分子标记辅助改良03S的稻瘟病和褐飞虱抗性
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摘要
稻瘟病是水稻的三大病害之一,褐飞虱是水稻生产上的首要农业害虫,两者都严重地影响了稻米生产。化学防治稻瘟病和褐飞虱不仅增加成本,而且污染环境,长期育种实践证明种植抗稻瘟病、褐飞虱水稻新品种,是防治稻瘟病和褐飞虱最经济、有效、环保的方法,而常规育种方法鉴定难、时间长、选择准确差、育种效率低,育种进展慢。利用分子标记辅助育种是解决抗性育种难题的有效途径之一。本文采用分子标记辅助选择技术,结合田间检测,改良了03S的稻瘟病和褐飞虱抗性。
以抗稻瘟病材料福伊B(携带抗稻瘟病基因Pid2)和RF48(携带抗稻瘟病基因Pi9)作为稻瘟病抗源供体,以03S为轮回亲本,分别杂交、连续回交,构建回交导入系群体,通过目标基因的前景选择和遗传背景的分子标记辅助选择,选择抗稻瘟病而遗传背景与03S相似的新株系。以分子标记RM3辅助选择,将来源于水稻品种福伊B的抗稻瘟病基因Pid2导入两系不育系03S,该标记辅助选择的准确率达到95.24%,再结合少量的田间检测,就能育成准确度极高的抗性株；以分子标记AP5659-5、AP5930辅助选择,将来源于水稻品种RF48的抗稻瘟病基因Pi9也导入到两系不育系03S中,这两个标记比以往用于辅助育种的标记Pb8准确率高,与田间抗性检测的结果完全吻合。采用了220个较均匀分布于全基因组的SSR标记分析了亲本03S和福伊B间的多态性,共有55个SSR标记表现出多态性,多态性的比例为25%,并利用这55个SSR标记,分析了回交后代相对于轮回亲本03S的背景恢复率,结果表明在BC3F1中,背景恢复率最高可达96.40%。
在改良褐飞虱抗性方面,分别采用了与抗褐飞虱基因Bph14, Bph15紧密连锁的SSR标记RM570、MS5对03S与B5的回交群体进行标记辅助前景选择。采用了220个较均匀分布于全基因组的SSR标记分析了亲本03S和B5间的多态性,共发现48个SSR标记表现出多态性,多态性比例达21.8%,并利用这48个SSR标记,分析了回交后代相对于轮回亲本03S的背景恢复率,结果表明在14个BC3F1群体的42个单株中,背景恢复率变化范围为81.20%-100%,平均值为93.16%,也明显高于理论值87.5%,通过背景恢复率的选择可加速抗性单株的背景恢复,使表型与轮回亲本更相似,提高育种效率和速度。
Blast and brown planthopper are destructive to the growth of rice, resulting in great loss of crop every year. Breeding and planting the rice cultivars with resistances to blast and brown planthopper is significative, not only to the rice production but also to the protection of envioroment. But the conventional breeding for developing rice cultivars with resistances to blast and brown planthopper is very difficult because of time consuming and cost. Marker Assisted Selection (MAS) gives a new way to develop the rice resistant to diseases and pests, owing to its effectiveness to identify the diseases and pest. In this thesis, we reported that the resistances of 03S to blast and brown planthopper was improved by MAS.
The recurrent parent 03S was crossed with the blast resistance donor parents carrying Pid2 and RF48 carrying Pi9 and backcrossed to construct the backcross introgression lines for selecting new lines with both resistances to blast and the genetical background similar to the 03S. The molecular marker RM3 was used to select the blast resistance gene Pid2 in the backcross BC2F1 population derived from 03S with FuyiB, the best accurate rate is 95.24% for selection.The markers of AP5659-5 and AP5930 were used to select the blast resistance gene Pi9 in the backcross BC2F1 population derived from 03S with RF48.The results show the selecting accurate rate is 100% and much better than using the marker Pb8. Among 220 SSR markers covering 12 chromsomes,55 SSR markers were detected having polymorphism between 03S and FuyiB, then the 55 markers were used to analyse the background recovery rates of the resistant lines derived from the BC3F1 population. Among the selected lines, the maximum of the background recovery rates is 96.40%.
To improve the resistance of 03S to brown planthopper, SSR markers RM570 and MS5 linking closely to brown planthopper resistance genes of Bph14 and Bph15 respectively were used to select the resistant genes in the backcross populations derived from B5(carrying the brown planthopper resistance gene Bph14 and Bph15) and 03S(the recurrent parent).48 out of 220 SSR markers covering 12 chromsomes were detected having polymorphism between 03S and FuyiB, The markers were used to analyse the background recovery rates of the resistant lines derived from the BC3F1 population. Among the selected lines, the maximum of the background recovery rates is 96.40%. The experiment results showing that the background recovery rates of the 42 resistant individuals derived from BC3F1 subpopulations vary from 81.20% to 100%, with the average of 93.16% much higher than the theoretical value 87.5%. Background selection can accelerates the recovery of the backgrounds of the lines selected for resistance, save time and enhance the efficiency for breeding.
以抗稻瘟病材料福伊B(携带抗稻瘟病基因Pid2)和RF48(携带抗稻瘟病基因Pi9)作为稻瘟病抗源供体,以03S为轮回亲本,分别杂交、连续回交,构建回交导入系群体,通过目标基因的前景选择和遗传背景的分子标记辅助选择,选择抗稻瘟病而遗传背景与03S相似的新株系。以分子标记RM3辅助选择,将来源于水稻品种福伊B的抗稻瘟病基因Pid2导入两系不育系03S,该标记辅助选择的准确率达到95.24%,再结合少量的田间检测,就能育成准确度极高的抗性株；以分子标记AP5659-5、AP5930辅助选择,将来源于水稻品种RF48的抗稻瘟病基因Pi9也导入到两系不育系03S中,这两个标记比以往用于辅助育种的标记Pb8准确率高,与田间抗性检测的结果完全吻合。采用了220个较均匀分布于全基因组的SSR标记分析了亲本03S和福伊B间的多态性,共有55个SSR标记表现出多态性,多态性的比例为25%,并利用这55个SSR标记,分析了回交后代相对于轮回亲本03S的背景恢复率,结果表明在BC3F1中,背景恢复率最高可达96.40%。
在改良褐飞虱抗性方面,分别采用了与抗褐飞虱基因Bph14, Bph15紧密连锁的SSR标记RM570、MS5对03S与B5的回交群体进行标记辅助前景选择。采用了220个较均匀分布于全基因组的SSR标记分析了亲本03S和B5间的多态性,共发现48个SSR标记表现出多态性,多态性比例达21.8%,并利用这48个SSR标记,分析了回交后代相对于轮回亲本03S的背景恢复率,结果表明在14个BC3F1群体的42个单株中,背景恢复率变化范围为81.20%-100%,平均值为93.16%,也明显高于理论值87.5%,通过背景恢复率的选择可加速抗性单株的背景恢复,使表型与轮回亲本更相似,提高育种效率和速度。
Blast and brown planthopper are destructive to the growth of rice, resulting in great loss of crop every year. Breeding and planting the rice cultivars with resistances to blast and brown planthopper is significative, not only to the rice production but also to the protection of envioroment. But the conventional breeding for developing rice cultivars with resistances to blast and brown planthopper is very difficult because of time consuming and cost. Marker Assisted Selection (MAS) gives a new way to develop the rice resistant to diseases and pests, owing to its effectiveness to identify the diseases and pest. In this thesis, we reported that the resistances of 03S to blast and brown planthopper was improved by MAS.
The recurrent parent 03S was crossed with the blast resistance donor parents carrying Pid2 and RF48 carrying Pi9 and backcrossed to construct the backcross introgression lines for selecting new lines with both resistances to blast and the genetical background similar to the 03S. The molecular marker RM3 was used to select the blast resistance gene Pid2 in the backcross BC2F1 population derived from 03S with FuyiB, the best accurate rate is 95.24% for selection.The markers of AP5659-5 and AP5930 were used to select the blast resistance gene Pi9 in the backcross BC2F1 population derived from 03S with RF48.The results show the selecting accurate rate is 100% and much better than using the marker Pb8. Among 220 SSR markers covering 12 chromsomes,55 SSR markers were detected having polymorphism between 03S and FuyiB, then the 55 markers were used to analyse the background recovery rates of the resistant lines derived from the BC3F1 population. Among the selected lines, the maximum of the background recovery rates is 96.40%.
To improve the resistance of 03S to brown planthopper, SSR markers RM570 and MS5 linking closely to brown planthopper resistance genes of Bph14 and Bph15 respectively were used to select the resistant genes in the backcross populations derived from B5(carrying the brown planthopper resistance gene Bph14 and Bph15) and 03S(the recurrent parent).48 out of 220 SSR markers covering 12 chromsomes were detected having polymorphism between 03S and FuyiB, The markers were used to analyse the background recovery rates of the resistant lines derived from the BC3F1 population. Among the selected lines, the maximum of the background recovery rates is 96.40%. The experiment results showing that the background recovery rates of the 42 resistant individuals derived from BC3F1 subpopulations vary from 81.20% to 100%, with the average of 93.16% much higher than the theoretical value 87.5%. Background selection can accelerates the recovery of the backgrounds of the lines selected for resistance, save time and enhance the efficiency for breeding.
引文
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