Studies On Expression Of Vdac, Ant And HL-sp1 Genes In Rice

Cytoplasmic male sterility (CMS) is a very common phenomenon in plant kingdom which plays an important role in genetic breeding. In most plants, CMS is in relation to mitochondrial genomic dysfunction. For instances, the mitochondria-associated genes, HL-sp1 and orfH79, are involved in the Honglian CMS (HL-CMS) in rice. The function of mitochondria is also regulated by nuclear genes. The voltage-dependent anion channel protein (VDAC), which across the mitochondrial outer membrane, coded by nuclear genome is involved in the transport of metabolites, including NADH and ATP, and plays an important role in mitochondrial metabolism and ion signaling. Adenine nucleotide translocase (ANT) coded by nuclear genome is an inner membrane transporter that facilitates the exchange of ATP and ADP and is associated in cellular energy metabolism by influencing mitochondrial oxidative phosphorylation. In this study, the expressions of HL-sp1, vdac and ant in rice were carried out. The results are as follow:1. The bioinformatics analysis on eight vdac genes, previously predicted based on Nipponbare genome in rice, and their expression products was first carried out. The expression models of eight vdac genes in the different tissues of HL-CMS sterile line Yuetai A (YTA) at different growth and development stages were analyzed by method of real-time PCR. The results showed that the expressions of osvdac1 and osvdac4 were much higher among the eight vdac genes during the whole growth stages of YTA. When being treated with abiotic stress of drought, 200 mmol/L NaCl and 360 mmol/L mannitol, the expression patterns of eight vdac genes in different tissues of YTA were greatly changed during the periods of treatment and stress recovery. And one of vdac genes, osvdac4, was then selected, and its 5'-UTR and 3'-UTR sequences was analyzed by method of SMARTer RACE. According to sequencing data, the complete 3'-UTR sequence of osvdac4 gene was confirmed.2. The expression patterns of three ant genes in the different tissues of YTA at different growth and development stages were analyzed by method of real-time PCR. The results indicated that the expression of osant1 and osant2 was higher at vegetative stages of YTA, but lower during reproductive stage. Osant3 was weakly expressed in most tissues of YTA at all growth and developmental stages.3. SMARTer RACE was used to amplify the complete 5'-UTR and 3'-UTR sequences of orf216 gene. According to sequencing data of amplified cDNA, the complete 3'-UTR sequence of orf216 gene and its co-transcription with its upstream genes was confirmed.