Amarasinghe Chami, Wang Jian-Hua, Liao Yu-Cai, Fernando W G Dilantha
Department of Plant Science, University of Manitoba, Winnipeg, MB R3T2N2, Canada; E-Mail:
Int J Mol Sci. 2011;12(9):6164-75. doi: 10.3390/ijms12096164. Epub 2011 Sep 20.
Positive-negative PCR assays based on the genes involved in the trichothecene biosynthesis pathway are useful in assessing the risk of trichothecene contamination in grain and are important in epidemiological studies. A single PCR detection method based on the structural gene sequence of TRI13 gene has been developed to predict the 3-ADON, 15-ADON and NIV chemotypes in China. The chemotypic differences are based on the deletions within the TRI13 gene. The objective of this study was to assess the reliability of using this single primer based on the TRI13 gene to differentiate the F. graminearum chemotypes in Canada. In this study, we found that, this single PCR detection method based on the deletions in the TRI13 gene cannot be used to differentiate the 3-ADON and 15-ADON chemotypes in the Canadian F. graminearum isolates; further sequence analysis of the PCR products confirmed that both Canadian 3-ADON and 15-ADON chemotypes have the 61 bp deletion in the TRI13 gene. This 61 bp deletion was absent in the Chinese 3-ADON isolates. Therefore these findings revealed that there are genetic differences between the examined 3-ADON F. graminearum isolates from Canada and China. The observed genetic differences between the 3-ADON chemotype populations in Canada and China may be resulted from a random mutation (insertion/deletion) that took place in one of the populations and accumulated due to genetic drift and/or selection.
基于单端孢霉烯生物合成途径相关基因的正负PCR检测方法,对于评估谷物中单端孢霉烯污染风险很有用,且在流行病学研究中具有重要意义。已开发出一种基于TRI13基因结构基因序列的单一PCR检测方法,用于预测中国的3-ADON、15-ADON和NIV化学型。化学型差异基于TRI13基因内的缺失。本研究的目的是评估使用基于TRI13基因的这一单一引物来区分加拿大禾谷镰刀菌化学型的可靠性。在本研究中,我们发现,这种基于TRI13基因缺失的单一PCR检测方法不能用于区分加拿大禾谷镰刀菌分离株中的3-ADON和15-ADON化学型;对PCR产物的进一步序列分析证实,加拿大的3-ADON和15-ADON化学型在TRI13基因中均有61 bp的缺失。中国的3-ADON分离株中不存在这种61 bp的缺失。因此,这些发现揭示了加拿大和中国检测的3-ADON禾谷镰刀菌分离株之间存在遗传差异。加拿大和中国3-ADON化学型群体之间观察到的遗传差异可能是由于其中一个群体发生了随机突变(插入/缺失),并因遗传漂变和/或选择而积累。