Is zinniol a true phytotoxin? Evaluation of its activity at the cellular level against <Emphasis Type="Italic">Tagetes erecta</Emphasis>

Zinniol, a non-host selective phytotoxin commonly produced by fungi of the Alternaria genus, has been reported as the metabolite responsible for the phytotoxicity of the lipophilic fraction of A. tagetica. While both the lipophilic fraction and zinniol have been shown to produce necrosis on leaves of susceptible marigold (Tagetes erecta) plants, the true role of zinniol in the infectious process remains uncertain. Using marigold cell cultures as a model, we evaluated the effects of zinniol and the lipophilic fraction at the cellular level and showed that pure zinniol is not markedly phytotoxic at concentrations known to induce necrosis in leaves of T. erecta. Moreover, the effects of zinniol on cell membranes and DNA fragmentation are less intense than those caused by the lipophilic fraction. These results suggest that zinniol may not play a significant role in the A. tagetica–T. erecta interaction and, consequently, its classification as a non-host selective phytotoxin is questionable.     

Differential effects of phytotoxic metabolites from <Emphasis Type="Italic">Alternaria tagetica</Emphasis> on <Emphasis Type="Italic">Tagetes erecta</Emphasis> cell cultures

Alternaria tagetica, a fungus that causes early blight in marigold (Tagetes erecta), produces two groups of phytotoxic metabolites: one hydrophilic and the other lipophilic that show phytotoxic activity when tested by the leaf-spot assay in T. erecta. We evaluated the cellular effects of the phytotoxic culture filtrate of A. tagetica and the hydrophilic and lipophilic fractions, then determined whether the filtrate or the fractions differentially induced pathogenesis-related mechanisms in the plant. The culture filtrate and the phytotoxic fractions had adverse effects on cell viability, fresh mass, and the number of cells, and induced ROS accumulation, lipid peroxidation and DNA damage on T. erecta cell suspension cultures, and these effects are related to pathogenic mechanisms attributed to phytotoxins. However, although exposure of marigold cells to the phytotoxic culture filtrate of A. tagetica triggered programmed cell death, the hydrophilic and the lipophilic phytotoxic fractions induced death that was more related to a toxic effect leading to necrosis. This study presents a complementary perspective in the search for the roles of metabolites, including phytotoxins, produced by phytopathogenic fungi during plant infection.     

中国-印尼玉米产业科技合作进展及展望

为了提升印度尼西亚国家玉米产业发展水平,服务国家“一带一路”、“科技服务外交”等国家战略,执行国家科技部对发展中国家科技援助项目,山东省农业科学院玉米研究所开展了印尼本土化玉米育种、新品种与新技术集成和玉米产业科技园区建设等项目内容。利用黄淮海优异种质与印尼富含热带资源种质组配了育种基础材料,育成的10份玉米优异杂交组合正在参加印尼国家玉米品种审定试验;实现了山东省第一次境外农业技术培训班和玉米现场观摩活动,高产栽培技术集成创造了印尼雨季玉米13.70 t/hm²的高产纪录;建成的50 hm²中国-印尼玉米产业科技园区,正在为印尼玉米产业的快速发展发挥重要作用。通过科企合作模式,发挥国内农业企业优势,将中国的玉米新品种、新技术引进到印度尼西亚,将会加速印尼玉米产业发展水平,为国家科技外交和“一带一路”战略服务。     

Natural zinniol derivatives from Alternaria tagetica. Isolation, synthesis, and structure-activity correlation

Two novel phytotoxins, 8-zinniol methyl ether (5) and 8-zinniol acetate (6), in addition to 6-(3',3'-dimethylallyloxy)-4-methoxy-5-methylphthalide (2), 5-(3',3'-dimethylallyloxy)-7-methoxy-6-methylphthalide (3), and the novel metabolites 8-zinniol 2-(phenyl)ethyl ether (4) and 7-zinniol acetate (7) have been identified as natural zinniol derivatives from the organic crude extract of Alternaria tagetica culture filtrates. Using zinniol as the starting material, phytotoxin 5 was synthesized, together with a number of synthetic intermediates (8-13). Both natural and synthetic zinniol derivatives were evaluated in the leaf-spot bioassay against marigold leaves (Tagetes erecta).     

Production of hydrophilic phytotoxins by <Emphasis Type="Italic">Mycosphaerella fijiensis</Emphasis>

We have established a reproducible strategy to purify hydrophilic phytotoxins present in the aqueous filtrate of Mycosphaerella fijiensis Morelet. The lyophilized culture filtrate is initially treated with activated charcoal, then successively purified using vacuum liquid chromatography and semipreparative high performance liquid chromatography. Phytotoxic activity was tested using a leaf-spot assay on healthy banana leaves. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Physiological effects of the hydrophilic phytotoxins produced by <Emphasis Type="Italic">Mycosphaerella fijiensis</Emphasis>, the causal agent of black sigatoka in banana plants

Although Mycosphaerella fijiensis, the causal agent of black sigatoka disease of banana, has been known to produce numerous lipophilic host-selective (HSTs) and nonhost selective phytotoxins (non-HSTs), only recently we have reported that the pathogen also produces hydrophilic phytotoxins. Here we examined the effect of light on the toxicity of the hydrophilic phytotoxins and estimated the electrolyte leakage and H₂O₂ and superoxide generation in detached banana leaves to study their mode of action at the cellular level. Nonhost plant species were also tested to determine whether the toxins are HSTs or non-HSTs. Our results suggest that the hydrophilic phytotoxins are non-HSTs, that their phytotoxicity is not light dependent, and that they may act at the plasma membrane by altering permeability through oxidative damage, by inducing ROS production as part of their mechanism of action.     

Phytoanticipins from banana (<Emphasis Type="Italic">Musa acuminata</Emphasis> cv. Grande Naine) plants,with antifungal activity against <Emphasis Type="Italic">Mycosphaerella fijiensis</Emphasis>, the causal agent of black Sigatoka

The previously detected antifungal activity against Mycosphaerella fijiensis of aqueous infusions of healthy banana (Musa acuminata cv. Grande Naine) leaves, suggested the production of phytoprotectants by the plant. The bioassay-guided VLC-purification of the lyophilized infusion of the leaves of 4-month old healthy banana (M. acuminata cv. Grande Naine) plants, resulted in the purification of a phytoanticipin with strong antifungal activity against M. fijiensis Morelet, the causal agent of black Sigatoka, the most destructive and devastating disease of bananas and plantains in the world. The LC-MS analysis of the purified phytoanticipin suggests a steroidal saponin structure with four sugar units attached to the C-3 position of a diosgenin-like aglycone. This represents the first report of phytoanticipins occurring in M. acuminata.