| Auxin, as the first discovered plant hormone, play a crucial role in the whole process of plant growth and development. ARF(Auxin response factor), as transcription factor, play a important role in the auxin signaling transduction pathway. By biding to the Auxin-responsive element (AUXRE) in the promoter region of the auxin-response genes, ARF can regulate the expression level of these genes. AUXIN/IAA gene family members encode proteins that can interact with the ARFs which lead to hetero-dimerization, by this way, AUXIN/IAAs can also regulate the expression of the auxin-response genes. The expression level of AUXIN/IAA, GH3and SAUR gene family members can be induced by IAA in several minutes and they are called primary auxin response genes. In Production, unsuitable temperature is always a limitation for improving the yield of tomato. Previous have shown that the Auxin-related genes play important role in the parthenocarpy of tomato. So, the genome wide analysis of auxin-related genes from tomato gene will be useful for the functional confirmation of those genes, it will also be helpful for the improvement of tomato yield under unsuitable temperature.In this study, we focus on the expression pattern of ARF and other auxin-related gene family members during the young tomato fruit development. We also constructed the Artifieial MirRNA plant expression vector of S1ARF5to investigate the role S1ARF5played in the tomato fruit developmet. The Purpose of this study is to provide the helpful tools for understanding molecular mechanism of the role of IAA played in the tomato fruit development. The major study results are following:(1) Amino acid sequences which were previous reported were used to TBLASTN against tomato genome to get the DNA sequences. These sequences were predicted by FGENESH. Based on the predicted results, Those gene were isolated by PCR-based methods or E-clone. The length of the nucleotide varied from1218bp (S1ARF12) to3372bp(S1ARF7). Those21genes were distributed on all the12chromosome excepted chromosome9. Most of these gene cantain DBD,MR and CTD domains, but S1ARF3,6-1,12-14,13-1and17lost the CTD domain. Based on the phylogenetic tree, all the21genes could be divided into three groups (group â… , â…¡ and â…¢). Expression analysis shown that S1ARFs could be detedted in all the parts of the plant and shown no marked difference. S1ARF genes shown different expression patterns during the flower development. With the development of ovary and young fruit, the expression level increased to a peak on the third or sixth day after flower opening and then markedly decreased on the nineth day. Noticeably, the expression level of S1ARF4was quite different with the other ARF genes, constantly increasing even after pollination during young fruit development.(2) Using the same metods,26AUX/IAA genes were isolated from the tomato genome. The length of the nucleotide varied from228bp (S1IAA13) to1050bp(S1IAA4). Those26genes were distributed on all the12chromosome excepted chromosome2,10and11. Most of these gene cantain â… , â…¡, â…¢ and â…£ domains, but S1IAA13,15,16,18,20and27lost the domain â…¢ and â…£. Based on the phylogenetic tree, all the26genes could be divided into two groups (group A and B). The expression levels of S1IAAs in different parts of plant were extremely different, during the flower development, all the S1IAA genes, except S1IAA16,20,23and24shown a same expression pattern, with a constant increase of expression levels during the flower development. With the development of ovary and young fruit, S1IAA genes shown different expression patterns. Most of expression levels of the S1IAAs could be induced by auxin, but the expression of S1IAA19and20were inhibited by auxin. Under the stress treatments, differen+genes shown different responses to these treatment.(3) The toamto genome contains32GH3genes, but about half of these genes were small genes, which was not reported by previous study. Those32genes were distributed on all the12chromosome excepted chromosome3,4,9and11. Based on the phylogenetic tree, all the32genes could be divided into two groups (group â… and â…¡). The expression levels of12selected S1GH3s in different parts of plant were extremely different. With the development of ovary and young fruit, S1GH3genes shown different expression patterns. During the ovary development, all the genes show the same expression patterns, with a constant increase. All the12selected S1GH3genes, excepted S1GH3.18, show no obvious change after axuin treatment.(4)The toamto genome contains99SAUR genes. The length of nucleotide varied from 186bp (SI SAUR30) to597bp (S1SAUR54). Those99genes were distributed on all the12chromosomes. Based on the phylogenetic tree, all the484SAUR gene from different specious were divided inth4major groups and further divided into16subgroups. The expression levels of11selected SISAURs in different parts of plant were extremely different. Different genes shown different expression patterns during the flower and ovary development. All the11selected SISAUR genes excepted S1SAUR36, S1SAUR39and S1SAUR58shown obvious change after auxin treatment. H-rich region were found from the peptide sequences of26SAUR genes from different specious. The expression level of S1SAUR58, which contain a high Histidine content, was inhibited by drought and heat but upregulated by salt treatment. Combine with the evidence that the expression of S1SAUR58also can be induces by IAA, we may put forward the hypotheses that S1SAUR58may play a specific role that connect the salt signaling and auxin signaling transduction pathways.(6) We constructed the Artificial MirRNA plant expression vector. First of all, using the miR164a of Arabidopsis as skeleton, We replaced the mature MirRNA with the specific sequence of S1ARF5. There is no report that miR164a can interaction with ARF which exclude the possibility that natural miR164a in tomato can interact with S1ARF5. Secondly, we linked the Artificial MirRNA sequence to pCAMBIA1301-35s and named it P35s:1031ArtmirRNA ARF5. PCAMBIA1301-35s was used as a check. Then the plant expression vector was introduced into tomato by the leaf discs method of Agrobacterium-mediate transformation. At last,80kanamycin resistant shoots and were obtained so for. The job is still going on. |
PDF Full Text Request