Chinese Journal OF Rice Science ›› 2016, Vol. 30 ›› Issue (1): 44-52.DOI: 10.16819/j.1001-7216.2016.5126
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Tuan LONG1,2, Bao-guang AN1,2, Xin-peng LI1,2, Wei ZHANG1,2, Jing-lin LI1,2, Yao-hua YANG1,2, Xiang ZENG1,2, Yong-zhong WU1,2, Pei-jin HUANG1,2,*()
Received:
2015-08-18
Revised:
2015-10-27
Online:
2016-01-10
Published:
2016-01-10
Contact:
Pei-jin HUANG
龙湍1,2, 安保光1,2, 李新鹏1,2, 张维1,2, 李京琳1,2, 杨瑶华1,2, 曾翔1,2, 吴永忠1,2, 黄培劲1,2,*()
通讯作者:
黄培劲
基金资助:
CLC Number:
Tuan LONG, Bao-guang AN, Xin-peng LI, Wei ZHANG, Jing-lin LI, Yao-hua YANG, Xiang ZENG, Yong-zhong WU, Pei-jin HUANG. Construction and Screening of an Irradiation-induced Mutant Library of indica Rice 93-11[J]. Chinese Journal OF Rice Science, 2016, 30(1): 44-52.
龙湍, 安保光, 李新鹏, 张维, 李京琳, 杨瑶华, 曾翔, 吴永忠, 黄培劲. 籼稻93-11辐射诱变突变体库的创建及其筛选[J]. 中国水稻科学, 2016, 30(1): 44-52.
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URL: http://www.ricesci.cn/EN/10.16819/j.1001-7216.2016.5126
引物 Primer | 序列(5'-3') Sequence (5'-3') | 目的基因 Target locus |
---|---|---|
704B2-1F | CAAAGATTGTCTCAAGGTTGGTAG | CYP704B2 |
704B2-1R | GGTATTAGGCAAGGAATTCAGTTG | |
704B2-2F | TCGAAGGACAGGACGGTGAC | CYP704B2 |
704B2-2R | TTTGAGCAAGAGAGGAAGGATC | |
704B2-3F | GCAAGAACTAACCAAAATTCAGG | CYP704B2 |
704B2-3R | GGTCAGACGGAGGTGGAGA | |
81A6-F1 | CAAACTTCCAACTTTCCCGT | CYP81A6 |
81A6-R1 | CTGACGCCTGAAACGCAG | |
81A6-F2 | GCAACGGAGTGAGTAGAAGTAATC | CYP81A6 |
81A6-R2 | CAGAAAGGTAAAACAGCAGAAGA | |
81A6-F3 | GGGCGAGAAGAAGAGCATG | CYP81A6 |
81A6-R3 | GGCACGACCTTGGGGAT | |
81A6-F4 | GGTGCGACGGCAATCTC | CYP81A6 |
81A6-R4 | GAGCAGAAAGCCCTTCCTC | |
1907_F | AGGTCGGGTTTGGGGTT | 1907株系中CYP704B2的突变位点 |
1907_R | GATGTTGGCAGCGTCGAA | CYP704B2 in the line 1907 |
2245_F | AGCTTCGGGGACGACAAGA | 2245株系中CYP704B2的突变位点 |
2245_R | TGCGTCATCGCCATGTACGT | CYP704B2 in the line 2245 |
3522-F | ATGAAATCTTAGTTCCACCCTCTTGCCG | 3522株系中CYP81A6的突变位点 |
3522-R | CTCCCTGACGATGCACTGGAGGT | CYP81A6 in the line 3522 |
3622-F | TGTTCGAGGTCTCCCTCAGCGTG | 3622株系中CYP81A6的突变位点 |
3622-R | CCTGCTTAAACTCCTGGGCTTCCA | CYP81A6 in the line 3622 |
Table 1 Primers used in this study.
引物 Primer | 序列(5'-3') Sequence (5'-3') | 目的基因 Target locus |
---|---|---|
704B2-1F | CAAAGATTGTCTCAAGGTTGGTAG | CYP704B2 |
704B2-1R | GGTATTAGGCAAGGAATTCAGTTG | |
704B2-2F | TCGAAGGACAGGACGGTGAC | CYP704B2 |
704B2-2R | TTTGAGCAAGAGAGGAAGGATC | |
704B2-3F | GCAAGAACTAACCAAAATTCAGG | CYP704B2 |
704B2-3R | GGTCAGACGGAGGTGGAGA | |
81A6-F1 | CAAACTTCCAACTTTCCCGT | CYP81A6 |
81A6-R1 | CTGACGCCTGAAACGCAG | |
81A6-F2 | GCAACGGAGTGAGTAGAAGTAATC | CYP81A6 |
81A6-R2 | CAGAAAGGTAAAACAGCAGAAGA | |
81A6-F3 | GGGCGAGAAGAAGAGCATG | CYP81A6 |
81A6-R3 | GGCACGACCTTGGGGAT | |
81A6-F4 | GGTGCGACGGCAATCTC | CYP81A6 |
81A6-R4 | GAGCAGAAAGCCCTTCCTC | |
1907_F | AGGTCGGGTTTGGGGTT | 1907株系中CYP704B2的突变位点 |
1907_R | GATGTTGGCAGCGTCGAA | CYP704B2 in the line 1907 |
2245_F | AGCTTCGGGGACGACAAGA | 2245株系中CYP704B2的突变位点 |
2245_R | TGCGTCATCGCCATGTACGT | CYP704B2 in the line 2245 |
3522-F | ATGAAATCTTAGTTCCACCCTCTTGCCG | 3522株系中CYP81A6的突变位点 |
3522-R | CTCCCTGACGATGCACTGGAGGT | CYP81A6 in the line 3522 |
3622-F | TGTTCGAGGTCTCCCTCAGCGTG | 3622株系中CYP81A6的突变位点 |
3622-R | CCTGCTTAAACTCCTGGGCTTCCA | CYP81A6 in the line 3622 |
Fig. 1. Different mutant phenotypes in M2 families. A, Monoculm; B, Dwarfism; C, Late heading; D, Malformed spikelet; E, Lesion mimic. Mutant plants or samples are on the left, and wild type plants or samples are on the right.
表型 Phenotype | 家系数 Number of lines | 频率a Frequency a /% |
---|---|---|
株高增加Increased plant height | 3 | 0.08 |
矮化 Dwarfism | 65 | 1.80 |
株型紧凑 Reduced tiller angle | 1 | 0.03 |
分蘖少 Reduced tiller number | 52 | 1.44 |
分蘖多 Increased tiller number | 2 | 0.06 |
穗型变异 Malformed panicle | 6 | 0.17 |
小穗变异 Malformed spikelet | 19 | 0.53 |
育性 Fertility | 99 | 2.74 |
叶色 Leaf color | 2 | 0.06 |
叶片变宽 Broad leaf | 2 | 0.06 |
假病斑 Lesion mimic | 12 | 0.33 |
早衰 Premature senescence | 1 | 0.03 |
早抽穗 Early heading | 12 | 0.33 |
迟抽穗 Late heading | 30 | 0.83 |
苯达松敏感 Bentazon-lethal | 2 | 0.15b |
总数 Total | 308 | 8.52 |
Table 2 Summery of mutant phenotypes in M2 and M3 families.
表型 Phenotype | 家系数 Number of lines | 频率a Frequency a /% |
---|---|---|
株高增加Increased plant height | 3 | 0.08 |
矮化 Dwarfism | 65 | 1.80 |
株型紧凑 Reduced tiller angle | 1 | 0.03 |
分蘖少 Reduced tiller number | 52 | 1.44 |
分蘖多 Increased tiller number | 2 | 0.06 |
穗型变异 Malformed panicle | 6 | 0.17 |
小穗变异 Malformed spikelet | 19 | 0.53 |
育性 Fertility | 99 | 2.74 |
叶色 Leaf color | 2 | 0.06 |
叶片变宽 Broad leaf | 2 | 0.06 |
假病斑 Lesion mimic | 12 | 0.33 |
早衰 Premature senescence | 1 | 0.03 |
早抽穗 Early heading | 12 | 0.33 |
迟抽穗 Late heading | 30 | 0.83 |
苯达松敏感 Bentazon-lethal | 2 | 0.15b |
总数 Total | 308 | 8.52 |
Fig. 2. Phenotypic and molecular characterization of two male-sterile mutants without pollen grains. A and B, I2-KI staining of pollen grains of a wild type plant (A) and a mutant (B) in the line 1907, bar = 100 μm; C and D, I2-KI staining of pollen grains of a wild type plant (C) and a mutant (D) in the line 2245, bar = 100 μm; E, The structure of CYP704B2. Black boxes represent the coding region; Lines represent introns; Triangles point to the mutation loci. “GGG” following the 794th nucleotide of CYP704B2 is substituted by “T” in mutants of the line 1907 (1907m), and “GG” following the 1267th nucleotide of CYP704B2 is deleted in mutants of the line 2245 (2245m); F and G, Genotyping of the CYP704B2 loci in the lines 1907 (F) and 2245 (G). The sizes (bp) of the PCR products are shown on the left. W, Wild type; H, Heterozygote; M, Mutant.
Fig. 3. Phenotypic and molecular characterization of two bentazon-lethal mutants. A and B, Phenotypes of the bentazon-lethal mutants (left) and wild type (right) plants in the lines 3522 (A) and 3622 (B) after bentazon spraying at tillering stage; C, The structure of CYP81A6. Black boxes represent the coding region; Lines represent introns; Triangles point to the mutation loci. A 23-bp deletion is found following the 1747th bp of the coding region of CYP81A6 in mutants of the line 3522 (3522m), and a 7-bp deletion is found following the 586th bp of the coding region of CYP81A6 in mutants of the line 3622 (3622m); D and E, Genotyping of the CYP81A6 loci in the lines 3522 (D) and 3622 (E). The sizes (bp) of the PCR products are shown on the left. W, Wild type; H, Heterozygote; M, Mutant.
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