中国水稻科学 ›› 2017, Vol. 31 ›› Issue (2): 149-156.DOI: 10.16819/j.1001-7216.2017.6137
韩保林, 张洪凯, 顾朝剑, 廖泳祥, 彭永彬, 张红宇, 徐培洲, 陈晓琼, 吴先军*()
出版日期:
2017-03-20
发布日期:
2017-03-10
通讯作者:
吴先军
基金资助:
Baolin HAN, Yu TAO, Hongkai ZHANG, Chaojian GU, Yongxiang LIAO, Yongbin PENG, Hongyu ZHANG, Peizhou XU, Xiaoqiong CHEN, Xianjun WU*()
Online:
2017-03-20
Published:
2017-03-10
Contact:
Xianjun WU
摘要:
目的 叶片是水稻理想株型的重要内容,叶片适度卷曲可以提高光合效率。对卷叶相关基因进行遗传分析和初步定位,为下一步的基因克隆与功能分析提供研究基础。方法 利用EMS诱变雄性不育保持系宜香1B获得一份稳定遗传的叶片向内卷曲突变体,暂命名为rl(t)。在成熟期,测定野生型和rl(t) 的主要农艺性状;在分蘖期,取野生型和rl(t) 叶片用FAA固定液固定进行石蜡切片,同时,用野生型和rl(t)剑叶测定叶绿素含量;在抽穗期,利用Li-6400便携式光合仪测定10株抽穗期的野生型和rl(t)的光合参数;将rl(t)与野生型及日本晴杂交,观察 F1 植株表型,对F2表型分离进行χ2 测验,对突变体进行遗传分析。以 rl(t)/日本晴的 F2 群体为材料,利用 BSA 法进行定位。结果 与野生型相比,突变体叶片向内卷曲明显,叶片更加直立,叶色变深,其他主要农艺性状均有不同程度降低。光合特性分析表明,突变体比野生型具有更高的光合色素含量,但光合效率没有明显差异。叶片组织切片观察表明,突变体中泡状细胞变小可能是导致叶片卷曲的主要原因。遗传分析表明,该突变体受一对隐性核基因控制,利用突变体与日本晴的F2群体进行基因定位,最终将该基因定位在第7染色体长臂InDel标记Ind3和Ind4间610 kb的物理区间。结论 rl(t)叶片内卷是由于近轴面泡状细胞面积减小。RL(t)定位区间内未见卷叶相关基因报道,推测RL(t)可能是一对新基因。
韩保林, 张洪凯, 顾朝剑, 廖泳祥, 彭永彬, 张红宇, 徐培洲, 陈晓琼, 吴先军. 水稻叶片内卷突变体rl(t)的鉴定与基因定位[J]. 中国水稻科学, 2017, 31(2): 149-156.
Baolin HAN, Yu TAO, Hongkai ZHANG, Chaojian GU, Yongxiang LIAO, Yongbin PENG, Hongyu ZHANG, Peizhou XU, Xiaoqiong CHEN, Xianjun WU. Identification and Gene Mapping of a Rolled Leaf Mutant rl(t) in Rice[J]. Chinese Journal OF Rice Science, 2017, 31(2): 149-156.
标记 Marker | 正向引物(5'-3') Forward primer(5'-3') | 反向引物(5'-3') Reverse primer(5'-3') | |
RM3394 | CCCTTACGTGCAGTACATTG | ATGCAGGCTACTTACTAGCG | |
Ind1 | CCAGTCAACAGGAGCATA | TTAATTATTAGCCTGCTCGA | |
Ind2 | TATAGATGAGCACGCAATCA | GTATGATCGGAAACGTCACT | |
Ind3 | TTGCAGGAGTACTGAAATGA | GCCAGTATTACCAGAGGATG | |
Ind4 | CAAGGTCATCAAGTTCGC | CTGACGGAGCCAGGATCT | |
Ind5 | TGGAAAAGAACTTCAATGCT | TTGAATCACCACAATTTAGC | |
Ind6 | GTCAACCGTCAAGAGATCAT | TGCTTTAGCAGAGTTGCATA | |
RM5752 | TTGCAATTAATTCGATCTCC | GCAGATCGATTCGTTAGTTC | |
ACL1 | CTGAAGCTGAACCTCTCGCTG | GGAGCATGACGTAGATGAAGCAG | |
RL14 | CTCTTTCAGGCATTCCATTGATG | CAACACCTTGTCAGCTTTCAAGC | |
ROC5 | CGCAAGAGGAAGAAGCGATAC | GCTCCAGTTGCGTCTTCATC | |
SRL1 | TGCATCTTTTGACCCAGCTAC | ACAACAAGGGTGCCCAGATAA | |
SLL1 | CAGGTGTCCAACCATGAGC | GCCTCTGTGATTGCCATCTAAT | |
CFL1 | TCGAGCTCAACTCCGAGGTC | ACGGACGACACGGAGGTGTA | |
NRL1 | TCAGTAGTGTAGTGGTGTCGAGTTCA | GCACTCCTTCATGTGAGCTTCA | |
NAL7 | CAAGAACATCACCGGCAAGA | CGATTTGATCAAGGACCATGCT | |
OSAGO7 | CCGCATCCCCTTGATGATT | GGCCAATTCATGCTTGCAA | |
SRL2 | TCCATCTGCGCAGCATTTCA | CTACTGGGCACGATATGCAG |
表1 基因定位分子标记引物及qRT-PCR引物序列
Table 1 The sequences of molecular markers used for gene mapping and qRT-PCR markers.
标记 Marker | 正向引物(5'-3') Forward primer(5'-3') | 反向引物(5'-3') Reverse primer(5'-3') | |
RM3394 | CCCTTACGTGCAGTACATTG | ATGCAGGCTACTTACTAGCG | |
Ind1 | CCAGTCAACAGGAGCATA | TTAATTATTAGCCTGCTCGA | |
Ind2 | TATAGATGAGCACGCAATCA | GTATGATCGGAAACGTCACT | |
Ind3 | TTGCAGGAGTACTGAAATGA | GCCAGTATTACCAGAGGATG | |
Ind4 | CAAGGTCATCAAGTTCGC | CTGACGGAGCCAGGATCT | |
Ind5 | TGGAAAAGAACTTCAATGCT | TTGAATCACCACAATTTAGC | |
Ind6 | GTCAACCGTCAAGAGATCAT | TGCTTTAGCAGAGTTGCATA | |
RM5752 | TTGCAATTAATTCGATCTCC | GCAGATCGATTCGTTAGTTC | |
ACL1 | CTGAAGCTGAACCTCTCGCTG | GGAGCATGACGTAGATGAAGCAG | |
RL14 | CTCTTTCAGGCATTCCATTGATG | CAACACCTTGTCAGCTTTCAAGC | |
ROC5 | CGCAAGAGGAAGAAGCGATAC | GCTCCAGTTGCGTCTTCATC | |
SRL1 | TGCATCTTTTGACCCAGCTAC | ACAACAAGGGTGCCCAGATAA | |
SLL1 | CAGGTGTCCAACCATGAGC | GCCTCTGTGATTGCCATCTAAT | |
CFL1 | TCGAGCTCAACTCCGAGGTC | ACGGACGACACGGAGGTGTA | |
NRL1 | TCAGTAGTGTAGTGGTGTCGAGTTCA | GCACTCCTTCATGTGAGCTTCA | |
NAL7 | CAAGAACATCACCGGCAAGA | CGATTTGATCAAGGACCATGCT | |
OSAGO7 | CCGCATCCCCTTGATGATT | GGCCAATTCATGCTTGCAA | |
SRL2 | TCCATCTGCGCAGCATTTCA | CTACTGGGCACGATATGCAG |
性状 Trait | 宜香1B Yixiang 1B | rl(t) | ||
抽穗期 Days to heading / d | 87.3±1.3 | 81.1 | ±1.4** | |
株高 Plant height / cm | 112.3±4.1 | 103.5 | ±2.8** | |
分蘖数 No. of tillers | 14.6±1.1 | 6.0 | ±0.8** | |
穗长 Panicle length / cm | 29.1±0.9 | 26.9 | ±0.9** | |
每穗实粒数 No. of filled grains per panicle | 196.7±5.2 | 134.3 | ±7.0** | |
每穗粒数 No. of grains per panicle | 212.4±7.2 | 173.6 | ±9.5** | |
结实率 Seed-setting rate / % | 93.0±1.0 | 77.0 | ±22.0** | |
千粒重 1000-grain weight / g | 30.1±0.2 | 21.9 | ±0.1** | |
粒长 Grain length / mm | 9.9±0.0 | 9.2 | ±0.0** | |
粒宽 Grain width / mm | 2.7±0.2 | 2.7 | ±0.0 | |
长宽比 Grain length-width ratio | 3.7±0.3 | 3.4 | ±0.0* | |
数据表示为平均数±标准差;样本数量n=10;**, *分别表示差异达0.01和0.05显著水平(t检验)。 Values are listed as means ± SD (n=10). **, * Significantly different at 0.01 and 0.05 levels, respectively(t test). |
表1 宜香1B与rl(t)的农艺性状比较
Table 2 Comparison of agronomic traits between rl(t) and its wild type.
性状 Trait | 宜香1B Yixiang 1B | rl(t) | ||
抽穗期 Days to heading / d | 87.3±1.3 | 81.1 | ±1.4** | |
株高 Plant height / cm | 112.3±4.1 | 103.5 | ±2.8** | |
分蘖数 No. of tillers | 14.6±1.1 | 6.0 | ±0.8** | |
穗长 Panicle length / cm | 29.1±0.9 | 26.9 | ±0.9** | |
每穗实粒数 No. of filled grains per panicle | 196.7±5.2 | 134.3 | ±7.0** | |
每穗粒数 No. of grains per panicle | 212.4±7.2 | 173.6 | ±9.5** | |
结实率 Seed-setting rate / % | 93.0±1.0 | 77.0 | ±22.0** | |
千粒重 1000-grain weight / g | 30.1±0.2 | 21.9 | ±0.1** | |
粒长 Grain length / mm | 9.9±0.0 | 9.2 | ±0.0** | |
粒宽 Grain width / mm | 2.7±0.2 | 2.7 | ±0.0 | |
长宽比 Grain length-width ratio | 3.7±0.3 | 3.4 | ±0.0* | |
数据表示为平均数±标准差;样本数量n=10;**, *分别表示差异达0.01和0.05显著水平(t检验)。 Values are listed as means ± SD (n=10). **, * Significantly different at 0.01 and 0.05 levels, respectively(t test). |
材料 Material | 净光合速率Pn /(µmol·m-2 s-1) | 气孔导度Gs /(µmol·m-2 s-1) | 胞间二氧化碳浓度Ci /(µmol·m-2 s-1) | 蒸腾速率Tr /(µmol·m-2 s-1) |
宜香1B Yixiang 1B | 18.62±0.14 | 0.51±0.00 | 295.92±5.68 | 4.59±0.06 |
rl(t) | 18.57±0.25 | 0.50±0.00 | 297.53±1.52 | 4.57±0.04 |
数据表示为平均数±标准差;样本数量n=10; **, *分别表示差异达0.01和0.05显著水平(t检验)。 Values are means ± SD (n=10). **, * Significantly different at 0.01 and 0.05 levels, respectively(t test). |
表3 野生型宜香1B与突变体rl(t)光合特性比较
Table 3 Comparison of the photosynthetic characters between Yixiang 1B and rl(t).
材料 Material | 净光合速率Pn /(µmol·m-2 s-1) | 气孔导度Gs /(µmol·m-2 s-1) | 胞间二氧化碳浓度Ci /(µmol·m-2 s-1) | 蒸腾速率Tr /(µmol·m-2 s-1) |
宜香1B Yixiang 1B | 18.62±0.14 | 0.51±0.00 | 295.92±5.68 | 4.59±0.06 |
rl(t) | 18.57±0.25 | 0.50±0.00 | 297.53±1.52 | 4.57±0.04 |
数据表示为平均数±标准差;样本数量n=10; **, *分别表示差异达0.01和0.05显著水平(t检验)。 Values are means ± SD (n=10). **, * Significantly different at 0.01 and 0.05 levels, respectively(t test). |
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