中山大学学报（自然科学版）
中山大學學報（自然科學版）
중산대학학보（자연과학판）
ACTA SCIENTIARUM NATURALIUM UNIVERSITATIS SUNYATSENI
2013年
5期
118-122
,共5页
周晓穗%陈佳林%张梅%王方海
週曉穗%陳佳林%張梅%王方海
주효수%진가림%장매%왕방해
MS-RDA%DNA甲基化%白背飞虱%翅二型
MS-RDA%DNA甲基化%白揹飛虱%翅二型
MS-RDA%DNA갑기화%백배비슬%시이형
MS-RDA%DNA methylation%Sogatella furcifera%wing dimorphism
甲基化敏感性代表性差异分析方法（MS-RDA）适用于特异、低丰度基因组DNA甲基化的筛查与鉴定，已大量应用于哺乳动物的疾病发生和植物发育机理方面的研究，但昆虫中尚未发现该方法的应用。从DNA提取、酶切、扩增子制备、消减杂交等方面对该方法进行了改进，并在水稻重要害虫白背飞虱上得到成功应用，获得了4条与雌、雄性别相关联的特异性甲基化差异条带，其中有3条序列已被GenBank收录，登陆号分别为JX847621、JX624161、JX472453，经BLAST比对，有3条未发现同源性序列，属于功能未知的新基因；同时还获得了2条与长短翅型相关联的特异性甲基化差异条带，也已被 GenBank 收录，登陆号分别为 JX514031、JX472454，同源性分析发现与多种动物的18S核糖体和28S核糖体RNA基因序列高度相似，因此有关核糖体基因的甲基化可能对白背飞虱的翅型分化起到一定的调控作用。
甲基化敏感性代錶性差異分析方法（MS-RDA）適用于特異、低豐度基因組DNA甲基化的篩查與鑒定，已大量應用于哺乳動物的疾病髮生和植物髮育機理方麵的研究，但昆蟲中尚未髮現該方法的應用。從DNA提取、酶切、擴增子製備、消減雜交等方麵對該方法進行瞭改進，併在水稻重要害蟲白揹飛虱上得到成功應用，穫得瞭4條與雌、雄性彆相關聯的特異性甲基化差異條帶，其中有3條序列已被GenBank收錄，登陸號分彆為JX847621、JX624161、JX472453，經BLAST比對，有3條未髮現同源性序列，屬于功能未知的新基因；同時還穫得瞭2條與長短翅型相關聯的特異性甲基化差異條帶，也已被 GenBank 收錄，登陸號分彆為 JX514031、JX472454，同源性分析髮現與多種動物的18S覈糖體和28S覈糖體RNA基因序列高度相似，因此有關覈糖體基因的甲基化可能對白揹飛虱的翅型分化起到一定的調控作用。
갑기화민감성대표성차이분석방법（MS-RDA）괄용우특이、저봉도기인조DNA갑기화적사사여감정，이대량응용우포유동물적질병발생화식물발육궤리방면적연구，단곤충중상미발현해방법적응용。종DNA제취、매절、확증자제비、소감잡교등방면대해방법진행료개진，병재수도중요해충백배비슬상득도성공응용，획득료4조여자、웅성별상관련적특이성갑기화차이조대，기중유3조서렬이피GenBank수록，등륙호분별위JX847621、JX624161、JX472453，경BLAST비대，유3조미발현동원성서렬，속우공능미지적신기인；동시환획득료2조여장단시형상관련적특이성갑기화차이조대，야이피 GenBank 수록，등륙호분별위 JX514031、JX472454，동원성분석발현여다충동물적18S핵당체화28S핵당체RNA기인서렬고도상사，인차유관핵당체기인적갑기화가능대백배비슬적시형분화기도일정적조공작용。
Methylation-sensitive representational difference analysis (MS-RDA)is suitable for screening and identification of specific and low abundances genomic DNA methylation.It has been applied in many researches on mammal disease occurrence and plant developmental mechanism,but no application was reported in insects.In this study,we developed a MS-RDA method for insects,which was improved from DNA extraction,restriction enzyme digestion,amplicon preparation,and competitive hybridization. The method was further applied successfully in Sogatella furcifera (Horvath),an important rice pest in China.Four differentially methylated DNA sequences related with female and male were obtained.Three of them had been accepted by GenBank,the accession numbers were JX847621,JX624161,JX472453, respectively.Three of them were not found homologous sequences by BLAST and should be belonging to the new genes with functions unknown.Meanwhile,two differentially methylated DNA sequences related with long and short wing were obtained.Their accession numbers of GenBank were JX514031、JX472454,respectively.They were found highly similar with the 18S ribosomal or 28S ribosomal RNA gene sequences of many kinds of animals by homology analysis,therefore ribosomal RNA gene methyla-tion maybe play some roles on wing dimorphism of S.furcifera.
