Rice (Oryza Sativa L.) DH PoPulation Construction And SSRs Markers Analysis

Microsatellites, also known as simple nucleotide repeats (SSRs), are tandem arrays of short nucleotide repeats from 1 to 6 bases per unit, which exhibit high level polymorphism at a locus and become one of the most widely used genetics markers. To study the genetics behaviors of two foreign genes, a double haploid (DH) population with 84 plant lines and three F2 populations were constructed, based on which the polymorphism of 145 SSRs markers and segregation patterns of 7 SSRs markers were investigated, and the linkage groups involving 5 SSRs nearby waxy gene were built based on cw3 and DH populations. The main results could be summarized as follows:1 Effects of S-3307 on the induction and differentiation of calli and the growth ofplantlets in rice anther cultureEffects of plant growth retardant S-3307 on anther culture of japonica rice varieties were studied by comparison to another plant growth retardant multi-effects triazole (MET or PP333). After S-3307 (lmg/L) was supplemented to the callus induction medium, the callus induction and green plantlet differentiation increased intensively, but anther would die when the concentration of S-3307 increased to lOmg/L. And supplemented S-3307 to the regeneration medium, the green plantlet formation were promoted significantly, especially with a low concentration of S-3307 (lmg/L). However, with the concentration of S-3307 increased, callus differentiation and green plantlets formation were inhibited dramatically and calli would die at 25mg/L. It was also noted that different genotypes show different responses to culture on S-3307-containing regeneration medium. Effects of S-3307 in growth medium were also investigated. 0.1-lmg/L S-3307 could strengthen plantlets and increase the survival rate of hardening plantlets.2 Homozygous Basta resistance rice lines obtained via anther cultureA new technology was developed for identifying hybrid offspring of transgenic rice with non-transgenic rice based on anther culture combining with resistant screening in order to produce objective homozygous lines. PPT, the active compound in Basta or bialaphos, was added to the anther culture calli induction and regeneration medium of FG202, hybrid of transgenic line PP2 of bar gene with non-transgenic line Ux58. With 0.05mg/L PPT in the medium, many calli and green plantlets were obtained, but the frequencies of "escape" plants (i.e. non-containing of bar gene) were not obviously different to the control according to PCR analysis and field resistance assay. With the increase of PPT concentration, the ratio of calli induction and frequency of green plantlet were decreased dramatically. PCR analysis and field resistance assay indicated that all plants from Img/L PPT-containing medium were PPT resistant and contained bar gene. When PPT concentration increased to 5mg/L, the calli induction and differentiation were restrained completely. 6 agronomic characters of 30 obtained double haploid lines were analyzed in two generations (H.2 and hs) and the variation coefficient of high heritability traits were below 10% in most lines (exceed 73%), which suggested that using the method could obtain homozygous resistance lines.3 The inheritance and field performance of foreign genes in DH populationA double haploid population with 84 plant lines were constructed through the high efficient anther culture method from FG211, which is the hybrid of transgenic line HH35 harboring two foreign genes (bar gene and Pinll gene which were constructed in one vector) with non-transgenic line Ux58. The inheritance and field performance of the foreign genes in two generations of DH lines were studied, and it seemed that the two foreign genes were coinheritance and the expression level of the genes were various in different generations. The resistance of Basta was increased in the lines with bar gene, but the extends of resistance were different as bar gene silenced in several positive lines. Potato proteinase inhibitor â…¡ (Pinâ…¡) is a kind of plant derived proteinase inhibitors that can pr...