Abstract

Genetic engineering of seed size and increasing biomass in crop plants has an important significant contribution to the world. Arabidopsis DA1 is one of the key factors that negatively control seed and organ size by restricting the period of cell proliferation, and the mutant of Arabidopsis DA1, da1-1 (DA1R358K) can dramatically increase the size of seed. However, it is not clear whether overexpression of Zmda1-1, the mutant of ZmDA1 which is homology of DA1 in Arabidopsis, has the same biological effect as da1-1 in Arabidopsis. Therefore, in this study, the plant expression vector harboring bothZmda1-1 driven by the corn ubiquitin promoter and a PAT selectable marker gene driven by 35S CAMV promoter was constructed and introduced into maize inbred line ‘ji444’ using pollen-tube-pathway method. Screened with herbicide phosphinothricin (PPT), 22 seedlings of 2563 transformed samples survived, and 21 independence lines of which were positive in polymerase chain reaction (PCR) analysis, and the transformation rate of T0 generation was about 0.82%. Further PCR-southern blotting results proved that theZmda1-1 had integrated into maize genome, and the Zmda1-1 had expression in low level by reverse transcription-polymerase chain reaction (RT-PCR) analysis. The seed mass of transgenic maize increased at an average of 33.6% of empty vector control lines, and the harvest yield was increased by 23.6 to 114.1% in different lines than empty vector control lines. The result suggests that Zmda1-1 can be used to engineer higher harvest yield in crops plant, thus providing the first successful example of increasing the harvest yield of maize by transgenic technology.

Key words: Transgenic maize, pollen-tube pathway, Zmda1-1, seed mass.

Abbreviation

PCR, Polymerase chain reaction; RT-PCR, reverse transcription-polymerase chain reaction; ARGOS, Arabidopsis thaliana auxin-inducible protein; ANT,AINTEGUMENTA; BB, BIG BROTHER; EOD1, enhancer of DA1; ABA, abscisic acid;ACT, actin gene; ORF, open reading frame; UIM, ubiquitin interaction conserved motifs;CK, control check; DREB, dehydration responsive element binding protein; BT, Bacillus thuringiensis.