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Funtion Identification of eIF1A Gene from Tamarix.s

Author: WangYouJu
Tutor: LiuGuiFeng
School: Northeast Forestry University
Course: Tree Genetics and Breeding
Keywords: eIF1A gene Salt tolerance Populus Functional verification
CLC: S793.5
Type: Master's thesis
Year: 2008
Downloads: 66
Quote: 0
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Abstract


The gene encoding the eukaryotic translation initiation factor（eIF1A） had been cloned from Tamarix.spp,a tree species with high tolerant ability to extreme environmental conditions including saline,drought and freezing.In this study,the eIF1A gene was heterogenously expressed in yeast to test its resistant abilities.Meanwhile,the eIF1A gene was transformed into Populus davidiana Dode×P.bolleana Lauche.genome by Agrobacterium-mediated transformation method.Then,the physiological traits were determined to examine the functions of the exogenous eIF1A gene in Populus davidiana Dode×P.bolleana under saline stress.The results showed that:（1） The recombinant plasmid pYES-eIF1A was constructed by inserting eIF1A gene to yeast expression vector pYES2.The recombinant vector was transformed into Saccharomyces cerevisiae （INVSc1 strain）.PCR results indicated that the recombinant vector had been transformed into the yeast successfully.Furthermore,Northern bloting indicated that the exogenous gene can be transcript correctly.（2） The recombinant yeast INVSc1（pYES-eIF1A） was treated under NaCl,Na2CO3,high temperature and low-tempertuate stress.The results revealed that the stress-resistance of recombinant yeast INVSc1（pYES-eIF1A） yeast was better than control strain.These indicated that the protin synthesizes function and the ability of stress-resistance of recombiant yeast had been enhanced due to over-expression of eIF1A gene.（3） eIF1A gene had been integrated into Populus davidiana Dode×P.bolleana Lauche via genome by Agrobacterium-mediated transformation systems.Twenty-one independent kanamycin-resistant shoots were obtained and fifteen shoots showed positive result in PCR assay.Then Northern blotting indicated that exogenous gene can be transcript correctly in transgenic poplar plants.（4） Contents of malondialdehyde（MDA）,activity of superoxide dismutase（SOD）,activity of peroxidase（POD）,content of chlorophyll and total soluble protein of the transgenic plants were examined and analysised under NaCl stress.The result shows:when the transgenic lines were treated with 1.2M NaCl,the content of chlorophyll of all lines decreased,but the content of chlorophyll of transgenic lines higher than nontransgenic lines;the activity of SOD and POD of transgenic lines were higher than control,and the tenth transgenic line was best,its activity of SOD and POD was 1.19 and 2.77 times than control lines;the content of soluble protein of all lines were higher than control.The sixth transgenic line was excellence and its content of soluted protein was 1.76 times than control lines;the content of MDA lower than control,content of MDA in control lines was 2.21 times than the tenth transgenic plant.All the results indicated that the constitutively expression of eIF1A gene can improve the

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CLC: > Agricultural Sciences > Forestry > Forest tree species > Broad-leaved shrubs > Tamarix
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