The Functional Complemention And RNAi Analysis Of EUI2(t) Gene In Rice

A newly found rice gene eui2 (t) controlling the elongation of uppermost internode ,aquired by mutation ,was finely mapped and cloned by Zhu (2003), and the candidate gene was concluded by comparing the DNA sequences among the four eui2 CO allelic mutants showing different nucleotide deletion exons in the same gene. Based on the studies of Zhu, the functional complementation and RNAi analysis for the condidate gene was conducted by mediated Agrobacterium tumefaciens in this research .The main results were as follows.1 The contructed vecter-pCAMBIA 1301-EUI2 was introduced into embryogenic calli from immature seeds of the populations with the the elongation uppermost internode phenotype in the F₂ and F3 of the cross Xieqingzao eB2 (ssp. indica) and Zhonghua14 (ssp. japonica). A total of 6597 embryogenic calli were co-cultivated with Agrobacterium tumefaciens and the 380 Hyg resistant calli were obtained with the frequency of 5.76%; The 109 calli which had the percentage of 28.68% in the 380 Hyg resistant calli obtained transgenic plants. The PCR for 23plants in 12transgenic lines suggested that the candidate gene of EUI2(t) was integrated into rice genome. But the only two positive plants (37-1,82-3) were identified with obviously contracted uppermost internode. Their T₁ plants had the segregation of the mutant type (longer uppermost internode) and the wild type with the rate 37:12 and 38:10, respectively.2 The vecter-pCAMBIA1301-EUI2-ihpRNAi was transfected into embryogenic calli from immature seeds of Minghui86 and Zhonghua11 by transformed Agrobacterium tumefaciens. The 34 Hyg resistant calli were obtained from 1304 embryogenic calli and the frequency of transformation was 2.60%; The five T₀ transgenic lines were received from 34 Hyg resistant calli; The frequency...