中国水稻科学 ›› 2019, Vol. 33 ›› Issue (5): 421-428.DOI: 10.16819/j.1001-7216.2019.9007
朱永生1,2, 白建林2, 谢鸿光1, 吴方喜1, 罗曦1, 姜身飞1, 何炜1, 陈丽萍1, 蔡秋华1, 谢华安1,2, 张建福1,2,*()
收稿日期:
2019-01-10
修回日期:
2019-03-18
出版日期:
2019-09-10
发布日期:
2019-09-10
通讯作者:
张建福
基金资助:
Yongsheng ZHU1,2, Jianlin BAI2, Hongguang XIE1, Fangxi WU1, Xi LUO1, Shenfei JIANG1, Wei HE1, Liping CHEN1, Qiuhua CAI1, Huaan XIE1,2, Jianfu ZHANG1,2,*()
Received:
2019-01-10
Revised:
2019-03-18
Online:
2019-09-10
Published:
2019-09-10
Contact:
Jianfu ZHANG
摘要:
【目的】 为了创制兼抗白背飞虱和褐飞虱的水稻恢复系,【方法】 分别以抗褐飞虱材料B5(携带褐飞虱抗性基因Bph14和Bph15)及携带白背飞虱抗性位点qsI-4的籼型恢复系福恢7011为供体亲本,以骨干恢复系福恢676为轮回亲本,应用低世代分离群体田间表型结合单株鉴定与高世代稳定株系室内筛选和分子标记辅助选择相结合的方法,并对抗虫株系及其测交后代进行考查和农艺性状分析。【结果】 选育出聚合Bph14、Bph15 和qsI-4的恢复系材料3份,携带2个抗虫基因的恢复系材料3份。其中6份恢复系的褐飞虱抗性鉴定结果均表现中抗以上。通过抗性鉴定和杂交后代农艺性状分析筛选出具有生产应用潜力的恢复系材料2份。【结论】 为褐飞虱和白背飞虱抗性聚合新种质的创制和应用提供了基础材料。
中图分类号:
朱永生, 白建林, 谢鸿光, 吴方喜, 罗曦, 姜身飞, 何炜, 陈丽萍, 蔡秋华, 谢华安, 张建福. 聚合白背飞虱和褐飞虱抗性基因创制杂交水稻恢复系[J]. 中国水稻科学, 2019, 33(5): 421-428.
Yongsheng ZHU, Jianlin BAI, Hongguang XIE, Fangxi WU, Xi LUO, Shenfei JIANG, Wei HE, Liping CHEN, Qiuhua CAI, Huaan XIE, Jianfu ZHANG. Breeding Restore Lines of Hybrid Rice by Pyramiding Genes for Resistance to White Backed Planthoppers and Brown Planthoppers[J]. Chinese Journal OF Rice Science, 2019, 33(5): 421-428.
基因 Gene | 标记性质 Characteristics | 染色体 Chromosome | 标记 Marker | 类型 Type | 标记引物序列 Primer sequence(5′-3′) | 解链温度 Melting temperature /℃ | 产物大小 Production size/bp |
---|---|---|---|---|---|---|---|
qSI-4 | 共显性 Codominant | 4 | RM8213 | SSR | F: AGCCCAGTGATACAAAGATG R: GCGAGGAGATACCAAGAAAG | 55 | 208 |
Bph14 | 共显性 Codominant | 3 | RI35 | InDel | F: CAATCATCAAGCACGCGTTA R: ATCGAAGCCACTTGGTGAAC | 58 | 325 |
Bph15 | 共显性 Codominant | 4 | Y15 | InDel | F: AGGAACAGTGACACGTAGCA R: GGAGAGTTCAGTTTGCCATCC | 55 | 460 |
表1 水稻褐飞虱与白背飞虱抗性基因连锁标记及引物序列
Table 1 Linkage markers of the white-backed and brown planthoppers resistance genes and primer sequences.
基因 Gene | 标记性质 Characteristics | 染色体 Chromosome | 标记 Marker | 类型 Type | 标记引物序列 Primer sequence(5′-3′) | 解链温度 Melting temperature /℃ | 产物大小 Production size/bp |
---|---|---|---|---|---|---|---|
qSI-4 | 共显性 Codominant | 4 | RM8213 | SSR | F: AGCCCAGTGATACAAAGATG R: GCGAGGAGATACCAAGAAAG | 55 | 208 |
Bph14 | 共显性 Codominant | 3 | RI35 | InDel | F: CAATCATCAAGCACGCGTTA R: ATCGAAGCCACTTGGTGAAC | 58 | 325 |
Bph15 | 共显性 Codominant | 4 | Y15 | InDel | F: AGGAACAGTGACACGTAGCA R: GGAGAGTTCAGTTTGCCATCC | 55 | 460 |
图1 抗稻飞虱水稻材料创制技术路线 MAS表示分子标记辅助选择; IIR表示抗虫鉴定
Fig. 1. Processes of rice planthopper resistance lines. MAS, Marker-assisted selection; IIR, Identification of insect resistance.
图2 对群体中抗虫单株进行抗虫基因Bph15的分子标记检测 M为DNA标记,1为阳性对照B5,其他为分离群体中的单株。
Fig. 2. Molecular marker analysis of insect-resistant gene Bph15 for insect-resistant individuals in segregating populations. M, DNA marker; 1, B5; 2-49 are individual plants of segregating population.
株系 Line | 抗虫基因 Insect-resistant gene | 抗性均值 Average resistance | 抗性水平 Resistance level | ||
---|---|---|---|---|---|
qsI-4 | Bph14 | Bph15 | |||
FJ717 | + | + | + | 2.93 | R |
FJ718 | + | + | + | 3.81 | R |
FJ719 | - | + | + | 3.08 | R |
FJ723 | + | - | + | 5.43 | MR |
FJ742 | + | + | - | 5.57 | MR |
FJ745 | + | + | + | 3.84 | R |
FJ747 | - | - | + | 6.67 | S |
B5 | - | + | + | 3.07 | R |
FH7011 | + | - | - | 6.33 | S |
FH676 | - | - | - | 8.38 | S |
TN1 | - | - | - | 7.87 | S |
表2 各个株系携带的抗虫基因及抗性水平鉴定
Table 2 Identification of insect-resistant genes and resistance levels of each line.
株系 Line | 抗虫基因 Insect-resistant gene | 抗性均值 Average resistance | 抗性水平 Resistance level | ||
---|---|---|---|---|---|
qsI-4 | Bph14 | Bph15 | |||
FJ717 | + | + | + | 2.93 | R |
FJ718 | + | + | + | 3.81 | R |
FJ719 | - | + | + | 3.08 | R |
FJ723 | + | - | + | 5.43 | MR |
FJ742 | + | + | - | 5.57 | MR |
FJ745 | + | + | + | 3.84 | R |
FJ747 | - | - | + | 6.67 | S |
B5 | - | + | + | 3.07 | R |
FH7011 | + | - | - | 6.33 | S |
FH676 | - | - | - | 8.38 | S |
TN1 | - | - | - | 7.87 | S |
图4 经田间筛选的稳定株系在室内通过人工接虫进行抗性鉴定 左图为接虫前的幼苗状态,右图为接虫7 d后的情况。
Fig. 4. Artificial resistance identification in the laboratory of the stable lines screened in the field. The picture on the left shows the seedling before identification and the picture on the right shows the seedling at 7 days after insect inoculation.
品系 Line | 株高 Plant height/cm | 单株穗数 Panicle number per plant | 每穗粒数 Grain number per panicle | 每穗实粒数 Filled grain number per panicle | 结实率 Seed-setting rate/% | 千粒重 1000-grain weight/g |
---|---|---|---|---|---|---|
FH676(CK) | 131.9±4.4 | 7.6±0.9 | 175.0±11.6 | 154.0±9.6 | 88.0±1.6 | 31.2±0.3 |
FJ717 | 127.4±1.2* | 8.3±1.2 | 173.6±13.6 | 159.0±12.8 | 91.6±1.9* | 30.7±0.3 |
FJ718 | 125.9±5.3** | 9.2±2.1** | 168.9±15.9* | 154.3±13.3 | 91.3±2.5* | 29.8±0.2 |
FJ719 | 133.9±6. 7 | 6.8±0.6* | 187.1±9.5* | 159.5±8.6 | 85.2±2.7* | 32.0±0.1 |
FJ745 | 126.9±5.2** | 7.4±1.3 | 169.5±12.3 | 151.4±10.2 | 89.3±0.4 | 30.1±0.4 |
表3 入选抗虫恢复系与轮回亲本的农艺性状比较分析(福建沙县,2016)
Table 3 Comparative analysis of agronomic traits between selected insect-resistant restorer lines and recurrent parents (Shaxian County, Fujian, 2016).
品系 Line | 株高 Plant height/cm | 单株穗数 Panicle number per plant | 每穗粒数 Grain number per panicle | 每穗实粒数 Filled grain number per panicle | 结实率 Seed-setting rate/% | 千粒重 1000-grain weight/g |
---|---|---|---|---|---|---|
FH676(CK) | 131.9±4.4 | 7.6±0.9 | 175.0±11.6 | 154.0±9.6 | 88.0±1.6 | 31.2±0.3 |
FJ717 | 127.4±1.2* | 8.3±1.2 | 173.6±13.6 | 159.0±12.8 | 91.6±1.9* | 30.7±0.3 |
FJ718 | 125.9±5.3** | 9.2±2.1** | 168.9±15.9* | 154.3±13.3 | 91.3±2.5* | 29.8±0.2 |
FJ719 | 133.9±6. 7 | 6.8±0.6* | 187.1±9.5* | 159.5±8.6 | 85.2±2.7* | 32.0±0.1 |
FJ745 | 126.9±5.2** | 7.4±1.3 | 169.5±12.3 | 151.4±10.2 | 89.3±0.4 | 30.1±0.4 |
杂交组合 Hybrid rice combination | 单株穗数 Panicle number per plant | 每穗粒数 Grain number per panicle | 每穗实粒数 Filled grain number per panicle | 结实率 Seed-setting rate/% | 千粒重 1000-grain weight/g | 理论产量 Theoretical yield /(kg∙hm-2) |
---|---|---|---|---|---|---|
II-32A/FH676(CK) | 9.7±0.9 | 190.5±13.6 | 177.6±9.6 | 93.2±1.4 | 28.2±0.9 | 9160.5±169.2 |
II-32A/FJ717 | 9.9±1.2 | 193.6±21.2 | 181.0±12.8 | 93.5±1.5 | 28.7±0.2 | 9749.3±210.2** |
II-32A/FJ718 | 10.1±2.1 | 188.3±5.6 | 175.7±13.3 | 93.3±2.0 | 28.1±1.1 | 9426.1±107.1* |
II-32A/FJ719 | 7.8±0.6 | 213.5±9.8 | 185.3±8.6 | 86.8±1.7 | 29.0±0.2 | 8878.3±192.5 |
II-32A/FJ745 | 10.4±1.3 | 194.5±18.4 | 173.8±10.2 | 89.3±1.3 | 27.2±0.4 | 9308.9±98.4 |
Y58S//FH676(CK) | 9.1±0.8 | 205.1±11.6 | 176.4±9.6 | 86.0±0.9 | 26.2±0.3 | 7950.9±214.5 |
Y58S//FJ717 | 8.3±0.2 | 210.3±14.6 | 185.5±12.8 | 88.2±0.9 | 26.7±1.4 | 7732.2±110.2 |
Y58S//FJ718 | 10.3±1.4 | 196.7±5.6 | 171.5±13.3 | 87.2±3.2 | 26.6±0.1 | 8886.5±183.1** |
Y58S//FJ719 | 7.8±0.8 | 223.5±109.9 | 184.8±8.6 | 82.7±1.7 | 28.0±0.9 | 7571.3±199.2 |
Y58S//FJ745 | 9.8±0.3 | 197.4±13.2 | 174.2±10.2 | 88.3±1.4 | 26.1±0.3 | 8397.5±210.1* |
表4 入选抗虫恢复系与轮回亲本的杂交后代农艺性状比较分析(福建沙县,2018)
Table 4 Comparative analysis of agronomic traits of hybrid progenies between selected insect-resistant restorer lines and recurrent parents(Shaxian County, Fujian, China, 2018).
杂交组合 Hybrid rice combination | 单株穗数 Panicle number per plant | 每穗粒数 Grain number per panicle | 每穗实粒数 Filled grain number per panicle | 结实率 Seed-setting rate/% | 千粒重 1000-grain weight/g | 理论产量 Theoretical yield /(kg∙hm-2) |
---|---|---|---|---|---|---|
II-32A/FH676(CK) | 9.7±0.9 | 190.5±13.6 | 177.6±9.6 | 93.2±1.4 | 28.2±0.9 | 9160.5±169.2 |
II-32A/FJ717 | 9.9±1.2 | 193.6±21.2 | 181.0±12.8 | 93.5±1.5 | 28.7±0.2 | 9749.3±210.2** |
II-32A/FJ718 | 10.1±2.1 | 188.3±5.6 | 175.7±13.3 | 93.3±2.0 | 28.1±1.1 | 9426.1±107.1* |
II-32A/FJ719 | 7.8±0.6 | 213.5±9.8 | 185.3±8.6 | 86.8±1.7 | 29.0±0.2 | 8878.3±192.5 |
II-32A/FJ745 | 10.4±1.3 | 194.5±18.4 | 173.8±10.2 | 89.3±1.3 | 27.2±0.4 | 9308.9±98.4 |
Y58S//FH676(CK) | 9.1±0.8 | 205.1±11.6 | 176.4±9.6 | 86.0±0.9 | 26.2±0.3 | 7950.9±214.5 |
Y58S//FJ717 | 8.3±0.2 | 210.3±14.6 | 185.5±12.8 | 88.2±0.9 | 26.7±1.4 | 7732.2±110.2 |
Y58S//FJ718 | 10.3±1.4 | 196.7±5.6 | 171.5±13.3 | 87.2±3.2 | 26.6±0.1 | 8886.5±183.1** |
Y58S//FJ719 | 7.8±0.8 | 223.5±109.9 | 184.8±8.6 | 82.7±1.7 | 28.0±0.9 | 7571.3±199.2 |
Y58S//FJ745 | 9.8±0.3 | 197.4±13.2 | 174.2±10.2 | 88.3±1.4 | 26.1±0.3 | 8397.5±210.1* |
杂交组合 Hybrid rice combination | 单株穗数 Panicle number per plant | 每穗粒数 Grain number per panicle | 每穗实粒数 Filled grain per panicle | 结实率 Seed-setting rate/% | 千粒重 1000-grain weight/g | 理论产量 Theoretical yield /(kg∙hm-2) |
---|---|---|---|---|---|---|
丰两优4号(CK) Fengliangyou 4(CK) | 8.9±0.7 | 191.4±8.3 | 159.5±6.4 | 83.4±2.2 | 28.5±0.1 | 7619.2±203.5 |
Ⅱ优明86 ⅡYouming 86 | 9.3±0.7 | 190.7±4.4 | 174.0±3.3 | 91.2±2.8 | 28.3±0.1 | 8683.0±219.3 |
泰丰A/FJ717 Taifeng A/FJ717 | 10.3±0.5 | 186.6±11.3 | 161.4±8.6 | 86.5±1.7 | 25.3±0.1 | 7965.3±184.8* |
泰丰A/FJ718 Taifeng A/FJ718 | 9.8±0.4 | 179.5±10.3 | 157.2±9.0 | 87.6±3.4 | 25.0±0.1 | 7241.6±114.7 |
泰丰A/FJ745 Taifeng A/FJ745 | 10.3±0.4 | 182.8±13.4 | 157.9±12.0 | 86.4±1.8 | 25.3±0.1 | 7795.3±217.6 |
泸香618A/FJ717 Luxiang 618A/FJ717 | 9.7±0.3 | 199.8±9.1 | 178.4±7.3 | 89.3±1.2 | 25.4±0.2 | 8279.0±117.9** |
泸香618A/FJ718 Luxiang 618A/FJ718 | 11.1±0.7 | 176.4±9.8 | 154.1±6.3 | 87.3±0.8 | 25.5±0.2 | 8214.2±254.7** |
泸香618A/FJ745 Luxiang 618A/FJ745 | 9.4±1.0 | 193.0±12.7 | 171.8±10.7 | 89.0±1.1 | 25.8±0.4 | 7857.7±247.7 |
表5 入选抗虫恢复系与轮回亲本的杂交后代农艺性状比较分析(海南三亚,2018)
Table 5 Comparative analysis of agronomic traits of hybrid progenies between selected insect-resistant restorer lines and recurrent parents(Sanya, Hainan, China, 2018).
杂交组合 Hybrid rice combination | 单株穗数 Panicle number per plant | 每穗粒数 Grain number per panicle | 每穗实粒数 Filled grain per panicle | 结实率 Seed-setting rate/% | 千粒重 1000-grain weight/g | 理论产量 Theoretical yield /(kg∙hm-2) |
---|---|---|---|---|---|---|
丰两优4号(CK) Fengliangyou 4(CK) | 8.9±0.7 | 191.4±8.3 | 159.5±6.4 | 83.4±2.2 | 28.5±0.1 | 7619.2±203.5 |
Ⅱ优明86 ⅡYouming 86 | 9.3±0.7 | 190.7±4.4 | 174.0±3.3 | 91.2±2.8 | 28.3±0.1 | 8683.0±219.3 |
泰丰A/FJ717 Taifeng A/FJ717 | 10.3±0.5 | 186.6±11.3 | 161.4±8.6 | 86.5±1.7 | 25.3±0.1 | 7965.3±184.8* |
泰丰A/FJ718 Taifeng A/FJ718 | 9.8±0.4 | 179.5±10.3 | 157.2±9.0 | 87.6±3.4 | 25.0±0.1 | 7241.6±114.7 |
泰丰A/FJ745 Taifeng A/FJ745 | 10.3±0.4 | 182.8±13.4 | 157.9±12.0 | 86.4±1.8 | 25.3±0.1 | 7795.3±217.6 |
泸香618A/FJ717 Luxiang 618A/FJ717 | 9.7±0.3 | 199.8±9.1 | 178.4±7.3 | 89.3±1.2 | 25.4±0.2 | 8279.0±117.9** |
泸香618A/FJ718 Luxiang 618A/FJ718 | 11.1±0.7 | 176.4±9.8 | 154.1±6.3 | 87.3±0.8 | 25.5±0.2 | 8214.2±254.7** |
泸香618A/FJ745 Luxiang 618A/FJ745 | 9.4±1.0 | 193.0±12.7 | 171.8±10.7 | 89.0±1.1 | 25.8±0.4 | 7857.7±247.7 |
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