小麦—长穗偃麦草杂种后代矮秆基因的鉴定和分子标记研究
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摘要
本研究以小麦－长穗偃麦草杂种后代材料山农31504-1等5个矮秆种质系以及相关亲本为材料，对其主要农艺性状特点和细胞学特点进行了鉴定，在此基础上，进一步利用种子醇溶蛋白A-PAGE、麦谷蛋白SDS-PAGE和RAPD技术对矮秆种质系及其携带的矮秆基因进行了鉴定和标记分析，同时对矮秆基因的遗传特点进行了研究，并利用中国春缺体－四体系将矮秆基因进行了定位。获得的主要结果如下：
1、 从小麦－长穗偃麦草矮秆双体异附加系山农31504和山农31505中分别选育鉴定出染色体数为2n＝42的矮秆材料山农31504-1和山农31505-1，同时利用这2个矮秆种质系和山农31504与山农049-3杂种高代材料山农L024、山农L026和山农L113与小麦育种中利用的7个主要矮源的农艺性状进行了比较。结果表明，山农31504-1等5个矮秆种质系株高仅50cm-60cm左右，具有良好的农艺性状，后期不早衰，是良好的矮源，在小麦遗传改良及遗传研究中具有重要利用价值。
2、 利用细胞遗传学方法对山农31504-1等5个矮秆种质系进行了鉴定，结果表明山农31504-1、山农31505-1、山农L024和山农L026为异易位系，山农L113为异代换系。种子醇溶蛋白A-PAGE、麦谷蛋白SDS-PAGE和RAPD分析结果表明，山农31504-1等5个矮秆种质系的谱带中，都含有长穗偃麦草的特征带，说明它们均含有渗入的长穗偃麦草遗传物质。
3、 利用异易位系山农31504-1分别与高秆材料山农298和中国春杂交，对山农31504-1的遗传特点进行了分析。结果证明，山农31504-1中的矮秆基因能够明显降低杂种的株高，其作用表现为部分显性，F2群体株高分离为矮秆、中秆和高秆三种类型，其分离比例符合1:2:1，证明矮秆种质系山农31504-1中所含的矮秆基因为部分显性单基因。通过对5个矮秆种质系进行GA3鉴定，5个矮秆种质系均表现不敏感，从而证明它们所含的矮秆基因为赤霉酸不敏感型。
利用中国春缺体－四体系和端体系与矮秆种质系山农31504-1杂交，进行染色体定位分析。结果表明，22个杂种F1中，只有N2AT2B/山农31504-1的杂种F1株高显著低于中亲值，其余杂种F1株高均高于中亲
4、 值。从而初步将矮秆种质系山农31504-1的矮秆基因定位于染色体2A上。
5、 利用分离群体分组法对矮秆种质系31504-1中的矮秆基因进行了RAPD分析。从300个随机引物中筛选出引物S152，可在矮秆DNA池中稳定的扩增出一条约2100bp的带，该带在高秆DNA池中没有出现，将其命名为S1522100，可作为矮秆基因的RAPD标记。该标记经多次重复表现稳定，它与矮秆基因的遗传距离为10.73±3.31cM。
Wheat-Elytrigia elongatum chromosome translocation and substitution lines were isolated by using morphological markers, cytological markers and biochemical markers. Meanwhile, in order to find the molecular marker of the dwarf gene, RAPD markers were used. The results are showed just as follows.
1. Two dwarf germplasm lines shannong31504-1 and shannong31505-1 with the chromosome number 2n=42 are selected from self progenies of short stalked Wheat-Elytrigia elongatum disomic addition lines Shannong31504 and Shannong 31505 These two dwarf germplasm lines and three dwarf material lines ShannongL024, ShannongL026, ShannongL113, which came from the hybrid between Shannong31504 and Shannong049-3, were compared with seven main dwarf sources in field characters. The results indicate that five dwarf germplasm lines have good field characters and no premature senility. They are very valuable in the area of plant breeding and genetics.
2. Cytogenetic analysis of five dwarf germplasm lines showed that Shannong31504-1, Shannong31505-1, ShannongL024 and ShannongL026 are translocation lines, ShannongL113 is substitution line. Further studies of seed Gliadin A-PAGE, Glutenin SDS-PAGE and RAPD assay all indicated that the five dwarf germplasm lines have the chromatin of Elytrigia elongatum.
3. Crosses between dwarf translocation line shannong31504-1 and high material Shannong298 and Chinese Spring were made respectively. The plant height of F1 hybrids were slightly over the mid-parent value, the proportion of the dwarf semi-dwarf and high individuals in the F2 segregating population was 1:2:1,showing that the dwarf trait was controlled by one partial dominant gene.
Cytogenetic method was used in order to location the dwarf gene. Twenty Chinese spring nullitetrasomic lines and two ditelosomic lines were crossed with the dwarf
4. translocation line shannong31504-1. The plant height in F1s of all crosses were over or near the mean heights of their two parents except that from shannong31504-1/CS N2AT2B in which the plant height of the F1 was significantly shorter than the mid-parent value, indicating that the Rht gene was located on the chromosome 2A.
5. The random amplified polymorphic DNA(RAPD) analysis was made in the translocation line shannong31504-1 using the method of "bulked segregate analysis(BSA)". From 300 random primers ,primer S152 was screened and could produce a 2100bp reproducible fragment only in the dwarf DNA pool, but absent in high DNA pool, the polymorphic fragment was designated as S1522100. Using F2 mapping population from a cross of shannong31504-1/ shannong298,the dwarf gene was shown to be linked to the marker S1522100 at a genetic distance of 10.73±3.31cM.
1、 从小麦－长穗偃麦草矮秆双体异附加系山农31504和山农31505中分别选育鉴定出染色体数为2n＝42的矮秆材料山农31504-1和山农31505-1，同时利用这2个矮秆种质系和山农31504与山农049-3杂种高代材料山农L024、山农L026和山农L113与小麦育种中利用的7个主要矮源的农艺性状进行了比较。结果表明，山农31504-1等5个矮秆种质系株高仅50cm-60cm左右，具有良好的农艺性状，后期不早衰，是良好的矮源，在小麦遗传改良及遗传研究中具有重要利用价值。
2、 利用细胞遗传学方法对山农31504-1等5个矮秆种质系进行了鉴定，结果表明山农31504-1、山农31505-1、山农L024和山农L026为异易位系，山农L113为异代换系。种子醇溶蛋白A-PAGE、麦谷蛋白SDS-PAGE和RAPD分析结果表明，山农31504-1等5个矮秆种质系的谱带中，都含有长穗偃麦草的特征带，说明它们均含有渗入的长穗偃麦草遗传物质。
3、 利用异易位系山农31504-1分别与高秆材料山农298和中国春杂交，对山农31504-1的遗传特点进行了分析。结果证明，山农31504-1中的矮秆基因能够明显降低杂种的株高，其作用表现为部分显性，F2群体株高分离为矮秆、中秆和高秆三种类型，其分离比例符合1:2:1，证明矮秆种质系山农31504-1中所含的矮秆基因为部分显性单基因。通过对5个矮秆种质系进行GA3鉴定，5个矮秆种质系均表现不敏感，从而证明它们所含的矮秆基因为赤霉酸不敏感型。
利用中国春缺体－四体系和端体系与矮秆种质系山农31504-1杂交，进行染色体定位分析。结果表明，22个杂种F1中，只有N2AT2B/山农31504-1的杂种F1株高显著低于中亲值，其余杂种F1株高均高于中亲
4、 值。从而初步将矮秆种质系山农31504-1的矮秆基因定位于染色体2A上。
5、 利用分离群体分组法对矮秆种质系31504-1中的矮秆基因进行了RAPD分析。从300个随机引物中筛选出引物S152，可在矮秆DNA池中稳定的扩增出一条约2100bp的带，该带在高秆DNA池中没有出现，将其命名为S1522100，可作为矮秆基因的RAPD标记。该标记经多次重复表现稳定，它与矮秆基因的遗传距离为10.73±3.31cM。
Wheat-Elytrigia elongatum chromosome translocation and substitution lines were isolated by using morphological markers, cytological markers and biochemical markers. Meanwhile, in order to find the molecular marker of the dwarf gene, RAPD markers were used. The results are showed just as follows.
1. Two dwarf germplasm lines shannong31504-1 and shannong31505-1 with the chromosome number 2n=42 are selected from self progenies of short stalked Wheat-Elytrigia elongatum disomic addition lines Shannong31504 and Shannong 31505 These two dwarf germplasm lines and three dwarf material lines ShannongL024, ShannongL026, ShannongL113, which came from the hybrid between Shannong31504 and Shannong049-3, were compared with seven main dwarf sources in field characters. The results indicate that five dwarf germplasm lines have good field characters and no premature senility. They are very valuable in the area of plant breeding and genetics.
2. Cytogenetic analysis of five dwarf germplasm lines showed that Shannong31504-1, Shannong31505-1, ShannongL024 and ShannongL026 are translocation lines, ShannongL113 is substitution line. Further studies of seed Gliadin A-PAGE, Glutenin SDS-PAGE and RAPD assay all indicated that the five dwarf germplasm lines have the chromatin of Elytrigia elongatum.
3. Crosses between dwarf translocation line shannong31504-1 and high material Shannong298 and Chinese Spring were made respectively. The plant height of F1 hybrids were slightly over the mid-parent value, the proportion of the dwarf semi-dwarf and high individuals in the F2 segregating population was 1:2:1,showing that the dwarf trait was controlled by one partial dominant gene.
Cytogenetic method was used in order to location the dwarf gene. Twenty Chinese spring nullitetrasomic lines and two ditelosomic lines were crossed with the dwarf
4. translocation line shannong31504-1. The plant height in F1s of all crosses were over or near the mean heights of their two parents except that from shannong31504-1/CS N2AT2B in which the plant height of the F1 was significantly shorter than the mid-parent value, indicating that the Rht gene was located on the chromosome 2A.
5. The random amplified polymorphic DNA(RAPD) analysis was made in the translocation line shannong31504-1 using the method of "bulked segregate analysis(BSA)". From 300 random primers ,primer S152 was screened and could produce a 2100bp reproducible fragment only in the dwarf DNA pool, but absent in high DNA pool, the polymorphic fragment was designated as S1522100. Using F2 mapping population from a cross of shannong31504-1/ shannong298,the dwarf gene was shown to be linked to the marker S1522100 at a genetic distance of 10.73±3.31cM.
引文
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