OsGA3ox通过合成不同活性GA调控水稻育性及株高
收稿日期: 2023-03-15
修回日期: 2023-06-14
网络出版日期: 2023-07-19
基金资助
国家自然科学基金面上项目(32071930);湖南省自然科学基金面上项目(2022JJ30421);湖南省自然科学基金面上项目(2021JJ30445);长沙市自然科学基金项目(kq2014076)
OsGA3ox genes regulate rice fertility and plant height by synthesizing diverse active GA
Received date: 2023-03-15
Revised date: 2023-06-14
Online published: 2023-07-19
Supported by
National Natural Science Foundation of China(32071930);Hunan Province Natural Science Foundation Project(2022JJ30421);Hunan Province Natural Science Foundation Project(2021JJ30445);Changsha City Natural Science Foundation Project(kq2014076)
赤霉素(gibberellin，GA)是一种重要的激素，参与调控植物多种生长发育过程。GA生物合成通路已基本阐明，其中赤霉素3β羟化酶(gibberellin 3β-hydroxylase，GA3ox)是多种活性GA合成的关键酶。水稻中有2个GA3ox基因(OsGA3ox1和OsGA3ox2)，其生理功能虽有初步研究，但它们在合成活性GA调控水稻发育过程中是如何分工协作尚不清楚。本研究通过CRISPR/Cas9技术获得基因编辑突变体ga3ox1和ga3ox2，发现ga3ox1花粉育性显著下降，而ga3ox2株高显著变矮，表明OsGA3ox1是花粉正常发育必需的，而OsGA3ox2是茎叶伸长必需的。组织表达分析表明，OsGA3ox1主要在未开的花中表达，OsGA3ox2主要在未伸长的叶中表达。进一步对野生型(WT)和两个ga3ox突变体未开的花、未伸长的叶及根中的GA进行检测分析，发现OsGA3ox1在花中催化GA9形成GA7与花粉育性密切相关；OsGA3ox2在未伸长的叶中催化GA20形成GA1调控株高；OsGA3ox1在根中催化GA19形成GA20，调控GA3的生成。总之，OsGA3ox1和OsGA3ox2响应发育信号，在不同组织分工协作合成内源GA，调控水稻生长和发育。本研究为阐明OsGA3ox1和OsGA3ox2在GA生物合成中的作用以及深入理解OsGA3ox的功能提供参考。
关键词： 水稻; 赤霉素; 赤霉素3β羟化酶基因; 功能分析
郝小花, 胡爽, 赵丹, 田连福, 谢子靖, 吴莎, 胡文俐, 雷晗, 李东屏 . OsGA3ox通过合成不同活性GA调控水稻育性及株高[J]. 遗传, 2023 , 45(9) : 845 -855 . DOI: 10.16288/j.yczz.23-058
Gibberellin (GA) is an important hormone, which is involved in regulating various growth and development. GA biosynthesis pathway and synthetase have been basically clarified. Gibberellin 3β hydroxylase (GA3ox) is the key enzyme for the synthesis of various active GA. There are two GA3ox genes (OsGA3ox1 and OsGA3ox2) in rice, and their physiological functions have been preliminarily studied. However, it is not clear how they work together to synthesize active GA to regulate rice development. In this study, the knockout mutants ga3ox1 and ga3ox2 were obtained by CRISPR/Cas9 technology. The pollen fertility of ga3ox1 decreased significantly, while the plant height of ga3ox2 decreased significantly. It shows that OsGA3ox1 is necessary for normal pollen development, while OsGA3ox2 is necessary for stem and leaf elongation. Tissue expression analysis showed that OsGA3ox1 was mainly expressed in unopened flowers, while OsGA3ox2 was mainly expressed in unexpanded leaves. The GA in different tissues of wild type (WT), and two ga3ox mutants were detected. It was found that pollen fertility is most closely related to the content of GA7, and plant height is most closely related to the content of GA1. It was found that OsGA3ox1 catalyzes GA9 to GA7 in flowers, which is closely related to pollen fertility; OsGA3ox2 catalyzes the GA20 to GA1 in unexpanded leaves, thereby regulating plant height; OsGA3ox1 catalyzes the GA19 to GA20 in roots, regulating the generation of GA3. OsGA3ox1 and OsGA3ox2 respond to developmental and environmental signals, and cooperate to synthesize endogenous GA in different tissues to regulate rice development. This study provides a reference for clarifying its role in GA biosynthesis pathway and further understanding the function of OsGA3ox.
Key words： Oryza sativa L.; gibberellin (GA); OsGA3ox; functional analysis
| [1] | Gao SP, Chu CC. Gibberellin metabolism and signaling: targets for improving agronomic performance of crops. Plant Cell Physiol, 2020, 61(11): 1902-1911. |
| [2] | Jin Y, Song XY, Chang HZ, Zhao YY, Cao CH, Qiu XB, Zhu J, Wang ET, Yang ZN, Yu N. The GA-DELLA- OsMS188 module controls male reproductive development in rice. New Phytol, 2022, 233(6): 2629-2642. |
| [3] | Mo WP, Tang WJ, Du YX, Jing YJ, Bu QY, Lin RC. PHYTOCHROME-INTERACTING FACTOR-LIKE14 and SLENDER RICE1 interaction controls seedling growth under salt stress. Plant Physiol, 2020, 184(1): 506-517. |
| [4] | Li LL, Zhang HH, Yang ZH, Wang C, Li SS, Cao C, Yao TS, Wei ZY, Li YJ, Chen JP, Sun ZT. Independently evolved viral effectors convergently suppress DELLA protein SLR1-mediated broad-spectrum antiviral immunity in rice. Nat Commun, 2022, 13(1): 6920. |
| [5] | Tang JQ, Tian XJ, Mei EY, He ML, Gao JW, Yu J, Xu M, Liu JL, Song L, Li XF, Wang ZY, Guan QJ, Zhao ZG, Wang CM, Bu QY. WRKY53 negatively regulates rice cold tolerance at the booting stage by fine-tuning anther gibberellin levels. Plant Cell, 2022, 4(11): 4495-4515. |
| [6] | Sun HW, Guo XL, Zhu XL, Gu PY, Zhang W, Tao WQ, Wang DJ, Wu YZ, Zhao QZ, Xu GH, Fu XD, Zhang YL. Strigolactone and gibberellin signaling coordinately regulate metabolic adaptations to changes in nitrogen availability in rice. Mol Plant, 2023, 16(3): 588-598. |
| [7] | Eriksson S, B?hlenius H, Moritz T, Nilsson O. GA4 is the active gibberellin in the regulation of LEAFY transcription and Arabidopsis floral initiation. Plant Cell, 2006, 18(9): 2172-2181. |
| [8] | Murakami Y. Endogenous gibberellins in the panicle of rice plants determined by the rice seedling bioassay. Jarq-Jpn Agr Res Q, 1983, 17(3): 154-160. |
| [9] | Kobayashi M, Sakurai A, Saka H, Takahashi N. Quantitative analysis of endogenous gibberellins in normal and dwarf cultivars of rice. Plant Cell Physiol, 1989, 30(7): 963-969. |
| [10] | Kobayashi M, Yamaguchi I, Murofushi N, Ota Y, Takahashi N. Fluctuation and localization of endogenous gibberellins in rice. Agr Biol Chem, 1988, 5(52): 1189-1194. |
| [11] | Hasegawa M, Nakajima M, Takeda K, Yamaguchi I, Murofushi N. Endogenous levels of gibberellins in normal and male sterile anthers of rice (Oryza sativa cv. Nihonmasari). Biosci Biotech Bioch, 1995, 59(9): 1716-1720. |
| [12] | Honda I, Iwasaki S, Sudo K, Kato H, Maruyama K, Hasegawa M, Yamaguchi I, Murofushi N, Yanagisawa T, Takahashi N. Semi quantification of gibberellins in the anthers of thermosensitive genetic male sterile rice (Oryza sativa L. cv.PL12). Biosci Biotech Bioch, 1997, 61(10): 1748-1750. |
| [13] | Sakamoto T, Miura K, Itoh H, Tatsumi T, Ueguchi-Tanaka M, Ishiyama K, Kobayashi M, Agrawal GK, Takeda S, Abe K, Miyao A, Hirochika H, Kitano H, Ashikari M, Matsuoka M. An overview of gibberellin metabolism enzyme genes and their related mutants in rice. Plant Physiol, 2004, 134(4): 1642-1653. |
| [14] | Ueguchi-Tanaka M, Ashikari M, Nakajima M, Itoh H, Katoh E, Kobayashi M, Chow TY, Hsing YIC, Kitano H, Yamaguchi I, Matsuoka M. GIBBERELLIN INSENSITIVE DWARF1 encodes a soluble receptor for gibberellin. Nature, 2005, 437(7059): 693-698. |
| [15] | Ueguchi-Tanaka M, Nakajima M, Katoh E, Ohmiya H, Asano K, Saji S, Hongyu X, Ashikari M, Kitano H, Yamaguchi I, Matsuoka M. Molecular interactions of a soluble gibberellin receptor, GID1, with a rice DELLA protein, SLR1, and gibberellin. Plant Cell, 2007, 19(7): 2140-2155. |
| [16] | Gomi K, Sasaki A, Itoh H, Ueguchi-Tanaka M, Ashikari M, Kitano H, Matsuoka M. GID2, an F-box subunit of the SCF E3 complex, specifically interacts with phosphorylated SLR1 protein and regulates the gibberellin- dependent degradation of SLR1 in rice. Plant J, 2004, 37(4): 626-634. |
| [17] | Olszewski N, Sun TP, Gubler F. Gibberellin signaling: biosynthesis, catabolism, and response pathways. Plant Cell, 2002, 14(Suppl): S61-S80. |
| [18] | Lo SF, Yang SY, Chen KT, Hsing YI, Zeevaart JAD, Chen LJ, Yu SM. A novel class of gibberellin 2-oxidases control semidwarfism, tillering, and root development in rice. Plant Cell, 2008, 20(10): 2603-2618. |
| [19] | Qin X, Liu JH, Zhao WS, Chen XJ, Guo ZJ, Peng YL. Gibberellin 20-oxidase gene OsGA20ox3 regulates plant stature and disease development in rice. Mol Plant Microbe Interact, 2013, 26(2): 227-239. |
| [20] | Spielmeyer W, Ellis MH, Chandler PM. Semidwarf(sd-1), “green revolution” rice, contains a defective gibberellin 20-oxidase gene. Proc Natl Acad Sci USA, 2002, 99(13): 9043-9048. |
| [21] | Itoh H, Ueguchi-Tanaka M, Sentoku N, Kitano H, Matsuoka M, Kobayashi M. Cloning and functional analysis of two gibberellin 3 beta-hydroxylase genes that are differently expressed during the growth of rice. Proc Natl Acad Sci USA, 2001, 98(5): 8909-8914. |
| [22] | Murakami Y. Dwarfing genes in rice and their relation to glbberellin biosynthesis. In: Carr DJ, Plant Growth Substances. Springer-Verlag, Berlin, Germany, eds. 1970, 166-174. |
| [23] | Hu SK, Hu XM, Hu J, Shang LG, Dong GJ, Zeng DL, Gou LB, Qian Q. Xiaowei, a new rice germplasm for large- scale indoor research. Mol Plant, 2018, 11(11): 1418-1420. |
| [24] | Kobayashi M, Kamiya Y, Sakurai A, Saka H, Takahashi N. Metabolism of gibberellins in cell-free extracts of anthers from normal and dwarf rice. Plant Cell Physiol, 1990, 31(2): 289-293. |
| [25] | Miao J, Guo DS, Zhang JZ, Huang QP, Qin GJ, Zhang X, Wan JM, Gu HY, Qu LJ. Targeted mutagenesis in rice using CRISPR-Cas system. Cell Res, 2013, 23(10): 1233-1236. |
| [26] | Li J, Huang Y, Tan H, Yang X, Tian LF, Luan S, Chen LB, Li DP. An endoplasmic reticulum magnesium transporter is essential for pollen development in Arabidopsis. Plant Sci, 2015, 231: 212-220. |
| [27] | Chen ML, Fu XM, Liu JQ, Ye TT, Hou SY, Huang YQ, Yuan BF, Wu Y, Feng YQ. Highly sensitive and quantitative profiling of acidic phytohormones using derivatization approach coupled with nano-LC-ESI-Q- TOF-MS analysis. J Chromatogr B, 2012, 905: 67-74. |
| [28] | Uozu S, Tanaka-Ueguchi M, Kitano H, Hattori K, Matsuoka M. Characterization of XET-related genes of rice. Plant Physiol, 2000, 122(3): 853-859. |
| [29] | Lee Y, Kende H. Expression of β-expansins is correlated with internodal elongation in deepwater rice. Plant Physiol, 2001, 127(2): 645-654. |
| [30] | Lee Y, Kende H. Expression of α-expansin and expansin- like genes in deepwater rice. Plant Physiol, 2002, 130(3): 1396-1405. |
| [31] | Hao XH, Xiang YT, Zeng M, Yang YW, Xie P, Li M, Li DP, Tian LF. Cytological characteristics and gene location of a rice Dwarf mutant. Life Sci Res, 2021, 25(1): 39-47. |
| [31] | 郝小花, 向玉婷, 曾孟, 杨友伟, 谢鹏, 李密, 李东屏, 田连福. 水稻矮杆突变体的细胞学特征及基因定位研究. 生命科学研究, 2021, 25(1): 39-47. |
| [32] | Lu YZ, Feng Z, Meng YL, Bian LY, Xie H, Mysore KS, Liang JS. SLENDER RICE1 and Oryza sativa INDETERMINATE DOMAIN2 regulating OsmiR396 are involved in stem elongation. Plant Physiol, 2020, 182(4): 2213-2227. |
| [33] | Kuroha T, Nagai K, Gamuyao R, Wang DR, Furuta T, Nakamori M, Kitaoka T, Adachi K, Minami A, Mori Y, Mashiguchi K, Seto Y, Yamaguchi S, Kojima M, Sakakibara H, Wu JZ, Ebana K, Mitsuda N, Ohme-Takagi M, Yanagisawa S, Yamasaki M, Yokoyama R, Nishitani K, Mochizuki T, Tamiya G, McCouch SR, Ashikari M. Ethylene-gibberellin signaling underlies adaptation of rice to periodic flooding. Science, 2018, 361(6398): 181-186. |
| [34] | Cheng JY, Hill CB, Shabala S, Li CD, Zhou MX. Manipulating GA-related genes for cereal crop improvement. Int J Mol Sci, 2022, 23(22): 14046. |
| [35] | Cheng XY, Huang YP, Tan Y, Tan L, Yin JH, Zou GX. Potentially useful dwarfing or semi-dwarfing genes in rice breeding in addition to the sd1 gene. Rice (N Y), 2022, 15(1): 66. |
/
| 〈 |
|
〉 |
www.chinagene.cn
备案号：京ICP备09063187号-4
总访问:,今日访问:,当前在线:
