中国水稻科学
中國水稻科學
중국수도과학
Chinese Journal of Rice Science
2015年
5期
457-466
,共10页
孙廉平%张迎信%张沛沛%杨正福%占小登%沈希宏%张振华%胡霞%轩丹丹%吴玮勋%曹立勇%程式华
孫廉平%張迎信%張沛沛%楊正福%佔小登%瀋希宏%張振華%鬍霞%軒丹丹%吳瑋勛%曹立勇%程式華
손렴평%장영신%장패패%양정복%점소등%침희굉%장진화%호하%헌단단%오위훈%조립용%정식화
水稻%开颖不育%基因定位%可变剪接
水稻%開穎不育%基因定位%可變剪接
수도%개영불육%기인정위%가변전접
rice%open hull male sterile%gene mapping%alternative splicing
在60 Co-γ射线辐射诱变籼稻中恢8015的突变体库内发现了一个花器官发育突变体,暂命名为开颖不育突变体ohms1(openhullandmalesterile1).ohms1突变体表现为颖花开裂,在雄蕊和柱头之间形成类似内外稃的结构,使得突变体的颖花形成类似“三齿稃”状的三个颖壳,小穗完全不育,花粉育性为60％~70％,但自交不结实.遗传分析和基因定位结果表明,ohms1受一对隐性单基因控制,位于第3染色体短臂 KY2和 KY29标记之间,物理距离约42 kb,该区域包含4个开放阅读框 ORFs.进一步序列分析发现,突变体中一个编码 MADS盒的基因 LOC_Os03g11614的第5内含子末位碱基由 A突变为 G.酶切实验和 cDNA测序证实,该基因的第5内含子未被剪切,致使该基因的第6外显子所编码的14个氨基酸完整缺失,但并未造成该蛋白 MADS结构域的改变或移码.qRT-PCR结果显示,突变体中OsMADS1基因的表达水平显著降低,水稻开花调控因子和内外稃发育调控基因的表达量也发生了显著变化.说明该基因对水稻花器官发育尤其是内外稃发育和小花原基的分化具有重要作用.
在60 Co-γ射線輻射誘變秈稻中恢8015的突變體庫內髮現瞭一箇花器官髮育突變體,暫命名為開穎不育突變體ohms1(openhullandmalesterile1).ohms1突變體錶現為穎花開裂,在雄蕊和柱頭之間形成類似內外稃的結構,使得突變體的穎花形成類似“三齒稃”狀的三箇穎殼,小穗完全不育,花粉育性為60％~70％,但自交不結實.遺傳分析和基因定位結果錶明,ohms1受一對隱性單基因控製,位于第3染色體短臂 KY2和 KY29標記之間,物理距離約42 kb,該區域包含4箇開放閱讀框 ORFs.進一步序列分析髮現,突變體中一箇編碼 MADS盒的基因 LOC_Os03g11614的第5內含子末位堿基由 A突變為 G.酶切實驗和 cDNA測序證實,該基因的第5內含子未被剪切,緻使該基因的第6外顯子所編碼的14箇氨基痠完整缺失,但併未造成該蛋白 MADS結構域的改變或移碼.qRT-PCR結果顯示,突變體中OsMADS1基因的錶達水平顯著降低,水稻開花調控因子和內外稃髮育調控基因的錶達量也髮生瞭顯著變化.說明該基因對水稻花器官髮育尤其是內外稃髮育和小花原基的分化具有重要作用.
재60 Co-γ사선복사유변선도중회8015적돌변체고내발현료일개화기관발육돌변체,잠명명위개영불육돌변체ohms1(openhullandmalesterile1).ohms1돌변체표현위영화개렬,재웅예화주두지간형성유사내외부적결구,사득돌변체적영화형성유사“삼치부”상적삼개영각,소수완전불육,화분육성위60％~70％,단자교불결실.유전분석화기인정위결과표명,ohms1수일대은성단기인공제,위우제3염색체단비 KY2화 KY29표기지간,물리거리약42 kb,해구역포함4개개방열독광 ORFs.진일보서렬분석발현,돌변체중일개편마 MADS합적기인 LOC_Os03g11614적제5내함자말위감기유 A돌변위 G.매절실험화 cDNA측서증실,해기인적제5내함자미피전절,치사해기인적제6외현자소편마적14개안기산완정결실,단병미조성해단백 MADS결구역적개변혹이마.qRT-PCR결과현시,돌변체중OsMADS1기인적표체수평현저강저,수도개화조공인자화내외부발육조공기인적표체량야발생료현저변화.설명해기인대수도화기관발육우기시내외부발육화소화원기적분화구유중요작용.
A rice floral organ development mutant was obtained from the 60 Co-γ-treated indica restorer line Zhonghui 8015 and termed asopenhullandmalesterile1 (ohms1).The mutant was characterized as open hull and lemma-and palea-like structure between anthers and stigma that made the spikelet of the mutant showed three“triodia-like”flower glumes.The mutant was self-sterile but 60%-70% pollen were fertile.Genetic analysis and gene mapping showed ohms1 was controlled by a single ressessive gene and the mutant gene was fine mapped to a region of 42 kb on the short arm of chromosome 3 between markers KY2 and KY2 9 .Sequence analysis of the four open reading frames(ORFs)in this region revealed that LOC_Os03g11614 was probably corresponding to ohms1,which encoded a MADS-box gene allelic with OsMADS1 ,and had a single nucleotide transformation (A to G)at the bottom of the fifth intron.Enzyme digestion and cDNA sequencing furtherly indicated that the variable splicing was responsible for the delection of the sixth exon of ohms1 ,but no structure changes in MADS domain or amino acid frameshift occurred.Additionally, real-time fluorescence quantitative PCR analysis showed that the expression level of OHMS1 decreased significantly and the expression level of rice flowering factors and floral glume development-related genes changed significantly.All those results demonstrated that OHMS1 may play an important role in rice floral organ development,particularly in floral glume development and floret primordium differentiation.
