 陈勤, pp. 77-80. (Zhang C.2021. Identification of jasmonic acid regulatory gene GH3 family and analysis of disease resistance and wounding in potato[D]. Thesis for Ph.D., Northwest A&F University, Supervisor: Chen Q, pp. 77-80.) [7] Arpat B, Magliano P, Wege S, et al.2012. Functional expression o f PHO1 to the Golgi and trans-Golgi network and its role in export of inorganic phosphate[J]. Plant Journal, 71(3): 479-491. [8] Bokszczanin K L, Fragkostefanakis S.2013. Perspectives on deciphering mechanisms unde rlying plant heat stress response and thermotolerance[J]. Frontiers in Plant Science, 4: 315-335. [9] Cao M, Liu H, Zhang C, et al.2020. Functional analysis of StPHT1;7, a Solanum tuberosum L. phosphate transporter gene, in growth and drought tolerance[J]. Plants, 9(10): 1384. [10] Ciereszko I, Johansson H, Hurry V, et al.2001. Phosphate status affects the gene expression, protein content and enzymatic activity of UDP glucose pyrophosphorylase in wild-type and pho mutants of Arabidopsis[J]. Planta, 212(4): 598-605. [11] Ferreira G C, Pratt R D, Pedersen P L.1989. Energy-linked anion transport. cloning, sequencing, and characterization of a full length cDNA encoding the rat liver mitochondrial proton/phosphate symporter[J]. Journal of Biological Chemistry, 264(26): 15628-15633. [12] Gong M, Van der Luit A H, Knight M R, et al. 1998. Heat-shock-induced changes in intracellular Ca2+ level in tobacco seedlings in relation to thermotolerance[J]. Plant Physiology, 1998, 116(1): 429-437. [13] Guo B, Irigoyen S, Fowler T B, et al.2008. Differential expression and phylogenetic analysis suggest specialization of plastid-localized members of the PHT4 phosphate transporter family for photosynthetic and heterotrophic tissues[J]. Plant Signaling and Behavior, 3(10): 784-790. [14] Guo C, Guo L, Li X, et al.2014. TaPT2, a high-affinity phosphate transporter gene in wheat (Triticum aestivum L.), is crucial in plant Pi uptake under phosphorus deprivation[J]. Acta Physiologiae Plantarum, 36(6): 1373-1384. [15] Karthikeyan A S, Varadarajan D K, Mukatira U T, et al.2002. Regulated expression of Arabidopsis phosphate transporters[J]. Plant Physiology, 130(1): 221-233. [16] Khan G A, Bouraine S, Wege S, et al.2014. Coordination between zinc and phosphate homeostasis involves the transcription factor PHR1, the phosphate exporter PHO1, and its homologue PHO1;H3 in Arabidopsis[J]. Journal of Experimental Botany, 65(3): 871-884. [17] Li W, Dong J, Cao M, et al.2019. Genome-wide identification and characterization of HD-ZIP genes in potato[J]. Gene, 697: 103-117. [18] Liu B, Zhao S, Wu X, et al.2017. Identification and characterization of phosphate transporter genes in potato[J]. Journal of Biotechnology, 264: 17-28. [19] Mudge S R, Rae A L, Diatloff E, et al.2002. Expression analysis suggests novel roles for members of Pht1 family of phosphate transporters in Arabidopsis[J]. Plant Journal, 31(3): 341-353. [20] Qin L, Guo Y, Chen L, et al.2012. Functional characterization of 14 Pht1 family genes in yeast and their expressions in response to nutrient starvation in soybean[J]. PLOS ONE, 7(10): e47726. [21] Rafael R B A, Fernández-Marcos M L, Cocco S, et al.2020. Increased phosphorus availability to corn resulting from the simultaneous applications of phosphate rock, calcareous rock, and biochar to an acid sandy soil[J]. Pedosphere, 30(6): 719-733. [22] Remy E, Cabrito T, Batista R A, et al.2012. The Pht1;9 and Pht1;8 transporters mediate inorganic phosphate acquisition by the Arabidopsis thaliana root during phosphorus starvation[J]. New Phytologist, 195(2): 356-371. [23] Ribot C, Wang Y, Poirier Y.2008a. Expression analyses of three members of the AtPHO1 family reveal differential interactions between signaling pathways involved in phosphate deficiency and the responses to auxin, cytokinin, and abscisic acid[J]. Planta, 227(5): 1025-1036. [24] Ribot C, Zimmerli C, Farmer E E, et al.2008b. Induction of the Arabidopsis PHO1;H10 gene by 12-oxo-phytodienoic acid but not jasmonic acid via a CORONATINE INSENSITIVE1-dependent pathway[J]. Plant Physiology, 147(2): 696-706. [25] Rouached H, Stefanovic A, Secco D, et al.2011. Uncoupling phosphate deficiency from its major effects on growth and transcriptome via PHO1 expression in Arabidopsis[J]. Plant Journal, 65(4): 557-570. [26] van der Graaff E, Schwacke R, Schneider A, et al.2006. Transcription analysis of Arabidopsis membrane transporters and hormone pathways during developmental and induced leaf senescences[J]. Plant Physiology, 141(2): 776-792. [27] Wang Y, Secco D, Poirier Y.2008. Characterization of the PHO1 gene family and the responses to phosphate deficiency of Physcomitrella patens[J]. Plant Physiology, 146(2): 646-656. [28] Zhou X, Zha M, Huang J, et al.2017. StMYB44 negatively regulates phosphate transport by suppressing expression of PHOSPHATE1 in potato[J]. Journal of Experimental Botany, 68(5): 1265-1281.