Lee Theresa, Han You-Kyoung, Kim Kook-Hyung, Yun Sung-Hwan, Lee Yin-Won
School of Agricultural Biotechnology and Research Center for New Bio-materials in Agriculture, Seoul National University, Suwon 441-744, Korea.
Appl Environ Microbiol. 2002 May;68(5):2148-54. doi: 10.1128/AEM.68.5.2148-2154.2002.
Gibberella zeae, a major cause of cereal scab, can be divided into two chemotypes based on production of the 8-ketotrichothecenes deoxynivalenol (DON) and nivalenol (NIV). We cloned and sequenced a Tri13 homolog from each chemotype. The Tri13 from a NIV chemotype strain (88-1) is located in the trichothecene gene cluster and carries an open reading frame similar to that of Fusarium sporotrichioides, whereas the Tri13 from a DON chemotype strain (H-11) carries several mutations. To confirm the roles of the Tri13 and Tri7 genes in trichothecene production by G. zeae, we genetically altered toxin production in 88-1 and H-11. In transgenic strains, the targeted deletion of Tri13 from the genome of 88-1 caused production of DON rather than NIV. Heterologous expression of the 88-1 Tri13 gene alone or in combination with the 88-1 Tri7 gene conferred on H-11 the ability to synthesize NIV; in the latter case, 4-acetylnivalenol (4-ANIV) also was produced. These results suggest that Tri13 and Tri7 are required for oxygenation and acetylation of the oxygen at C-4 during synthesis of NIV and 4-ANIV in G. zeae. These functional analyses of the Tri13 and Tri7 genes provide the first clear evidence for the genetic basis of the DON and NIV chemotypes in G. zeae.
玉米赤霉,谷物赤霉病的主要致病菌,可根据8-酮单端孢霉烯脱氧雪腐镰刀菌烯醇(DON)和雪腐镰刀菌烯醇(NIV)的产生分为两种化学型。我们从每种化学型中克隆并测序了一个Tri13同源基因。来自NIV化学型菌株(88-1)的Tri13位于单端孢霉烯基因簇中,其开放阅读框与拟分枝孢镰刀菌的相似,而来自DON化学型菌株(H-11)的Tri13携带多个突变。为了证实Tri13和Tri7基因在玉米赤霉产生单端孢霉烯中的作用,我们对88-1和H-11中的毒素产生进行了基因改造。在转基因菌株中,从88-1基因组中靶向缺失Tri13导致产生DON而非NIV。单独或与88-1 Tri7基因联合异源表达88-1 Tri13基因赋予H-11合成NIV的能力;在后一种情况下,还产生了4-乙酰雪腐镰刀菌烯醇(4-ANIV)。这些结果表明,Tri13和Tri7是玉米赤霉合成NIV和4-ANIV过程中C-4位氧的氧化和乙酰化所必需的。对Tri13和Tri7基因的这些功能分析为玉米赤霉中DON和NIV化学型的遗传基础提供了首个明确证据。