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In order to explore the epigenetic phenomenon in the the bamboo plant growth and development process, using methylation-sensitive amplification polymorphism (MSAP) and high performance liquid chromatography (HPLC) of different physiological age bamboo (Phyllostachys heterocycla var. Pubescens and) , flowering and non-flowering filial the bamboo (Bambusa. multiplex.) same physiology origin, under different habitats, the yellow stalks Ukrainian feeding chicken bamboo (Ph.vivax f.aureocaulis), young leaves of genomic DNA methylation detection the following conclusions: 1, bamboo of genomic DNA methylation level physiological age increases with an upward trend. Select five years, 31 years and> 60 years old the Phyllostachys points, with 35 pairs of primers: in Moso Bamboo nutrition growth process, with increasing physiological age of the genome simultaneously MSAP analysis of DNA methylation and demethylation phenomenon, which occurred methylation variable sites (52.3%) is much larger than the demethylation of variable sites (10.3%), so that the genomic DNA methylation rate increased with age upward trend, analysis of variance showed: same stand bamboo trees (including the same age of the age of the bamboo shoots and bamboo shoots bamboo monoclonal) genomic DNA methylation levels not significantly different, but different physiological age difference between Phyllostachys pubescens Stand reached a very significant level (P lt; 0.001). 2, screening six primer combinations MSAP technology of bamboo of genomic DNA methylation and physiological age regression models. Statistical analysis of experiments using 35 primer pairs screened with Mao Zhusheng reasonable age to increase the level of DNA methylation increased, and the difference between each adjacent two age reached a very significant level of six pairs of primer combinations (E CA / HM CAC , E CA / HM CTC, E CA / HM CTG, E AG / HM CTA, E AG / HM CTC, E AA / HM CTA). The filtered six pairs of primers to principal component analysis by the cumulative contribution ratio is greater than 90%, and a characteristic root is greater than the one of these two conditions to determine the weight of each pair of primers in contracting the consensus Mao Zhusheng age, to obtain a comprehensive index, i.e. : Y = 0.173x1 0.172x2 0.172x3 0.178x4 0.176x5 0.180x6, physiological age changes on behalf of the Moso bamboo with genomic DNA methylation levels. Then select 2 years, 6 years, 13 years, 18 years, 32 years and> 60 years old, 6 ages pubescens, the six pairs of primers MSAP detection. The model fit statistics, to get the Mao Zhusheng reasonable age to their genomic DNA methylation rate quadratic regression model: Z = 17.999 0.542y2 0.003y, R2 = 0.970. This field pubescens points physiological age determination has important practical significance. 3, filial bamboo from vegetative growth turn reproductive growth of genomic DNA methylation levels were significantly lower when using HPLC techniques on genomic DNA methylation filial bamboo flowering around the study found that: filial bamboo in the process of growth and development of non-flowering , DNA methylation rate showed irregular fluctuations phenomenon. Analysis of variance: whether it is the same Clumps different periods, or the same period of different the Clumps DNA methylation was no significant difference (P = 0.109> 0.05 and P = 0.622> 0.05). Methylation of the filial bamboo (same clones) flowering and non-flowering monoclonal MSAP detection results found: the flowering bamboo strains of DNA methylation variation bands accounted for 33.17% of the total number of bands, which occurred the same plexus sites (22.28%) is much larger than the demethylated sites (1.98%), flowering plants DNA methylation was significantly lower than that of flowering plants, the difference reached a significant level (P lt; 0.001). Flowering and non-flowering bamboo strains of the same samples using HPLC and MSAP technology to detect DNA methylation, found that: HPLC detection of total methylation rate (flowering 26.69% 30.61%; not flowering) were significantly higher than the MSAP detection results ( flowering 9.00% 12.42%); not flowering, but the trend is consistent. HPLC techniques capable of measuring the level of overall genomic DNA methylation MSAP technology can only be related to the characteristics of the analysis of genome-wide cytosine methylation level. 4, the same physiological origin, under different habitats yellow feeding chicken bamboo stalks Ukraine with decreasing geographical latitude, gradually reduce the level of DNA methylation. Yellow stalks Ukraine feeding chicken the Ukrainian feeding chicken bamboo variants, and An Ji Zhubo Park in Zhejiang Province in 1982 found that the propagation is now introduced to many regions of the country due to high ornamental value. Collected from Beijing Liangxiang, Nanjing, Jiangsu, Zhejiang Anji, Nanchang, Jiangxi, Sichuan Changning, Nanxiong, a total of six regions the yellow stalks Ukrainian feeding chicken bamboo genomic DNA. First, using the AFLP technique on six areas of the samples carried the genetic diversity analysis showed that: the same clonal origin, different habitats under the yellow stalks Ukraine feeding chicken bamboo leaves piece genomic DNA sequence no significant difference; but by MSAP detection found: different regions of DNA The total rate of methylation and methylation rate: Beijing Liangxiang (33.62% and 19.57%, respectively), Nanjing, Jiangsu, 33.19% and 19.27%), the Zhejiang Anji (31.19% and 8.60%), Nanchang (30.93% and 18.01%), Sichuan Changning (29.50% and 16.32%) the Nanxiong (28.78% and 14.49%, respectively), reduce the level of DNA methylation with geographic latitude shown a reducing trend. In addition, the screening of Mao Zhusheng reasonable age six pairs of primer combinations used in the yellow stalks Ukraine feeding chicken bamboo MSAP detection found: yellow stalks Ukraine, in different parts of feeding chicken bamboo MSAP amplified bands were consistent, no significant differences, in which the same asexual origins, different habitats under the yellow stalks Ukrainian feeding chicken bamboo genomic DNA is not significant methylation variation. From another point of validation, screening six primer pairs bamboo plant physiological age in determining feasibility.
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