WIWAM葉綠素熒光成像助力禾谷鐮刀菌感染小麥穗篩選生物防治細菌
近日，來自歐洲的科學家利用WIWAM XY（Pathoviewer）高通量植物表型成像平臺發表了題為A Novel In Planta Enrichment Method Employing Fusarium graminearum-Infected Wheat Spikes to Select for Competitive Biocontrol Bacteria，文章發表于知名期刊Toxins 2022,14(3), 222。
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
SMO構建WIWAM XY高通量植物表型成像平臺
利用禾谷鐮刀菌感染的小麥穗篩選競爭性生防細菌的一種新的植物內富集方法
這項工作介紹了一種在小麥中發現新的抗赤霉病細菌生防劑的替代工作流程。與隔離物收集的大規模測試不同，我們從不同的接種物開始，在灌漿期從田間種植的小麥穗中提取微生物組。在感染禾谷鐮刀菌PH1的分離小麥穗上，產生了四個不同的微生物群落，這些微生物群落暴露在3個14天的非培養實驗富集物中。我們發現，一個細菌群落在3個周期后減少了感染癥狀，隨后選擇該細菌群落通過限制稀釋來分離細菌。所有94個分離株均在體外和植物試驗中進行了測試，并選擇了14個分離株在分離的小麥穗上進行了進一步測試。結果似乎表明，我們的富集方法導致細菌在FHB控制方面具有不同的作用模式。波斯歐文氏菌（Erwinia persicina）分離物C3的疾病嚴重程度（Fv/Fm）顯著降低，波斯歐文氏菌（Erwinia persicina）C3和假單胞菌(Pseudomonassp.)B3的真菌生物量（cGFP）顯著降低。然而，這兩種治療的真菌毒素分析顯示，DON水平沒有降低。盡管如此，安那提泛球菌(Pantoea ananatis)H3和H11以及波斯歐文氏桿菌(Erwinia persicina)H2能夠將DON濃度降低50%以上，盡管這些影響在統計學上并不顯著。最后，波斯歐文氏菌(Erwinia persicina)H2也顯示出DON對植物毒性較小的DON-3G的顯著更高的糖基化。據報道，通過富集循環分離的細菌屬在開放棲息地發育的微生物群落中占主導地位，這表明分離的細菌可以降低禾谷鐮刀菌在穗葉層上的感染壓力。
關鍵詞：小麥；禾谷鐮刀菌；生物防治；病理生物群落；獨立培養；微生物組工程；連續通過；實驗富集；葉層
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
WIWAM植物表型成像系統由比利時SMO公司與Ghent大學VIB研究所研制生產，整合了LED植物智能培養、自動化控制系統、葉綠素熒光成像測量分析、植物熱成像分析、植物近紅外成像分析、植物高光譜分析、植物多光譜分析、植物CT斷層掃描分析、自動條碼識別管理、RGB真3D成像等多項先進技術，以優化的方式實現大量植物樣品以優化的方式實現大量植物樣品——從擬南芥、水稻、玉米到各種其它植物的生理生態與形態結構成像分析，用于高通量植物表型成像分析測量、植物脅迫響應成像分析測量、植物生長分析測量、生態毒理學研究、性狀識別及植物生理生態分析研究等。
PathoViewer是一款高通量植物表型成像平臺（屬于WIWAM XY系列），使用了高分辨率圖像在可控條件下對植物進行表型成像。該系統配備有6MP-16 bit的相機系統以及各種光學濾波輪，有廣泛用途，例如測量葉綠素熒光、紅色熒光蛋白、綠色熒光蛋白、自熒光、RGB改進花青素反射指數以及葉綠素指數等。系統為自動系統，用于測量植物或植物部分的生物或非生物脅迫。
PathoViewer是一款高通量植物表型成像平臺（屬于WIWAM XY系列），使用了高分辨率圖像在可控條件下對植物進行表型成像。該系統配備有6MP-16 bit的相機系統以及各種光學濾波輪，有廣泛用途，例如測量葉綠素熒光、紅色熒光蛋白、綠色熒光蛋白、自熒光、RGB改進花青素反射指數以及葉綠素指數等。系統為自動系統，用于測量植物或植物部分的生物或非生物脅迫。
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