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Acta Agronomica Sinica ›› 2018, Vol. 44 ›› Issue (02): 169-176.doi: 10.3724/SP.J.1006.2018.00169

• Orginal Article • Previous Articles     Next Articles

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 Online:2018-02-12 Published:2017-10-27
  • Contact: Xian-Cheng YAN,Li-Kai CHEN,Tao GUO,Zhi-Qiang CHEN E-mail:guo.tao@vip.163.com;zqchen@scau.edu.cn
  • 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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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

Table 1

Comparison of main agronomic traits between wild type and mf2 mutant"

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**

Fig. 1

Phenotypes of mf2 and the wild type A: phenotype of wild type (left) and mf2 (right) individual plant during the filling stage; B: grains of wild type (lower) and mutant mf2 (upper); C, D: spikelet of wild-type, D is the C removing the lemma and palea; E-J: spikelets of the mutant mf2, where F, H, and J are E, G, and I, respectively removing hulls. Scale bars = 100 mm (A) or 1mm (C-J)."

Fig. 2

Ratio of the floret and grain number per spikelet of mf2 mutant"

Fig. 3

Micrographs of wild-type and mf2 spikelets I: observation of wild type and mf2 spikelets paraffin sections. A: wild type spikelet; B, C: mf2 spikelets containing hull-like structure; D: partial enlargement of the vascular bundle, red arrowheads indicate vascular bundles. II: scanning electron micrographs of wild type and mf2 young spikelets. A, B: wild type young spikelets at a time when stamen primordia are emerging; C, D: mf2 young spikelets at the same time of A and B, asterisk indicates a extra palea primordium in D."

Table 2

Genetic analysis of mf2 mutant"

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

Fig. 4

Fine mapping of MF2"

Table 3

Primer sequence used for MF2 mapping"

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

Table 4

Candidate genes of MF2 mapped region"

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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