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作物学报 ›› 2018, Vol. 44 ›› Issue (02): 169-176.doi: 10.3724/SP.J.1006.2018.00169

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水稻花器官数目突变体mf2的鉴定和基因定位

严贤诚**, 陈立凯**, 罗玉花, 罗文龙, 王慧, 郭涛*(), 陈志强*()   

  1. 华南农业大学国家植物航天育种工程技术研究中心, 广东广州 510642;
  • 收稿日期:2017-05-27 接受日期:2017-09-10 出版日期:2018-02-12 网络出版日期:2017-10-27
  • 通讯作者: 严贤诚,陈立凯,郭涛,陈志强
  • 作者简介:

    第一作者联系方式: 严贤诚, E-mail: 18659186232@163.com; 陈立凯, E-mail: leeking1113@163.com; ** 同等贡献(Contributed equally to this work)

  • 基金资助:
    本研究由国家重点研发计划项目(2016YFD0102102), 国家现代农业产业技术体系建设专项(CARS-01-12)和广东省应用型研发项目(2015B020231011)资助

Identification and Gene Mapping of a Floral Organ Number Mutant mf2 in Rice (Oryza sativa)

Xian-Cheng YAN**, Li-Kai CHEN**, Yu-Hua LUO, Wen-Long LUO, Hui WANG, Tao GUO*(), Zhi-Qiang CHEN*()   

  1. National Engineering Research Center of Plant Space Breeding, South China Agricultural University, Guangzhou 510642, Guangzhou, China;
  • Received:2017-05-27 Accepted:2017-09-10 Published:2018-02-12 Published online:2017-10-27
  • Contact: Xian-Cheng YAN,Li-Kai CHEN,Tao GUO,Zhi-Qiang CHEN
  • Supported by:
    This study was supported by the National Key Research and Development Program of China (2016YFD0102102), the China Agricultural Research System (CARS-01-12), and the Applied Research and Development Funds in Guangdong (2015B020231011).

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摘要：

水稻的花器官发育影响着水稻的产量与品质。本研究通过12C重离子诱变航恢7号获得一个水稻花器官突变体multi-floret 2 (mf2), 其稃片、浆片、雄蕊、雌蕊增多, 多数小穗内具2~3朵类似小花。mf2内外稃不能很好勾合, 而且形状和维管束的数目都产生了一定程度的变化。电镜扫描幼穗发现花器官的变异在幼穗分化期的各花器官原基分化时就已形成。另外, 该突变体的抽穗期推迟, 株高降低, 穗数增多, 表明其营养生长也受到一定的影响。遗传分析表明mf2突变体表型受单隐性核基因控制。利用SSR、InDel分子标记将MF2定位于第1染色体的标记SSR39108和InD39210之间, 区间大小约为102 kb。

关键词: 水稻, 重离子诱变, 花器官变异, 基因定位

Abstract:

A rice floral organ mutant mf2 (multi-floret 2) was identified from Hanghui 7, an indica restorer line in rice (Oryza sativa L.), which was treated with heavy ion irradiation. The glumes, lodicules, stamens, pistil of the mutant increased, and most of the spikelets had 2-3 flowers. The palea and lemma of mf2 mutant were dehiscent, the shape and number of vascular bundles were different from those of the wild-type. Scanning electron microscope analysis revealed that the flowers variation of mf2 had been formed in the differentiation of the primordium during the spike differentiation stage. The heading date of the mf2 mutant was delayed, the plant height was decreased, but the number of panicles was increased, showing that the vegetative growth of the mutant was also affected. Genetic analysis showed that the mf2 mutant phenotype was controlled by a recessive nuclear gene. There are 321 recessive individuals from the F2 segregation population were used to fine map MF2. Finally, MF2 was located in a 102 kb physical interval between markers SSR39108 and InDel39210 on chromosome 1.

Key words: rice (Oryza sativa L.), heavy ion mutagenesis, floral mutants, gene mapping

表1

野生型和突变体mf2主要农艺性状调查"

材料
Material		 抽穗期
Heading period
(d)		 株高
Plant height
(cm)		 单株穗重
Single plant spike weight (g)		 穗数
Panicle number per plant		 结实率
Seed-setting rate (%)		 千粒重
1000-grain weight (g)
野生型Wild-type 70.62±0.28 104.60±0.90 27.73±0.76 7.33±0.58 86.49±0.45 21.47±0.18
突变体mf2 79.33±2.16* 92.44±1.37** 10.66±1.70** 13.20±0.86** 12.36±1.82** 13.64±0.10**

图1

突变体mf2及其野生型的表型 A: 野生型(左)和突变体mf2 (右)灌浆期植株表型; B: 野生型(下)和突变体mf2 (上)籽粒; C, D: 野生型颖花, D为C去掉内外稃后的形态; E~J: 突变体mf2的颖花, 其中F、H、J分别为E、G、I去掉稃片后的形态。Scale bar=100 mm (A); Scale bars = 1 mm (C~J)。"

图2

mf2突变体小穗小花数和籽粒数所占比例"

图3

突变体mf2及其野生型小穗显微结构 I: 野生型和mf2稃片石蜡切片观察。A: 野生型小穗; B, C: mf2小穗含类稃结构; D: 维管束部分局部放大。箭头表示维管束。II: 野生型和mf2幼穗电镜观察。A, B: 野生型幼穗, 雄蕊原基出现; C, D: mf2幼穗, 雄蕊原基出现, D中具额外的稃片原基(星号表示)。"

表2

mf2突变体遗传分析"

组合
Cross		 F1表型
Phenotype of F1		 F2表型 Phenotype of F2 χ2(3:1) χ20.05
正常表型Wild type 突变表型Mutant type 总数Total
02428/mf2 野生型 Wild type 357 126 483 0.249 3.84
Francis/mf2 野生型 Wild type 1091 321 1412 3.748

图4

MF2基因的精细定位"

表3

定位MF2所设计的引物序列"

标记
Marker		 正向引物
Forward primer (5°-3°)		 反向引物
Reverse primer (5°-3°)
InD38433 AGGAGAAGCTCCCAGTCGTT AATCCAAGTATCCCTTGCAAAC
InD38713 AACAGCAACGGTAACTTTCACA ACGCTTAGGAAGCTATGAGGTG
SSR39108 CATGTCTTGCAAGCTAACCAAA CTATATATGCCACGACGAGCAG
InD39167 GCAAGTAAACCAACCACACGTA CCATCCATCAATCCGAACTACT
InD39210 TTTAAGGGGTTTCTAGCTGCTG ATAGCTTTGGACCTGTTTTGGA
InD39410 TGAACTCAGGCCCAGTTTAGTT TTGAGCCTCCAGTAGTTCCTTC

表4

MF2定位区间内的候选基因"

基因号
Gene ID		 位置
Location		 基因注释
Description
Os01g67364 39114681-39117532 Ty3-gypsy家族反转录转座子蛋白 Retrotransposon protein, putative, Ty3-gypsy subclass, expressed
Os01g67370 39129736-39134858 表达蛋白 Expressed protein
Os01g67380 39136361-39136861 表达蛋白 Expressed protein
Os01g67390 39136937-39139380 花粉Ole-e-1变应原和扩展蛋白家族的蛋白前体
POEI32-Pollen Ole-e-I allergen and extensin family protein precursor, expressed
Os01g67400 39140444-39140762 假定蛋白 Hypothetical protein
Os01g67410 39141130-39145479 AP2/EREBP转录因子 AP2/EREBP transcription factor BABY BOOM, putative, expressed
Os01g67420 39158153-39165729 脂肪酶 Lipase, putative, expressed
Os01g67430 39177169-39178676 脂肪酶 Lipase, putative, expressed
Os01g67440 39179769-39180065 转座子蛋白 Transposon protein, putative, unclassified, expressed
Os01g67450 39195689-39196972 脂肪酶 Lipase, putative, expressed
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