中国水稻科学
中國水稻科學
중국수도과학
CHINESE JOURNAL OF RICE SCIENCE
2014年
6期
581-588
,共8页
张亚东%梁彦丽%郑佳%丁丹%赵春芳%陈涛%赵庆勇%朱镇%周丽慧%姚姝%赵凌%于新%王才林
張亞東%樑彥麗%鄭佳%丁丹%趙春芳%陳濤%趙慶勇%硃鎮%週麗慧%姚姝%趙凌%于新%王纔林
장아동%량언려%정가%정단%조춘방%진도%조경용%주진%주려혜%요주%조릉%우신%왕재림
水稻%极端粒形%GW2基因%序列分析%粒形
水稻%極耑粒形%GW2基因%序列分析%粒形
수도%겁단립형%GW2기인%서렬분석%립형
rice%extreme materials%GW2%sequence analysis%grain shape
粒形是影响水稻产量的重要因子,对粒形相关基因的发掘和利用一直倍受重视.在已克隆的粒形基因中,GW2是一个控制粒宽的重要基因.通过同源克隆的方法,获得极端大粒水稻 TD70和极端小粒水稻 Kasalath 的 GW2基因 cDNA片段,分析序列差异,设计功能标记,检测240个来源于 TD70/Kasalath 的重组自交系(RILs)和部分籼稻骨干恢复系、江苏省主栽粳稻品种的基因类型及其效应.结果表明,TD70的 GW2与大粒粳稻 WY3完全一致,与已知序列(GenBank：EF447275)相似性达99．77％,在碱基的第114位,T 突变为 C,但并不改变氨基酸性质；在第316位缺失碱基 A,造成316位以后的所有碱基前移一位,氨基酸均发生变化,在碱基的第346、第347、第348位形成 TAA 的终止密码子,提前结束了TD70的 GW2蛋白的表达.而 Kasalath 的 GW2 序 列 仅 在碱基的第114位,由 T 突变为 C,但并不改变氨基酸性质；GW2 Kasalath 的316位没有突变,表达没有提前终止；设计的 dCAPs 标记,产物经内切酶 Apo I 酶切,TD70为21bp 和30bp 两个片段,而 Kasalath 条带为51 bp；240个 RILs 中含 GW2 TD70基因的株系有82个,含 GW2 Kasalath 基因的株系有158个,两者在粒宽和粒重表型上存在显著差异；8个籼稻骨干恢复系和10个江苏省主栽粳稻品种中都不存在 GW2 TD70基因.GW2 TD70基因对籽粒宽度有一定的调控作用,水稻育种中可以利用 GW2 TD70基因来提高籽粒粒重.
粒形是影響水稻產量的重要因子,對粒形相關基因的髮掘和利用一直倍受重視.在已剋隆的粒形基因中,GW2是一箇控製粒寬的重要基因.通過同源剋隆的方法,穫得極耑大粒水稻 TD70和極耑小粒水稻 Kasalath 的 GW2基因 cDNA片段,分析序列差異,設計功能標記,檢測240箇來源于 TD70/Kasalath 的重組自交繫(RILs)和部分秈稻骨榦恢複繫、江囌省主栽粳稻品種的基因類型及其效應.結果錶明,TD70的 GW2與大粒粳稻 WY3完全一緻,與已知序列(GenBank：EF447275)相似性達99．77％,在堿基的第114位,T 突變為 C,但併不改變氨基痠性質；在第316位缺失堿基 A,造成316位以後的所有堿基前移一位,氨基痠均髮生變化,在堿基的第346、第347、第348位形成 TAA 的終止密碼子,提前結束瞭TD70的 GW2蛋白的錶達.而 Kasalath 的 GW2 序 列 僅 在堿基的第114位,由 T 突變為 C,但併不改變氨基痠性質；GW2 Kasalath 的316位沒有突變,錶達沒有提前終止；設計的 dCAPs 標記,產物經內切酶 Apo I 酶切,TD70為21bp 和30bp 兩箇片段,而 Kasalath 條帶為51 bp；240箇 RILs 中含 GW2 TD70基因的株繫有82箇,含 GW2 Kasalath 基因的株繫有158箇,兩者在粒寬和粒重錶型上存在顯著差異；8箇秈稻骨榦恢複繫和10箇江囌省主栽粳稻品種中都不存在 GW2 TD70基因.GW2 TD70基因對籽粒寬度有一定的調控作用,水稻育種中可以利用 GW2 TD70基因來提高籽粒粒重.
립형시영향수도산량적중요인자,대립형상관기인적발굴화이용일직배수중시.재이극륭적립형기인중,GW2시일개공제립관적중요기인.통과동원극륭적방법,획득겁단대립수도 TD70화겁단소립수도 Kasalath 적 GW2기인 cDNA편단,분석서렬차이,설계공능표기,검측240개래원우 TD70/Kasalath 적중조자교계(RILs)화부분선도골간회복계、강소성주재갱도품충적기인류형급기효응.결과표명,TD70적 GW2여대립갱도 WY3완전일치,여이지서렬(GenBank：EF447275)상사성체99．77％,재감기적제114위,T 돌변위 C,단병불개변안기산성질；재제316위결실감기 A,조성316위이후적소유감기전이일위,안기산균발생변화,재감기적제346、제347、제348위형성 TAA 적종지밀마자,제전결속료TD70적 GW2단백적표체.이 Kasalath 적 GW2 서 렬 부 재감기적제114위,유 T 돌변위 C,단병불개변안기산성질；GW2 Kasalath 적316위몰유돌변,표체몰유제전종지；설계적 dCAPs 표기,산물경내절매 Apo I 매절,TD70위21bp 화30bp 량개편단,이 Kasalath 조대위51 bp；240개 RILs 중함 GW2 TD70기인적주계유82개,함 GW2 Kasalath 기인적주계유158개,량자재립관화립중표형상존재현저차이；8개선도골간회복계화10개강소성주재갱도품충중도불존재 GW2 TD70기인.GW2 TD70기인대자립관도유일정적조공작용,수도육충중가이이용 GW2 TD70기인래제고자립립중.
Grain shape is one of the most important influencing factor to rice grain yield.Among all of the cloned grain shape genes,GW2 is an important one controlling grain width.We obtained the sequences of the important grain shape gene GW2 using cDNA cloning strategy from rice variety TD70 and Kasalath and design its functional marker based on gene sequence difference to analyze genotype and phenotype of recombinant inbred lines(RILs)from TD70/Kasalath cross,some indica restorer lines and japonica rice cultivars in Jiangsu Province.GW2 TD70 shared 100% identity to GW2 WY3 and 99.77% identity to GW2 sequence(GenBank:EF447275)and there exist a single nucleotide mutation at site 1 14 without changing amino acid,a single nucleotide deletion at site 3 1 6 which generate termination codon at site 346, 347 and 348,thus the expression of GW2 protein stopped in advance.The sequence of GW2 Kasalath showed 99.92% identi-ty to GW2 which is known,and there exists a single nucleotide change at site 1 14 without the variation in amino acid. Comparing with GW2 in TD70,the gene in Kasalath express the intact protein without advanced stop.For GW2 gene, the dCAPS marker was designed according to a base deletion of A in the 3 1 6 site of sequence.Digested by Apo I,TD70 showed 21 bp and 30 bp fragments and Kasalath showed 5 1 bp fragment.Among the 240 RILs,82 contained GW2 TD70 and 1 58 contain GW2 Kasalath ,there are significant differences between their phenotypic value.No GW2 TD70 gene exit in 10 japonica rice cultivars from Jiangsu Province and 8 indica restorer lines.GW2 gene derived from TD70 may have regula-tory effects on grain width,and we can use it to improve grain weight in rice breeding.
