Welcome to Acta Agronomica Sinica,

Acta Agron Sin ›› 2017, Vol. 43 ›› Issue (01): 51-62.doi: 10.3724/SP.J.1006.2017.00051


Gene Mapping and Cloning of a Premature Leaf Senescence Mutant psls1 in Rice

HUANG Ya-Min1,**,ZHU Shan-Shan1,**,ZHAO Zhi-Chao1,**,PU Zhi-Gang2,LIU Tian-Zhen1, LUO-Sheng1,ZHANGXin1,*   

  1. 1National Key Facility for Crop Gene Resources and Genetic Improvement/Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, China; 2Biotechnology Institute of Nuclear Technology, Sichuan Academy of Agricultural Sciences, Chengdu 610066?, China
  • Received:2016-03-18 Revised:2016-06-20 Online:2017-01-12 Published:2016-07-04
  • Contact: 张欣,E-mail: zhangxin02@caas.cn ** 同等贡献(Contributed equally to this work) E-mail:zhangxin02@caas.cn
  • Supported by:

    This study was supported by the National Natural Science Foundation of China (91535302) and the Major Project of China on New Varieties of GMO Cultivation (2015ZX08010-004).


Leaf early senescence directly reduces its photosynthetic capacity, decreasing crop yield and grain quality. It is of great importance to identify novel mutants and characterize their molecular and physiological mechanisms. In this paper, we reported gene mapping and cloning of a psls1 mutant (premature senescence leaf with spots) in rice. The mutant showed early senescence symptoms, including decreased chlorophyll content, over-accumulated H2O2, yellow-withered leaf and increased dead cell from bottom to top of plant after the 7-leaf stage. Moreover, the plant height, tiller number, panicle length, and the number of grains per panicle were significantly lower in psls1. We observedthe degradation of chloroplast, unclean thylakoid and numerous osmiophilic granules in psls1 leavesby transmission electron microscope (TEM). Genetic analysis demonstrated that the phenotype of psls1 was determined by a single recessive gene. Using genetic population derived from psls1 × IRAT129, the psls1 locus was mapped in a 89 kb region flanked by markers zs-3 and zs-8 on chromosome 7, containing 12 putative open reading frames (ORFs). Sequence analysis revealed a single-base substitution occurred in the genomic sequence of LOC_Os07g46460, which led to a 59 bp deletion in its cDNA, and therefore, predicted LOC_Os07g46460 (PSLS1), encoding a ferredoxin-dependent glutamate synthase as the candidate gene. The mRNA expression level of mutated PSLS1 decreased sharply, resulting in reduction of glutamate synthase activity and abnormal amino acid metabolism in psls1. Under low nitrogen treatment, the senescence phenotype of psls1 could occur as early as at the 3-leaf stage. These results indicated that psls1 senescence phenotype might be associated with the loss of glutamate synthase activity and the abnormal amino acid metabolism.

Key words: Rice, Leaf senescence, Gene-mapping, PSLS1 gene, Glutamate synthase (Fd-GOGAT)

[1]刘道宏. 植物叶片的衰老. 植物生理学通讯, 1983, (2): 12–19
Liu D H. Plant leaf senescence. Plant Physiol Commun, 1983, (2): 12–19 (in Chinese)
[2]Grbic V, Bleecker A B. Ethylene regulates the timing of leaf senescence in Arabidopsis. Plant J, 1995, 8: 595–602
[3]Alonso J M, Hirayama T, Roman G, Nourizadeh S, Ecker J R. EIN2, a bifunctional transducer of ethylene and stress responses in Arabidopsis. Science, 1999, 284: 2148–2152
[4]Li Z H, Peng J Y, Wen X, Guo H W. ETHYLENE-INSENSITIVE3 is a senescence-associated gene that accelerates age-dependent leaf senescence by directly repressing miR164 transcription in Arabidopsis. Plant Cell, 2013, 25: 3311–3328
[5]Liang C Z, Wang Y Q, Zhu Y N, Tang J Y, Hu B, Liu L C, Ou S J, Wu H K, 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. Proc Acad Natl Sci USA, 2014, 111: 10013–10018
[6]Danisman S, van der Wa.l F, Dhondt S, Waites R, de Folter S, Bimbo A, van Dijk A, Muino J M, Cutri L, Dornelas M C, Angenent G C, Immink R G H. Arabidopsis class I and class II TCP transcription factors regulate jasmonic acid metabolism and leaf development antagonistically. Plant Physiol, 2012, 159: 1511–1523
[7]Rivero R M, Kojima M, Gepstein A, Sakakibara H, Mittler R, Gepstein S, Blumwald E. Delayed leaf senescence induces extreme drought tolerance in a flowering plant. Proc Natl Acad Sci USA, 2007, 104: 19631–19636
[8]Morris K, MacKerness S A H, Page T, John C F, Murphy A M, Carr J P, Buchanan-Wollaston V. Salicylic acid has a role in regulating gene expression during leaf senescence. Plant J, 2000, 23: 677–685
[9]FukaoT,Yeung E, Bailey-Serres J. The submergence tolerance gene SUB1A delays leaf senescence under prolonged darkness through bormonal regulation in rice. Plant Physiol, 2012, 160: 1795–1807
[10]Han M H, Kim C Y, Lee J, Lee S K, Jeon J S. OsWRKY42 represses OsMT1d and induces reactive oxygen species and leaf senescence in rice. Mol Cell, 2014, 37: 532–539
[11]Rao Y C,Yang Y L, Xu J, Li X J, Leng Y J, Dai L P, Huang L C, Shao G S, Ren D Y,Hu J, Guo L B, Pan J W, Zeng D L. EARLY SENESCENCE1encodes a SCAR-LIKE PROTEIN2 that affects water loss in rice. Plant Physiol, 2015, 169: 1225–1239
[12]Gao Q S, Yang Z F, Zhou Y, Yin Z T, Qiu J, Liang G H, Xu C W. Characterization of an Abc1 kinase family gene OsABC1-2 conferring enhanced tolerance to dark-induced stress in rice. Gene, 2012, 498: 155–163
[13]Kusaba M, Ito H, Morita R, Iida S, Sato Y, Fujimoto M, Kawasa-ki 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. Plant Cell, 2007, 19: 1362–1375
[14]Park S Y, Yu J W, Park J S, Li J J, Yoo S C, Lee N Y, Lee S K, Jeong S W, Seo H, Koh H J, Jeon J S, Park Y, Paek N C. The senescence-induced stay green protein regulates chlorophyll degradation. Plant Cell, 2007, 19: 1649–1664
[15]Yoshida S, Ito M, Nishida I, Akira W. Identification of a novel gene HYS /CPR5 that has a repressive role in the induction of leaf senescence and pathogen defence responses in Arabidopsis thaliana. Plant J, 2002, 29: 427–437
[16]Aki T, Konishi M, Kikuchi T, Fujimori T, YoneyamaT,Yanagisawa S. Distinct modulations of the hexokinase1-mediated glucose response and hexokinase1-independent processes by HYS1/CPR5 in Arabidopsis. J Exp Bot, 2007, 58: 3239–3248
[17]Lea P J, Miflin B J. Alternative route for nitrogen assimilation in higher plants. Nature, 1974, 251: 614–616
[18]Lam H M, Coschigano K T, Oliveira I C, OliveiraM, Coruzzi G M. The molecular-genetics of nitrogen assimilation into amino acids in higher plants. Annu Rev Plant Physiol Plant Mol Biol, 1996, 47: 569–593
[19]Somerville C R, Ogren W L. Inhibition of photosynthesis in Arabidopsis mutants lacking leaf glutamate synthase activity. Nature, 1980, 286: 257–259
[20]Suzuki A, Rothstein S. Structure and regulation of ferredoxin-dependent glutamate synthase from Arabidopsis thaliana: cloning of cDNA, expression in different tissues of wild-type and gltS mutant strains, and light induction. Eur J Biochem, 1997, 243: 708–718
[21]Coschigano K T, Melo-Oliveira R, Lim J, Coruzzi G M. Arabidopsis gls mutants and distinct Fd-GOGAT genes: implications for photorespiration and primary nitrogen assimilation. Plant Cell, 1998, 10: 741–752
[22]Suzuki A, Vidal J, Gadal P. Glutamate synthase isoforms in rice: immunological studies of enzymes in green leaf, etiolated leaf, and root tissues. Plant Physiol, 1982, 70: 827–832
[23]Zhao X Q, Shi W M. Expresssionanalysisof the glutamine synthetase and glutamate synthase gene families in young rice (Oryza sativa) seedlings. Plant Sci, 2006, 170: 748–754
[24]卢永恩, 罗风, 杨猛, 李香花, 练兴明. 抑制表达谷氨酸合酶基因对水稻碳氮代谢的影响.生命科学, 2011, 41: 481–493
Lu Y N, Luo F, Yang M, Li X H, Lian X M. Suppression of glutamate synthase genes significantly affects carbon and nitrogenmetabolism in rice (Oryza sativa L.). Sci China Life Sci, 2011, 41: 481–493 (in Chinese)
[25]Mattana M, Biazzi E, Bertani A, Coraggio I. Characterization of the Ferredoxin-Gogat gene (OsGog2 clone) expression in rice. Biol Plant, 2006, 50:187–192
[26]Wu Z, Zhang X, He B, Diao L, Sheng S, Wang J, Guo X, Su N, Wang L, Jiang L, Wang C, Zhai H, Wan J. A chlorophyll-deficient rice mutant with impaired chlorophyllide esterification in chlorophyll biosynthesis. Plant Physiol, 2007, 45: 29–40
[27]Thordal-Christensen H, Zhang Z, Wei Y, Collinge D B. Sub-cellular localization of H2O2 in plants. H2O2 accumulation in papillaeand hypersensitive response during the barley-powdery mildew interaction. Plant J, 1997,11: 1187–1194
[28]Dietrich R A, Delaney T P, Uknes S J, Ward E R, Ryals J A, Dangl J L. Arabidopsis mutants simulating disease resistance response. Cell, 1994, 77: 565–577
[29]Dong H, Fei G L,Wu C Y, Wu F Q, Sun Y Y, Chen M J, Ren Y L,Zhou K N, Cheng Z J,Wang J L, Jiang L, Zhang X, Guo X P,Lei C L,Su N, Wang H Y, Wan J M. A rice Virescent-yellow leaf mutant reveals new insights into the role and assenbly of plastid caseinolytic protease in higher plants. Plant Physiol, 2013, 162: 1867–1880
[30]孙玉莹. 水稻叶片早衰基因PSL2的图位克隆及功能初步分析.中国农业科学院硕士学位论文,北京,2013
Sun Y Y. Map-based Cloning and Basic Functional Analysis of Presenescing Leaf Gene PSL2 in Rice(Oryza sativa). MS Thesis of Chinese Academy of Agricultural Sciences, Beijing, China, 2013
[31]Liu K, Liu L L, Ren Y L,Wang Z Q, Zhou K N, Liu X,Wang D, Zheng M, Cheng Z J, Lin Q B,Wang J L,Wu F Q, Zhang X,Guo X P, Wang C M, Zhai H Q, JiangL, Wan J M. Dwarf and tiller-enhancing 1 regulates growth and development by influencing boron uptake in boron limited condition in rice. Plant Sci, 2015, 236: 18–28
[32]Tamura W, Kojima S, Toyokawa A, Watanabe H, Tabuchi K M, Hayakawa T, Yamaya T. Disruption of a novel NADH-glutamate synthase2 gene caused marked reduction in spikelet number of rice. Plant Sci, 2011, 2: 1–9
[33]Crawford N M, Ford B G. Molecular and developmental biology of inorganic nitrogen nutrition. The Arabidopsis Book, 2002[2016-03-07] http://www.aspb.org/publications/
[34]吴巍, 赵军. 植物对氮素吸收利用的研究进展.中国农学通报, 2010, 26(13): 75–78
Wu W, Zhao J. Advances on plants’ nitrogen assimilation and utilization. Chin Agric Sci Bull, 2010, 26(13): 75–78 (in Chinese with English abstract)
[35]Keys A J, Bird I F, Cornelius M J, Lea P J, Wallsgrove R M, Miflin B J. Photorespiratory nitrogen cycle. Nature, 1978, 275: 741–743
[36]Blackwell R D, Murray A J S, Lea P J. The isolation and characterization of photorespiratory mutants of barley and pea. In: BigginsJ eds. In Progress in Photosynthesis Research. RhodeIsland: Springer Netherlands, 1987, pp 625–628
[37]Somerville C R, Ogren W L. Inhibition of photosynthesis in Arabidopsis mutants lacking leaf glutamate synthase activity. Nature, 1980, 286: 257–259

[1] TIAN Tian, CHEN Li-Juan, HE Hua-Qin. Identification of rice blast resistance candidate genes based on integrating Meta-QTL and RNA-seq analysis [J]. Acta Agronomica Sinica, 2022, 48(6): 1372-1388.
[2] ZHENG Chong-Ke, ZHOU Guan-Hua, NIU Shu-Lin, HE Ya-Nan, SUN wei, XIE Xian-Zhi. Phenotypic characterization and gene mapping of an early senescence leaf H5(esl-H5) mutant in rice (Oryza sativa L.) [J]. Acta Agronomica Sinica, 2022, 48(6): 1389-1400.
[3] ZHOU Wen-Qi, QIANG Xiao-Xia, WANG Sen, JIANG Jing-Wen, WEI Wan-Rong. Mechanism of drought and salt tolerance of OsLPL2/PIR gene in rice [J]. Acta Agronomica Sinica, 2022, 48(6): 1401-1415.
[4] ZHENG Xiao-Long, ZHOU Jing-Qing, BAI Yang, SHAO Ya-Fang, ZHANG Lin-Ping, HU Pei-Song, WEI Xiang-Jin. Difference and molecular mechanism of soluble sugar metabolism and quality of different rice panicle in japonica rice [J]. Acta Agronomica Sinica, 2022, 48(6): 1425-1436.
[5] YAN Jia-Qian, GU Yi-Biao, XUE Zhang-Yi, ZHOU Tian-Yang, GE Qian-Qian, ZHANG Hao, LIU Li-Jun, WANG Zhi-Qin, GU Jun-Fei, YANG Jian-Chang, ZHOU Zhen-Ling, XU Da-Yong. Different responses of rice cultivars to salt stress and the underlying mechanisms [J]. Acta Agronomica Sinica, 2022, 48(6): 1463-1475.
[6] YANG Jian-Chang, LI Chao-Qing, JIANG Yi. Contents and compositions of amino acids in rice grains and their regulation: a review [J]. Acta Agronomica Sinica, 2022, 48(5): 1037-1050.
[7] DENG Zhao, JIANG Nan, FU Chen-Jian, YAN Tian-Zhe, FU Xing-Xue, HU Xiao-Chun, QIN Peng, LIU Shan-Shan, WANG Kai, YANG Yuan-Zhu. Analysis of blast resistance genes in Longliangyou and Jingliangyou hybrid rice varieties [J]. Acta Agronomica Sinica, 2022, 48(5): 1071-1080.
[8] YANG De-Wei, WANG Xun, ZHENG Xing-Xing, XIANG Xin-Quan, CUI Hai-Tao, LI Sheng-Ping, TANG Ding-Zhong. Functional studies of rice blast resistance related gene OsSAMS1 [J]. Acta Agronomica Sinica, 2022, 48(5): 1119-1128.
[9] ZHU Zheng, WANG Tian-Xing-Zi, CHEN Yue, LIU Yu-Qing, YAN Gao-Wei, XU Shan, MA Jin-Jiao, DOU Shi-Juan, LI Li-Yun, LIU Guo-Zhen. Rice transcription factor WRKY68 plays a positive role in Xa21-mediated resistance to Xanthomonas oryzae pv. oryzae [J]. Acta Agronomica Sinica, 2022, 48(5): 1129-1140.
[10] WANG Xiao-Lei, LI Wei-Xing, OU-YANG Lin-Juan, XU Jie, CHEN Xiao-Rong, BIAN Jian-Min, HU Li-Fang, PENG Xiao-Song, HE Xiao-Peng, FU Jun-Ru, ZHOU Da-Hu, HE Hao-Hua, SUN Xiao-Tang, ZHU Chang-Lan. QTL mapping for plant architecture in rice based on chromosome segment substitution lines [J]. Acta Agronomica Sinica, 2022, 48(5): 1141-1151.
[11] WANG Ze, ZHOU Qin-Yang, LIU Cong, MU Yue, GUO Wei, DING Yan-Feng, NINOMIYA Seishi. Estimation and evaluation of paddy rice canopy characteristics based on images from UAV and ground camera [J]. Acta Agronomica Sinica, 2022, 48(5): 1248-1261.
[12] KE Jian, CHEN Ting-Ting, WU Zhou, ZHU Tie-Zhong, SUN Jie, HE Hai-Bing, YOU Cui-Cui, ZHU De-Quan, WU Li-Quan. Suitable varieties and high-yielding population characteristics of late season rice in the northern margin area of double-cropping rice along the Yangtze River [J]. Acta Agronomica Sinica, 2022, 48(4): 1005-1016.
[13] CHEN Yue, SUN Ming-Zhe, JIA Bo-Wei, LENG Yue, SUN Xiao-Li. Research progress regarding the function and mechanism of rice AP2/ERF transcription factor in stress response [J]. Acta Agronomica Sinica, 2022, 48(4): 781-790.
[14] WANG Lyu, CUI Yue-Zhen, WU Yu-Hong, HAO Xing-Shun, ZHANG Chun-Hui, WANG Jun-Yi, LIU Yi-Xin, LI Xiao-Gang, QIN Yu-Hang. Effects of rice stalks mulching combined with green manure (Astragalus smicus L.) incorporated into soil and reducing nitrogen fertilizer rate on rice yield and soil fertility [J]. Acta Agronomica Sinica, 2022, 48(4): 952-961.
[15] QIN Qin, TAO You-Feng, HUANG Bang-Chao, LI Hui, GAO Yun-Tian, ZHONG Xiao-Yuan, ZHOU Zhong-Lin, ZHU Li, LEI Xiao-Long, FENG Sheng-Qiang, WANG Xu, REN Wan-Jun. Characteristics of panicle stem growth and flowering period of the parents of hybrid rice in machine-transplanted seed production [J]. Acta Agronomica Sinica, 2022, 48(4): 988-1004.
Full text



No Suggested Reading articles found!