摘要
甜瓜是葫芦科重要的经济作物之一。大多数甜瓜是雄全同株材料,而雌雄异花同株材料除了在育种过程中能减少大量复杂的去雄工序外,其F1果实发育也要强于雄全同株材料。因此,培育雌雄异花同株材料,成为重要的育种目标之一。本试验调查各杂交组合分离后代、回交后代的开花类型及分离情况,对甜瓜雌雄异花同株遗传规律进行了分析,同时筛选出与甜瓜雌雄异花同株性状连锁的分子标记,探索了利用分子标记分析甜瓜雌雄异花同株性状的可能性,为利用分子标记辅助选择甜瓜育种,加快育种进程,开辟了新的技术途径。
本试验选择亲本材料分别为:母本3-2-2,来自中国,薄皮甜瓜,雌雄异花同株材料;父本TopMark,来源美国,厚皮网纹甜瓜,雄全同株材料;检验亲本:WI998,来源美国,网纹甜瓜,全雌株材料。常规手段配制杂交组合3-2-2×TopMark,借鉴刘威、路绪强试验中配置的杂交组合WI998×TopMark、WI998×3-2-2,调察各杂交组合F1、F2、BC1P1、BC1P2开花类型及分离比率,确定各亲本基因型。从3-2-2×TopMark组合F1代中取出350粒种子,通过连续自交,得到F1S6代重组自交系群体(RILs)共152株,用于构建遗传图谱。
调查3-2-2×TopMark组合F2代的开花类型,结果表明:雌雄异花同株性状主要受一对主效基因控制;研究雌花率结果显示:雌花率受到一对主效基因控制,同时存在微效基因,主效基因遗传率为83.3%,微效多基因遗传率为8.125 %。调查WI998×3-2-2组合的F1、F2、BC1P1、BC1P2群体确定该组合杂交后代是由两对基因控制;调查WI998×TopMark杂交组合F1、F2、BC1P1、BC1P2群体确定该杂交组合后代由两对基因、一对修饰基因控制。综合分析上述三个杂交组合,确定雌雄异花同株材料3-2-2的基因型为AA,雄全同株材料TopMark基因型为aa,本试验中3-2-2×TopMark的杂交后代只有一对基因表达。控制甜瓜性别分化共有三对基因,M基因为微效基因,试验结果提出存在另一微效基因F,协同M基因控制三性花株与雌全同株性别表达。
筛选428对SSR分子标记(其中包括380对EST-SSR标记,48对g-SSR标记),得到94对具有多态性的标记,其片段大小分布在150~300bp,多态性比例占21.29%;筛选256对EcorI/MseI AFLP引物组合,得到22对AFLP多态性引物,共含有146个AFLP多态性位点,其片段大小分布在100~600bp之间。利用3-2-2×TopMark杂交组合得到152株重组自交系群体(F1S6),构建甜瓜遗传图谱。该图谱包括70个SSR标记,100个AFLP标记及1个形态标记,图谱由17个连锁群构成,覆盖基因组总长度为1222.9cM,最长连锁群为LG1(197.4 cM),最短的连锁群为LG5(11.0 cM),标记之间平均距离为7.19 cM,标记间最大连锁距离为34.4 cM,标记间最小连锁距离为0;10个标记与A基因在同一连锁群,该连锁群覆盖基因组长度55.7 cM,找到两个与雌雄异花同株性状连锁的分子标记MU13328-3、E33M43-1,与A基因的遗传距离分别为4.8 cM和6.0 cM。
本试验分析了雌雄异花同株遗传规律,确定了甜瓜雌雄异花同株材料的基因型,不仅对甜瓜性别基因的研究提供了方法与指导,同时构建了较饱和的甜瓜永久遗传连锁图谱,对今后深入开展甜瓜雌雄异花同株基因的分离与图位克隆具有重要的理论价值。
Melon is an important economic group in cucubit. Most of melon were andromonoecious, monoecy can not only reduce the hand pollination but also can improve the quality of F1 hybrids than other andromonoecious accessions. So, the monoecious become to an important goal for melon breeding. Investigation sex expression on different generations from different crosses to analysis the inheritance of monoecious and identify the molecular markers which linkage to monoecious character. The possibility of Markers-assisted selection was explored in monoecious analysis which could speed up seed breeding process and develop a new method on melon breeding.
Parental lines were: female parent 3-2-2 from China, thin-skinned melon, monoecious; male parent TopMark, from USA, thick-skinned and netted melon, andromonoecious; tested parent WI998, from USA, netted melon, gynoecious. Investigated the sex type and expression of F1, F2 and BC generations from crosses 3-2-2×TopMark, WI998×TopMark, WI998×3-2-2, respectively to confirm the genotype of different sex performance on melon. Using 350 F1 generations from 3-2-2×TopMark to constructe recombination inbreed lines, acqurie 152 F1S6 generations to constructe genetic map.
Sex expression was controlled by a domiance gene with two mine genes and modify gene after investigated the sex performance of F2 and BC generations from cross 3-2-2×TopMark, main-gene heridity was 83.3% and mine-gene heridity was 8.125%. Ensured the performance was controlled by two dominance gene after investigated the F2 generation from WI998×3-2-2, and three gene controlled sex performance of cross WI998×TopMark. Segregation of every generation were investigated and confirmed the genotype of 3-2-2(monoecious)was AA, and the genotype of TopMark (andomonoecious)was aa. The M was modify gene and exist another modify gene F to coorperate with M gene to controlle Trimonoecious and Gynomonoecious.
Selected 428 SSR markers which included 380 pairs of EST–SSR markers and 48 pairs g-SSR markers, 94 primers were polymophic which the size from 150~300bp; selected 22 primers were polimophic in 256 AFLP EcorI/MseI primers, and 146 polymophic sits from 100~600bp.
Using RILs genrations of 3-2-2×TopMark to construct genetic map, a total of 70 SSR markers, 100 AFLP markers and one agronomic traits was grouped in to 17 large linkage groups which spanned 1222.9cM with an average marker interval of 7.19 cM. The longest linkage group was LG1(spanning 197.4 cM)and the shortest group was LG3(spanning 11.0 cM). Ten markers in the same linkage group with A gene and the group spanning 55.7 cM, SSR marker MU13328-3 and AFLP marker e33m43-1 were linkage to A gene with 4.8 cM and 6.0 cM, respectively.
Identified the genotype of monoecious and studied on inheritance of monoecious will not only increase the saturation of genetic map on molecular markers but also have the theoretics value on gene seperation and mapping clone of monoecious melon.
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