| Kenaf is Malvaceae Hibiscus, Annual herbaceous short-day plant bast fiber, With strong resistance to environmental stress, adaptability, high yield and fiber quality characteristics, and a major fiber materials of Hemp and Paper Industry in China, But the early flowering hybrid varieties has seriously affected the fiber output. In this study, space mutation Fuhong 952 (light insensitive) and control varieties fuhong 952 was used as material, In Hainan showed three types of mutant:Buds falling off(Gâ… types), Not flowering bud(Gâ…¡types), flowering but Fruitless(Gâ…¢types), through plant physiology, genomic differences, proteomics light insensitive mutant with short light-induced infertility and genetic law respectively to study its Molecular mechanism, The results are as follows:1,kenaf Photoperiod Observation:Kenaf light insensitive mutant under the short-day conditions in Hainan Province's winter, the mutant is not Bloom until 155 days, but the control varieties can flower in 120 days,12 hours of artificial short day treatment in Fuzhou's winter, the control varieties can flower in 20 days, the mutant 30days, Budding time delayed 10days.2,hybrid with light insensitive of kenaf varieties Fuhong 952 mutant and light-sensitive cytoplasmic maintainer line L23B, Hainan 130 days short day conditions induced differentiation and growth of buds, That mutant buds appeared three types, that is buds yellow off, squaring non-flowering, when the flowers but not fruit, F2 group Separation ratio was 3:1 (light sensitive with light insensitive),the results showed that, light Insensitive and light sensitivity were controlled by a pair of gene,photoperiod insensitivity Characters was Recessive gene, The results can further light insensitive kenaf Cloning and provide an important molecular mechanism of genetic material.3,Measured in the artificial short day photoperiod treatment and control of light insensitive kenaf Fuhong 952 in three different periods of leaf soluble sugar, soluble protein and soluble amino acid content of the dynamic changes, The results show that With the different stages of development of kenaf, that is one split leaf, three splits leaf, five splits leaf, Leaf soluble sugar, soluble protein, soluble amino acids all showed a low, high, low variation, the Change of varieties fuhong 952 was larger than that of light insensitive plants, Show that the light insensitive kenaf induced in the light cycle shown photoperiod insensitivity and its leaves are closely related to the physiological metabolism.4,From the 24 polymorphisms RAPD primers were selected and 3 primers can distinguish light insensitive mutant of the control kenaf varieties, Get four specific bands, Sequencing showed that the size of the four fragments were 612bp,723bp, 743bp,1178bp,the results can prepare for further SCAR markers development.5,eight universal Primer Pairs of chloroplast genome were used to amplify chloroplast DNA (mtDNA) non-coding regions in kenaf varieties,the results shows that those primers can used to amplify mitochondrial gene spacer regions and introns, and Detected only two variable sites, the results also showed that mtDNA in the light insensitive kenaf between mutants and controls was very conservative.6,eight universal Primer Pairs of Mitochondria genome were used to amplify chloroplast DNA (mtDNA) non-coding regions in kenaf varieties, approximately 20.4kb,8.1% of the chloroplast genome was analyzed. The products were digested by four restriction enzymes. The results showed that all the primes can amplify bands of size between 1.5kb-2.0Kb, The amplification products both can be digested by restriction enzymes, only primers cp4 has digested Polymorphic fragments, but there is no difference between control varieties and the mutant, These results indicate that the mutant gene of kenaf light-insensitive was in genome DNA, and has no relationship to Mitochondria and chloroplast genome DNA.7,To establish a research system of kenaf in proteomics, we use ligh-insensitive kenaf mutants flowering and normal varieties Fuhong varieties 952 as materials. established and optimized the proteins firstly bidirectional electrophoresis system, the protein electrophoreses (bidirectional to dimensional electrophoresis,2-DE) technology. In the sample preparation, different protein samples and dyeing methods in comparative study, etc. Research results show that:(1) kenaf leaf total protein with TCA acetone extraction method was more effective than Phenol extraction, Protein extraction rate of TCA acetone extraction was 20.56%, Phenol extraction was of 4.90%.(2) the protein Lysate contain 0.2 M thiourea can get more points of different protein, which were mainly basic protein.(3) Of the silver stain and examination of the second MaSiLiang blue staining to SDS-PAGE glue staining, the results show that the highest sensitivity silver stain, but poor, repeatability MaSiLiang G250 blue staining method is more sensitive than R250, therefore suggest using test MaSiLiang G250 blue, can get ideal result.(4) Using the improved method extracting varieties fuhong 952 and kenaf are insensitive to kenaf blades, total protein electrophoresis, two-way to dye colloid pictures using PDQUEST software for two-way electrophoresis of further 2-DE protein map comparison and analysis of the difference, get nine protein points, including five proteins in mutant were down-regulated expression, Ribulose 1, 5-biphosphate carboxylase large subunit (spot3) in the mutant was up-regulated expression, Ribulose 1,5-biphosphate carboxylase large subunit (spot2) and a hypothetical protein (spot7) were not expressed in varieties fuhong 952, but in the mutants, H(+)-transporting ATP synthase (Spot4) was only expressed in varieties of 952 reference, these differences may be relevant with the light sensitivity of kenaf, The results of the light insensitive kenaf proteomics analysis lays the foundation of functional genome... |
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