| Rice (Oryza sativa L) is one of important crops, and is also investigative mode in molecular biology, isolation and cloning of rice functional genes has been the basic of exploiting genetic engineering to improve rice species. At present, indica-japonica hybrid rice has shown stronger yield potential, but its fertility is sensitive to low temperature, this character is the main factor to restrict its expansion planting.Yayou 2 is a typical indica-japonica hybrid between 02428 (japonica) and 3037(indica). The fertility of this hybrid is sensitive to low temperature fluctuations (low temperature sensitive sterility, LTSS). So the study on it is very practical to indica-japonica hybrid breeding in rice.The method mRNA differential display was used to study LTSS of Yayou2, and established gene cloning system, acquiring the LTSS related genes of Yayou2, and offering theoretical gist for exploiting indica-japonica hybrid heterosis.The result of cytological observation is shown that the chromosomes are abnormal in meiosis stage when young spikes of Yayou2 are affected by low temperature.Totally 216 primer combinations were used to isolate LTSS related cDNA fragments, 35 differential fragments were obtained. The different of these fragments showed that: The genes, which expressed in contrast, were restrained in treatment; the genes, which expressed in treatment, were restrained in contrast; expressions of some fragments were stronger in contrast, but others were stronger in treatment. These differential fragments may play an important role in the fertility of indica-japonica hybrid. These fragments were reamplificated, 26 fragments, which can be reamplificated, were cloned, and 11 fragments of these were sequenced and compared with BLAST software, the results were as follows:Protein coded by DF1 has identity with SNF2 domain/helicase domain containing protein-like of rice, and has varied degree identity with glycine rich protein of Daucus carota , hydroxyproline-rich glycoprotein precursor of common tobacco, Serine/threonine protein kinase of yeast; Possible protein coded by DF2 has identity (72%) with auxin-regulated protein of Arabidopsis thaliana, and has some identity with auxin-regulated protein of soybean> putative Nt-gh3 deduced protein of rice, catalase of maize; Possible protein coded by DF3 has some identity with related to TPR-containing protein, histidine-rich glycoprotein, zinc finger protein, CDC28/cdc2 related protein kinase; Possibly products of DF4 has high identity with peptidase Ml family protein of Arabidopsis thaliana; Possibly products of DF9 has some identity with protein kinase C, Serine/threonine protein kinase, RRM type RNA binding protein;Sequence of DFU has 98% identity with sequence of chromosome 4 of rice, this fragment may be come from chromosome 4 of rice, and this fragment has some identity with chromosome 7,8, 10 of rice, and chloroplast of wheat;Sequence of DF12 has identity (99%) with chromosome 3 of rice, it may come from chromosome 3 of rice, and it has also some identity with chromosome 1 2 4 7 8 10 12, it has more copies in rice genome. Its product has identity with retrotransposons polyprotein and copy -type po! polyprotein of maize; The sequence of DF 13 has the identity with chromosome 1 of rice and some bases of DF13 have identity with protein phosphatase; The sequence of DF 14 has 99% identity with chromosome 3 of rice, it may be come from chromosome 3,and products has identity with calcium-dependent protein kinase putative leukotriene A-4 hydroiase of rice calcium-dependent protein kinase and cold-active aminopeptidase.Sequence of DF1 has partial identity with DF3; sequence of DF12 has identity with DF16, and sequence of DF 14 is complemented with DF15 each other. 9 differential fragments come from contrast material, but DF4 and DF9 come from treatment material. DF4 and DF9 are induced by low temperature, but the other 9 differential fragments are suppressed by low temperature.DF13 and DF14 were made into probe, Hindlll digested rice genome, and So... |
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