草业学报
草業學報
초업학보
PRATACULTURAL SCIENCE
2015年
4期
164-173
,共10页
唐晓梅%王艳%马东伟%程海洋%杨宏%代娅%陶向%马欣荣
唐曉梅%王豔%馬東偉%程海洋%楊宏%代婭%陶嚮%馬訢榮
당효매%왕염%마동위%정해양%양굉%대아%도향%마흔영
高羊茅%干旱胁迫%DNA 甲基化%MSAP%转座子
高羊茅%榦旱脅迫%DNA 甲基化%MSAP%轉座子
고양모%간한협박%DNA 갑기화%MSAP%전좌자
tall fescue%drought stress%DNA methylation%MSAP%transposon
DNA 甲基化是真核细胞基因组重要的一种表观遗传调控。DNA 甲基化影响基因表达,在植物逆境胁迫适应中起着重要作用。高羊茅是禾本科单子叶羊茅属植物,具有良好的耐寒、耐热性,大量用作运动场草坪和防护草坪,也可用做牧草。干旱是限制高羊茅分布和产量的主要因素之一。本研究利用甲基化敏感扩增多态性技术(MSAP),比较了高羊茅在干旱胁迫15 d 后基因组胞嘧啶甲基化的变化。在干旱胁迫下,植株生长受到严重抑制。MSAP 分析显示,10对选择性扩增引物,共扩增出475个 CCGG 位点,其中131个位点发生了甲基化或去甲基化变化,占27.58%。对照组和干旱处理组,总甲基化水平分别是43.16%和42.11%,显示干旱胁迫诱导高羊茅总的甲基化率下降了1.05%。对差异条带进行归类分析,并克隆、测序,获得13条不同变化类型的序列。序列同源分析显示,大多数序列是参与胁迫应答的基因片段。其中2个片段,Fa6和 Fa7为干旱诱导下发生甲基化的位点,分别与小麦和大麦的一个逆转录转座子具有较高的同源性,在干旱胁迫下它们的甲基化程度增加,表明干旱诱导二者进一步甲基化。甲基化的转座子在维持基因组稳定性具有重要作用。综上,干旱胁迫诱导的甲基化的变化,可能参与高羊茅对环境的适应性调节。
DNA 甲基化是真覈細胞基因組重要的一種錶觀遺傳調控。DNA 甲基化影響基因錶達,在植物逆境脅迫適應中起著重要作用。高羊茅是禾本科單子葉羊茅屬植物,具有良好的耐寒、耐熱性,大量用作運動場草坪和防護草坪,也可用做牧草。榦旱是限製高羊茅分佈和產量的主要因素之一。本研究利用甲基化敏感擴增多態性技術(MSAP),比較瞭高羊茅在榦旱脅迫15 d 後基因組胞嘧啶甲基化的變化。在榦旱脅迫下,植株生長受到嚴重抑製。MSAP 分析顯示,10對選擇性擴增引物,共擴增齣475箇 CCGG 位點,其中131箇位點髮生瞭甲基化或去甲基化變化,佔27.58%。對照組和榦旱處理組,總甲基化水平分彆是43.16%和42.11%,顯示榦旱脅迫誘導高羊茅總的甲基化率下降瞭1.05%。對差異條帶進行歸類分析,併剋隆、測序,穫得13條不同變化類型的序列。序列同源分析顯示,大多數序列是參與脅迫應答的基因片段。其中2箇片段,Fa6和 Fa7為榦旱誘導下髮生甲基化的位點,分彆與小麥和大麥的一箇逆轉錄轉座子具有較高的同源性,在榦旱脅迫下它們的甲基化程度增加,錶明榦旱誘導二者進一步甲基化。甲基化的轉座子在維持基因組穩定性具有重要作用。綜上,榦旱脅迫誘導的甲基化的變化,可能參與高羊茅對環境的適應性調節。
DNA 갑기화시진핵세포기인조중요적일충표관유전조공。DNA 갑기화영향기인표체,재식물역경협박괄응중기착중요작용。고양모시화본과단자협양모속식물,구유량호적내한、내열성,대량용작운동장초평화방호초평,야가용주목초。간한시한제고양모분포화산량적주요인소지일。본연구이용갑기화민감확증다태성기술(MSAP),비교료고양모재간한협박15 d 후기인조포밀정갑기화적변화。재간한협박하,식주생장수도엄중억제。MSAP 분석현시,10대선택성확증인물,공확증출475개 CCGG 위점,기중131개위점발생료갑기화혹거갑기화변화,점27.58%。대조조화간한처리조,총갑기화수평분별시43.16%화42.11%,현시간한협박유도고양모총적갑기화솔하강료1.05%。대차이조대진행귀류분석,병극륭、측서,획득13조불동변화류형적서렬。서렬동원분석현시,대다수서렬시삼여협박응답적기인편단。기중2개편단,Fa6화 Fa7위간한유도하발생갑기화적위점,분별여소맥화대맥적일개역전록전좌자구유교고적동원성,재간한협박하타문적갑기화정도증가,표명간한유도이자진일보갑기화。갑기화적전좌자재유지기인조은정성구유중요작용。종상,간한협박유도적갑기화적변화,가능삼여고양모대배경적괄응성조절。
DNA methylation is one of the most important mechanisms in the epigenetic regulation in eukaryotic cells.DNA methylation affects gene expression and plays an important role in plants adapting to adverse envi-ronments.Tall fescue (Festuca arundinacea ),a gramineous monocot,has superior resistance to cold and heat. It is widely used for sports turf,erosion control,and as a forage grass.However,the distribution and yield of tall fescue is constrained by drought stress.In this paper,the growth was measured and the DNA methylation profile of tall fescue (cv.Barlexas)evaluated using methylation-sensitive amplification polymorphism (MSAP) after 15 d drought treatment.The growth of the plants was seriously restricted under water-deficit challenge. Genome-wide,a total of 475 CCGG sites were detected by MSAP using 10 selective primer pairs in tall fescue seedlings.Among these sites,131 sites (27.58%)showed drought-induced methylation and demethylation. The total methylation level was 43.16% in control and 42.11% in drought-treatment,which indicates a 1.05%decrease when exposed to drought.The sequences of 13 amplified fragments were successfully obtained,repre-senting the differential types of methylation changes.Homologous analysis showed that most of the fragments were relevant to stress.Two sequences (Fa6 and Fa7)with enhanced methylation induced by drought were ho-mologous to a retrotransposon of Triticum aestivum and Hordeum vulgare ,respectively,showing drought in-duced methylation of the retrotransposons.The methylation of transposons is vital in maintaining DNA stabili-ty.In summary,drought stress decreased the total DNA methylation level in the tall fescue genome,and the altered methylation in response to stress was probably involved in environmental stress acclimation.