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基于端粒法的黄连木古树树龄测定

Determining telomere length and estimating the age of ancient trees using the telomere method in Pistacia chinensis

  • 摘要: 以不同树龄黄连木叶片为试验材料,通过qPCR法测定端粒长度,分析黄连木端粒长度与树龄之间的相关性。结果表明:黄连木叶片的端粒长度与树龄呈正相关,在此基础上,建立了黄连木端粒长度与树龄的相关数学模型L(x) = −4 × 10−11 x2 + 2 × 10−4 x + 16.707(R2 = 0.9528)。从Hinf Ⅰ、Taq Ⅰ、BamH Ⅰ、BstX Ⅰ、Xho Ⅰ、Sal Ⅰ、Hind Ⅲ及EcoR Ⅰ共8种限制性内切酶中筛选合适的限制性内切酶,采用端粒末端限制性片段(TRF)分析法测定黄连木叶片的端粒长度,使用MATLAB等软件处理分析数据,结果表明限制性内切酶Taq Ⅰ和Hinf Ⅰ对黄连木叶片基因组DNA具有较好酶切效果,二者均可证明黄连木叶片的端粒长度随着树龄的增长逐渐增加,与qPCR法测定结果一致,其中Hinf Ⅰ的检测精度更高。

     

    Abstract: Leaves of Pistacia chinensis at various developmental stages were collected and used as experimental materials to investigate the correlation between telomere length and tree age. Telomere length was quantified using quantitative polymerase chain reaction (qPCR), and the results revealed a positive correlation between telomere length and tree age in P. chinensis leaves. Based on these findings, a mathematical model was developed to describe the relationship between telomere length and tree age: L(x) = −4 × 10−11 x2 + 2 × 10−4 x + 16.707 (R2 = 0.9528), where x represents telomere length and L(x) denotes tree age. Additionally, eight restriction endonucleases—Hinf I, Taq I, BamH I, BstX I, Xho I, Sal I, Hind III, and EcoR I—were evaluated to identify suitable enzymes for telomere length analysis. Terminal restriction fragment (TRF) analysis was employed to measure telomere length, and data were processed and analyzed using MATLAB software. The results indicated that Taq Ⅰ and Hinf Ⅰ effectively digested genomic DNA extracted from P. chinensis leaves. Both enzymes confirmed a gradual increase in telomere length with increasing tree age, consistent with the qPCR results, with Hinf Ⅰ demonstrating higher detection accuracy.

     

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