玉米<i>ZmC<sub>4</sub>NADP-ME</i>基因进化及功能分析

doi:10.7668/hbnxb.20195463

华北农学报 ›› 2025, Vol. 40 ›› Issue (2): 10-17. doi: 10.7668/hbnxb.20195463

所属专题： 玉米；苹果；生物技术；

• 作物遗传育种·种质资源·生物技术 • 上一篇下一篇

玉米ZmC₄NADP-ME基因进化及功能分析

陈华峰¹^,²^,³, 张家宁⁴, 张晓¹, 袁月¹, 刘秀峰¹, 刘丹¹^,²^,

¹ 天津市农业科学院农作物研究所,天津市农作物遗传育种重点实验室,天津 300384
² 天津中天大地科技有限公司,天津 300384
³ 山东农业大学,山东泰安 066000
⁴ 河北科技师范学院,河北省作物逆境生物学重点实验室,河北秦皇岛 066000

收稿日期:2024-09-27 出版日期:2025-05-12
通讯作者:
刘丹(1989—),女,河北沧州人,正高级工程师,副研究员,博士,主要从事作物基因挖掘与功能验证研究。
作者简介:
陈华峰(1987—),男,山东泰安人,正高级工程师,博士,主要从事玉米遗传育种研究。
陈华峰、张家宁为同等贡献作者。
基金资助:
天津市青年基金项目(23JCQNJC00640); 河北科技师范学院科学研究基金项目(2024YB020); 津甘双地科技特派员项目(24CXNL009); 2024年度中国博士后科学基金会与天津市联合资助项目(2024T023TJ)

Evolution and Functional Analysis of Maize Gene ZmC₄NADP-ME

CHEN Huafeng¹^,²^,³, ZHANG Jianing⁴, ZHANG Xiao¹, YUAN Yue¹, LIU Xiufeng¹, LIU Dan¹^,²^,

¹ Tianjin Crop Research Institute,Tianjin Academy of Agriculture Sciences, Tianjin Key Laboratory of Crop Genetics and Breeding,Tianjin 300384,China
² Tianjin Zhongtiandadi Technology Company Limited,Tianjin 300384,China
³ Shandong Agricultural University,Tai'an 066000,China
⁴ Hebei Normal University of Science & Technology,Hebei Key Laboratory of Crop Stress Biology,Qinhuangdao 066000,China

Received:2024-09-27 Published:2025-05-12

摘要/Abstract

摘要：

为进一步了解光合作用关键基因的作用,获得了玉米光合作用暗反应限速酶编码基因ZmC₄NADP-ME的功能缺失突变体(zmC₄nadp-me)。通过构建不同物种同源基因的进化树,表明ZmC₄NADP-ME及其同源基因在大多数植物中都是以多拷贝的形式存在,且不同同源基因间的表达模式不同。对zmC₄nadp-me的表型分析发现,与野生型相比,zmC₄nadp-me整株呈黄绿色,光照条件下,苗期叶片很快干枯死亡。对zmC₄nadp-me叶片的叶绿素荧光分析发现,Y(Ⅱ)及电子传递速率ETR(Ⅱ)显著降低,Y(NPQ)变化较小;但Y(NO)值显著升高;对zmC₄nadp-me的PS Ⅰ吸收能力(P700)进行测定后发现,zmC₄nadp-me电子传递速率ETR(Ⅰ)和实际光电子效率Y(Ⅰ)均出现了大幅下降,并且伴随着光照强度的增强,差距越来越显著;光照强度小于870 μmol/(m²·s)时,zmC₄nadp-me的Y(ND)大于对照,当光照强度大于227 μmol/(m²·s),zmC₄nadp-me的Y(NA)也开始逐渐大于WT。以上结果表明,ZmC₄NADP-ME对植物生长发育是必需的,当该基因被破坏后,PSⅡ遭到了严重胁迫,而且在任何光照强度下,植株都无法通过提高Y(NPQ)来缓解这种抑制。同时,说明当光照低于一定强度时,来自PSⅠ电子供体侧的抑制可能是造成PSⅠ抑制的原因之一,而随着光照的增强,PSⅠ电子受体侧的抑制逐渐成为抑制PSⅠ的重要因素。

关键词: 玉米, 光合作用, 苹果酸酶, 光合系统Ⅰ, 光合系统Ⅱ

Abstract:

To explore the function of key genes in photosynthesis, a functional knockout mutant (zmC₄nadp-me) of ZmC₄NADP-ME, the gene encoding the rate-limiting enzyme of the dark reaction of photosynthesis in maize, was obtained. Evolutionary tree analysis showed that ZmC₄NADP-ME and its homologous genes exist in multiple copies in most plants, with diverse expression patterns. Phenotypic analysis revealed that the entire zmC₄nadp-me plant was yellow-green, and its seedling-stage leaves dried up and died rapidly under light. Chlorophyll fluorescence analysis indicated that Y(Ⅱ) and electron transport rate ETR(Ⅱ) of photosystem Ⅱ (PSⅡ) in zmC₄nadp-me decreased significantly, with little change in Y(NPQ), while the Y(NO) increased notably. Measurement of the absorption capacity (P700) of photosystem Ⅰ (PSⅠ) found that both the electron transport rate (ETR(Ⅰ)) and the actual photoelectron efficiency (Y(Ⅰ)) of zmC₄nadp-me dropped substantially, and the gap widened with increasing light intensity. Under specific light intensities, Y(ND) and Y(NA) of zmC₄nadp-me were greater than those of the wild type (WT). In conclusion, ZmC₄NADP-ME is essential for plant growth and development. Disruption of this gene severely stresses PSⅡ, and the plant can't alleviate this stress by increasing Y(NPQ). Meanwhile, at low light intensities, the inhibition of PSⅠ may originate from the electron donor side of PSⅠ, and as the light intensity increases, the inhibition from the electron acceptor side of PSⅠ becomes a key factor.

Key words: Maize, Photosynthesis, Malate enzyme, PS Ⅰ, PS Ⅱ

陈华峰, 张家宁, 张晓, 袁月, 刘秀峰, 刘丹. 玉米ZmC₄NADP-ME基因进化及功能分析[J]. 华北农学报, 2025, 40(2): 10-17. doi: 10.7668/hbnxb.20195463.

CHEN Huafeng, ZHANG Jianing, ZHANG Xiao, YUAN Yue, LIU Xiufeng, LIU Dan. Evolution and Functional Analysis of Maize Gene ZmC₄NADP-ME[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(2): 10-17. doi: 10.7668/hbnxb.20195463.

http://www.hbnxb.net/CN/Y2025/V40/I2/10

图/表 5

图1 玉米ZmC₄NADP-ME及其同源基因分析 A.玉米ZmC₄NADP-ME及其同源基因在染色体上的分布及共线性;B.玉米ZmC₄NADP-ME及其同源基因编码蛋白序列相似性;C.玉米ZmC₄NADP-ME及其同源基因在各组织中的表达水平;D.玉米ZmC₄NADP-ME及其同源基因在叶肉细胞和维管束鞘细胞中的表达水平。M.叶肉细胞;BS.维管束鞘细胞。

Fig.1 Analysis of ZmC₄NADP-ME and its homologous genes in maize A.The distribution and collinearity of ZmC₄NADP-ME and its homologous genes on maize chromosome; B.Maize ZmC₄NADP-ME and its homologous gene protein sequence similarity; C.Expression level of ZmC₄NADP-ME and its homologous genes in different tissues of maize; D.Expression level of ZmC₄NADP-ME and its homologous genes in mesophyll cells and bundle sheath cells of maize.M.Mesophyll cells;BS.Bundle sheath cells.

图2 ZmC₄NADP-ME及其同源基因进化树分析

Fig.2 Phylogenetic tree analysis of ZmC₄NADP-ME and its homologous genes

图3 玉米ZmC₄NADP-ME 突变体表型鉴定 A.玉米ZmC₄NADP-ME基因结构及mu插入位置,黑色方块代表外显子, 黑色线条代表内含子,TSS表示转录起始位点;B.玉米野生型(WT)和突变体zmC₄nadp-me表型。

Fig.3 Phenotype identification of maize ZmC₄NADP-ME mutant A.Maize ZmC₄NADP-ME gene structure and mu insertion site,black squares represent exons and black lines represent introns,TSS represents transcription start site; B.Maize wild-type (WT) and mutant zmC₄nadp-me phenotype.

图4 zmC₄nadp-me和野生型(WT)幼苗PSⅡ光响应曲线

Fig.4 PSⅡ light response curves of zmC₄nadp-me and wild-type (WT) seedlings

图5 zmC₄nadp-me和野生型(WT)幼苗PSⅠ光响应曲线

Fig.5 PSⅠ light response curves of zmC₄nadp-me and wild-type (WT) seedlings

参考文献 30

[1]	王金香, 张阿良, 秦敏, 曹江梅, 陈文, 郭瑞瑞. 镉胁迫对玉米幼苗光合特性及活性氧代谢的影响[J]. 天津农业科学, 2023, 29(1):1-6.doi:10.3969/j.issn.1006-6500.2023.01.001.
	Wang J X, Zhang A L, Qin M, Cao J M, Chen W, Guo R R. Effects of cadmium stress on photosynthetic characteristics and active oxygen metabolism of maize seedling[J]. Tianjin Agricultural Sciences, 2023, 29(1):1-6.
[2]	Swift J, Luginbuehl L H, Hua L, Schreier T B, Donald R M, Stanley S, Wang N, Lee T A, Nery J R, Ecker J R, Hibberd J M. Exaptation of ancestral cell-identity networks enables C4 photosynthesis[J]. Nature, 2024, 636(8041):143-150.doi:10.1038/s41586-024-08204-3.
[3]	Aubry S, Brown N J, Hibberd J M. The role of proteins in C3 plants prior to their recruitment into the C4 pathway[J]. Journal of Experimental Botany, 2011, 62(9):3049-3059.doi:10.1093/jxb/err012.
[4]	Langdale J A, Zelitch I, Miller E, Nelson T. Cell position and light influence C4 versus C3 patterns of photosynthetic gene expression in maize[J]. The EMBO Journal, 1988, 7(12): 3643-3651. doi:10.1002/j.1460-2075.1988.tb03245.x.
[5]	Langdale J A.C 4 cycles:past,present,and future research on C4 photosynthesis[J]. The Plant Cell, 2011, 23(11):3879-3892.doi:10.1105/tpc.111.092098. pmid: 22128120
[6]	Zhao H L, Wang Y, Lyu M A, Zhu X G. Two major metabolic factors for an efficient NADP-malic enzyme type C4 photosynthesis[J]. Plant Physiology, 2022, 189(1):84-98.doi:10.1093/plphys/kiac051.
[7]	Bellasio C, Lundgren M R. The operation of PEPCK increases light harvesting plasticity in C4 NAD ME and NADP ME photosynthetic subtypes:a theoretical study[J]. Plant,Cell & Environment, 2024, 47(6):2288-2309.doi:10.1111/pce.14869.
[8]	Badia M B, Mans R, Lis A V, Tronconi M A, Arias C L, Maurino V G, Andreo C S, Drincovich M F, van Maris A J A, Gerrard Wheeler M C. Specific Arabidopsis thaliana malic enzyme isoforms can provide anaplerotic pyruvate carboxylation function in Saccharomyces cerevisiae[J]. The FEBS Journal, 2017, 284(4):654-665.doi:10.1111/febs.14013.
[9]	Arias C L, Pavlovic T, Torcolese G, Badia M B, Gismondi M, Maurino V G, Andreo C S, Drincovich M F, Gerrard Wheeler M C, Saigo M. NADP-dependent malic enzyme 1 participates in the abscisic acid response in Arabidopsis thaliana[J]. Frontiers in Plant Science, 2018, 9:1637.doi:10.3389/fpls.2018.01637.
[10]	付振艳, 苟小清, 张正斌, 肖向文, 王晓军. 小麦TaNADP-ME2基因重组植物表达载体构建及对水稻的遗传转化[J]. 华北农学报, 2013, 28(3):58-61.doi:10.3969/j.issn.1000-7091.2013.03.011.
	Fu Z Y, Gou X Q, Zhang Z B, Xiao X W, Wang X J. The construction of plant overexpression vector of wheat TaNADP-ME2 gene and genetic transformation of rice[J]. Acta Agriculturae Boreali-Sinica, 2013, 28(3):58-61.
[11]	Badia M B, Maurino V G, Pavlovic T, Arias C L, Pagani M A, Andreo C S, Saigo M, Drincovich M F, Gerrard Wheeler M C. Loss of function of Arabidopsis NADP-malic enzyme 1 results in enhanced tolerance to aluminum stress[J]. The Plant Journal, 2020, 101(3):653-665.doi:10.1111/tpj.14571.
[12]	Brown N J, Palmer B G, Stanley S, Hajaji H, Janacek S H, Astley H M, Parsley K, Kajala K, Quick W P, Trenkamp S, Fernie A R, Maurino V G, Hibberd J M. C acid decarboxylases required for C photosynthesis are active in the mid-vein of the C species Arabidopsis thaliana,and are important in sugar and amino acid metabolism[J]. The Plant Journal, 2010, 61(1): 122-133. doi:10.1111/j.1365-313x.2009.04040.x.
[13]	Detarsio E, Maurino V G, Alvarez C E, Müller G L, Andreo C S, Drincovich M F. Maize cytosolic NADP-malic enzyme (ZmCytNADP-ME):a phylogenetically distant isoform specifically expressed in embryo and emerging roots[J]. Plant Molecular Biology, 2008, 68(4):355-367.doi:10.1007/s11103-008-9375-8.
[14]	Voll L M, Zell M B, Engelsdorf T, Saur A, Wheeler M G, Drincovich M F, Weber A P M, Maurino V G. Loss of cytosolic NADP-malic enzyme 2 in Arabidopsis thaliana is associated with enhanced susceptibility to Colletotrichum higginsianum[J]. New Phytologist, 2012, 195(1): 189-202. doi:10.1111/j.1469-8137.2012.04129.x.
[15]	Wheeler M C G, Tronconi M A, Drincovich M F, Andreo C S, Fluügge U I, Maurino V G. A comprehensive analysis of the NADP-malic enzyme gene family of Arabidopsis[J]. Plant Physiology, 2005, 139(1):39-51.doi:10.1104/pp.105.065953.
[16]	Suzuki K, Ohmori Y, Ratel E. High root temperature blocks both linear and cyclic electron transport in the dark during chilling of the leaves of rice seedlings[J]. Plant and Cell Physiology, 2011, 52(9):1697-1707.doi:10.1093/pcp/pcr104. pmid: 21803813
[17]	牛永昆, 李军乔, 曲俊儒, 李晨芹, 王鑫慈, 田甜. 铁胁迫对蕨麻叶绿素荧光及生理特性的影响[J]. 天津农业科学, 2022, 28(2):1-6.doi:10.3969/j.issn.1006-6500.2022.02.001.
	Niu Y K, Li J Q, Qu J R, Li C Q, Wang X C, Tian T. Effects of iron stress on chlorophyll fluorescence and physiological characteristics of Potentilla anserina L.[J]. Tianjin Agricultural Sciences, 2022, 28(2):1-6.
[18]	Tikkanen M, Grebe S. Switching off photoprotection of photosystem Ⅰ a novel tool for gradual PS Ⅰ photoinhibition[J]. Physiologia Plantarum, 2018, 162(2):156-161.doi:10.1111/ppl.12618.
[19]	Maxwell K, Johnson G N. Chlorophyll fluorescence-a practical guide[J]. Journal of Experimental Botany, 2000, 51(345):659-668.doi:10.1093/jxb/51.345.659. pmid: 10938857
[20]	姜晓君, 蒋英. 不同LED光质对黄瓜叶片叶绿素荧光、光合参数及SPAD值的影响[J]. 天津农业科学, 2019, 25(9):7-9.doi:10.3969/j.issn.1006-6500.2019.09.002.
	Jiang X J, Jiang Y. Effects of different LED light quality on chlorophyll fluorescence,photosynthetic parameters and SPAD in cucumber leaves[J]. Tianjin Agricultural Sciences, 2019, 25(9):7-9.
[21]	Earl H J, Ennahli S. Estimating photosynthetic electron transport via chlorophyll fluorometry without Photosystem Ⅱ light saturation[J]. Photosynthesis Research, 2004, 82(2):177-186.doi:10.1007/s11120-004-1454-3.
[22]	Chen H F, Qin L, Tian J G, Wang X L. Identification and evolutionary analysis of the GOLDEN 2-LIKE gene family in foxtail millet[J]. Tropical Plant Biology, 2022, 15(4):301-318.doi:10.1007/s12042-022-09324-8.
[23]	Hall B G. Building phylogenetic trees from molecular data with MEGA[J]. Molecular Biology and Evolution, 2013, 30(5):1229-1235.doi:10.1093/molbev/mst012. pmid: 23486614
[24]	Wang Y P, Tang H B, DeBarry J D, Tan X, Li J P, Wang X Y, Lee T H, Jin H Z, Marler B, Guo H, Kissinger J C, Paterson A H. MCScanX:a toolkit for detection and evolutionary analysis of gene synteny and collinearity[J]. Nucleic Acids Research, 2012, 40(7):e49.doi:10.1093/nar/gkr1293.
[25]	Dong P F, Tu X Y, Chu P Y, Lyu P T, Zhu N, Grierson D, Du B J, Li P H, Zhong S L. 3D chromatin architecture of large plant genomes determined by local A/B compartments[J]. Molecular Plant, 2017, 10(12):1497-1509.doi:10.1016/j.molp.2017.11.005. pmid: 29175436
[26]	Seader V H, Thornsberry J M, Carey R E. Utility of the Amborella trichopoda expansin superfamily in elucidating the history of angiosperm expansins[J]. Journal of Plant Research, 2016, 129(2):199-207.doi:10.1007/s10265-015-0772-1.
[27]	Project A G, Albert V A, Barbazuk W B, de Pamphilis C W, Der J P, Leebens-Mack J, et al. The Amborella genome and the evolution of flowering plants[J]. Science, 2013, 342(6165):1241089.doi:10.1126/science.1241089.
[28]	Zhang Y L, Giuliani R, Zhang Y J, Zhang Y, Araujo W L, Wang B C, Liu P, Sun Q, Cousins A, Edwards G, Fernie A, Brutnell T P, Li P H. Characterization of maize leaf pyruvate orthophosphate dikinase using high throughput sequencing[J]. Journal of Integrative Plant Biology, 2018, 60(8):670-690.doi:10.1111/jipb.12656.
[29]	Studer A J, Gandin A, Kolbe A R, Wang L, Cousins A B, Brutnell T P. A limited role for carbonic anhydrase in C4 photosynthesis as revealed by a ca1ca2 double mutant in maize[J]. Plant Physiology, 2014, 165(2):608-617.doi:10.1104/pp.114.237602. pmid: 24706552
[30]	Kalaji H M, Jajoo A, Oukarroum A, Brestic M, Zivcak M, Samborska I A, Cetner M D, Łukasik I, Goltsev V, Ladle R J, Dᶏbrowski P, Ahmad P. The use of chlorophyll fluorescence kinetics analysis to study the performance of photosynthetic machinery in plants[M]. Amsteradam:Elsevier, Emerging Technologies and Management of Crop Stress Tolerance, 2014, 2:347-384.doi:10.1016/B978-0-12-800875-1.00015-6.

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玉米ZmC4NADP-ME基因进化及功能分析

Evolution and Functional Analysis of Maize Gene ZmC4NADP-ME

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玉米ZmC₄NADP-ME基因进化及功能分析

Evolution and Functional Analysis of Maize Gene ZmC₄NADP-ME