小麦K型细胞质雄性不育育性恢复基因的标记定位和效应分析
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摘要
小麦K型细胞质雄性不育系是非常有应用潜力的一种不育系,现已证明小麦K型雄性不育系的主效恢复基因有两对,分别是位于1BS上的Rfk1和2BL上的Rfk2,挖掘与恢复基因紧密连锁的分子标记,对恢复基因进行基因定位和搞清两对恢复基因的效应和作用方式,将有助于选育出恢复度高而稳定的恢复系。本文选用了不诱发单倍体的K型不育系豫麦3号与恢复系豫麦2号构建的遗传背景为豫麦3号的恢复基因Rfk1和Rfk2的近等位基因系,豫麦2号/豫麦3号F5重组近交系,以及5个K型不育系K豫麦3号、K豫农43、K豫教1号、K豫农93151和K豫麦21,对两个恢复基因Rfk1和Rfk2进行了分子标记的定位和效应分析。主要结论如下:
1.利用所筛选的与恢复基因连锁的SSR标记对豫麦2号/豫麦3号F5重组近交系进行标记鉴定,根据杂交组合的育性分离和豫麦2号/豫麦3号F5重组近交系的带型对Rfk1和Rfk2进行基因定位,1BS染色体上的5个SSR标记TAGLGAP、Xgwm374、Xgwm18、Xgwm11、BARC061与恢复基因Rfk1的遗传距离分别为10.1 cM、7.1 cM、3.0 cM、3.0 cM、13.4 cM,该恢复基因位于标记Xgwm18和Xgwm11之间。恢复基因Rfk2与引物cfd26和Xgwm526遗传距离为5.2和18.8 cM。
2.根据豫麦2号/豫麦3号F5重组近交系群体带型和重组近交系/K豫麦3号杂交F1群体育性恢复情况对重组近交系/K豫麦3号杂交F1群体的基因型进行判定,只携带Rfk1恢复基因的单株有69株,平均国内法恢复度为64.92%,国际法为106.68%;只携带Rfk2恢复基因的单株有56株,平均国内法恢复度为57.82%,国际法为93.35%;同时携带2个恢复基因的单株有66株,平均国内法恢复度为73.98%,国际法为117.37%。因此,可以初步证明Rfk1的恢复力较强,Rfk2的恢复力较弱,2个恢复基因有累加效应。同时,还可以看出,育性恢复度较低的单株(<45%),大多携带恢复基因Rfk2。
3.利用5个K型不育系K豫麦3号、K豫农43、K豫教1号、K豫农93151和K豫麦21与Rfk1和Rfk2的近等位基因系组配杂交组合,进一步明确恢复基因Rfk1和Rfk2的效应大小,结果表明Rfk1对5个不育系的国内法、国际法恢复度均高于Rfk2,进一步证明恢复基因Rfk1的效应大于Rfk2。
Wheat male sterile line with Ae.kotschyi cytoplasm has the best potential application and was controlled by two pairs of major fertility restorer genes;The two pairs of major fertility restorer genes were located in Rfk1 for 1BS and Rfk2 for 2BL of wheat;Good restoring lines would be selected and hybrid wheat varieties would be developed by mapping restorer genes and analysising the effect of two restorer genes;In this study, the near-isogenic lines (NILs) were constructed using sterile line Yumai 3 with Ae.kotschyi cytoplasm which don’t induced haploid, restorer line Yumai 2 which have two pairs of major restorer genes of Rfk1和Rfk2;The F5 group of Yumai3/Yumai2 NILs and 5 sterile lines of Yumai3,Yunong43,Yujiao1,Yunong93151 and Yumai21;That materials wered be Markered and effect analysised,the results showed as follows:
1,By using the SSR makers linkaged to restorer genes,which mapped and identified genes of Rfk1 and Rfk2 in F5 groups of Yumai3/Yumai2 NILs;5 SSR makers of TAGLGAP、Xgwm374、Xgwm18、Xgwm11、and BARC061wered identified in 1BS,which markers to Rfk1 distance are 10.1 cM、7.1 cM、3.0 cM、3.0 cM and13.4 cM, respectively. The Rfk1 mapped between Xgwm18 and Xgwm11;The genetic distance between Rfk2 and primer cfd267 was 5.2,Rfk2 and primer Xgwm526 was 18.8 cM.
2. By identified genotypes in F5 groups of Yumai3/Yumai2 NILs and restoration performance in F1 groups of NILS/ Yumai3,which judged genotypes of F1 groups of NILS/ Yumai3;There are 69 plants carried Rfk1 restoring genes , which average restoring degrees was 64.92% with domestic method and 106.68% with international method; There are 56 plants carried Rfk2 restoring genes, which average restoring degrees is 57.82% with domestic method , 93.35% with international method;There are 66 plants carried two restoring genes, which average restoring degrees is 73.98% with domestic method and 117.37% with international method.Therefore, the restoring ability of Rfk1 is stroger than Rfk2 ,the 2 restoring genes have accumulative effect. At the same time , the low restoring ability (<45%) plants always carry Rfk2 gene.
3, The hybridized combination NILs of 5 sterile lines with Ae.kotschyi cytoplasm Yunong43,Yujiao1,Yunong93151 and Yumai21 wered used in identifying the effect of Rfk1 and Rfk2 ;The result show the average restoring degrees of Rfk1 on 5 sterile lines was higher than Rfk2,Hence,the effect of Rfk1 is bigger than Rfk2.
1.利用所筛选的与恢复基因连锁的SSR标记对豫麦2号/豫麦3号F5重组近交系进行标记鉴定,根据杂交组合的育性分离和豫麦2号/豫麦3号F5重组近交系的带型对Rfk1和Rfk2进行基因定位,1BS染色体上的5个SSR标记TAGLGAP、Xgwm374、Xgwm18、Xgwm11、BARC061与恢复基因Rfk1的遗传距离分别为10.1 cM、7.1 cM、3.0 cM、3.0 cM、13.4 cM,该恢复基因位于标记Xgwm18和Xgwm11之间。恢复基因Rfk2与引物cfd26和Xgwm526遗传距离为5.2和18.8 cM。
2.根据豫麦2号/豫麦3号F5重组近交系群体带型和重组近交系/K豫麦3号杂交F1群体育性恢复情况对重组近交系/K豫麦3号杂交F1群体的基因型进行判定,只携带Rfk1恢复基因的单株有69株,平均国内法恢复度为64.92%,国际法为106.68%;只携带Rfk2恢复基因的单株有56株,平均国内法恢复度为57.82%,国际法为93.35%;同时携带2个恢复基因的单株有66株,平均国内法恢复度为73.98%,国际法为117.37%。因此,可以初步证明Rfk1的恢复力较强,Rfk2的恢复力较弱,2个恢复基因有累加效应。同时,还可以看出,育性恢复度较低的单株(<45%),大多携带恢复基因Rfk2。
3.利用5个K型不育系K豫麦3号、K豫农43、K豫教1号、K豫农93151和K豫麦21与Rfk1和Rfk2的近等位基因系组配杂交组合,进一步明确恢复基因Rfk1和Rfk2的效应大小,结果表明Rfk1对5个不育系的国内法、国际法恢复度均高于Rfk2,进一步证明恢复基因Rfk1的效应大于Rfk2。
Wheat male sterile line with Ae.kotschyi cytoplasm has the best potential application and was controlled by two pairs of major fertility restorer genes;The two pairs of major fertility restorer genes were located in Rfk1 for 1BS and Rfk2 for 2BL of wheat;Good restoring lines would be selected and hybrid wheat varieties would be developed by mapping restorer genes and analysising the effect of two restorer genes;In this study, the near-isogenic lines (NILs) were constructed using sterile line Yumai 3 with Ae.kotschyi cytoplasm which don’t induced haploid, restorer line Yumai 2 which have two pairs of major restorer genes of Rfk1和Rfk2;The F5 group of Yumai3/Yumai2 NILs and 5 sterile lines of Yumai3,Yunong43,Yujiao1,Yunong93151 and Yumai21;That materials wered be Markered and effect analysised,the results showed as follows:
1,By using the SSR makers linkaged to restorer genes,which mapped and identified genes of Rfk1 and Rfk2 in F5 groups of Yumai3/Yumai2 NILs;5 SSR makers of TAGLGAP、Xgwm374、Xgwm18、Xgwm11、and BARC061wered identified in 1BS,which markers to Rfk1 distance are 10.1 cM、7.1 cM、3.0 cM、3.0 cM and13.4 cM, respectively. The Rfk1 mapped between Xgwm18 and Xgwm11;The genetic distance between Rfk2 and primer cfd267 was 5.2,Rfk2 and primer Xgwm526 was 18.8 cM.
2. By identified genotypes in F5 groups of Yumai3/Yumai2 NILs and restoration performance in F1 groups of NILS/ Yumai3,which judged genotypes of F1 groups of NILS/ Yumai3;There are 69 plants carried Rfk1 restoring genes , which average restoring degrees was 64.92% with domestic method and 106.68% with international method; There are 56 plants carried Rfk2 restoring genes, which average restoring degrees is 57.82% with domestic method , 93.35% with international method;There are 66 plants carried two restoring genes, which average restoring degrees is 73.98% with domestic method and 117.37% with international method.Therefore, the restoring ability of Rfk1 is stroger than Rfk2 ,the 2 restoring genes have accumulative effect. At the same time , the low restoring ability (<45%) plants always carry Rfk2 gene.
3, The hybridized combination NILs of 5 sterile lines with Ae.kotschyi cytoplasm Yunong43,Yujiao1,Yunong93151 and Yumai21 wered used in identifying the effect of Rfk1 and Rfk2 ;The result show the average restoring degrees of Rfk1 on 5 sterile lines was higher than Rfk2,Hence,the effect of Rfk1 is bigger than Rfk2.
引文
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