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    武霞, 张一南, 赵楠, 张莹, 赵瑞, 李金克, 周晓阳, 陈少良. 过表达胡杨PeAnn1负调控拟南芥的抗旱性[J]. 北京林业大学学报, 2020, 42(6): 14-25. DOI: 10.12171/j.1000-1522.20200031
    引用本文: 武霞, 张一南, 赵楠, 张莹, 赵瑞, 李金克, 周晓阳, 陈少良. 过表达胡杨PeAnn1负调控拟南芥的抗旱性[J]. 北京林业大学学报, 2020, 42(6): 14-25. DOI: 10.12171/j.1000-1522.20200031
    Wu Xia, Zhang Yinan, Zhao Nan, Zhang Ying, Zhao Rui, Li Jinke, Zhou Xiaoyang, Chen Shaoliang. Overexpression of PeAnn1 from Populus euphratica negatively regulates drought resistance in transgenic Arabidopsis thaliana[J]. Journal of Beijing Forestry University, 2020, 42(6): 14-25. DOI: 10.12171/j.1000-1522.20200031
    Citation: Wu Xia, Zhang Yinan, Zhao Nan, Zhang Ying, Zhao Rui, Li Jinke, Zhou Xiaoyang, Chen Shaoliang. Overexpression of PeAnn1 from Populus euphratica negatively regulates drought resistance in transgenic Arabidopsis thaliana[J]. Journal of Beijing Forestry University, 2020, 42(6): 14-25. DOI: 10.12171/j.1000-1522.20200031

    过表达胡杨PeAnn1负调控拟南芥的抗旱性

    Overexpression of PeAnn1 from Populus euphratica negatively regulates drought resistance in transgenic Arabidopsis thaliana

    • 摘要:
      目的Annexins是原核生物和真核生物中普遍存在的一大类膜联蛋白家族,能够参与氧化胁迫、热胁迫、干旱胁迫和盐胁迫等许多胁迫响应过程。但在胡杨中,对膜联蛋白家族在抗逆性中的作用情况还缺乏了解。本文研究胡杨Anneixn1在植物耐受渗透胁迫和干旱中的作用。
      方法本研究对渗透胁迫诱导的PeAnn1基因表达进行检测,对PeAnn1进行相关生物信息学分析,与毛白杨、拟南芥、大豆和水稻膜联蛋白基因家族成员进行序列比对和系统进化树分析,以过表达PeAnn1拟南芥(PeAnn1-OE1和PeAnn1-OE2)、Annexin1突变体(atann1)和野生型拟南芥(WT)为实验材料,利用不同浓度甘露醇处理(0、150、200、250、300 mmol/L)模拟渗透胁迫,并对各株系进行土壤干旱和复水处理,测定了不同处理株系的萌发率、根长、叶绿素含量及荧光参数、过氧化氢含量、抗氧化酶活性和基因表达等指标,分析了不同基因型拟南芥的抗旱性。
      结果短期渗透胁迫处理诱导了胡杨叶片中PeAnn1基因的上调表达。PeAnn1基因序列与毛白杨PtAnn1相似度最高,与PtAnn1亲缘关系较近。在甘露醇培养基上,过表达PeAnn1拟南芥的生存率和根长生长受到明显的抑制,且随着甘露醇浓度的升高,差异显著(P < 0.05)。土壤干旱8 d后测得的转基因拟南芥的叶绿素SPAD值、PSⅡ最大光量子效率(Fv/Fm)、实际光合量子产量(ΦPSⅡ)和相对电子传递速率(ETR)均显著低于野生型和突变体(P < 0.05)。复水后,PeAnn1转基因拟南芥光合参数的恢复程度也较低。在渗透胁迫下,转基因植株抗氧化物酶SOD、POD、CAT的活性以及编码基因的表达量显著低于野生型和突变体,不能清除过多活性氧,导致氧化伤害。
      结论以上结果表明,过表达PeAnn1降低了拟南芥对水分逆境的抗性。

       

      Abstract:
      ObjectiveAnnexins is a large class of annexin families common in prokaryotes and eukaryotes, and participates in many stress response processes such as oxidative stress, heat stress, drought stress and salt stress. However, the role of Populus euphratica annexin family genes in adapting to adverse conditions is still less known. This paper studies the role of P. euphratica Anneixn1 in osmotic stress and drought resistance.
      MethodThe mannitol-induced expression of PeAnn1 was examined in P. euphratica leaves. PeAnn1 sequence was compared with annexin genes from P. tomentosa, Arabidopsis thaliana, Glycine soja, and Oryza sative. Phylogenetic tree of annexins was constructed using MEGA 6 software. PeAnn1-overexpressed A. thaliana (PeAnn1-OE1 and PeAnn1-OE2), Annexin1 mutant (atann1) and wild-type A. thaliana (WT) were used in this study. Plants of each genotype were subjected to increasing osmotic stress caused by different concentrations of mannitol (0, 150, 200, 250, 300 mmol/L), drought, and subsequent water recovery. During the period of water stress, the germination rate, root length, chlorophyll content, and fluorescence parameters, H2O2, activity of antioxidant enzymes and expression of encoding genes were examined.
      ResultThe short-term mannitol treatment up-regulated the expression of PeAnn1 in P. euphratica leaves. The PeAnn1 sequence displayed higher similarity to P. tomentosa Ann1 (PtAnn1) than other plant species. Phylogenetic analysis revealed that the PeAnn1 was highly homologous to the PtAnn1. The survival rate and root growth of A. thaliana plants were inhibited with increasing concentrations of mannitol, and the inhibition was more pronounced in PeAnn1-overexpressing lines as compared to the WT and mutant (P < 0.05). All the measured photosynthesis parameters, such as chlorophyll content, the maximum photon efficiency of PSⅡ (Fv/Fm), the actual photosynthetic quantum yield (ΦPSⅡ), and the relative electron transfer rate (ETR) were lower in the transgenic plants than in the WT and mutant (P < 0.05). After rehydration, the recovery degree of photosynthetic parameters of PeAnn1-transgenic A. thaliana was also lower than the WT. Under osmotic stress, the activities of antioxidant enzymes, such as SOD, POD, and CAT were significantly lower in PeAnn1-overexpressed plants than those of the WT and mutant. In accordance, the expression of these antioxidant enzyme genes showed a same trend as the activities, which could not remove reactive oxygen species, causing oxidative damage.
      ConclusionThe above results indicate that overexpression of PeAnn1 decreases the capacity of A. thaliana in the adaptation to water deficit conditions.

       

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