锌对不同发育时期子代果蝇基因组DNA甲基化的影响

doi:10.3724/SP.J.1003.2012.06090

摘要/Abstract

摘要：

为了初步探讨微量元素锌对不同发育阶段子代果蝇基因组DNA甲基化的影响及可能机制, 以黑腹果蝇(Drosophila melanogaster)Canton-S品系为实验材料, 应用改良的AFLP分子标记技术分析了果蝇基因组DNA甲基化多态性, 以及不同浓度(2.5%、5%、10%)锌处理亲代果蝇后, 不同发育时期的子代果蝇基因组DNA甲基化的改变。结果显示: 胚胎、三龄幼虫、蛹期、雌及雄蝇DNA甲基化多态条带比率均存在显著性差异; 亲本果蝇经锌处理后, 其5-8 h胚胎DNA甲基化多态条带比率在3个实验组中均显著高于对照组; 锌浓度为2.5%时, 子代果蝇DNA甲基化所受影响最大, 在5-8 h胚胎、12-15 h胚胎、18-21 h胚胎及雌蝇中, DNA甲基化多态条带比率均显著高于对照组。结果表明胚胎期DNA甲基化多样性最高, 同时适当浓度锌处理, 可提高子代果蝇基因组DNA甲基化多态性。

关键词: 果蝇, 锌, AFLP, DNA甲基化

Abstract

To examine the features of DNA methylation diversity, we used a zinc treated Canton-S strain of Drosophila melanogaster as a model organism in this experiment. Our results showed significant differences in DNA methylation polymorphisms among embryos, third instar larvae, pupae, and female and male individuals. After zinc treatment (at a concentration of 2.5%, 5%, and 10%) to the parental generation, the percentage of DNA methylation polymorphic bands of offspring (5-8 h embryo) in the experimental groups F₁-1, F₁-2, F₁-3 were significantly higher than the control group. The experimental group F₁-2 (zinc concentration 2.5%) in 5-8 h embryo was affected most by zinc treatment of the parental generation, and DNA methylation polymorphic band ratios in 12-15 h embryo, 18-21 h embryo and female D. melanogaster were significantly higher than the control groups. Our results indicate that the embryo stage contains the highest DNA methylation diversity, and appropriate concentration of zinc material can increase DNA methylation diversity of D. melanogaster offspring.

Key words: Drosophila, zinc, AFLP, DNA methylation

郭欣欣, 毛雪, 张敏 (2012) 锌对不同发育时期子代果蝇基因组DNA甲基化的影响. 生物多样性, 20, 710-715. DOI: 10.3724/SP.J.1003.2012.06090.

Xinxin Guo, Xue Mao, Min Zhang (2012) Effect of zinc on DNA methylation diversity at different developmental stages of filial generation in Drosophila. Biodiversity Science, 20, 710-715. DOI: 10.3724/SP.J.1003.2012.06090.

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链接本文: https://www.biodiversity-science.net/CN/10.3724/SP.J.1003.2012.06090

https://www.biodiversity-science.net/CN/Y2012/V20/I6/710

图/表 4

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接头及引物 Adapter and primer	序列 Sequence
E-adapter 1	5′-CTCGTAGACTGCGTACC
E-adapter 2	CTGACGCATGGTTAA-5′
A-adapter 1	5′-GACGATGAGTCCTGAG
A-adapter 2	GCTACTCAGGACTC-5′
E00	5′-GACTGCGTACCAATTC
A00	5′-GATGAGTCCTGAGCT
E+3	E-AAC, E-AAG, E-ACA, E-ACT, E-ACC, E-AGC, E-ACG, E-AGG
A+3	A-CAA, A-CAC, A-CAG, A-CAT, A-CTA, A-CTC, A-CTG, A-CTT

接头及引物 Adapter and primer	序列 Sequence
E-adapter 1	5′-CTCGTAGACTGCGTACC
E-adapter 2	CTGACGCATGGTTAA-5′
A-adapter 1	5′-GACGATGAGTCCTGAG
A-adapter 2	GCTACTCAGGACTC-5′
E00	5′-GACTGCGTACCAATTC
A00	5′-GATGAGTCCTGAGCT
E+3	E-AAC, E-AAG, E-ACA, E-ACT, E-ACC, E-AGC, E-ACG, E-AGG
A+3	A-CAA, A-CAC, A-CAG, A-CAT, A-CTA, A-CTC, A-CTG, A-CTT

引物组合 Primer combination	总条带数 Total bands	多态条带 No. of polymorphic bands (%)
E-AAC/A-CTT	19	14(73.68±0.44)
E-ACT/A-CTA	24	17(70.83±0.43)
E-ACT/A-CTC	18	11(61.11±0.42)
E-ACT/A-CTG	15	5(33.33±0.42)
E-AGC/A-CAC	23	17(73.91±0.50)
E-AGC/A-CAT	17	10(58.82±0.47)
E-ACG/A-CAA	16	8(50.00±0.43)
E-AGG/A-CAC	16	9(56.25±0.43)
E-ACC/A-CAC	14	11(78.51±0.46)
E-AAC/A-CTC	16	8(50.00±0.42)
E-AGC/A-CTT	18	9(50.00±0.51)
E-ACG/A-CTA	22	18(81.82±0.45)
总计 Total	218	137(62.84±0.48)

引物组合 Primer combination	总条带数 Total bands	多态条带 No. of polymorphic bands (%)
E-AAC/A-CTT	19	14(73.68±0.44)
E-ACT/A-CTA	24	17(70.83±0.43)
E-ACT/A-CTC	18	11(61.11±0.42)
E-ACT/A-CTG	15	5(33.33±0.42)
E-AGC/A-CAC	23	17(73.91±0.50)
E-AGC/A-CAT	17	10(58.82±0.47)
E-ACG/A-CAA	16	8(50.00±0.43)
E-AGG/A-CAC	16	9(56.25±0.43)
E-ACC/A-CAC	14	11(78.51±0.46)
E-AAC/A-CTC	16	8(50.00±0.42)
E-AGC/A-CTT	18	9(50.00±0.51)
E-ACG/A-CTA	22	18(81.82±0.45)
总计 Total	218	137(62.84±0.48)

不同发育时期 Developmental stages	不同处理组多态条带比率(%) Percentage of polymorphic bands of different treatments
不同发育时期 Developmental stages	对照 Control group	实验组F₁-1 Experimental group F₁-1	实验组F₁-2 Experimental group F₁-2	实验组F₁-3 Experimental group F₁-3
0-3 h胚胎 0-3 h embryo	27.52±0.42^a	28.72±0.42^a	29.94±0.36^a	28.35±0.40^a
5-8 h胚胎 5-8 h embryo	28.44±0.59^a	31.74±0.59^a*	33.52±0.50^a*	32.36±0.62^a*
12-15 h胚胎 12-15 h embryo	28.02±0.44^a	29.92±0.44^a	31.05±0.49^a*	31.01±0.44^a*
18-21 h胚胎 18-21 h embryo	26.31±0.42^a	29.21±0.42^a	29.93±0.28^a*	28.97±0.71^a
三龄幼虫 Third instar larvae	11.95±0.34^b	12.87±0.34^b	13.65±0.65^b	13.60±0.39^b
蛹 Pupae	13.02±0.56^b	12.53±0.56^b	13.01±0.42^b	12.98±0.42^b
雌蝇 Adult female	16.49±0.49^c	18.79±0.49^c	19.41±0.43^c *	18.80±0.53^c
雄蝇 Adult male	19.96±0.62^d	19.56±0.60^c	20.03±0.55^c	20.01±0.28^c