Construction ofZmPti1Sense and RNAi Expression Vectors and Their Transformation into Maize

doi:10.7668/hbnxb.2008.05.001

Abstract

Abstract: ZmPti1(AY708048)cloned in our lab encodes a maize pti - like kinase.In order to efficiently identify the functions of Zm Pti1 gene,two plant - transformation vectors,pBPC - ZmPti1 - bar(sense expression vector)and pBPC - PtS - GFP - PtAs - bar(RNAi vector)were prepared.And promoters used in these two constructs were maize ubiquitin promoter. These two vectors contain herbicide resistance gene(bar gene)that can facilitate large scale selection of transformant in farm land.Sense expression and RNAi vectors of Zm Pti1 were delivered into maize inbred lines178 by pollen tube path - way method respectively,and 33 transformed plants were obtained through glufosinate screening,PCR analysis and PCR - southern identification.Among these transformants,fifteen plants are Zm Pti1 overexpresssion lines,and 18 plants are Zm Pti1 RNAi lines.Furthermore,the transformation efficiency in our experiment is related to the introducing time.In T 0 generation,overexpression lines are more tolerant to disease stress than that of knockout lines.It indicates that Zm Pti1 is involved in disease resistant signal pathway.The transformants obtained can be used to study the mechanism of defense response for Zm Pti1 in maize.And the results demonstrate that the approach of pollen tube pathway is simple and effi - cient for maize transformation.

Key words: ZmPti1gene, Sense expression vector, RNAi vector, Pollen tube pathway, Transgenic plants

CLC Number:

S513.035.3

ZHANG Sen-yan, WU Zhong-yi, ZHANG Xiu-hai, LIU Zhan-lei, WANG Yong-qin, HUANG Cong-lin. Construction ofZmPti1Sense and RNAi Expression Vectors and Their Transformation into Maize[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2008, 23(5): 1-5. doi: 10.7668/hbnxb.2008.05.001.

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