中国水稻科学 ›› 2023, Vol. 37 ›› Issue (3): 225-232.DOI: 10.16819/j.1001-7216.2023.220506
• 研究报告 • 下一篇
沈雨民, 陈明亮, 熊焕金, 熊文涛, 吴小燕, 肖叶青*()
收稿日期:
2022-05-09
修回日期:
2022-12-02
出版日期:
2023-05-10
发布日期:
2023-05-16
通讯作者:
*email: xiaoyq1965@163.com
基金资助:
SHEN Yumin, CHEN Mingliang, XIONG Huanjin, XIONG Wentao, WU Xiaoyan, XIAO Yeqing*()
Received:
2022-05-09
Revised:
2022-12-02
Online:
2023-05-10
Published:
2023-05-16
Contact:
*email: xiaoyq1965@163.com
摘要:
【目的】水稻花器官的正常发育对水稻的成功繁殖和产量至关重要。对水稻花器官发育相关基因进行遗传定位,可以完善和丰富其花器官发育调控网络,对水稻育种具有指导意义。【方法】从育种材料中分离鉴定出一个内外稃发育异常的突变体blg1,对blg1突变体和对照株系花器官形态进行了观察。将该突变体与Dular杂交,构建F2群体,对突变表型进行遗传分析,并对BLG1基因进行了精细定位和候选基因分析。【结果】表型观察显示,blg1突变体表现为内外稃不能正常闭合、稃间弯曲严重、籽粒变小等。遗传分析表明,blg1的突变性状由一对隐性核基因控制。通过精细定位,将目标基因定位于第5染色体长臂末端InD3和InD5两个标记之间,物理距离为50 kb,包含8个开放阅读框。通过分析,LOC_Os05g48760可能是BLG1基因的候选基因。【结论】BLG1是一个控制水稻内外稃发育的新基因。
沈雨民, 陈明亮, 熊焕金, 熊文涛, 吴小燕, 肖叶青. 水稻内外稃异常发育突变体blg1 (beak like grain 1)的表型分析与精细定位[J]. 中国水稻科学, 2023, 37(3): 225-232.
SHEN Yumin, CHEN Mingliang, XIONG Huanjin, XIONG Wentao, WU Xiaoyan, XIAO Yeqing. Phenotypic Analysis and Fine Mapping of blg1(beak like grain 1), a Rice Mutant with Abnormal Palea and Lemma Development[J]. Chinese Journal OF Rice Science, 2023, 37(3): 225-232.
分子标记 Marker | 前引物 (5’-3’) Forward sequence (5’-3’) | 后引物 (5’-3’) Reverse sequence (5’-3’) |
---|---|---|
S1 | TAATTGCGGTCTCGTGCC | AACCACCTCAACTACTGCCG |
S2 | CGCCTGCAAAAAAGGTAGAG | GATCAAAGGAACCCCCGTAG |
S3 | GCCGGTAGATGCAGTATTGC | GTGCACAGTCACGCAGAAAG |
S4 | TGCTGTCTCCACTTCCCTTC | AAGGAGGCGTGCTTAACAGG |
S5 | CTTGTCTTGGCTTGGCTAGG | ATCCACGCAGATAGACCTGC |
InD1 | AAAACGATCATAGTTGCTTGC | GGAGTTCGGGCTCAAGTA |
InD2 | GGGGCTTATTCACTTTGT | TTCTTTCCTCCTGGCTCT |
InD3 | CCCTTTTATCCTCAGTAGT | TACAAGCAGTTGTTTGCA |
InD4 | GGCGACCGAGAAGTGTAG | CGAGATAGCGAAAGCGAC |
InD5 | CGGGTCAGTGTTATCGTC | CATCAATGAAGGGCAAAT |
表1 精细定位引物
Table 1. Molecular markers used for fine mapping.
分子标记 Marker | 前引物 (5’-3’) Forward sequence (5’-3’) | 后引物 (5’-3’) Reverse sequence (5’-3’) |
---|---|---|
S1 | TAATTGCGGTCTCGTGCC | AACCACCTCAACTACTGCCG |
S2 | CGCCTGCAAAAAAGGTAGAG | GATCAAAGGAACCCCCGTAG |
S3 | GCCGGTAGATGCAGTATTGC | GTGCACAGTCACGCAGAAAG |
S4 | TGCTGTCTCCACTTCCCTTC | AAGGAGGCGTGCTTAACAGG |
S5 | CTTGTCTTGGCTTGGCTAGG | ATCCACGCAGATAGACCTGC |
InD1 | AAAACGATCATAGTTGCTTGC | GGAGTTCGGGCTCAAGTA |
InD2 | GGGGCTTATTCACTTTGT | TTCTTTCCTCCTGGCTCT |
InD3 | CCCTTTTATCCTCAGTAGT | TACAAGCAGTTGTTTGCA |
InD4 | GGCGACCGAGAAGTGTAG | CGAGATAGCGAAAGCGAC |
InD5 | CGGGTCAGTGTTATCGTC | CATCAATGAAGGGCAAAT |
图1 blg1突变体及对照株系Z1803的花器官表型分析 A和B分别为blg1和对照Z1803的小穗(标尺=2 cm);C、D、E分别为blg1的小花、外稃和内稃(标尺=3 mm);F、G、H分别为Z1803的小花、外稃和内稃(标尺=3 mm);I和K分别为blg1和Z1803的内部花器官(标尺=1.5 mm);J和L分别为blg1和Z1803的花粉染色(标尺=100 μm);M和N分别为blg1和Z1803的小花横切面石蜡切片(标尺=1 mm)。
Fig. 1. Phenotypic analysis of floret organs in blg1 mutant and Z1803. A and B, Spikelets of blg1 and Z1803, respectively(Bar=2 cm); C, D, and E, Floret, lemma, palea of blg1, respectively(Bar=3 mm); F, G and H, Floret, lemma, palea of Z1803, respectively(Bar=3 mm); I and K, Internal floral organs of blg1 and Z1803, respectively(Bar=1.5 mm); J and L, Pollen staining of blg1 and Z1803, respectively(Bar=100 μm); M and N, Paraffin section of floret transection of blg1 and Z1803, respectively(Bar=1 mm).
图2 blg1突变体成熟籽粒表型 A―Z1803籽粒;B~H―blg1突变体籽粒;I~P―分别为A~H的糙米;其中A~D为南昌种植籽粒,E~H为三亚种植籽粒表型。A~H,标尺=3 mm;I~P, 标尺=2 mm。
Fig. 2. Phenotype of mature grains of blg1. A, A mature grain of Z1803; B-H, Mature grains of blg1; I-P, Brown rice of A-H, respectively; A-D, Grains of the mutant planted in Nanchang City, Jiangxi Province; E-H, Grains of the mutant planted in Sanya City, Hainan Province. A-H, Bar=3 mm; I-P, Bar=2 mm.
基因 | 功能注释 |
---|---|
Gene | Function annotation |
LOC_Os05g48700 | 赤霉素2-β-双加氧酶 Gibberellin 2-beta-dioxygenase |
LOC_Os05g48710 | 表达蛋白 Expressed protein 转座子蛋白Transposon protein |
LOC_Os05g48720 | 转座子蛋白 Transposon protein |
LOC_Os05g48730 | 表达蛋白 Expressed protein |
LOC_Os05g48740 | 表达蛋白 Expressed protein |
LOC_Os05g48750 | 3-脱氧-甘露醇辛酸胞苷酰转移酶 3-deoxy-manno-octulosonate cytidylyltransferase |
LOC_Os05g48760 | 含有DUF1421结构域的功能未知蛋白质 Protein of unknown function containing DUF1421 domain |
LOC_Os05g48770 | 表达蛋白 Expressed protein |
表2 精细定位区间内8个候选基因
Table 2. Eight candidate genes within the fine mapping region.
基因 | 功能注释 |
---|---|
Gene | Function annotation |
LOC_Os05g48700 | 赤霉素2-β-双加氧酶 Gibberellin 2-beta-dioxygenase |
LOC_Os05g48710 | 表达蛋白 Expressed protein 转座子蛋白Transposon protein |
LOC_Os05g48720 | 转座子蛋白 Transposon protein |
LOC_Os05g48730 | 表达蛋白 Expressed protein |
LOC_Os05g48740 | 表达蛋白 Expressed protein |
LOC_Os05g48750 | 3-脱氧-甘露醇辛酸胞苷酰转移酶 3-deoxy-manno-octulosonate cytidylyltransferase |
LOC_Os05g48760 | 含有DUF1421结构域的功能未知蛋白质 Protein of unknown function containing DUF1421 domain |
LOC_Os05g48770 | 表达蛋白 Expressed protein |
图4 基因LOC_Os05g48760部分序列的测序结果 A―基因LOC_Os05g48760的部分序列;B―5个SNP位点在基因LOC_Os05g48760上的位置及突变类型。
Fig. 4. Sequencing result of partial sequence of LOC_Os05g48760. A, Partial sequence of LOC_Os05g48760; B, Location and mutation type of 5 SNPs on gene LOC_Os05g48760.
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