欧亚国际

丹麦Videometer开发出的多光谱成像系统用途范围极广，也可用于高通量植物表型分析和研究

利用高通量酶活性特征图谱和多光谱成像技术同时筛选大麦抗白粉病性

分子标记分析在植物育种过程中可进行快速和先进的预选和抗性筛选。在表型鉴定过程中，光学传感器已被证明具有确定和评估育种材料基因型功能的潜力。因此，特定抗病性状的生物标记物为在早期植物-病原相互作用中校准光学传感器方法提供了有价值的信息。在这种情况下，结合生理、代谢表型和表型特征，可以在育种过程中有效地识别和量化相关基因型。实验是用H. vulgare近等基因系（易感、霉变位点o（mlo）和霉变位点a（Mla）抗性）。每天在接种后0-8天在高通量设备中进行大麦植株的多光谱成像，多光谱为10个波长，波长范围为400-1000 nm。同时，利用半高通量方法分析了转化酶同功酶活性的时间动态，转化酶同功酶是将运输糖蔗糖不可逆地分解为己糖单体的关键特异性酶。细胞壁、胞浆和液泡转化酶的活性揭示了接种后0-120小时（hai）敏感基因型和具有mlo和基于Mla的抗性基因型的活性特征的特定动力学。这些模式可用于区分相互作用类型，并揭示了小麦白粉菌f.sp.hordei（Bgh）分生孢子对已达到0.5 hai的特定转化酶活性的早期影响。在白粉病的早期发病过程中，蓝波和690nm处的反射强度增加。抗Mla植物在680nm和710nm处的反射率增加，近红外波段的反射率从3 dai开始降低。应用支持向量机分类作为有监督机器学习方法，建立了白粉病大麦组织和过敏反应点的像素级识别和定量。这使得大麦白粉病相互作用的自动识别成为可能。该研究通过高通量多光谱成像进行植物抗性表型研究。转化酶分析和多光谱成像的结合证明是一个互补的验证系统。这将有助于更深入地理解光学数据及其在抗病性筛查中的应用。

Screening of Barley Resistance Against Powdery Mildew by Simultaneous High-Throughput Enzyme Activity Signature Profiling and Multispectral Imaging

Matheus T. Kuska1*†, Jan Behmann1*†, Dominik K. Großkinsky2, Thomas Roitsch3 and Anne-Katrin Mahlein1,4

1Institute for Crop Science and Resource Conservation-Plant Diseases and Plant Protection, University of Bonn, Bonn, Germany

2Department of Plant and Environmental Sciences, Copenhagen Plant Science Centre, University of Copenhagen, Frederiksberg, Denmark

3Department of Plant and Environmental Sciences, Copenhagen Plant Science Centre, University of Copenhagen, Taastrup, Denmark

4Institute of Sugar Beet Research (IfZ), Göttingen, Germany

Molecular marker analysis allow for a rapid and advanced pre-selection and resistance screenings in plant breeding processes. During the phenotyping process, optical sensors have proved their potential to determine and assess the function of the genotype of the breeding material. Thereby, biomarkers for specific disease resistance traits provide valuable information for calibrating optical sensor approaches during early plant-pathogen interactions. In this context, the combination of physiological, metabolic phenotyping and phenomic profiles could establish efficient identification and quantification of relevant genotypes within breeding processes. Experiments were conducted with near-isogenic lines of H. vulgare (susceptible, mildew locus o (mlo) and Mildew locus a (Mla) resistant). Multispectral imaging of barley plants was daily conducted 0–8 days after inoculation (dai) in a high-throughput facility with 10 wavelength bands from 400 to 1,000 nm. In parallel, the temporal dynamics of the activities of invertase isoenzymes, as key sink specific enzymes that irreversibly cleave the transport sugar sucrose into the hexose monomers, were profiled in a semi high-throughput approach. The activities of cell wall, cytosolic and vacuole invertase revealed specific dynamics of the activity signatures for susceptible genotypes and genotypes with mlo and Mla based resistances 0–120 hours after inoculation (hai). These patterns could be used to differentiate between interaction types and revealed an early influence of Blumeria graminis f.sp. hordei (Bgh) conidia on the specific invertase activity already 0.5 hai. During this early powdery mildew pathogenesis, the reflectance intensity increased in the blue bands and at 690 nm. The Mla resistant plants showed an increased reflectance at 680 and 710 nm and a decreased reflectance in the near infrared bands from 3 dai. Applying a Support Vector Machine classification as a supervised machine learning approach, the pixelwise identification and quantification of powdery mildew diseased barley tissue and hypersensitive response spots were established. This enables an automatic identification of the barley-powdery mildew interaction. The study established a proof-of-concept for plant resistance phenotyping with multispectral imaging in high-throughput. The combination of invertase analysis and multispectral imaging showed to be a complementing validation system. This will provide a deeper understanding of optical data and its implementation into disease resistance screening. 

地衣芽孢杆菌FMCH001通过在正常和干旱条件下的生长刺激提高水分利用效率

由于气候变化造成的生物和非生物胁迫造成的农业损失不断增加，这对全世界的粮食安全构成了挑战。在有限的水资源条件下维持作物生产力的一个有希望的策略是使用促进植物生长的根际细菌（PGPR）。本文研究了地衣芽孢杆菌（FMCH001）对玉米生长和生理的影响，研究了其 (Zea mays L. cv. Ronaldinho)在充足水分和干旱胁迫条件下的表现。盆栽试验在自动化高通量表型分析平台PhenoLab和温室条件下进行。物候实验结果表明，接种B与未接种的植物相比，地衣FMCH001表现出根干重（DW）和植物水分利用效率（WUE）增加。在温室试验中，与未接种对照植物相比，接种植物的根和茎干干重显著增加15%以上。此外，FMCH001植物的水分利用效率在水分充足和干旱胁迫的植物中都增加了46%。鉴别了应FMCH001处理的11种碳水化合物和8种抗氧化酶的根和枝叶活性。这表明，与未接种的植物相比，FMCH001处理的植物根中过氧化氢酶（CAT）具有更高的抗氧化活性。无论水分状况如何，都观察到较高的CAT活性。这些结果表明，形成革兰氏阳性孢子 B. licheniformis在正常和干旱胁迫条件下均可作为提高植物水分利用效率的生物刺激剂。

Bacillus licheniformis FMCH001 Increases Water Use Efficiency via Growth Stimulation in Both Normal and Drought Conditions

Akhtar, Saqib Saleem; Amby, Daniel Buchvaldt; Hegelund, Josefine Nymark; Fimognari,

Lorenzo; Großkinsky, Dominik K.; Westergaard, Jesper Cairo; Müller, Renate; Moelbak, Lars; Liu, Fulai; Roitsch, Thomas Saqib Saleem Akhtar1, Daniel Buchvaldt Amby1, Josefine Nymark Hegelund1*, Lorenzo Fimognari2, Dominik K. Großkinsky1†, Jesper Cairo Westergaard1, Renate Müller1,Lars Moelbak2, Fulai Liu1 and Thomas Roitsch1,3

1Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Taastrup, Denmark

2Plant Health Innovation, Chr-Hansen A/S, Hørsholm, Denmark

3Department of Adaptive Biotechnologies, Global Change Research Institute, Czech Academy of Sciences, Brno, Czechia

Increasing agricultural losses due to biotic and abiotic stresses caused by climate change challenge food security worldwide. A promising strategy to sustain crop productivity under conditions of limited water availability is the use of plant growth promoting rhizobacteria (PGPR). Here, the effects of spore forming Bacillus licheniformis (FMCH001) on growth and physiology of maize (Zea mays L. cv. Ronaldinho) under well-watered and drought stressed conditions were investigated. Pot experiments were conducted in the automated high-throughput phenotyping platform PhenoLab and under greenhouse conditions. Results of the PhenoLab experiments showed that plants inoculated with B. licheniformis FMCH001 exhibited increased root dry weight (DW) and plant water use efficiency (WUE) compared to uninoculated plants. In greenhouse experiments, root and shoot DW significantly increased by more than 15% in inoculated plants compared to uninoculated control plants. Also, the WUE increased in FMCH001 plants up to 46% in both well-watered and drought stressed plants. Root and shoot activities of 11 carbohydrate and eight antioxidative enzymes were characterized in response to FMCH001 treatments. This showed a higher antioxidant activity of catalase (CAT) in roots of FMCH001 treated plants compared to uninoculated plants. The higher CAT activity was observed irrespective of the water regime. These findings show that seed coating with Gram positive spore forming B. licheniformis could be used as biosestimulants for enhancing plant WUE under both normal and drought stress conditions.