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Expression of Pinellia pedatisecta Lectin Gene in Transgenic Wheat Enhances Resistance to Wheat Aphids

机译:半夏pedatisecta凝集素基因在转基因小麦中的表达增强了对小麦蚜虫的抗性

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摘要

Wheat aphids are major pests during the seed filling stage of wheat. Plant lectins are toxic to sap-sucking pests such as wheat aphids. In this study, Pinellia pedatisecta agglutinin (ppa), a gene encoding mannose binding lectin, was cloned, and it shared 92.69% nucleotide similarity and 94% amino acid similarity with Pinellia ternata agglutinin (pta). The ppa gene, driven by the constitutive and phloem-specific ribulose bisphosphate carboxylase small subunit gene (rbcs) promoter in pBAC-rbcs-ppa expression vector, was transferred into the wheat cultivar Baofeng104 (BF104) by particle bombardment transformation. Fifty-four T0 transgenic plants were generated. The inheritance and expression of the ppa gene were confirmed by PCR and RT-PCR analysis respectively, and seven homozygous transgenic lines were obtained. An aphid bioassay on detached leaf segments revealed that seven ppa transgenic wheat lines had lower aphid growth rates and higher inhibition rates than BF104. Furthermore, two-year aphid bioassays in isolated fields showed that aphid numbers per tiller of transgenic lines were significantly decreased, compared with wild type BF104. Therefore, ppa could be a strong biotechnological candidate to produce aphid-resistant wheat.
机译:小麦蚜虫是小麦种子灌浆期的主要害虫。植物凝集素对诸如小麦蚜虫之类的汁液吸收害虫有毒。在本研究中,克隆了编码甘露糖结合凝集素的基因半夏凝集素(ppa),并且与半夏凝集素(pta)具有92.69%的核苷酸相似性和94%的氨基酸相似性。由pBAC-rbcs-ppa表达载体中的组成型韧皮部特异性韧皮部双核糖双磷酸羧化酶小亚基基因(rbcs)启动子驱动的ppa基因通过粒子轰击转化转移到小麦品种Baofeng104(BF104)中。产生了五十四个T0转基因植物。通过PCR和RT-PCR分析分别证实了ppa基因的遗传和表达,获得了7个纯合的转基因品系。对分离的叶段进行的蚜虫生物测定显示,与BF104相比,七个ppa转基因小麦品系的蚜虫生长速率和抑制率更高。此外,在隔离田地中进行的两年蚜虫生物测定显示,与野生型BF104相比,转基因品系每耕作的蚜虫数量明显减少。因此,ppa可能是生产抗蚜虫小麦的强大生物技术候选人。

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