
Chinese Journal OF Rice Science >
Identification and Gene Cloning of a Long Sterile Lemma and Small Grain Mutant lsg8 in Rice (Oryza sativa L.)
Received date: 2024-10-11
Revised date: 2024-12-02
Online published: 2025-11-19
【Objective】 The aim of this study was to identify and clone a gene responsible for long sterile lemma and small grain size in rice, and to explore the genetic basis and molecular mechanism controlling these traits. 【Method】 A stably inherited mutant displaying long sterile lemmata and small grains was obtained from the japonica rice WYJ27 through ethyl methanesulfonate (EMS) mutagenesis. This mutant was named as lsg8 (long sterile lemma and small grain on chromosome 8). The agronomic traits of both the wild-type and the mutant were observed and recorded. The epidermal cells of the glume were observed using scanning electron microscopy (SEM). An F2 segregating population was generated by crossing lsg8 with the indica rice variety IR36, and used for genetic analysis and gene mapping. Candidate genes were further identified through gene sequencing and expression analysis. RT-qPCR was performed to analyze the relative expression levels of genes regulating cell expansion and floral development. 【Result】 Compared with the wild type, the mutant lsg8 exhibited a significantly longer sterile lemma, significantly decreased grain width and grain thickness, and consequently a reduced 1000-grain weight. In addition, the plant height, panicle length, the length of the first, second, and fourth internodes from the top, the number of primary branches, the number of secondary branches, the number of grains per panicle, and the seed setting rate of lsg8 were significantly lower than those of the wild type WYJ27. SEM observation of the outer epidermis of the glume showed that the cell length and width of the mutant lsg8 were significantly shorter and narrower than those of the wild type. Genetic analysis suggested that the mutant phenotype is controlled by a single recessive nuclear gene. Through map-based cloning, the LSG8 gene was mapped to a region between markers M5 and M6 on chromosome 8. The physical distance between these two markers is approximately 276 kb, containing 42 open reading frames (ORFs). Sequencing analysis revealed a single nucleotide difference in ORF18 (LOC_Os08g06480) between the wild type and the lsg8 mutant, indicating that ORF18 is likely the candidate gene controlling the long sterile lemma and small grain phenotype. RT-qPCR analysis revealed that LSG8 is expressed in various tissues at different developmental stages, with the highest expression in panicles at the mature stage and the lowest in leaf sheaths at the mature stage. Furthermore, significant alterations were observed in the relative expression levels of genes related to cell expansion, sterile lemma development, and floral organ identity in the lsg8 mutant. 【Conclusion】 The lsg8 mutant is a new allele of the previously reported ASP1 gene. The mutation in OsLSG8 results in a long sterile lemma and small grain phenotype, playing an important role in maintaining the morphogenesis of rice sterile lemma and grain morphology.
LU Shuai, TAO Tao, LIU Ran, ZHOU Wenyu, CAO Lei, YANG Qingqing, ZHANG Mingqiu, REN Xinzhe, YANG Zhidi, XU Fuxiang, HUAN Haidong, GONG Yuanhang, ZHANG Haocheng, JIN Sukui, CAI Xiuling, GAO Jiping, LENG Yujia . Identification and Gene Cloning of a Long Sterile Lemma and Small Grain Mutant lsg8 in Rice (Oryza sativa L.)[J]. Chinese Journal OF Rice Science, 2025 , 39(6) : 813 -824 . DOI: 10.16819/j.1001-7216.2025.241003
| [1] | 朱玲, 陈晓琼, 杜康兮, 韩保林, 冉秀华, 张红宇, 徐培洲, 吴先军. 水稻长护颖突变体基因的克隆与表达分析[J]. 中国农业科学, 2015, 48(11): 2085-2095. |
| Zhu L, Chen X Q, Du K X, Han B L, Ran X H, Zhang H Y, Xu P Z, Wu X J. Gene cloning and expression analysis of long empty glumes mutants in rice[J]. Scientia Agricultura Sinica, 2015, 48(11): 2085-2095. (in Chinese with English abstract) | |
| [2] | 符德保, 李燕, 肖景华, 张启发, 吴昌银. 中国水稻基因组学研究历史及现状[J]. 生命科学, 2016, 28(10): 1113-1121. |
| Fu D B, Li Y, Xiao J H, Zhang Q F, Wu C Y. The history and current status of rice genomics research in China[J]. Chinese Bulletin of Life Sciences, 2016, 28(10): 1113-1121. (in Chinese with English abstract) | |
| [3] | 张必东, 林泓, 朱思颖, 李忠成, 庄慧, 李云峰. 水稻颖壳异常突变体ah1的鉴定与候选基因分析[J]. 中国农业科学, 2024, 57(3): 429-441. |
| Zhang B D, Lin H, Zhu S Y, Li Z C, Zhuang H, Li Y F. Identification and candidate gene analysis of the ABNORMAL HULL 1 (ah1) mutant in rice (Oryza sativa L.)[J]. Scientia Agricultura Sinica, 2024, 57(3): 429-441. (in Chinese with English abstract) | |
| [4] | 罗曦, 魏林燕, 郑燕梅, 魏毅东, 连玲, 谢华安, 吴方喜. 水稻护颖发育相关基因的研究进展[J]. 南京农业大学学报2021, 44(3): 412-420. |
| Luo X, Wei L Y, Zheng Y M, Wei Y D, Lian L, Xie H A, Wu F X. Research advances on developmental genes of sterile lemmas in rice[J]. Journal of Nanjing Agricultural University, 2021, 44(3): 412-420. (in Chinese with English abstract) | |
| [5] | Ren D Y, Hu J. FZP determines grain size and sterile lemma fate in rice[J]. Journal of Experimental Botany, 2018, 69(20): 4853-4866. |
| [6] | Yoshida A, Suzaki T, Tanaka W, Hirano H Y. The homeotic gene long sterile lemma (G1) specifies sterile lemma identity in the rice spikelet[J]. Proceedings of the National Academy of Sciences of the United States of Ameica, 2009, 106 (47): 20103-20108. |
| [7] | Liu M, Li H, Su Y, Li W, Shi C. G1/ELE functions in the development of rice lemmas in addition to determining identities of empty glumes[J]. Frontiers in Plant Science, 2016, 7: 1006. |
| [8] | Kobayashi K, Maekawa M, Miyao A, Hirochika H, Kyozuka J. PANICLE PHYTOMER2 (PAP2), encoding a SEPALLATA subfamily MADS-box protein, positively controls spikelet meristem identity in rice[J]. Plant Cell Physiology, 2010, 51(1): 47-57. |
| [9] | Gao X, Liang W, Yin C, Ji S, Wang H, Su X, Guo C, Kong H, Xue H, Zhang D. The SEPALLATA-Like Gene OsMADS34 is required for rice inflorescence and spikelet development[J]. Plant Physiology, 2010, 153(2): 728-740. |
| [10] | Li W Q, Yoshida A, Takahashi M, Maekawa M, Kojima M, Sakakibara H, Kyozuka J. SAD1, an RNA polymerase Ⅰ subunit A34.5 of rice, interacts with mediator and controls various aspects of plant development[J]. The Plant Jouranl, 2015, 81: 282-291. |
| [11] | Xu Q K, Yu H P, Xia S S, Cui Y J, Yu X Q, Liu H, Zeng D L, Hu J, Zhang Q, Gao Z Y, Zhang G H, Zhu L, Shen L, Guo L B, Rao Y C, Qian Q, Ren D Y. The C2H2 zinc-finger protein LACKING RUDIMENTARY GLUME 1 regulates spikelet development in rice[J]. Science Bulletin, 2020, 65: 753-764. |
| [12] | Zhuang H, Wang H L, Zhang T, Zeng X Q, Chen H, Wang Z W, Zhang J, Zheng H, Tang J, Ling Y H, Yang Z L, He G H, Li Y F. NONSTOP GLUMES1 encodes a C2H2 zinc finger protein that regulates spikelet development in rice[J]. The Plant Cell, 2020, 32: 392-413. |
| [13] | Li H G, Xue D W, Gao Z Y, Yan M X, Xu W Y, Xing Z, Huang D N, Qian Q, Xue Y B. A putative lipase gene EXTRA GLUME1 regulates both empty-glume fate and spikelet development in rice[J]. The Plant Journal, 2009, 57(4): 593-605. |
| [14] | Yoshida A, Ohmori Y, Kitano H, Taguchi-Shiobara F, Hirano H Y. ABERRANT SPIKELET AND PANICLE1, encoding a TOPLESS-related transcriptional co-repressor, is involved in the regulation of meristem fate in rice[J]. The Plant Journal, 2012, 70(2): 327-339. |
| [15] | Yang J G, Lee S Y, Hang R L, Kim S R, Lee Y S, Cao X F, Amasino R, An G. OsVIL2 functions with PRC2 to induce flowering by repressing OsLFL1 in rice[J]. The Plant Journal, 2013, 73: 566-578. |
| [16] | Zhang J R, Tang W, Huang Y L, Niu X L, Zhao Y, Han Y, Liu Y S. Down-regulation of a LBD-like gene, OsIG1, leads to occurrence of unusual double ovules and developmental abnormalities of various floral organs and megagametophyte in rice[J]. Journal of Experimental Botany, 2015, 66(1): 99-112. |
| [17] | Lee D Y, Lee J, Moon S, Park S Y, An G. The rice heterochronic gene SUPERNUMERARY BRACT regulates the transition from spikelet meristem to floral meristem[J]. The Plant Journal, 2007, 49(1): 64-78. |
| [18] | Lee D Y, An G. Two AP2 family genes, SUPERNUMERARY BRACT (SNB) and OsINDETERMINATE SPIKELET1 (OsIDS1) synergistically control inflorescence architecture and floral meristem establishment in rice[J]. The Plant Journal, 2012, 69(3): 445-461. |
| [19] | Ren D Y, Li Y F, Zhao F M, Sang X C, Shi J Q, Wang N, Guo S, Ling Y H, Zhang C W, Yang Z L, He G H. MULTI-FLORET SPIKELET1, which encodes an AP2/ERF protein, determines spikelet meristem fate and sterile lemma identity in rice[J]. Plant Physiology, 2013, 162(2): 872-884. |
| [20] | Fan C C, Xing Y Z, Mao H L, Lu T T, Han B, Xu C G, Li X H, Zhang Q F. GS3, a major QTL for grain length and weight and minor QTL for grain width and thickness in rice, encodes a putative transmembrane protein[J]. Theoretical and Applied Genetics, 2006, 112(6): 1164-1171. |
| [21] | Mao H L, Sun S Y, Yao J L, Wang C R, Yu S B, Xu C G, Li X H, Zhang Q F. Linking differential domain functions of the GS3 protein to natural variation of grain size in rice[J]. Proceedings of the National Academy of Sciences of the United States of USA, 2010, 107: 19579-19584. |
| [22] | Sun S Y, Wang L, Mao H L, Shao L, Li X H, Xiao J. H, Ouyang Y D, Zhang Q F. A G-protein pathway determines grain size in rice[J]. Nature Communications, 2018, 9(1): 851. |
| [23] | Qi P, Lin Y S, Song X J, Shen J B, Huang W, Shan J X, Zhu M Z, Jiang L, Gao J P, Lin H X. The novel quantitative trait locus GL3.1 controls rice grain size and yield by regulating Cyclin-T1;3[J]. Cell Research, 2012, 22: 1666-1680. |
| [24] | Zhang X J, Wang J F, Huang J, Lan H X, Wang C L, Yin C F, Wu Y Y, Tang H J, Qian Q, Li J Y, Zhang H S. Rare allele of OsPPKL1 associated with grain length causes extra-large grain and a significant yield increase in rice[J]. Proceedings of the National Academy of Sciences of the United States of America, 2012, 109: 21534-21539. |
| [25] | Yu J P, Xiong H Y, Zhu X Y, Zhang H L, Li H H, Miao J L, Wang W S, Tang Z S, Zhang Z Y, Yao G X, Zhang Q, Pan Y H, Wang X, Rashid M A R, Li J J, Gao Y M, Li Z K, Yang W C, Fu X D, Li Z C. OsLG3 contributing to rice grain length and yield was mined by Ho-LAMap[J]. BMC Biology, 2017, 15: 28. |
| [26] | Ying J Z, Ma M, Bai C, Huang X H, Liu J L, Fan Y Y, Song X J. TGW3, a major QTL that negatively modulates grain length and weight in rice[J]. Molecular Plant, 2018, 11: 750-753. |
| [27] | Hu Z J, Lu S J, Wang M J, He H H, Sun L, Wang H R, Liu X H, Jiang L, Sun J L, Xin X Y, Kong W, Chu C C, Xue H W, Yang J S, Luo X J, Liu J X. A novel QTL qTGW3 encodes the GSK3/SHAGGY-Like kinase OsGSK5/OsSK41 that interacts with OsARF4 to negatively regulate grain size and weight in rice[J]. Molecular Plant, 2018, 11: 736-749. |
| [28] | Song X J, Kuroha T, Ayano M, Furuta T, Nagai K, Komeda N, Segami S, Miura K, Ogawa D, Kamura T, Suzuki T, Higashiyama T, Yamasaki M, Mori H, Inukai Y, Wu J Z, Kitano H, Sakakibara H, Jacobsen S E, Ashikari M. Rare allele of a previously unidentified histone H4 acetyltransferase enhances grain weight, yield, and plant biomass in rice[J]. Proceedings of the National Academy of Sciences USA, 2015, 112: 76-81. |
| [29] | Wang A H, Hou Q Q, Si L Z, Huang X H, Luo J H, Lu D F, Zhu J J, Shangguan Y Y, Miao J H, Xie Y F, Wang Y C, Zhao Q, Feng Q, Zhou C C, Li Y, Fan D L, Lu Y Q, Tian Q L, Wang Z X, Han B. The PLATZ transcription factor GL6 affects grain length and number in rice[J]. Plant Physiology, 2019, 180: 2077-2090. |
| [30] | Ishimaru K, Hirotsu N, Madoka Y, Murakami N, Hara N, Onodera H, Kashiwagi T, Ujiie K, Shimizu B I, Onishi A, Miyagawa H, Katoh E. Loss of function of the IAA-glucose hydrolase gene TGW6 enhances rice grain weight and increases yield[J]. Nature Genetics, 2013, 45(6): 707-711. |
| [31] | Wang S K, Li S, Liu Q, Wu K, Zhang J Q, Wang S S, Wang Y, Chen X B, Zhang Y, Gao C X, Wang F, Huang H X, Fu X D. The OsSPL16-GW7 regulatory module determines grain shape and simultaneously improves rice yield and grain quality[J]. Nature Genetics, 2015, 47: 949-954. |
| [32] | Wang Y X, Xiong G S, Hu J, Jiang L, Yu H, Xu J, Fang Y X, Zeng L J, Xu E B, Xu J, Ye W J, Meng X B, Liu R F, Chen H Q, Jing Y H, Wang Y H, Zhu X D, Li J Y, Qian Q. Copy number variation at the GL7 locus contributes to grain size diversity in rice[J]. Nature Genetics, 2015, 47: 944-948. |
| [33] | Si L Z, Chen J Y, Huang X H, Gong H, Luo J H, Hou Q Q, Zhou T Y, Lu T T, Zhu J J, Shangguan Y Y, Chen E W, Gong C X, Zhao Q, Jing Y F, Zhao Y, Li Y, Cui L L, Fan D L, Lu Y Q, Weng Q J, Wang Y C, Zhan Q L, Liu K Y, Wei X H, An K, An G, Han B. OsSPL13 controls grain size in cultivated rice[J]. Nature Genetics, 2016, 48: 447-456. |
| [34] | Song X J, Huang W, Shi M, Zhu M Z, Lin H X. A QTL for rice grain width and weight encodes a previously unknown RING-type E3 ubiquitin ligase[J]. Nature Genetics, 2007, 39: 623-630. |
| [35] | Hao J, Wang D, Wu Y, Huang K, Duan P G, Li N, Xu R, Zeng D L, Dong G J, Zhang B L, Zhang L M, Inzé D, Qian Q, Li Y H. The GW2-WG1-OsbZIP47 pathway controls grain size and weight in rice[J]. Molecular Plant, 2021, 8: 1266-1280. |
| [36] | Ruan B P, Shang L G, Zhang B, Hu J, Wang Y X, Lin H, Zhang A P, Liu C L, Peng Y L, Zhu L, Ren D Y, Shen L, Dong G J, Zhang G H, Zeng D L, Guo L B, Qian Q, Gao Z Y. Natural variation in the promoter of TGW2 determines grain width and weight in rice[J]. New Phytologist, 2020, 227: 629-640. |
| [37] | Li Y B, Fan C C, Xing Y Z, Jiang Y H, Luo L J, Sun L, Shao D, Xu C J, Li X H, Xiao J H, He Y Q, Zhang Q F. Natural variation in GS5 plays an important role in regulating grain size and yield in rice[J]. Nature Genetics, 2011, 43: 1266-1269. |
| [38] | Shomura A, Izawa T, Ebana K, Ebitani T, Kanegae H, Konishi S, Yano M. Deletion in a gene associated with grain size increased yields during rice domestication[J]. Nature Genetics, 2008, 40: 1023-1028. |
| [39] | Weng J F, Gu S H, Wan X Y, Gao H, Guo T, Su N, Lei C L, Zhang X, Cheng Z J, Guo X P, Wang J L, Jiang L, Zhai H Q, Wan J M. Isolation and initial characterization of GW5, a major QTL associated with rice grain width and weight[J]. Cell Research, 2008, 18: 1199-1209. |
| [40] | Duan P G, Xu J S, Zeng D L, Zhang B L, Geng M F, Zhang G Z, Huang K, Huang L J, Xu R, Ge S, Qian Q, Li Y H. Natural variation in the promoter of GSE5 contributes to grain size diversity in rice[J]. Molecular Plant, 2017, 10(5): 685-694. |
| [41] | Wang S K, Wu K, Yuan Q B, Liu X Y, Liu Z B, Lin X Y, Zeng R Z, Zhu H T, Dong G J, Qian Q, Zhang G Q, Fu X D. Control of grain size, shape and quality by OsSPL16 in rice[J]. Nature Genetics, 2012, 44: 950-954. |
| [42] | Rogers S O, Bendich A J. Extraction of DNA from milligram amounts of fresh, herbarium and mummified plant tissues[J]. Plant Molecular Biology, 1985, 5: 69-76. |
| [43] | Howarth W O. The Gramineae: A study of cereal, bamboo and grasses[J]. Nature, 1935, 136(3435): 317-319. |
| [44] | Kellogg E A. The evolutionary history of Ehrhartoideae, Oryzeae, and Oryza[J]. Rice, 2009, 2: 1-14. |
| [45] | Terrell E E, Peterson P M, Wergin W P. Epidermal features and spikelet micromorphology in Oryza and related genera (Poaceae: Oryzeae)[J]. Smithsonian Contributions to Botany, 2001, 91: 1-50. |
| [46] | Wu T K, Ali A, Wang J H, Song J H, Fang Y Q, Zhou T T, Luo Y, Zhang H Y, Chen X Q, Liao Y X, Liu Y T, Xu P Z, Wu J J. A homologous gene of OsREL2/ASP1, ASP-LSL regulates pleiotropic phenotype including long sterile lemma in rice[J]. BMC Plant Biology, 2021, 21: 390. |
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