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Construction and Application of Efficient<i>Ac-Ds</i>Transposon Tagging Vectors in Rice
Qu, Shaohong,Jeon, Jong-Seong,Ouwerkerk, Pieter B.F.,Bellizzi, Maria,Leach, Jan,Ronald, Pamela,Wang, Guo-Liang Wiley (Blackwell Publishing) 2009 Journal of integrative plant biology Vol.51 No.11
<P>Transposons are effective mutagens alternative to T-DNA for the generation of insertional mutants in many plant species including those whose transformation is inefficient. The current strategies of transposon tagging are usually slow and labor-intensive and yield low frequency of tagged lines. We have constructed a series of transposon tagging vectors based on three approaches: (i) AcTPase controlled by glucocorticoid binding domain/VP16 acidic activation domain/Gal4 DNA-binding domain (GVG) chemical-inducible expression system; (ii) deletion of AcTPase via Cre-lox site-specific recombination that was initially triggered by Ds excision; and (iii) suppression of early transposition events in transformed rice callus through a dual-functional hygromycin resistance gene in a novel Ds element (HPT-Ds). We tested these vectors in transgenic rice and characterized the transposition events. Our results showed that these vectors are useful resources for functional genomics of rice and other crop plants. The vectors are freely available for the community.</P>
Guo-Liang Wang,Shaohong Qu,Zhengjie Yuan,Yu Zhang,Guojuan Xu,Dongling Bi,Haiyan Qu,Xiaowei Zou,Xiaoqing Gao,Haihe Yang,Haiyan He,Xuli Wang,Jiandong Bao,Shimin Zuo,Xuebiao Pan,Bo Zhou 한국식물학회 2018 Journal of Plant Biology Vol.61 No.3
Rice sheath blight (SB), caused by Rhizoctoniasolani (R. solani), is a major threat to rice productionworldwide. The molecular mechanisms of the SB resistancein rice are poorly understood. The transcriptomes of the SBresistantrice cultivar YSBR1 and the susceptible cultivarLemont were analyzed after R. solani infection. A total of7624 differentially expressed genes (DEGs) were identifiedat one or more timepoints in a cultivar. 5526 and 5618 DEGswere differentially expressed in Lemont and YSBR1,respectively. YSBR1 exhibited stronger and earlier transcriptionalresponse to R. solani than Lemont. Gene ontology enrichmentanalysis revealed that genes that encode cell wall-modifyingand glycosyl-degrading enzymes or anti-microbial proteinswere specifically induced in YSBR1 at 6 hpi. MapMananalysis revealed that more DEGs related with cell wall, β-glucanses, respiratory burst, phenylpropanoids and ligninwere highly induced by R. solani in YSBR1 than in Lemont. The results also showed that receptor-like kinases and jasmonicacid signaling may play important roles in host resistance toR. solani. This study highlights potential candidate genes andsignaling pathways involved in rice sheath resistance and canhelp to further clarify the mechanistic events underlyingresistance and susceptibility to R. solani.
Hyun-Bi Kim,전종성,Jung-Il Cho,Nayeon Ryoo,Shaohong Qu,Guo-Liang Wang 한국분자세포생물학회 2012 Molecules and cells Vol.33 No.1
The development of rapid and efficient strategies to gen-erate selectable marker-free transgenic plants could help increase the consumer acceptance of genetically modified (GM) plants. To produce marker-free transgenic plants without conditional treatment or the genetic crossing of offspring, we have developed a rapid and convenient DNA excision method mediated by the Cre/loxP recombination system under the control of a -46 minimal CaMV 35S promoter. The results of a transient expression assay showed that -46 minimal promoter::Cre recombinase (-46::Cre) can cause the loxP-specific excision of a selectable marker, thereby connecting the 35S promoter and -glucuronidase (GUS) reporter gene. Analysis of stable transgenic Arabidopsis plants indicated a positive correlation between loxP-specific DNA excision and GUS expression. PCR and DNA gel-blot analysis further revealed that nine of the 10 tested T1 transgenic lines carried both excised and non-excised constructs in their genomes. In the subsequent T2 generation plants, over 30% of the individuals for each line were marker-free plants harboring the excised construct only. These results demonstrate that the -46::Cre fusion construct can be efficiently and easily utilized for producing marker-free transgenic plants.