马尾松PAP1基因在烟草上的遗传转化及功能分析

发布时间：2018-07-26 17:46

【摘要】：植物紫色酸性磷酸酶(purple acid phosphatase,PAP)在植物响应低磷胁迫过程中起重要作用,为了验证马尾松(Pinus massoniana)紫色酸性磷酸酶(PmPAP1)基因在磷吸收上的功能,本研究通过农杆菌(Agrobacterium tumefaciens)介导法转化烟草(Nicotiana tabacum),获得25个转基因株系,RT-PCR及qRTPCR分析表明,PmPAP1在13个株系中高表达;对其中2个高表达株系(Line-10和Line-13)进行不同浓度磷处理,结果表明,缺磷可以诱导转基因植株中PmPAP1基因在根和叶中上调表达;低磷条件下,转基因植株根系及叶片酸性磷酸酶(acid phosphatase,APase)活性分别为野生型(wild type,WT)的1.27和1.25倍;转基因植株总磷与无机磷含量分别比WT提高了28.32%、45.48%(Line-10)与36.45%、62.31%(Line-13)。除此之外,缺磷条件下转基因植株过氧化物酶(peroxidase,POD)、超氧化物歧化酶(superoxide dismutase,SOD)均较WT显著提高(P0.05),而丙二醛(malondialdehyde,MDA)含量则显著下降(P0.05)。磷胁迫对植株生长影响表明,低磷条件下转基因植株总生物量、根系生物量及根冠比均显著提高(P0.05)。而在正常和高磷条件下,转基因与野生型烟草的酶活性、丙二醛含量、磷含量和生物量等与均无明显差异。因此,高表达PmPAP1基因能显著提高烟草植株在低有效磷下磷的利用能力,进而增强耐低磷胁迫能力。本研究为深入了解马尾松耐低磷的分子机制与创制耐低磷烟草新种质提供了理论依据。
[Abstract]:Plant purple acid phosphatase (purple acid) plays an important role in response to low phosphorus stress. In order to verify the function of (Pinus massoniana) purple acid phosphatase (PmPAP1) gene in phosphorus uptake of Pinus massoniana. RT-PCR and qRTPCR analysis of 25 transgenic lines obtained by Agrobacterium tumefaciens (Agrobacterium tumefaciens) mediated transformation of tobacco (Nicotiana tabacum), showed that PmPAP1 was overexpressed in 13 lines, and 2 of them (Line-10 and Line-13) were treated with different concentrations of phosphorus. Phosphorus deficiency could induce the expression of PmPAP1 gene in the roots and leaves of transgenic plants, and the activity of acid phosphatase apase in roots and leaves of transgenic plants was 1.27 and 1.25 times higher than that of wild type (wild type WT) under the condition of low phosphorus. Compared with WT, the total phosphorus and inorganic phosphorus contents of transgenic plants were increased by 45.48% (Line-10) and 62.31% (Line-13), respectively. In addition, the content of peroxidase (POD) and superoxide dismutase (superoxide) in transgenic plants were significantly higher than that in WT (P0.05), while the content of malondialdehyde (malondialdehyde) decreased significantly (P0.05). The effects of phosphorus stress on plant growth showed that the total biomass root biomass and root-shoot ratio of transgenic plants were significantly increased under low phosphorus condition (P0.05). However, there was no significant difference in enzyme activity, malondialdehyde content, phosphorus content and biomass between transgenic tobacco and wild type tobacco under normal and high phosphorus conditions. Therefore, the high expression of PmPAP1 gene could significantly improve the phosphorus utilization ability of tobacco plants under low available phosphorus, and then enhance the ability of tolerance to low phosphorus stress. This study provided a theoretical basis for understanding the molecular mechanism of low phosphorus tolerance of Pinus massoniana and for creating new low phosphorus tolerant tobacco germplasm.
【作者单位】：贵州大学农业生物工程研究院/生命科学学院/山地植物资源保护与种质创新教育部重点实验室;贵州大学林学院;
【基金】：国家高技术研究发展计划(863)项目(No.2011AA10020301) 贵州省重大专项(No.20126011-1)
【分类号】：S791.248

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