欧亚国际

最近，来自欧洲的科学家利用WIWAM XY（Pathoviewer）高通量植物表型成像平台发表了题为A Novel In Planta Enrichment Method Employing Fusarium graminearum-Infected Wheat Spikes to Select for Competitive Biocontrol Bacteria，文章发表于知名期刊Toxins 2022,14(3), 222。

SMO构建WIWAM XY（Pathoviewer）高通量植物表型成像平台

WIWAM植物表型成像系统由比利时SMO公司与Ghent大学VIB研究所研制生产，整合了LED植物智能培养、自动化控制系统、叶绿素荧光成像测量分析、植物热成像分析、植物近红外成像分析、植物高光谱分析、植物多光谱分析、植物CT断层扫描分析、自动条码识别管理、RGB真3D成像等多项先进技术，以优化的方式实现大量植物样品以优化的方式实现大量植物样品——从拟南芥、水稻、玉米到各种其它植物的生理生态与形态结构成像分析，用于高通量植物表型成像分析测量、植物胁迫响应成像分析测量、植物生长分析测量、生态毒理学研究、性状识别及植物生理生态分析研究等。

PathoViewer是一款高通量植物表型成像平台（属于WIWAM XY系列），使用了高分辨率图像在可控条件下对植物进行表型成像。该系统配备有6MP-16 bit的相机系统以及各种光学滤波轮，有广泛用途，例如测量叶绿素荧光、红色荧光蛋白、绿色荧光蛋白、自荧光、RGB改进花青素反射指数以及叶绿素指数等。系统为自动系统，用于测量植物或植物部分的生物或非生物胁迫。 

Pathoviewer特点

Pathoviewer组合了RGB, 叶绿素荧光、花青素、NIR和GFP/RFP图像处理，对生物和非生物胁迫的影响进行成像。这些相机也可为研究人员采集植物科学相关信息，另外还可对表型特征可视化。

Pathoviewer能测量的最重要参数是Fv/FM值。该值通过叶绿素荧光获取，是指示测量植物光合性能的参数。荧光可用于测量PSII光合作用中的多种参数如线性电子流，CO2同化以及变化。

WIWAM XY高通量表型成像平台

利用禾谷镰刀菌感染的小麦穗筛选竞争性生防细菌的一种新的植物内富集方法

摘要

这项工作介绍了一种在小麦中发现新的抗赤霉病细菌生防剂的替代工作流程。与隔离物收集的大规模测试不同，我们从不同的接种物开始，在灌浆期从田间种植的小麦穗中提取微生物组。在感染禾谷镰刀菌PH1的分离小麦穗上，产生了四个不同的微生物群落，这些微生物群落暴露在3个14天的非培养实验富集物中。我们发现，一个细菌群落在3个周期后减少了感染症状，随后选择该细菌群落通过限制稀释来分离细菌。所有94个分离株均在体外和植物试验中进行了测试，并选择了14个分离株在分离的小麦穗上进行了进一步测试。结果似乎表明，我们的富集方法导致细菌在FHB控制方面具有不同的作用模式。波斯欧文氏菌（Erwinia persicina）分离物C3的疾病严重程度（Fv/Fm）显著降低，波斯欧文氏菌（Erwinia persicina）C3和假单胞菌(Pseudomonassp.)B3的真菌生物量（cGFP）显著降低。然而，这两种治疗的真菌毒素分析显示，DON水平没有降低。尽管如此，安那提泛球菌(Pantoea ananatis)H3和H11以及波斯欧文氏杆菌(Erwinia persicina)H2能够将DON浓度降低50%以上，尽管这些影响在统计学上并不显著。最后，波斯欧文氏菌(Erwinia persicina)H2也显示出DON对植物毒性较小的DON-3G的显著更高的糖基化。据报道，通过富集循环分离的细菌属在开放栖息地发育的微生物群落中占主导地位，这表明分离的细菌可以降低禾谷镰刀菌在穗叶层上的感染压力。

关键词：小麦；禾谷镰刀菌；生物防治；病理生物群落；独立培养；微生物组工程；连续通过；实验富集；叶层

A Novel In Planta Enrichment Method Employing Fusarium graminearum-Infected Wheat Spikes to Select for Competitive Biocontrol Bacteria

Abstract

This work introduces an alternative workflow for the discovery of novel bacterial biocontrol agents in wheat against Fusarium head blight (FHB). Unlike the mass testing of isolate collections, we started from a diverse inoculum by extracting microbiomes from ears of field-grown plants at grain filling stage. Four distinct microbial communities were generated which were exposed to 3 14-day culture-independent experimental enrichments on detached wheat spikes infected with F. graminearum PH1. We found that one bacterial community reduced infection symptoms after 3 cycles, which was chosen to subsequently isolate bacteria through limiting dilution. All 94 isolates were tested in an in vitro and in planta assay, and a selection of 14 isolates was further tested on detached ears. The results seem to indicate that our enrichment approach resulted in bacteria with different modes-of-action in regard to FHB control. Erwinia persicina isolate C3 showed a significant reduction in disease severity (Fv/Fm), and Erwinia persicina C3 and Pseudomonas sp. B3 showed a significant reduction in fungal biomass (cGFP). However, the mycotoxin analysis of both these treatments showed no reduction in DON levels. Nevertheless, Pantoea ananatis H3 and H11 and Erwinia persicina H2 were able to reduce DON concentrations by more than 50%, although these effects were not statistically significant. Lastly, Erwinia persicina H2 also showed a significantly greater glucosylation of DON to the less phytotoxic DON-3G. The bacterial genera isolated through the enrichment cycles have been reported to dominate microbial communities that develop in open habitats, showing strong indications that the isolated bacteria can reduce the infection pressure of F. graminearum on the spike phyllosphere. 

Keywords:Triticum aestivum; Fusarium graminearum; biocontrol; pathobiome; culture independent; microbiome engineering; successive passage; experimental enrichment; phyllosphere