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Genetic improvement of clubroot resistance for the elite temperature-sensitive pol CMS line 616A in Brassica napus
Fu-cai WANG, Yi XU, Guang-sheng YANG, Zhao-yang WANG
CHINESE JOURNAL OF OIL CROP SCIENCES ›› 2024, Vol. 46 ›› Issue (5) : 999-1007.
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Genetic improvement of clubroot resistance for the elite temperature-sensitive pol CMS line 616A in Brassica napus
To improve rapeseed resistance to clubroot, Huashuang 5R and Huayouza 62R (carrying PbBa8.1 locus and CRb locus, respectively) were used as donor parents for transferring the resistant genes into the elite temperature-sensitive pol CMS line 616A, by combining with successive backcross and marker-assisted selection. By the process, three improved temperature-sensitive pol CMS lines 616A PbBa8.1, 616A CRb and 616A PbBa8.1 + CRb were obtained, with genetic background recovery rates of 95.45%, 94.80% and 92.58%, respectively. Subsequently, three clubroot resistant new hybrids Shengguang 128CR PbBa8.1, Shengguang 128CR CRb and Shengguang 128CR PbBa8.1 + CRb were derived from these three improved lines and 621R (the male parent of Shengguang 128). Clubroot resistance of the improved lines and the hybrids was identified by using race 4, the dominant physiological race in China. Results showed that 616A CRb, 616A PbBa8.1 + CRb and their hybrids (Shengguang 128CR CRb and Shengguang 128CR PbBa8.1 + CRb ) had immunity to race 4. The improved line 616A PbBa8.1 and the hybrid Shengguang 128CR PbBa8.1 were resistant to race 4; however, their resistance levels were lower than those of CRb-containing lines and hybrids. Evaluation of agronomic traits showed that no significant differences were observed between the improved lines and 616A, neither between the improved hybrids and Shengguang 128. The traits included plant height, number of the branch, silique length, seeds per silique and thousand seeds weight. Therefore, it is possible to use the three improved lines for breeding rapeseed hybrids with clubroot resistance.
Brassica napus L. / 616A / clubroot resistance / PbBa8.1 resistance locus / CRb resistance locus / molecular marker-assisted selection {{custom_keyword}} /
Table 1 Primer sequences of markers used in this study表1 引物序列信息 |
| 标记 Marker | 标记类型 Marker type | 左引物 Forward primer sequence（5'-3'） | 右引物 Reverse primer sequence （5'-3'） |
|---|---|---|---|
| A3-50 | InDel | GCTTTTCTTCCATACGTTTGC | CCAGGGCGGAGGTTCAAATC |
| A08-300 | InDel | GTAGTGCGGGCCACAAAAT | CACAATGGAGTGTTGAAATTCACT |
Table 2 Methods for trait evaluation表2 性状考察方法 |
| 性状名称Trait | 性状考察方法 Trait evaluation methods |
|---|---|
| 角果长 Silique length | 主花序中间部位10个角果的平均角果长 Mean length of silique from 10 siliques of middle parts on the main inflorescence |
| 每角果粒数 Seeds per silique | 主花序中间部位10个角果的平均籽粒数 Mean seeds per silique from 10 siliques of middle parts on the main inflorescence |
| 千粒重 Thousand seeds weight | 选取主花序中间10个角果的全部籽粒进行称重 All seeds of 10 pods in the middle of main inflorescence were weighed |
| 株高 Plant height | 从子叶节到植株主花序顶端的长度 The length from cotyledon node to the top of main inflorescence |
| 分枝数 | 主茎上着生的具有一个以上有效角果的第一次分枝数 |
| Effective branch number | Number of first branches with more than one effective pod on main stem |
Fig. 2 Genotype identification of the target genes in BC4F2 population图2 BC4F2群体目标基因型鉴定结果 |
Table 3 Foreground selection in the populations of DCF1 and BC1F1-BC4F2表3 DCF1、BC1F1-BC4F2各世代群体的前景选择 |
| 世代 Generation | 检测株数 No. of plants tested | 中选单株数 Selected plants | 基因型 Genotype |
|---|---|---|---|
| DCF1 | 6 | PbBa8.1pbba8.1/CRbcrb/RfpRfp | |
| BC1F1 | 48 | 8 | PbBa8.1pbba8.1/CRbcrb/RfpRfp |
| BC2F1 | 48 | 13 | PbBa8.1pbba8.1/CRbcrb/RfpRfp |
| BC3F1 | 24 | 2 | PbBa8.1pbba8.1/CRbcrb/RfpRfp |
| BC4F1 | 36 | 6 | PbBa8.1pbba8.1/CRbcrb/Rfprfp |
| BC4F2 | 600 | 4 | PbBa8.1PbBa8.1/rfprfp |
| BC4F2 | 600 | 4 | CRbCRb/rfprfp |
| BC4F2 | 600 | 8 | PbBa8.1PbBa8.1/CRbCRb/rfprfp |
Fig. 3 The fertility performance of the clubroot resistance improved pol TCMSline 616A PbBa8.1 + CRb and its restore line 616R图3 改良不育系616A PbBa8.1 + CRb 及其恢复系616R育性表现 |
Table 4 Disease index of improved lines and their parent 616A表4 616A改良株系及其亲本的病情指数 |
| 材料 Material | 鉴定株数 Identified plants | 平均感病株数 Infected plants | 平均病情指数 Disease index |
|---|---|---|---|
| 616A | 42.0 | 42.0±0 A | 94.62±0.6 A |
| 616A PbBa8.1 | 43.0 | 14.0±2.6 B | 25.67±1.3 B |
| 616A CRb | 41.0 | 0 C | 0 C |
| 616A PbBa8.1+CRb | 41.0 | 0 C | 0 C |
Fig. 4 Comparison of clubroot resistance between the improved lines and their parent 616A图4 改良株系与亲本616A根肿病抗性比较 |
Table 5 Disease index of the check hybrid shengguang128 and the improved hybrids表5 对照杂交种圣光128与改良杂交种的病情指数 |
| 材料 Materials | 鉴定株数 Identified plants | 感病株数 Infected plants | 病情指数 Disease index |
|---|---|---|---|
| 圣光128 | 40.0 | 40.0±0.8 A | 94.38±0.4 A |
| 圣光128CR PbBa8.1 | 41.0 | 8.0±0.3 B | 17.22±0.9 A |
| 圣光128CR CRb | 40.0 | 0 B | 0 B |
| 圣光128CR PbBa8.1+CRb | 40.0 | 0 B | 0 B |
Fig. 5 The comparison of resistance to clubroot between the improved hybrids and the check hybrid Shengguang128图5 改良杂交种与对照杂交种圣光128根肿病抗性比较 |
Table 6 Main agronomic traits of the improved lines of 616A表6 616A改良株系主要农艺性状考察 |
| 材料 Material | 角果长/cm SL/cm | 每角果粒数 NSS | 千粒重/g TSW/g | 株高/cm PH/cm | 分枝数 BN |
|---|---|---|---|---|---|
| 616A | 5.3±0.3 a | 20.0±1.2 a | 3.5±0.75 a | 135.1±3.5 a | 6.1±1.0 a |
| 616A PbBa8.1CRb | 5.4±0.6 a | 21.2±2.6 a | 3.3±0.4 a | 131.2±5.1 a | 6.1±0.9 a |
| 616A CRb | 5.5±0.5 a | 20.8±1.2 a | 3.3±0.4 a | 136.2±6.1 a | 6.0±1.0 a |
| 616A PbBa8.1 | 5.3±0.4 a | 20.5±2.0 a | 3.4±0.5 a | 132.7±8.0 a | 6.0±1.2 a |
Table 7 The evaluation of main agronomic traits of improved hybrids表7 改良杂交种主要农艺性状评价 |
| 材料 Material | 角果长/cm SL/cm | 每角果粒数 NSS | 千粒重/g TSW/g | 株高/cm PH/cm | 分枝数 BN |
|---|---|---|---|---|---|
| 圣光128 | 5.6±0.2 a | 21.5±2.0 a | 2.8±0.5 a | 141.4±5.2 a | 6.2±1.1 a |
| 圣光128CR PbBa8.1CRb | 5.7±0.4 a | 22.1±1.9 a | 2.7±0.4 a | 138.9±7.9 a | 6.1±1.0 a |
| 圣光128CR CRb | 5.5±0.3 a | 20.8±2.2 a | 2.7±0.3 a | 142.7±5.4 a | 6.3±0.8 a |
| 圣光128CR PbBa8.1 | 5.6±0.3 a | 21.1±1.1 a | 2.7±0.4 a | 143.9±6.3 a | 6.2±1.5 a |
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Fig. 1 Genetic improvement route of pol TCMS line 616A for resistance against clubroot disease
Table 1 Primer sequences of markers used in this studyTable 2 Methods for trait evaluationFig. 2 Genotype identification of the target genes in BC4F2 populationTable 3 Foreground selection in the populations of DCF1 and BC1F1-BC4F2Fig. 3 The fertility performance of the clubroot resistance improved pol TCMSline 616A PbBa8.1 + CRb and its restore line 616RTable 4 Disease index of improved lines and their parent 616AFig. 4 Comparison of clubroot resistance between the improved lines and their parent 616ATable 5 Disease index of the check hybrid shengguang128 and the improved hybridsFig. 5 The comparison of resistance to clubroot between the improved hybrids and the check hybrid Shengguang128Table 6 Main agronomic traits of the improved lines of 616ATable 7 The evaluation of main agronomic traits of improved hybrids/
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