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High-throughput analysis of anthocyanins in horticultural crops using probe electrospray ionization tandem mass spectrometry (PESI/MS/MS)

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Horticulture Research 2023年第4期10卷 249-258页

作者： Misaki Ishibashi Kei Zaitsu Ikue Yoshikawa Shungo Otagaki Shogo Matsumoto Akira Oikawa katsuhiro shiratake Graduate School of Bioagricultural Sciences Nagoya UniversityChikusaNagoyaAichi 464-8601Japan Graduate School of Agriculture Kyoto UniversitySakyoKyoto 606-8502Japan Faculty of Biology-Oriented Science and Technology Kindai UniversityNishimitaniKinokawaWakayama 649-6493Japan Faculty of Agriculture Meijo UniversityTenpakuNagoyaAichi 468-8502Japan

Plant secondary metabolites exhibit various horticultural traits.Simple and rapid analysis methods for evaluating these metabolites are in demand in breeding and consumer markets dealing with horticultural crops.We applied probe electrospray ionization(PESI)to evaluate secondary metabolite levels in horticultural crops.PESI does not require pre-treatment and separation of samples,which makes it suitable for high-throughput analysis.In this study,we targeted anthocyanins,one of the primary pigments in horticultural crops.Eighty-one anthocyanins were detected in approximately 3 minutes in the selected reaction-monitoring mode.Tandem mass spectrometry(MS/MS)could adequately distinguish between the fragments of anthocyanins and flavonols.Probe sampling,an intuitive method of sticking a probe directly to the sample,could detect anthocyanins qualitatively on a micro-area scale,such as achenes and receptacles in strawberry fruit.Our results suggest that PESI/MS/MS can be a powerful tool to characterize the profile of anthocyanins and compare their content among cultivars.

关键词： MS/MS ionization dealing

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PbMC1a/1b regulates lignification during stone cell development in pear(Pyrus bretschneideri)fruit

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Horticulture Research 2020年第1期7卷 1915-1927页

作者： Xin Gong Zhihua Xie Kaijie Qi Liangyi Zhao Yazhou Yuan Jiahui Xu Weikang Rui katsuhiro shiratake Jianping Bao Shahrokh Khanizadeh Shaoling Zhang Shutian Tao College of Horticulture Nanjing Agricultural UniversityNanjingChina Laboratory of Horticultural Science Nagoya UniversityNagoyaJapan College of Plant Science Tarim UniversityAla’er CityChina ELM Consulting Inc. St-LazareCanada Eastern Cereal and Oilseed Research Centre Agriculture and Agri-Food CanadaOttawaCanada State Key Laboratory of Crop Genetics and Germplasm Enhancement Nanjing Agricultural UniversityNanjingChina

Programmed cell death(PCD)and secondary cell wall(SCW)thickening in pear fruit are accompanied by the deposition of cellulose and lignin to form stone cells.Metacaspase is an important protease for development,tissue renewal and PCD.The understanding of the molecular mechanism whereby pear(Pyrus)metacaspase promotes PCD and cell wall lignification is still limited.In this study,the Metacaspases gene family(PbMCs)from P.bretschneideri was identified.PbMC1a/1b was associated with lignin deposition and stone cell formation by physiological data,semiquantitative real-time polymerase chain reaction(RT-PCR)and quantitative RT-PCR(qRT-PCR).Relative to wildtype(WT)Arabidopsis,the overexpression of PbMC1a/1b increased lignin deposition and delayed growth,thickened the cell walls of vessels,xylary fibers and interfascicular fibers,and increased the expression of lignin biosynthetic genes.Yeast two-hybrid(Y2H),bimolecular fluorescence complementation(BiFC)and GST pull-down assays indicated that the PbMC1a/1b protein physically interacted with PbRD21.Simultaneously,the transient expression of PbMC1a/1b and PbRD21 led to significant changes in the expression of genes and lignin contents in pear fruits and flesh calli.These results indicate that PbMC1a/1b plays an important role in cell wall lignification,possibly by interacting with PbRD21 to increase the mRNA levels of some lignin synthesis-associated genes and promote the formation of stone cells in pear fruit.

关键词： fibers walls transient

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