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Studies on in Vitro Flowering and Proteomics in Tagetes Patula L.

Author: JiangShunRi
Tutor: LaiZhongXiong
School: Fujian Agriculture and Forestry University
Course: Cell Biology
Keywords: Maidenhair In vitro flowering Proteomics Cyclophilin protein gene Clone
CLC: S681.9
Type: Master's thesis
Year: 2011
Downloads: 70
Quote: 5
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Abstract


In this study, maidenhair (Tagetes patula L.) plantlets were test material, maidenhair regeneration system, in vitro flowering, the proteome and maidenhair pro ring Protein Gene the following research: (1) establish maidenhair regeneration system; ② maidenhair test flowering research to establish the maidenhair efficient flowering system; ③ maidenhair various stages of material separation of proteins, to obtain a high-resolution two-dimensional electrophoresis of proteins (2D) maps, and 2D analysis software ImageMaster 2D Platinum 6.0 ( GE Healthcare) protein differential expression analysis, and identified by mass spectrometry and functional analysis of protein expression on the part of the difference of each stage of the maidenhair flower bud differentiation; ④ protein spectrum identification results of maidenhair cyclophilin protein gene was cloned. The main findings are as follows:. Maidenhair Establishment of Regeneration System maidenhair plantlets material, respectively, to study its bud induction, shoot proliferation, rooting and the best conditions for leaf callus induction. The test results show that: the best bud induction medium: MS 0.2 mg · L -1 6-BA 0.2mg · L -1 NAA 30g · L - 1 sugar; Best bud proliferation medium: MS 0.2 mg · L -1 6-BA 0.05mg L -1 NAA; optimal rooting medium: 1/2MS 0.1 mg · L -1 NAA; leaves callus induction optimum medium: MS of 1.0 mg · L -1 2,4 - D 1.0 mg · L -1 6-BA. 2. Maidenhair in vitro flowering research Irradiation on Seed Germination from material after growth 35d is test material, the time from a different light, different concentrations of sugar, paclobutrazol concentration of different quality, different hormone combinations and different factors (MS medium sugar, KT, GA3) of L9 (43) orthogonal test of the maidenhair in vitro flowering. MS medium containing paclobutrazol mass concentration test results show that: the single factor test short day (8h/16h), MS basic medium sugar mass concentration of 45g · L -1 0.6mg · L -1 , the most suitable for the further development of the maidenhair induction of flower buds and flower buds. Multi-factor test of 6-BA/NAA (performed at 0.2/0.05 mg · L when -1 ) 4 most suitable patula flower bud induction, but not conducive to flower buds further development, the conditions induced under the the flower buds final atrophy death. Orthogonal experiment 1/4MS 45g · L -1 sugar 0.4mg L -1 KT 0.6mg · L -1 of GA3 most suitable for the further development of the maidenhair bud induction and flower buds, under processing conditions the maidenhair can blossom and flowering a week or so in advance. Patula flower bud differentiation stage material proteomics research ① patula flower bud differentiation process protein component, MW and PI changes maidenhair in vitro flowering research based on maidenhair different culture plants of the number of days for the material carried flower buds differentiation proteomics research. Patula flower bud differentiation stage protein number, isoelectric point (pI) and molecular weight change results are as follows: ① the number of changes: maidenhair plantlets flower bud differentiation process, whether it is in white or in the Blu-ray light conditions, changes in the protein spots W2 stage are showing an increasing trend, the average protein spots can be detected up to 928 protein spots; followed: B2 phase proteins 803 761; W1 phase proteins; B1 phase protein spots 631; V0 stage the least protein spots, 607. ② Isoelectric point (pI) Change: patula Plantlets flower bud differentiation process in the protein pI value range is mainly between 5 to 6, regardless of the white light according to or in the under blue illumination conditions and different pI value range, with maidenhair plantlets growth and development, protein spots showed an upward trend. This trend illustrates the the B1 stage and W1 stage is a very crucial period in the maidenhair bud morphological build process, but also illustrates the pI value in the the maidenhair flower buds form of build process in the range of 5 to 6 protein plays a very important role. ③ a molecular weight (MW) changes: patula the Bud Differentiation W2 stage (white conditions), and Blu-ray under the conditions of stages (B2) of these two periods, the molecular weight of the protein spots with the other stage material of 20.1KD to 66kD between compared to maximum, and molecular weight in 30KD to 45KD between, protein points up, which illustrates the the maidenhair flower bud differentiation process, molecular weight of 30KD to 45KD proteins participate in the bud morphological building plays a very important role. ② patula flower bud differentiation process related proteins identified by mass spectrometry and its function analysis of 75 select patula flower bud differentiation stage differentially expressed proteins identified by mass spectrometry, 34 protein spots were credible match, protein identification success rate about 45.3%. Successfully identified 34 proteins, their functions can be divided into five functional groups, the largest proportion of the energy and glucose metabolism related proteins 15, 44%; redox stress response protein, 2, accounting for 6 %; protein synthesis, folding, assembly and decomposition associated protein 2 (6%); cells and signal transduction related proteins, three (3%); functionally unknown protein 14 (41%). Proportion of energy and glucose metabolism related proteins column maximum, which may be involved in the flower buds of the build process of patula flower bud differentiation process requires a lot of energy and matter, and thus metabolism, the type and quantity of protein is more . The high proportion of unknown proteins, and many of the small molecular weight proteins, these proteins are usually associated with signal transduction and gene regulation protein. Therefore, the inference that these proteins of unknown function in the process of patula flower bud differentiation plays a very important role. 4. Maidenhair pro ring protein gene cloning maidenhair seed germination after 35 days of growth (photoperiod: 12h / d; growth temperature: 25 ± 2 ℃), transferred to MS medium for 25 days (photoperiod: 8h / d; growth temperature: 20 ± 2 ℃) plantlets were material. The first successful cloning of a maidenhair cyclophilin protein gene of a partial cDNA sequence and a full-length cDNA sequences, were named as the CyP-1 and of CyP-2. The CyP-1 sequences are 716 nucleotides, CYP-2 sequence of 834 nucleotides, encoding 171 amino acids. Cloned into the CyP-1 sequence CyP-2 sequence homology of nucleotide sequence analysis showed that, of CyP-of CyP-2 and Gerbera (Gerbera EU126917.1) cyclophilin protein gene homologies were 99%, 89%.

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CLC: > Agricultural Sciences > Gardening > Ornamental Horticulture ( flowers and ornamental trees) > Annual and biennial flower class > Other
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