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强光胁迫对转玉米C4型ZmPEPC+ZmPPDK基因小麦光合和生理特性的影响

方宇辉，齐学礼，李艳，张煜，彭超军，华夏，陈艳艳，郭瑞，胡琳，许为钢*   

  1. 河南省作物分子育种研究院 / 河南省小麦生物学重点实验室 / 神农种业实验室，河南郑州 450002
  • 收稿日期:2023-12-01 修回日期:2024-01-30 接受日期:2024-01-30 网络出版日期:2024-02-20
  • 基金资助:
    本研究由河南省农业科学院自主创新基金项目(2023ZC090), 河南省科技攻关项目(232102110203), 神农种业实验室“一流课题”项目(SN01-2022-01), 农业生物育种重大项目(2023ZD0402302)和财政部和农业农村部国家现代农业产业技术体系建设专项(CARS-03-7)资助。

Effects of high light stress on photosynthesis and physiological characteristics of wheat with C4-type ZmPEPC+ZmPPDK gene#br#

FANG Yu-Hui,QI Xue-Li, LI Yan,ZHANG Yu,PENG Chao-Jun,HUA Xia,CHEN Yan-Yan,GUO Rui,HU Lin,XU Wei-Gang*   

  1. Institute of Crop Molecular Breeding, Henan Academy of Agricultural Sciences / Key Laboratory for Wheat Biology of Henan Province / Shennong Laboratory, Zhengzhou, Henan 450002, China
  • Received:2023-12-01 Revised:2024-01-30 Accepted:2024-01-30 Published online:2024-02-20
  • Supported by:
    This study was supported by the Independent Innovation Foundation of Henan Academy of Agricultural Sciences (2023ZC090), the Henan Provincial Science and Technology Research Project (232102110203), the ‘First-class Project’ of Shennong Laboratory (SN01-2022-01), the Major Project of Agricultural Biological Breeding (2023ZD0402302), and the China Agriculture Research System of MOF and MARA (CARS-03-7).

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摘要：

为研究转玉米C4型PEPC (磷酸烯醇式丙酮酸羧化酶基因)和PPDK(丙酮酸磷酸双激酶基因)双基因小麦对强光胁迫的光合和生理响应，以转ZmPEPC+ZmPPDK基因小麦株系PCK30和PCK60及其野生型对照材料(WT)为试验材料，鉴定了外源基因在转基因小麦中的表达量，在抽穗期和灌浆期测定正常光强(NL)和强光胁迫(HL)处理下转基因小麦的光合酶活性、叶绿素含量、气体交换参数、叶绿素荧光参数、活性氧物质含量和抗氧化酶活性。结果表明，2个转基因株系在转录水平上高效表达了PEPC和PPDK基因。在不同时期NL和HL处理下，转基因小麦的PEPC、PPDK、NADP-ME和Rubisco的酶活均显著高于WT，且HL处理下高出WT的幅度更明显。与NL处理相比，转基因小麦和WT的叶绿素含量在HL处理下显著降低，但转基因株系的下降幅度更小，且HL处理下转基因株系的叶绿素含量显著高于WT。两种水平处理下，转基因小麦PCK30、PCK60的净光合速率(Pn)均显著高于WT，且HL处理下高出幅度更明显，抽穗期增幅分别为15.26%和17.57%，灌浆期为13.41%和15.82%。气孔导度、Fv/Fm、qp的变化趋势与Pn一致，胞间二氧化碳浓度的变化趋势与Pn相反。转基因株系在HL处理下产生的活性氧物质和丙二醛含量显著低于WT，而抗氧化酶变化趋势与之相反。连续两年田间小区产量试验，转基因小麦PCK30和PCK60平均比WT高8.37%和10.16%。PEPC和PPDK在小麦中的过表达增强了小麦内源的光合酶、光化学效率和抗氧化酶活性，增强了强光下的叶片细胞膜的稳定性，保护了光合机构，维持了较高的光合效率，从而提高了转基因小麦的耐强光胁迫能力。

关键词: 小麦, 玉米C4光合基因, 强光胁迫, 光合, 生理特性

Abstract:

To study the photosynthetic and physiological response of maize C4-type PEPC (phosphoenolpyruvate carboxylase gene) and PPDK (pyruvate phosphate double kinase gene) dual-gene wheat to high light, the ZmPEPC + ZmPPDK wheat lines PCK30 and PCK60 and their wild type control material (WT) were used as experimental materials. The relative expression levels of exogenous genes in transgenic wheat were identified. The photosynthetic enzyme activity, chlorophyll content, gas exchange parameters, chlorophyll fluorescence parameters, active oxygen content, and antioxidant enzyme activity of transgenic wheat were measured under normal light intensity (NL) and high light stress (HL) at heading and grain-filling stages. The results showed that the two transgenic lines expressed the PEPC and PPDK genes efficiently at the transcriptional level. The enzyme activity of PEPC, PPDK, NADP-ME, and Rubisco of transgenic wheat was significantly higher than WT under NL or HL stress at different stage, and the increase of WT was more obvious under HL stress. Compared with NL stress, the chlorophyll content of transgenic wheat and WT decreased significantly under HL stress, but the decrease of transgenic lines was smaller, and the chlorophyll content of transgenic lines was significantly higher than WT under HL stress. Under two treatment stresses, the net photosynthetic rate (Pn) of PCK30 and PCK60 was significantly higher than WT, and the higher range was more obvious under HL stress. The increases were 15.26% and 17.57% at heading stage and 13.41% and 15.82% at grain-filling stage, respectively. The variation trend of stomatal conductance, Fv/Fm and qp was consistent with Pn, while the variation trend of intercellular carbon dioxide concentration was opposite to Pn. The content of reactive oxygen species and malondialdehyde produced by transgenic lines under HL stress was significantly lower than WT, while the trend of antioxidant enzymes was opposite. In two consecutive years of field plot yield experiments, PCK30 and PCK60 were 8.37% and 10.16% higher than WT on average. The overexpression of PEPC and PPDK in wheat enhanced the endogenous photosynthetic enzyme, photochemical efficiency, and antioxidant enzyme activities of wheat, enhanced the stability of leaf cell membrane under high light stress, protected the photosynthetic apparatus, and maintained a high photosynthetic efficiency, thus improving the tolerance of transgenic wheat to high light stress.

Key words: wheat, maize C4-type photosynthetic gene, high light stress, photosynthesis, physiological characteristics

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