水稻叶片衰老基因LPS1的克隆与功能研究
收稿日期: 2021-03-04
修回日期: 2021-04-12
网络出版日期: 2021-09-10
基金资助
国家科技重大专项(2016ZX08009003-003-008);国家自然科学基金资助项目(31971921);广西水稻遗传育种重点实验室开放基金资助项目;中国水稻生物学国家重点实验室开放项目(20200102)
Cloning and Functional Analysis of Leaf Senescence Gene LPS1 in Oryza sativa
Received date: 2021-03-04
Revised date: 2021-04-12
Online published: 2021-09-10
【目的】早衰突变体是研究早衰机制的良好载体,对于探究早衰的遗传机理与作用机制及提高水稻的产量和品质具有重要作用。【方法】本研究利用EMS诱变获得了一个早衰突变体lps1,并对该突变体及其野生型进行表型观察、细胞学及组织化学分析、生理生化分析、遗传分析、基因定位和激素处理。【结果】lps1的叶片从3叶期开始发黄,成熟期株高、有效分蘖数、结实率、千粒重等极显著降低。电镜观察发现lps1叶表面光滑,硅质化突起和叶绿体数目减少、片层结构紊乱。生理生化分析表明lps1中有大量的活性氧积累,同时伴有蛋白质的降解、细胞膜的损伤以及大规模的细胞死亡。遗传分析表明该早衰表型受单隐性核基因控制,并且其在第5染色体上编码了一个泛素结合酶。亚细胞定位结果证明LPS1蛋白在细胞质与核中均有表达。外源激素处理发现,lps1对外源激素的处理更为敏感,且LPS1突变促进了ABA合成相关基因的表达。【结论】LPS1 突变使水稻ABA合成信号途径异常,进而引发H2O2等一系列与衰老相关生理指标的异常变动,导致lps1过早衰老,最终造成水稻产量严重降低。
褚晓洁, 芦涛, 叶涵斐, 王盛, 林晗, 吴先美, 何瑞, 严钢, 王跃星, 李三峰, 路梅, 胡海涛, 杨窑龙, 饶玉春 . 水稻叶片衰老基因LPS1的克隆与功能研究[J]. 中国水稻科学, 2021 , 35(5) : 427 -438 . DOI: 10.16819/j.1001-7216.2021.210303
【Objective】 As a good carrier, the premature aging mutant plays an important role in exploring the genetic mechanism and mechanism of action on premature aging, as well as improving grain yield and quality of rice.【Method】 In this study, a premature senescence mutant lps1 was obtained by EMS mutagenesis, and phenotypic observation, cytological and histochemical analysis, physiological and biochemical analysis, genetic analysis, gene localization and hormone treatment were performed on wild-type and mutants.【Result】 The leaves of lps1 began to turn yellow during the three-leaf stage, and the plant height, effective tiller number, seed-setting rate, and thousand-grain weight were extremely significantly reduced in the maturing stage. Electron microscopy observation found that the surface of lps1 leaves was smooth, the numbers of silicified protrusions and chloroplasts decreased, and the lamella structure was disordered. Physiological and biochemical analysis showed that a large amount of reactive oxygen species accumulated in mutant lps1, accompanied by protein degradation, cell membrane damage and large-scale cell death. Genetic analysis showed that the progeria phenotype was controlled by a single recessive nuclear gene, which encodes an ubiquitin conjugating enzyme on chromosome 5. The subcellular localization results prove that LPS1 protein is expressed in both the cytoplasm and the nucleus. Exogenous hormone treatment demonstrated that the lps1 was more sensitive to exogenous hormone treatment, and the LPS1 mutation promoted the expression of genes related to ABA synthesis.【Conclusion】 LPS1 mutation makes the rice ABA synthesis signal pathway abnormal, which in turn triggers abnormal changes in a series of aging-related physiological indicators such as H2O2, leading to premature senescence of lps1, and ultimately resulting in a serious decrease in rice yield.
Key words： rice; premature senescence; genetic analysis; subcellular localization; hormone
| [1] | Cao J, Jiang F, Sodmergen, Cui K M. Time-course of programmed cell death during leaf senescence in Eucommia ulmoides[J]. Journal of Plant Research, 2003, 116(1): 7-12. |
| [2] | Vicky B W. The molecular biology of leaf senescence[J]. Journal of Experimental Botany, 1997, 48(2): 181-199. |
| [3] | Lim P O, Kim H J, Nam H G. Leaf senescence[J]. Annual Review of Plant Biology, 2007, 58(1): 115-136. |
| [4] | Zhang H S, Zhou C J. Signal transduction in leaf senescence[J]. Plant Molecular Biology, 2013, 82(6): 539-545. |
| [5] | Panda D, Sarkar R K. Natural leaf senescence: Probed by chlorophyll fluorescence, CO2 photosynthetic rate and antioxidant enzyme activities during grain filling in different rice cultivars[J]. Physiology and Molecular Biology of Plants, 2013, 19(3): 43-51. |
| [6] | Li Z H, Zhang Y, Zou D, Zhao Y, Wang H L, Zhang Y, Xia X L, Luo J C, Zhang Z. LSD 3.0: A comprehensive resource for the leaf senescence research community[J]. Nucleic Acids Research, 2020, 48(D1): D1069-D1075. |
| [7] | Mao C J, Lu S C, Lü B, Zhang B, Shen J B, He J M, Luo L Q, Xi D D, Chen X, Ming F. A rice NAC transcription factor promotes leaf senescence via ABA biosynthesis[J]. Plant Physiology, 2017, 174(3): 1747-1763. |
| [8] | Liang C Z, Wang Y Q, Zhu Y N, Tang J Y, Hu B, Liu L C, Ou S J, Sun X H, Chu J F, Chu C C. OsNAP connects abscisic acid and leaf senescence by fine-tuning abscisic acid biosynthesis and directly targeting senescence-associated genes in rice[J]. Proceedings of the National Academy of Sciences of the United States of America, 2014, 111(27): 10013-10018. |
| [9] | Han M, Kim C Y, Lee J, Lee S K, Jeon J S. OsWRKY42 represses OsMT1d and induces reactive oxygen species and leaf senescence in rice[J]. Molecular Cells, 2014, 37(7): 532-539. |
| [10] | Uji Y, Akimitsu K, Gomi K. Identification of OsMYC2-regulated senescence-associated genes in rice[J]. Planta, 2017, 245(6): 1241-1246. |
| [11] | Lu G W, Casaretto J A, Ying S, Mahmood K, Liu F, Bi Y M, Rothstein S J. Overexpression of OsGATA12 regulates chlorophyll content, delays plant senescence and improves rice yield under high density planting[J]. Plant Molecular Biology, 2017, 94(1): 215-227. |
| [12] | Liu W Z, Fu Y P, Hu G C, Si H M, Wu C, Sun Z X. Identification and fine mapping of a thermo-sensitive chlorophyll deficient mutant in rice (Oryza sativa L.)[J]. Planta, 2007, 226(3): 785-795. |
| [13] | Liu C H, Zhu H T, Xing Y, Tan J J, Chen X H, Zhang J J, Peng H F, Xie Q J, Zhang Z M. Albino Leaf 2 is involved in the splicing of chloroplast group I and II introns in rice[J]. Journal of Experimental Botany, 2016, 67(18): 5339-5347. |
| [14] | Kusaba M, Ito H, Morita R, Iida S, Sato Y, Fujimoto M, Kawasaki S, Tanaka R, Hirochika H, Nishimura M, Tanaka A. Rice NON-YELLOW COLORING1 is involved in light-harvesting complex II and grana degradation during leaf senescence[J]. Plant Cell, 2007, 19(4): 1362-1375. |
| [15] | Rong H, Tang Y Y, Zhang H, Wu P Z, Chen Y P, Li M R, Wu G J, Jiang H W. The Stay-Green Rice like (SGRL) gene regulates chlorophyll degradation in rice[J]. Journal of Plant Physiology, 2013, 170(15): 1367-1373. |
| [16] | Fang C Y, Zhang H, Wan J, Wu Y Y, Chen W, Wang S C, Wang W S, Zhang H W, Zhang F, Qu L H, Liu X Q, Zhou D X, Luo J. Control of leaf senescence by an MeOH-jasmonates cascade that is epigenetically regulated by OsSRT1 in rice[J]. Molecular Plant, 2016, 9(10): 1366-1378. |
| [17] | Xu W Y, Kong Z S, Li M N, Yang W Q, Xue Y B. A novel nuclear-localized CCCH-type ainc finger protein, OsDOS, is involved in delaying leaf senescence in rice[J]. Plant Physiology, 2006, 141(4): 1376-1388. |
| [18] | Chen Y, Xu Y Y, Luo W, Li W X, Chen N, Zhang D J, Chong K. The F-Box protein OsFBK12 targets OsSAMS1 for degradation and affects pleiotropic phenotypes, including leaf senescence, in rice[J]. Plant Physiology, 2013, 163(4): 1673-1685. |
| [19] | Zhao Y, Chan Z L, Gao J H, Xing L, Cao M J, Yu C M, Hu Y L, You J, Shi H T, Zhu Y F, Gong Y H, Mu Z X, Wang H Q, Deng X, Wang P C, Bressan R A, Zhu J K. ABA receptor PYL9 promotes drought resistance and leaf senescence[J]. Proceedings of the National Academy of Sciences of The United States of America, 2016, 113(7): 1949-1954. |
| [20] | Yang X, Gong P, Li K Y, Huang F D, Cheng F M, Pan G. A single cytosine deletion in the OsPLS1 gene encoding vacuolar-type H+-ATPase subunit A1 leads to premature leaf senescence and seed dormancy in rice[J]. Journal of Experimental Botany, 2016, 67(9): 2761-2776. |
| [21] | Zhou Y, Liu L, Huang W F, Yuan M, Zhou F, Li X H, Lin Y J. Overexpression of OsSWEET5 in rice causes growth retardation and precocious senescence[J]. PloS One, 2014, 9(4): e94210. |
| [22] | Singh S, Singh A, Nandi A K. The rice OsSAG12-2 gene codes for a functional protease that negatively regulates stress-induced cell death[J]. Journal of Biosciences, 2016, 41(3): 445-453. |
| [23] | Wu L W, Ren D Y, Hu S K, Li G M, Dong G J, Jiang L, Hu X M, Ye W J, Cui Y T, Zhu J, Zhang G H, Gao Z Y, Zeng D L, Qian Q, Guo L B. Down-regulation of a nicotinate phosphoribosyltransferase gene OsNaPRT1 leads to withered leaf tips[J]. Plant Physiology, 2016, 171(2): 1085-1098. |
| [24] | Lee R H, Hsu J H, Huang H J, Lo S F, Grace S C. Alkaline α-galactosidase degrades thylakoid membranes in the chloroplast during leaf senescence in rice[J]. New Phytology, 2009, 184(3): 596-606. |
| [25] | Kang K, Kim Y S, Park S, Back K. Senescence-induced serotonin biosynthesis and its role in delaying senescence in rice leaves[J]. Plant Physiology, 2009, 150(3): 1380-1393. |
| [26] | Kudo T, Makita N, Kojima M, Tokunaga H, Sakakibara H. Cytokinin activity of cis-zeatin and phenotypic alterations induced by overexpression of putative cis-zeatin glucosyltransferase in rice[J]. Plant Physiology, 2012, 160(1): 319-331. |
| [27] | Wiseman B R. Plant-resistance to insects in integrated pest-management[J]. Plant Disease, 1994, 78(9): 927-932. |
| [28] | Liang C Z, Chu C C. Towards understanding abscisic acid-mediated leaf senescence[J]. Science China: Life Sciences, 2015, 58(5): 506-508. |
| [29] | Pan H R, Liu S M, Tang D Z. HPR1, a component of the THO/TREX complex, plays an important role in disease resistance and senescence in Arabidopsis[J]. Plant Journal, 2012, 69(5): 831-843. |
| [30] | Hu Y N, Jiang Y J, Han X, Wang H P, Pan J J, Yu D Q. Jasmonate regulates leaf senescence and tolerance to cold stress: Crosstalk with other phytohormones[J]. Journal of Experimental Botany, 2017, 68(6): 1361-1369. |
| [31] | Dani K G S, Fineschi S, Michelozzi M, Loreto F. Do cytokinins, volatile isoprenoids and carotenoids synergically delay leaf senescence[J]. Plant Cell Environment, 2016, 39(5): 1103-1111. |
| [32] | Kim J I, Murphy A S, Baek D, Lee S W, Yun D J, Bressan R A, Narasimhan M L. YUCCA6 over-expression demonstrates auxin function in delaying leaf senescence in Arabidopsis thaliana[J]. Journal of Experimental Botany, 2011, 62(11): 3981-3992. |
| [33] | Chen L G, Xiang S Y, Chen Y L, Li D B, Yu D Q. Arabidopsis WRKY45 interacts with the DELLA protein RGL1 to positively regulate age-triggered leaf senescence[J]. Molecular Plant, 2017, 10, 1174-1189. |
| [34] | Riefler M, Novak O, Strnad M, Schmülling T. Arabidopsis cytokinin receptor mutants reveal functions in shoot growth, leaf senescence, seed size, germination, root development, and cytokinin metabolism[J]. Plant Cell, 2006, 18(1): 40-54. |
| [35] | Wang T, Li C X, Wu Z H, Jia Y C, Wang H, Sun S Y, Mao C Z, Wang X L. Abscisic acid regulates auxin homeostasis in rice root tips to promote root hair elongation[J]. Frontier in Plant Science, 2017, 8: 1121-1138. |
| [36] | Xu N, Chu Y L, Chen H L, Li X X, Wu Q, Jin L, Wang G X, Huang J L. Rice transcription factor OsMADS25 modulates root growth and confers salinity tolerance via the ABA-mediated regulatory pathway and ROS scavenging[J]. PLoS Genet, 2018, 14(10): e1007662. |
| [37] | Chang Y, Nguyen B H, Xie Y J, Xiao B Z, Tang N, Zhu W L, Mou T M, Xiong L Z. Co-overexpression of the constitutively active form of OsbZIP46 and ABA-activated protein kinase SAPK6 improves drought and temperature stress resistance in rice[J]. Frontier in Plant Science, 2017, 8: 1102-1117. |
| [38] | Promchuea S, Zhu Y J, Chen Z Z, Zhang J, Gong Z Z. ARF2 coordinates with PLETHORAs and PINs to orchestrate ABA-mediated root meristem activity in Arabidopsis[J]. Journal of Integrative Plant Biology, 2017, 59(1): 30-43. |
| [39] | Sun L R, Wang Y B, He S B, Hao F S. Mechanisms for abscisic acid inhibition of primary root growth[J]. Plant Signaling & Behavior, 2018, 13(9): e1500069. |
| [40] | Hung K T, Yi T H, Kao C H. Hydrogen peroxide is involved in methyl jasmonate-induced senescence of rice leaves[J]. Physiology Plantarum, 2006, 127(2): 293-303. |
| [41] | 李兆伟. 水稻叶片早衰突变体的糖代谢基因表达与抗氧化生理调控[D]. 杭州: 浙江大学, 2014. |
| [41] | Li Z W. The expression alteration of various genes related to sugar metabolism in senescing leaves and its antioxidation modulation for esl mutant[D]. Hangzhou: Zhejiang University, 2014. |
| [42] | Lichtenthaler H K. Chlorophylls and carotenoids: Pigments of photosynthetic biomembranes[J]. Methods in Enzymology, 1987, 148: 350-382. |
| [43] | 游均, 方玉洁, 熊立仲. 活性氧检测[J/OL].Bio-101, 2018: e1010170. |
| [43] | You J, Fang Y J, Xiong L Z. Reactive oxygen detection [J/OL]. Bio-101, 2018: e1010170. (in Chinese) |
| [44] | Rao Y C, Jiao R, Wang S, Wu X M, Ye H F, Pan C Y, Li S F, Xin D D, Zhou W Y, Dai G X, Hu J, Ren D Y, Wang Y X. SPL36 encodes a receptor-like protein kinase that regulates programmed cell death and defense responses in rice[J]. Rice, 2021, 14(1): 34-47. |
| [45] | 沈恒胜, 陈君琛, 黄进华, 汤葆莎. 水稻叶表皮硅体显微结构及其分布[J]. 福建农业大学学报, 2005, 34(2): 137-140. |
| [45] | Shen H S, Chen J C, Huang J H, Tang B S. Microtructure and distribution of silica bodies rice epidermis[J]. Fujian Academy of Agricultural Sciences, 2005, 34(2): 137-140. (in Chinese with English abstract) |
| [46] | 杨秉耀, 陈新芳, 刘向东, 郭海滨. 水稻不同品种叶表面硅质细胞的扫描电镜观察[J]. 电子显微学报, 2006, 25(2): 68-72. |
| [46] | Yang B Y, Chen X F, Liu X D, Guo H B. Scanning electron microscopic observation of silicon cells on the leaf surface of different rice varieties[J]. Journal of Chinese Electron Microscopy Society, 2005, 25(2): 68-72. |
| [47] | Christopher E. A possible mechanism of biological silicification in plants[J]. Frontier in Plant Science, 2015, 6: 853-859. |
| [48] | Yang L, Han Y Q, Li P, Li F, Ali S, Hou M L. Silicon amendment is involved in the induction of plant defense responses to a phloem feeder[J]. Scientific Report, 2017, 7(1): 4232-4240. |
| [49] | Hideg E, Kálai T, Kós P B, Asada K, Hideg K. Singletoxygen in plants: Its significance and possible detection with double (fluorescent and spin) indicator reagents[J]. Photochemistry and Photobiology, 2006, 82(5): 1211-1218. |
| [50] | 华春, 王仁雷. 杂交稻及其三系叶片衰老过程中SOD、CAT活性和MDA含量的变化[J]. 西北植物学报, 2003, 23(3): 406-409. |
| [50] | Hua C, Wang R L. Changes of SOD and CAT activities and MDA content during senescence of hybrid rice and three lines leaves[J]. Acta Botanica Boreali- Occidentalia Sinica, 2003, 23(3): 406-409. |
| [51] | Hu B, Zhu C G, Li F, Tang J Y, Wang Y Q, Liu L C, Che R H, Chu C C. LEAF TIP NECROSIS1 plays a pivotal role in the regulation of multiple phosphate starvation responses in rice[J]. Plant Physiology, 2011, 156(3): 1101-1115. |
| [52] | Lee I C, Hong S W, Whang S S, Lim P O, Nam H G, Koo J C. Age-dependent action of an ABA-inducible receptor kinase, RPK1, as a positive regulator of senescence in Arabidopsis leaves[J]. Plant and Cell Physiology, 2011, 52(4): 651-662. |
/
| 〈 |
|
〉 |