Igawa Tomoko, Takahashi-Ando Naoko, Ochiai Noriyuki, Ohsato Shuichi, Shimizu Tsutomu, Kudo Toshiaki, Yamaguchi Isamu, Kimura Makoto
Plant & Microbial Metabolic Engineering Research Unit and Laboratory for Remediation Research, Discovery Research Institute (DRI), RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
Appl Environ Microbiol. 2007 Mar;73(5):1622-9. doi: 10.1128/AEM.01077-06. Epub 2007 Jan 5.
Maize is subject to ear rot caused by toxigenic Aspergillus and Fusarium species, resulting in contamination with aflatoxins, fumonisins, trichothecenes, and zearalenone (ZEN). The trichothecene group and ZEN mycotoxins are produced by the cereal pathogen Fusarium graminearum. A transgenic detoxification system for the elimination of ZEN was previously developed using an egfp::zhd101 gene (gfzhd101), encoding an enhanced green fluorescent protein fused to a ZEN-degrading enzyme. In this study, we produced a transgenic maize line expressing an intact copy of gfzhd101 and examined the feasibility of transgene-mediated detoxification in the kernels. ZEN-degrading activity has been detected in transgenic kernels during seed maturation (for a period of 6 weeks after pollination). The level of detoxification activity was unaltered after an additional storage period of 16 weeks at 6 degrees C. When the seeds were artificially contaminated by immersion in a ZEN solution for 48 h at 28 degrees C, the total amount of the mycotoxin in the transgenic seeds was uniformly reduced to less than 1/10 of that in the wild type. The ZEN in the transgenic maize kernels was also efficiently decontaminated under conditions of lower water activity (aw) and temperature; e.g., 16.9 microg of ZEN was removed per gram of seed within 48 h at an aw of 0.90 at 20 degrees C. F. graminearum infection assays demonstrated an absence of ZEN in the transgenic maize seeds, while the mycotoxin accumulated in wild-type kernels under the same conditions. Transgene-mediated detoxification may offer simple solutions to the problems of mycotoxin contamination in maize.
玉米易受产毒曲霉和镰刀菌属引起的穗腐病影响,导致被黄曲霉毒素、伏马毒素、单端孢霉烯族毒素和玉米赤霉烯酮(ZEN)污染。单端孢霉烯族毒素和ZEN霉菌毒素由谷物病原菌禾谷镰刀菌产生。先前利用编码与ZEN降解酶融合的增强型绿色荧光蛋白的egfp::zhd101基因(gfzhd101)开发了一种用于消除ZEN的转基因解毒系统。在本研究中,我们培育了一个表达完整gfzhd101拷贝的转基因玉米品系,并研究了转基因介导的籽粒解毒的可行性。在种子成熟期间(授粉后6周内),已在转基因籽粒中检测到ZEN降解活性。在6摄氏度下额外储存16周后,解毒活性水平未改变。当种子在28摄氏度下浸入ZEN溶液中人工污染48小时后,转基因种子中霉菌毒素的总量均匀降低至野生型的1/10以下。转基因玉米籽粒中的ZEN在较低水分活度(aw)和温度条件下也能有效去除;例如,在20摄氏度下aw为0.90时,每克种子在48小时内可去除16.9微克ZEN。禾谷镰刀菌感染试验表明转基因玉米种子中不存在ZEN,而在相同条件下霉菌毒素在野生型籽粒中积累。转基因介导的解毒可能为解决玉米中霉菌毒素污染问题提供简单的解决方案。