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Screening Of Arabidopsis Salt-sensitive Mutant Mns1 Mns2 Suppressors

Posted on:2020-09-26Degree:MasterType:Thesis
Country:ChinaCandidate:F F WangFull Text:PDF
GTID:2480305771956139Subject:Botany
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Salt stress is one of the main abiotic stresses and greatly affects crop growth and yield.Currently,two main technologies are used to control soil salinization,transforming soil by chemicals and physical methods and cultivating the salt tolerant varieties.The cost of the former is high and the addition of a large amount of chemical substances increase risk of secondary salinization of soil.Therefore,it becomes a very important alternative to cultivate salt-tolerant crop varieties.However,plant tolerance against salinity is mediated by a sophisticated network and multiple signaling pathways are involved in the process.Thus,the effect of improving plant salt tolerance by altering the expression of a single gene is often insignificant.N-glycosylation is an important protein post-translational modification process in eukaryotes and involved in various biological processes.Our previous studies reveal that the mutations in ? 1,2-mannosidase(Golgi ? 1,2-mannosidase I,MNS1 and MNS2),responsible for N-glycan trimming in Golgi,cause Arabidopsis hypersensitivity to salt.To further inverstigate the mechanism of N-glycan-mediated plant response to salt stress,I used mnsl mns2 as background to construct a genetic mutant library by activation tagging strategy.I screened a mutant strain(1703-119)exhibiting partially rescued salt-sensitive phenotype.The T-DNA insertion site on SGAM1 gene(suppressors of Golgi alpha-1,2-mannosidase I)was determined by Adaptor PCR,hiTAIL-PCR and whole genome sequencing.The down-regulation of SGAM1 was confirmed by RT-PCR and qRT-PCR.The phenotypic rescue of SGAM1 overexpression in 1703-119 to salt sensivity and the improved phenotype of mnsl mns2 to salt stress by knockdown of SGAM1 expression confimed that SGAM1 is a functional suppressor gene of mnsl mns2.The possible molecular mechanism was also discussed.This study provides a theoretical basis for further revealing the mechanism of N-glycan-mediated plant response to salt stress.
Keywords/Search Tags:Salt stress, N-glycan, Activation Tagging, Adaptor-PCR, hi-TAIL-PCR, CRISPR/Cas9
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