甘蓝型油菜苗期耐湿性和抗旱性相关QTL分析
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摘要
湿害和干旱是作物主要的逆境胁迫之一,也是我国油菜产区影响最终产量和品质的主要逆境因子之一。本研究利用具耐湿性和抗旱性差异的两份材料为双亲构建了一个由F_1小孢子培养得到的150个DH系,通过两个环境下的田间试验,考察株高等5个性状,结合分子标记分析,对性状及耐湿系数和抗旱系数进行QTL定位,分析甘蓝型油菜苗期的耐湿性和抗旱性。主要结果如下:
1、两亲本比较发现,母本(No2127-17×275B F_4)比父本(沪油15×5900 F_4)表现出更强的耐湿性和抗旱性。
2、以183个SSR标记和157个AFLP标记构建了全长为1489.4 cM的遗传连锁图谱,标记间的平均距离为4.4 cM。采用共同的SSR标记,将本图谱与甘蓝型油菜通用图谱进行了初步对应。
3、卡方测验表明,在DH群体中,86个标记(25.3%)表现显著偏分离(P＜0.01),且大部分偏向父本。所有定位的340个位点的等位基因在DH群体中的分布为父本等位基因占52.7%,母本等位基因占47.3%,近似为1:1,略微偏向父本。
4、DH群体各性状变异丰富,大部分表现出明显的超亲分离,同时表现为正态分布。
5、应用复合区间作图法,对株高、根长、地上部干重、根干重和总干重5个性状进行了QTL定位。对照和湿害胁迫下总共检测到45个QTL,其中对照条件下检测到28个,湿害胁迫条件下共检测到26个,在对照和湿害胁迫下同时检测到的QTL有9个;另外,由耐湿系数(WTC)检测到11个QTL。
6、应用复合区间作图法,对5个性状进行了QTL定位。对照和干旱胁迫下总共检测到50个QTL,其中对照条件下检测到28个,干旱胁迫条件下共检测到31个,在对照和干旱胁迫下同时检测到的QTL有9个;另外,由抗旱系数(DRC)检测到19个QTL。
7、检测到的各性状QTL以及其耐湿系数QTL和抗旱系数QTL有重叠分布,说明耐湿性和抗旱性的遗传基础有着紧密联系,可以在育种上将二者加以聚合。
Waterlogging and drought are serious environmental stresses on the growth of crop, which are the main abiotic constraints in oilseed production. In this study, we employed two B.napus lines which are different in tolerance to waterlongging and drought stress. The population with 150 doubled haploid (DH) lines was derived from the microspore culture of F_1. Testing experiments with 3 replicates following a randomized complete block design were implemented in two environments. Molecular markers were used to analyze the genetic model associated with 5 traits, WTC (waterlogging tolerance coefficient) and DRC (drought resistance coefficient) of all the traits. The main results are summarized as follow:
1. Comparing two parents, the female parent (No2127-17×275B F4) is more tolerant to waterlogging and drought stress than the male parent (Huyoul 5×5900 F_4).
2. A genetic linkage map, spanning 1489.4 cM with an average interval of 4.4 cM, was constructed using 183 simple sequence repeat (SSR) markers and 157 Amplified fragment length polymorphism (AFLP) markers. Based on the public information of microsatellites (SSR markers), our map was aligned with reference maps.
3. In the DH population, 86 markers (25.3%) skewed from the expected 1:1 genotypic segregation by Chi-square test, and most of them skewed towards the male parent. The allele frequencies of the male parent and the female parent in all 340 informative loci were 52.7% and 47.3% respectively. The allele ratio approximated to the expected 1:1.
4. Most traits in DH population showed transgressive segregation and demonstrated the normal distribution.
5. Using composite interval mapping method, a total of 45 QTL were detected for five traits, which are plant height, root length, shoot dry weight, root dry weight and total dry weight, under control and waterlogging condition. Twenty eight QTL were detected under control and 26 QTL were detected under waterlogging condition. There are 9 QTL detected under both control and waterlogging condition. The other 11 QTL were detected by WTC.
6. The entire genome was searched for QTL conferring significant effects on all scored traits by composite interval mapping in DH population. A total of 50 QTL were detected for five traits under control and drought condition. Twenty eight QTL were detected under control and 31 QTL were detected under drought condition. There are 9 QTL detected under both control and waterlogging condition The other 19 QTL related to DRC were identified.
7. QTL detected for five traits, WTC and DRC are clustering. All clusters detected indicated that the waterlogging tolerance and drought resistance may have strong relationship, which implied that waterlogging tolerance and drought resistance can be pyramided in breeding in B. napus.
1、两亲本比较发现,母本(No2127-17×275B F_4)比父本(沪油15×5900 F_4)表现出更强的耐湿性和抗旱性。
2、以183个SSR标记和157个AFLP标记构建了全长为1489.4 cM的遗传连锁图谱,标记间的平均距离为4.4 cM。采用共同的SSR标记,将本图谱与甘蓝型油菜通用图谱进行了初步对应。
3、卡方测验表明,在DH群体中,86个标记(25.3%)表现显著偏分离(P＜0.01),且大部分偏向父本。所有定位的340个位点的等位基因在DH群体中的分布为父本等位基因占52.7%,母本等位基因占47.3%,近似为1:1,略微偏向父本。
4、DH群体各性状变异丰富,大部分表现出明显的超亲分离,同时表现为正态分布。
5、应用复合区间作图法,对株高、根长、地上部干重、根干重和总干重5个性状进行了QTL定位。对照和湿害胁迫下总共检测到45个QTL,其中对照条件下检测到28个,湿害胁迫条件下共检测到26个,在对照和湿害胁迫下同时检测到的QTL有9个;另外,由耐湿系数(WTC)检测到11个QTL。
6、应用复合区间作图法,对5个性状进行了QTL定位。对照和干旱胁迫下总共检测到50个QTL,其中对照条件下检测到28个,干旱胁迫条件下共检测到31个,在对照和干旱胁迫下同时检测到的QTL有9个;另外,由抗旱系数(DRC)检测到19个QTL。
7、检测到的各性状QTL以及其耐湿系数QTL和抗旱系数QTL有重叠分布,说明耐湿性和抗旱性的遗传基础有着紧密联系,可以在育种上将二者加以聚合。
Waterlogging and drought are serious environmental stresses on the growth of crop, which are the main abiotic constraints in oilseed production. In this study, we employed two B.napus lines which are different in tolerance to waterlongging and drought stress. The population with 150 doubled haploid (DH) lines was derived from the microspore culture of F_1. Testing experiments with 3 replicates following a randomized complete block design were implemented in two environments. Molecular markers were used to analyze the genetic model associated with 5 traits, WTC (waterlogging tolerance coefficient) and DRC (drought resistance coefficient) of all the traits. The main results are summarized as follow:
1. Comparing two parents, the female parent (No2127-17×275B F4) is more tolerant to waterlogging and drought stress than the male parent (Huyoul 5×5900 F_4).
2. A genetic linkage map, spanning 1489.4 cM with an average interval of 4.4 cM, was constructed using 183 simple sequence repeat (SSR) markers and 157 Amplified fragment length polymorphism (AFLP) markers. Based on the public information of microsatellites (SSR markers), our map was aligned with reference maps.
3. In the DH population, 86 markers (25.3%) skewed from the expected 1:1 genotypic segregation by Chi-square test, and most of them skewed towards the male parent. The allele frequencies of the male parent and the female parent in all 340 informative loci were 52.7% and 47.3% respectively. The allele ratio approximated to the expected 1:1.
4. Most traits in DH population showed transgressive segregation and demonstrated the normal distribution.
5. Using composite interval mapping method, a total of 45 QTL were detected for five traits, which are plant height, root length, shoot dry weight, root dry weight and total dry weight, under control and waterlogging condition. Twenty eight QTL were detected under control and 26 QTL were detected under waterlogging condition. There are 9 QTL detected under both control and waterlogging condition. The other 11 QTL were detected by WTC.
6. The entire genome was searched for QTL conferring significant effects on all scored traits by composite interval mapping in DH population. A total of 50 QTL were detected for five traits under control and drought condition. Twenty eight QTL were detected under control and 31 QTL were detected under drought condition. There are 9 QTL detected under both control and waterlogging condition The other 19 QTL related to DRC were identified.
7. QTL detected for five traits, WTC and DRC are clustering. All clusters detected indicated that the waterlogging tolerance and drought resistance may have strong relationship, which implied that waterlogging tolerance and drought resistance can be pyramided in breeding in B. napus.
引文
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