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作物学报 ›› 2019, Vol. 45 ›› Issue (1): 1-9.doi: 10.3724/SP.J.1006.2019.82032

• 作物遗传育种·种质资源·分子遗传学 •    下一篇

水稻圆粒基因RS的鉴定和定位

陈雅萍,缪荣,刘喜,陈本佳,兰杰,马腾飞,王益华,刘世家,江玲()   

  1. 南京农业大学作物遗传与种质创新国家重点实验室 / 江苏省植物基因工程技术研究中心, 江苏南京210095
  • 收稿日期:2018-06-07 接受日期:2018-10-08 出版日期:2018-11-01 网络出版日期:2018-11-09
  • 通讯作者: 江玲
  • 基金资助:
    本研究由国家重点研发计划项目(2016YFD0100101-08);国家自然科学基金重点项目(91535302);国家级大学生创新性实验计划项目(201710307014)

Identification and mapping of round seed related gene in rice (Oryza sativa L.)

Ya-Ping CHEN,Rong MIAO,Xi LIU,Ben-Jia CHEN,Jie LAN,Teng-Fei MA,Yi-Hua WANG,Shi-Jia LIU,Ling JIANG()   

  1. State Key Laboratory of Crop Genetics and Germplasm Enhancement / Research Center of Jiangsu Plant Gene Engineering, Nanjing Agricultural University, Nanjing 210095, Jiangsu, China
  • Received:2018-06-07 Accepted:2018-10-08 Published:2018-11-01 Published online:2018-11-09
  • Contact: Ling JIANG
  • Supported by:
    This study was supported by the National Key Research and Development Program of China(2016YFD0100101-08);the Key Projects of National Natural Science Foundation(91535302);the National University Student Innovation Program(201710307014)

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摘要：

阐明水稻籽粒大小相关基因的遗传和分子机制对水稻产量形成具有重要意义。利用甲基磺酸乙酯(ethyl methanesulfonate, EMS)诱变粳稻品种“宁粳3号”筛选获得圆粒突变体round seed (rs)。遗传分析表明, 突变体rs圆粒表型由单隐性核基因控制。颖壳扫描电镜观察发现, rs籽粒变圆主要是细胞数目改变导致的。在突变体rs中, 细胞周期相关基因的表达较野生型显著升高。将RS定位在第3染色体短臂标记RM3413与N3-5之间, 物理距离约589 kb。RS突变影响BR信号途径, 改变了粒型相关基因的表达。本研究有助于阐明水稻籽粒发育的分子机制。

关键词: 水稻, 圆粒突变体, 表型分析, 基因定位

Abstract:

It is significant to clarify the genetic and molecular mechanism of genes related to rice grain size for rice yield. A mutant round seed (rs) was screened from japonica variety “Ningjing 3” mutagenized by ethyl methane sulfonate (EMS). Genetic analysis revealed that the phenotype of rs was controlled by a single recessive nuclear gene. Scanning electron microscope observation indicated that the change of cell number was responsible for the mutant phenotype of rs. Compared with the wild type, the expression of cell cycle related-genes increased significantly in rs. RS was located in the 589 kb region of chromosome 3, between markers RM3413 and N3-5. The RS mutation affected BR signal pathway, and changed the expression of grain size-related genes. This research contributes to elucidating the molecular mechanism of rice grain development.

Key words: rice (Oryza sativa L.), round seed mutant, phenotypic analysis, gene mapping

表1

用于精细定位的分子标记"

标记
Marker		 正向引物序列
Forward primer sequence (5°→3°)		 反向引物序列
Reverse primer sequence (5°→3°)
N3-5 GTCTGTGCGCTCCTTGTTCTAGC CCTGGACCAATTTGTATGGTTGG
N3-9 GATGCAGTAGGAACACCAAACAGC ATCGAGTACCAAGTGCCTGTGC
N3-10 GGTTTGGGAGCCCATAATCT CTGGGCTTCTTTCACTCGTC
RM3413 TTAGAGGAGATGATGGTGCAACG AGCAGCCATTGAATGTGTTTGG
RM14282 CCCAAACACAAACACAAAGAGAGC AACACGCAGGTCCTCTTGAACC
I3-2 TACTTTAATTTTGCAGCTC TTTTACCCCACTCCATCT

表2

野生型和rs的农艺性状比较"

性状 Trait 野生型 Wild type 突变体 rs
粒长 Grain length (mm) 7.50±0.009 6.39±0.023**
粒宽 Grain width (mm) 3.13±0.021 3.58±0.003**
长/宽比 Length/width ratio 2.42±0.013 1.80±0.009**
粒厚 Grain thickness (mm) 1.14±0.025 1.28±0.062**
千粒重 1000-grain weight (g) 25.45±0.890 22.51±0.252**
株高 Plant height (cm) 91.40±2.633 90.50±2.121
穗长 Panicle length (cm) 17.35±0.803 17.19±0.709
分蘖数 Number of tillers 11.00±2.160 10.1±2.025
结实率 Seed setting rate (%) 72.69±2.430 56.90±4.270**

表3

突变体rs的遗传分析"

杂交组合
Cross-combination		 总株数
Total number
of plants		 正常表现
Wild-type
phenotype		 突变表现
Mutant
phenotype		 χ2(3:1)
rs/WT 174 135 39 0.62
WT/rs 207 161 46 0.85

图 1

野生型和突变体rs的表型 A: 野生型和rs在抽穗期植株表现, 比例尺为10 cm。B, C: 野生型和rs糙米籽粒表现。比例尺为6 mm。D, F: 野生型和rs粒长、粒宽与千粒重比较。**代表突变体与野生型之间在P = 0.01水平上差异显著。"

图2

颖壳组织学分析 A, B: 扫描电镜观察颖壳外表皮。比例尺为200 μm。C, D: 颖壳外表皮细胞长度与宽度的比较。E, F: 颖壳外表皮细胞数目的比较。 *和**分别表示在P= 0.05与P=0.01水平上差异显著。"

图3

细胞周期相关基因(CycA2;1、CycD4、E2F2、CycT1、CDKB、CDKA1、CycA2;2、Cdc20和CycA2;3)在野生型与rs中的表达分析 数据结果为3次生物重复平均值, *和**分别表示在P = 0.05与0.01水平上差异显著。"

图4

RS基因的精细定位 黑色圆圈表示着丝粒; 粗竖线表示共分离标记; 标记下方的数字为极端个体的数目。"

图5

野生型和突变体rs 对2,4-epiBL处理的响应 A: 不同2,4-epiBL浓度处理, 上面为野生型, 下面为rs。B: 不同2,4-epiBL浓度处理叶夹角的变化。C: 野生型和rs中BR合成和信号转导途径基因的表达水平。**分别代表突变体与野生型之间差异达0.01显著水平。"

图6

粒型相关基因在野生型和突变体rs中的表达 *与**分别表示在P= 0.05与0.01水平上差异显著。*P= 0.05; **P =0.01."

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