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A systematic analysis of apple root resistance traits to Pythium ultimum infection and the underpinned molecular regulations of defense activation

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Horticulture Research 2020年第1期7卷 1878-1888页

作者： Yanmin Zhu melody saltzgiver USDA-ARS Tree Fruit Research LaboratoryWenatcheeWA 98801USA

Apple replant disease(ARD),caused by a pathogen complex,significantly impacts apple orchard establishment.The molecular regulation on ARD resistance has not been investigated until recently.A systematic phenotyping effort and a series of transcriptomic analyses were performed to uncover the underpinned molecular mechanism of apple root resistance to P.ultimum,a representative member in ARD pathogen complex.Genotype-specific plant survival rates and biomass reduction corresponded with microscopic features of necrosis progression patterns along the infected root.The presence of defined boundaries separating healthy and necrotic sections likely caused delayed necrosis expansion in roots of resistant genotypes compared with swift necrosis progression and profuse hyphae growth along infected roots of susceptible genotypes.Comprehensive datasets from a series of transcriptome analyses generated the first panoramic view of genome-wide transcriptional networks of defense activation between resistant and susceptible apple roots.Earlier and stronger molecular defense activation,such as pathogen perception and hormone signaling,may differentiate resistance from susceptibility in apple root.Delayed and interrupted activation of multiple defense pathways could have led to an inadequate resistance response.Using the panel of apple rootstock germplasm with defined resistant and susceptible phenotypes,selected candidate genes are being investigated by transgenic manipulation including CRISPR/Cas9 tools for their specific roles during apple root defense toward P.ultimum infection.Individual apple genes with validated functions regulating root resistance responses can be exploited for developing molecular tools for accurate and efficient incorporation of resistance traits into new apple rootstocks.

关键词： roots separating likely

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A Phenotyping Protocol for Detailed Evaluation of Apple Root Resistance Responses Utilizing Tissue Culture Micropropagated Apple Plants

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American Journal of Plant Sciences 2018年第11期9卷 2183-2204页

作者： Yanmin Zhu melody saltzgiver Jingxian Zhao United States Department of Agriculture Agricultural Research Service Tree Fruit Research Laboratory Wenatchee WA USA Hebei Academy of Forestry Shijiazhuang China

In the post-genomics era, reliable phenotypes are considered the bottleneck for unraveling the genetic control over the biology of interest. Phenotyping resistance response of roots to infection by soilborne pathogen is more challenging compared to that of plant aerial parts. In additional to the hidden nature and small stature of fine roots where infection occurs, extra obstacles exist for rosaceae tree crops such as apple. Due to self-incompatible reproduction and high-level heterozygosity of apple genome, genetically identical apple plants cannot be produced through apple seed germination. Here we report an established phenotyping protocol which includes a streamlined tissue culture procedure for micropropagation of uniform apple plants, standardized inoculation procedure using Pythium ultimum, and multilayered evaluating methods on apple root resistance traits. Because of the implementation of tissue culture based micropropagation procedure, constant availability of the uniform plants with defined genetic background, equivalent age and non-contaminated roots overcame a longstanding barrier of systematic and detailed phenotypic characterization of apple root resistance traits. Repeated infection assays by root-dipping inoculation demonstrated the reproducible and wide-range plant survival rates, from single-digit to over 90% survived plants for a given genotype. Genotype-specific values due to P. ultimum inoculation on shoot and root biomass reduction, maximum root lengths, leaf number and cumulative leaf areas were quantified between mock-inoculated and P. ultimum infected plants. Use of a glass-box container offered enhanced accessibility and minimized invasiveness for continuous and non-disruptive observation on the necrosis progression patterns along inoculated roots. With the assistance of a dissecting microscope, the genotype-specific resistance responses along the infected apple roots were captured and analyzed in detail. This reported phenotyping protocol represents a major development and should be ea

关键词： Tissue Culture Micropropagation Apple Rootstocks Root Resistance Responses Phenotyping Methods

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