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Functional Analysis Of RcNHX1 Gene In Castor (Ricinus Communis L.)

Posted on:2018-09-09Degree:MasterType:Thesis
Country:ChinaCandidate:Z Z FengFull Text:PDF
GTID:2323330512482583Subject:Crop Science
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Soil salinization has been one of the most difficult agricultural problems in the world.It has seriously affected the development of global agricultural economy and the protection of ecological environment.The use of genetic engineering for the selection of salt-tolerant crop varieties has become an economically effective way to utilize and improve saline soil.In recent years,with the development of the castor industry,people on the castor production and processing of the increasing importance,and castor salt-related genes to explore and use is rare.To identify the potential genes involved in salt stress response,we reported a vacuole Na~+/H~+ antiporter RcNHX1,isolated from Tongbi 5 castor inbred line using RACE(Rapid amplification of cDNA ends)PCR method.Full length cDNA that consisted of a single open reading frame encoded a putative polypeptide of 544 amino acid.Subcellular localization of RcNHX1 indicated that RcNHX1 was mainly localized in the cell membrane and vacuole membrane.Overexpression vector and RNAi interference vector were successfully constructed.We obtained stable RcNHX1 overexpression transgenic lines by using Agrobacterium-mediated cotyledon transformation.PCR and Southern blotting showed that two stable inheritable overexpression lines were obtained.We assayed phenotype analysis for salt tolerance in overexpressed RcNHX1 castor plants under seedling stage.The results displayed that shoot weight and dry weight were significantly increased in transgenic plants compared to wild type after 150 mmol/L NaCl treatment for 14 days.The content of proline in transgenic plants was significantly higher than that of wild type,while the content of malondialdehyde and the relative electrolyte permeability were significantly decreased under the salt stress.Moreover,compared to wild type,the transgenic plants accumulated more content of Na~+ and K~+ in roots and leaves under the salt stress,which may provide the possible mechanism for salt resistance.
Keywords/Search Tags:Ricinus communis L., Salt stress, RcNHX1, Genetic transformation
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