Key Laboratory of Crop Heterosis and Utilization (MOE)/Beijing Key Laboratory of Crop Genetic Improvement, College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China.
Molecules. 2018 Mar 24;23(4):748. doi: 10.3390/molecules23040748.
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, agglutinin (), a gene encoding mannose binding lectin, was cloned, and it shared 92.69% nucleotide similarity and 94% amino acid similarity with agglutinin (). The gene, driven by the constitutive and phloem-specific ribulose bisphosphate carboxylase small subunit gene () 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 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 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, could be a strong biotechnological candidate to produce aphid-resistant wheat.
小麦蚜虫是小麦灌浆期的主要害虫。植物凝集素对麦蚜等刺吸式害虫有毒。本研究克隆了凝集素（）基因，该基因编码甘露糖结合凝集素，与凝集素（）共享 92.69%的核苷酸相似性和 94%的氨基酸相似性。该基因在 pBAC-rbcs-ppa 表达载体中，由组成型和韧皮部特异性核酮糖-1,5-二磷酸羧化酶小亚基基因（）启动子驱动，通过粒子轰击转化法转入小麦品种宝丰 104（BF104）。共获得 54 个 T₀ 代转基因植株。通过 PCR 和 RT-PCR 分析分别验证了 基因的遗传和表达，获得了 7 个纯合的转基因系。对离体叶片段的蚜虫生物测定表明，与 BF104 相比，7 个 转基因小麦系的蚜虫生长率较低，抑制率较高。此外，在隔离田间进行的两年蚜虫生物测定表明，与野生型 BF104 相比，转基因系的每个分蘖上的蚜虫数量明显减少。因此， 可能是培育抗蚜虫小麦的有力生物技术候选基因。
