National Key Facility for Crop Gene Resources and Genetic Improvement/Key Laboratory of Biology and Genetic Improvement of Triticeae Crops of the Agriculture Ministry, Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, China.
Funct Integr Genomics. 2013 Aug;13(3):403-9. doi: 10.1007/s10142-013-0332-5. Epub 2013 Jul 10.
Take-all, caused by soil-borne fungus Gaeumannomyces graminis var. tritici (Ggt), is a devastating root disease of wheat (Triticum aestivum) worldwide. Breeding resistant wheat cultivars is the most promising and reliable approach to protect wheat from take-all. Currently, no resistant wheat germplasm is available to breed cultivars using traditional methods. In this study, gene transformation was carried out using Snakin-1 (SN1) gene isolated from potato (Solanum tuberosum) because the peptide shows broad-spectrum antimicrobial activity in vitro. Purified SN1 peptide also inhibits in vitro the growth of Ggt mycelia. By bombardment-mediated method, the gene SN1 was transformed into Chinese wheat cultivar Yangmai 18 to generate SN1 transgenic wheat lines, which were used to assess the effectiveness of the SN1 peptide in protecting wheat from Ggt. Genomic PCR and Southern blot analyses indicated that the alien gene SN1 was integrated into the genomes of five transgenic wheat lines and heritable from T₀ to T₄ progeny. Reverse transcription-PCR and Western blot analyses showed that the introduced SN1 gene was transcribed and highly expressed in the five transgenic wheat lines. Following challenging with Ggt, disease test results showed that compared to segregants lacking the transgene and untransformed wheat plants, these five transgenic wheat lines expressing SN1 displayed significantly enhanced resistance to take-all. These results suggest that SN1 may be a potentially transgenic tool for improving the take-all resistance of wheat.
全蚀病，由土壤传播的真菌禾顶囊壳菌小麦变种（Ggt）引起，是一种世界性的小麦毁灭性根部病害。培育具有抗性的小麦品种是保护小麦免受全蚀病侵害最有前途和可靠的方法。目前，由于缺乏可用的抗全蚀病小麦种质资源，传统的品种选育方法无法使用。本研究利用从马铃薯（Solanum tuberosum）中分离出的 Snakin-1（SN1）基因进行基因转化，因为该肽在体外具有广谱的抗菌活性。纯化的 SN1 肽也能抑制 Ggt 菌丝的体外生长。通过基因枪介导的方法，将 SN1 基因转化到中国小麦品种扬麦 18 中，生成 SN1 转基因小麦株系，用于评估 SN1 肽在保护小麦免受 Ggt 侵害方面的有效性。基因组 PCR 和 Southern blot 分析表明，外源基因 SN1 已整合到五个转基因小麦株系的基因组中，并可从 T₀ 代到 T₄ 代遗传。反转录 PCR 和 Western blot 分析表明，引入的 SN1 基因在这五个转基因小麦株系中转录并高度表达。在受到 Ggt 侵袭后，病害测试结果表明，与缺乏转基因的分离株和未转化的小麦植株相比，这五个表达 SN1 的转基因小麦株系对全蚀病的抗性显著增强。这些结果表明，SN1 可能是一种提高小麦抗全蚀病的潜在转基因工具。
