Zhejiang Provincial Key Laboratory of Crop Genetic Resources, Institute of Crop Science, Zhejiang University, Hangzhou 310058, Zhejiang, China.
Department of Molecular Phytopathology, Christian-Albrechts-University of Kiel, Hermann Rodewald Str. 9, D-24118 Kiel, Germany.
Int J Mol Sci. 2019 Apr 8;20(7):1740. doi: 10.3390/ijms20071740.
Beet cyst nematode ( Schm.) is one of the most damaging pests in sugar beet growing areas around the world. The and genes confer resistance to the beet cyst nematode, and both were cloned from sugar beet translocation line (A906001). The translocation line carried the locus from chromosome 1 including gene and resistance gene analogs (RGA), which confer resistance to . In this research, and genes were transferred into oilseed rape to obtain different transgenic lines by mediated transformation method. The gene was pyramided into the same plants by crossing homozygous and plants to identify the function and interaction of and genes. In vitro and in vivo cyst nematode resistance tests showed that and plants could be infested by beet cyst nematode (BCN) juveniles, however a large fraction of penetrated nematode juveniles was not able to develop normally and stagnated in roots of transgenic plants, consequently resulting in a significant reduction in the number of developed nematode females. A higher efficiency in inhibition of nematode females was observed in plants expressing pyramiding genes than in those only expressing a single gene. Molecular analysis demonstrated that and gene expressions in oilseed rape constitutively activated transcription of plant-defense related genes such as (non-expresser of ), (enhanced disease resistance 1) and (suppressor of the allele of ). Transcript of gene in transgenic and plants were slightly up-regulated, while its expression was considerably enhanced in gene pyramiding plants. The expression of gene did not change significantly among transgenic , and gene pyramiding plants and wild type. The expression of gene was slightly up-regulated in transgenic and plants compared with the wild type, however, its expression was not changed in gene pyramiding plant and had no interaction effect. gene expression was significantly up-regulated in transgenic and genes pyramiding plants, but almost no expression was found in transgenic plants. These results show that nematode resistance genes from sugar beet were functional in oilseed rape and conferred BCN resistance by activation of a CC-NBS-LRR R gene mediated resistance response. The gene pyramiding had enhanced resistance, thus offering a novel approach for the BCN control by preventing the propagation of BCN in oilseed rape. The transgenic oilseed rape could be used as a trap crop to offer an alternative method for beet cyst nematode control.
甜菜胞囊线虫（Schm.）是世界范围内甜菜种植区最具破坏性的害虫之一。和基因赋予了对甜菜胞囊线虫的抗性，这两个基因都从甜菜易位系（A906001）中克隆得到。该易位系携带 1 号染色体上的基因座，包括基因和抗性基因类似物（RGA），赋予了对的抗性。在这项研究中，通过介导转化方法将和基因导入油菜中，获得了不同的转基因系。通过杂交纯合和植株，将基因叠加到同一植株中，以鉴定和基因的功能和相互作用。体外和体内胞囊线虫抗性试验表明，和植株可以被甜菜胞囊线虫（BCN）幼虫侵袭，然而，大量穿透的线虫幼虫不能正常发育，并在转基因植物的根部停滞，从而导致发育的线虫雌性数量显著减少。表达叠加基因的植物对线虫雌性的抑制效率更高。分子分析表明，油菜中表达的和基因持续激活了植物防御相关基因的转录，如（非表达）、（增强的疾病抗性 1）和（的等位基因抑制物）。转基因和植株中的基因转录略有上调，而在基因叠加植株中其表达显著增强。在转基因、和基因叠加植物和野生型中，基因的表达没有显著变化。在转基因和基因叠加植物中，基因的表达略有上调，而在基因叠加植物中没有变化，也没有相互作用。在转基因和基因叠加植物中，基因的表达显著上调，但在转基因植物中几乎没有表达。这些结果表明，来自甜菜的线虫抗性基因在油菜中具有功能，并通过激活 CC-NBS-LRR R 基因介导的抗性反应赋予 BCN 抗性。基因叠加增强了抗性，从而通过防止 BCN 在油菜中的繁殖来提供控制 BCN 的新方法。转基因油菜可作为诱捕作物，提供控制甜菜胞囊线虫的替代方法。
