Ntitative Analysis Of Endogenous Hormones And Expression Of Flowering Related Genes In The Process Of Regenerating Floral Buds On In Vitro Cultures Of Perianth In Sinningia Speciosa

Phytohormones have been shown to participate in all the stages of plant development including seed germination, stem elongation, flower and fruit development and so on. Sinningia speciosa is a day-neutral plant. Vernalization is not requried for flowering. It need three to four months from seeding to flowering. Results of our previous study showed that the in vitro cultures of perianth can directly regenerate floral buds in Sinningia speciosa. To explore the role of phytohormones and genes related to flower development in floral bud differentiation in S. speciosa, we detect the relative expression amount of SsSOCla and SsSPY in perianth and young leaf segments via real-time fluorescent quantitative polymerase chain reaction(FQ-PCR); we also detect the content of endogenesis phytohormones in them via enzyme-linked immunosorbent assays(ELISA).Main research results are as follows:1. Primers were designed according to the conserved sequences in SsActin^SsSPY and SsSOCla of Sinningia speciosa. SsActin gene was used as the reference gene for SsSPY and SsSOCla gene expression analysis by real-time quantitative PCR technology during the process of Sepal, petal and young leaf segments cultured in vitro. The results showed that the expression level of SsSOCla tends to ascend in the initial stage, sepal and petal segments reach the peak when cultured for15days, and the value of petal segments is12.04. While the peak during the culturing of young leaf segments is5.82, when cultured for18days. The expression level of SsSPY tends to ascend in the initial stage. Until cultured for10days, petal and young leaf segments reach the peak, that is7.46and6.19respectively. Then it decreased. The peak in sepal segments arised at15days, and the relative expression amount is only5.94.3. we detect the content of different phytohormones at different stages of sepal, petal and young leaf segments cultured in vitro via enzyme-linked immunosorbent assay. The results showed the content of abscisic acid(ABA) was higher than others. And young leaf segments had the highest level, it could reach2622050pg/g when cultured for18days. The changing patterns of zeatin riboside(ZR) was divergent after cultured for15days, it went up in sepal and young leaf segments, but maitained around1600pg/g in petal segments. Indoleacetic acid(IAA) has the lowest level. Generally speaking, it was higher in petal and sepal segments, it could be0.30pg/g and0.25pg/g separately, as well as0.20pg/g in young leaf segments. The content of gibberellin(GA) changed obviously in petal segments, from1956.83pg/g at initial stage to4550.57pg/g at cultured for10days, then maitained at high level. While it kept above3000pg/g in young leaf segments during the whole process.