Chinese Journal OF Rice Science ›› 2020, Vol. 34 ›› Issue (2): 115-124.DOI: 10.16819/j.1001-7216.2020.9122
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Changjian WANG, Long CHEN, Liping DAI, Xueli LU, Jinli HE, Long YANG, Jiang HU, Li ZHU, Guojun DONG, Guangheng ZHANG, Zhenyu GAO, Deyong REN, Guang CHEN, Lan SHEN, Qiang ZHANG, Longbiao GUO, Qian QIAN*(), Dali ZENG*()
Received:
2019-11-15
Revised:
2019-12-27
Online:
2020-03-10
Published:
2020-03-10
Contact:
Qian QIAN, Dali ZENG
About author:
#These authors contributed equally to this work
王昌健, 陈龙, 代丽萍, 路雪丽, 贺金立, 杨龙, 胡江, 朱丽, 董国军, 张光恒, 高振宇, 任德勇, 陈光, 沈兰, 张强, 郭龙彪, 钱前*(), 曾大力*()
通讯作者:
钱前,曾大力
作者简介:
#共同第一作者
基金资助:
CLC Number:
Changjian WANG, Long CHEN, Liping DAI, Xueli LU, Jinli HE, Long YANG, Jiang HU, Li ZHU, Guojun DONG, Guangheng ZHANG, Zhenyu GAO, Deyong REN, Guang CHEN, Lan SHEN, Qiang ZHANG, Longbiao GUO, Qian QIAN, Dali ZENG. Identification and Fine Mapping of Defective Pistil and Stamens 2 in Rice[J]. Chinese Journal OF Rice Science, 2020, 34(2): 115-124.
王昌健, 陈龙, 代丽萍, 路雪丽, 贺金立, 杨龙, 胡江, 朱丽, 董国军, 张光恒, 高振宇, 任德勇, 陈光, 沈兰, 张强, 郭龙彪, 钱前, 曾大力. 水稻花器官发育基因DPS2的鉴定和精细定位[J]. 中国水稻科学, 2020, 34(2): 115-124.
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URL: http://www.ricesci.cn/EN/10.16819/j.1001-7216.2020.9122
引物名称 | 前引物 | 后引物 |
---|---|---|
Primer name | Forward primer(5'-3') | Reverse primer(5'-3') |
BY-4 | GGAGGTGACTACGAAGGCATT | GAAGCTTATCCTTTTCACTTTCG |
BY19 | GAACTCCAAGTCCACCCTGAT | GATAAATTCTGTTGGTGGGAGATTG |
BY5 | GTTCACCAATAATCACCACAAGAGC | TAGCACCCCAGCAAGGACTCG |
BY6-2 | CTCGGCTCTCTTTGCTCGG | TGCCGACGAGGTCACCGCC |
BY7-3 | TATTTAGCCCTGGTTGAATGGT | TACCTAAGCTAAGTGGAAAGA |
BY20 | GACCAGAGGTGGACCAAGGA | GTGAATGGAAAGTGTGTGAAAGATC |
BY10-1 | GACTATATAGATATGTCCTCGGAAG | TTACCTATAACGAACCAAAGGC |
BY13 | GGAAACAATATACTATGGTGTGGAT | CGTGGGTGGAGTTTAGAATG |
Table 1 Part of primers used in the research.
引物名称 | 前引物 | 后引物 |
---|---|---|
Primer name | Forward primer(5'-3') | Reverse primer(5'-3') |
BY-4 | GGAGGTGACTACGAAGGCATT | GAAGCTTATCCTTTTCACTTTCG |
BY19 | GAACTCCAAGTCCACCCTGAT | GATAAATTCTGTTGGTGGGAGATTG |
BY5 | GTTCACCAATAATCACCACAAGAGC | TAGCACCCCAGCAAGGACTCG |
BY6-2 | CTCGGCTCTCTTTGCTCGG | TGCCGACGAGGTCACCGCC |
BY7-3 | TATTTAGCCCTGGTTGAATGGT | TACCTAAGCTAAGTGGAAAGA |
BY20 | GACCAGAGGTGGACCAAGGA | GTGAATGGAAAGTGTGTGAAAGATC |
BY10-1 | GACTATATAGATATGTCCTCGGAAG | TTACCTATAACGAACCAAAGGC |
BY13 | GGAAACAATATACTATGGTGTGGAT | CGTGGGTGGAGTTTAGAATG |
Fig. 1. Phenotypic comparison of dps2 mutant and wild type CJ06 at heading and maturity stage. A and B, Plants of Chunjiang 06(CJ06) and dps2 mutant at heading date; C and D, Panicle of CJ06 and dps2 at heading date; E, Heading date of CJ06 and dps2 in Lingshui, Hainan and Hangzhou, Zhejiang; F and G, Plants of CJ06 and dps2 at maturity stage; H and I, Panicle of CJ06 and dps2 at maturity stage; J, Seed-setting rate of CJ06 and dps2 in Lingshui and Hangzhou. Mean±SD(n=3). * and ** The difference between the wild type and mutant is significant at 0.05 and 0.01 level, respectively according to Student’s t test. In A, B, F, G, Bar=10 cm; In C, D, H, I, Bar=2 cm.
Fig. 2. Stamens phenotypic observation of wild type Chunjiang 06(CJ06) and dps2 mutant. A, Glumous flower of CJ06; B to D, Glumous flower of dps2 mutant; E, F, Anthers of the wild type and dps2; G, H, The outmost surface on epidermis anthers of the wild type and dps2; I, J, Pollens of CJ06 and dps2 mutant. Lo, Lodicule; An, Anther; F, Filament. From A to D, Bar=1 mm; In E, F, Bar=25 μm; In G, H, Bar=10 μm; In I, J, Bar=50 μm.
Fig. 3. Transverse section of anthers of dps2 mutant and wild type Chunjiang 06(CJ06) at different stages. A to C, CJ06; D to I, dps2 mutant; A, D, G, Cross-section of anther at the stage 9; B, E, H, Cross-section of anther at the stage 10; C, F, I, Cross-section of anther at the stage 12; Ep, Epidermis; En, Endothecium; T, Tapetum; DT, Degeneration tapetum; Ms, Microsporocyte; Msp, Microspore; Mp, Mature pollen. Bar=10 μm.
Fig. 4. Pistils phenotypic observation of wild type Chunjiang 06(CJ06) and dps2 mutant. A, Pistils of wild type; B, C, Pistils of dps2 mutant; D, Embryo sac of wild type; E, F, Embryo sac of dps2 mutant. Sti, Stigma; ov, Ovary. Scale bars: 1 mm in A, B and C, 50 μm in D to F.
杂交组合 Hybridized combination | F1 | F2 | χ2(3: 1) | χ2(0.05) | ||||
---|---|---|---|---|---|---|---|---|
正常表型 Wild-type phenotype | 突变表型 dps2 phenotype | 正常表型 Wild-type phenotype | 突变表型 dps2 phenotype | |||||
DPS2dps2/日本晴 DPS2dps2/Nipponbare | 7 | 0 | 236 | 79 | 0.001 | 3.84 | ||
日本晴/DPS2dps2 Nipponbare/DPS2dps2 | 4 | 0 | 133 | 43 | 0.030 | 3.84 | ||
DPS2dps2/9311 | 5 | 0 | 160 | 42 | 1.908 | 3.84 | ||
9311/ DPS2dps2 | 4 | 0 | 127 | 35 | 0.996 | 3.84 |
Table 2 Genetic analysis of the dps2 locus
杂交组合 Hybridized combination | F1 | F2 | χ2(3: 1) | χ2(0.05) | ||||
---|---|---|---|---|---|---|---|---|
正常表型 Wild-type phenotype | 突变表型 dps2 phenotype | 正常表型 Wild-type phenotype | 突变表型 dps2 phenotype | |||||
DPS2dps2/日本晴 DPS2dps2/Nipponbare | 7 | 0 | 236 | 79 | 0.001 | 3.84 | ||
日本晴/DPS2dps2 Nipponbare/DPS2dps2 | 4 | 0 | 133 | 43 | 0.030 | 3.84 | ||
DPS2dps2/9311 | 5 | 0 | 160 | 42 | 1.908 | 3.84 | ||
9311/ DPS2dps2 | 4 | 0 | 127 | 35 | 0.996 | 3.84 |
Fig. 5. Location of DPS2 on rice chromosome 4. A, Preliminary localization of DPS2 gene; B, Fine mapping of DPS2 gene; C, ORFs between markers BY6-2 and BY7-3.
登录号 | 位置 | 功能注释 |
---|---|---|
Gene accession | Location/bp | Functional annotation |
LOC_Os04g08290 | 4 427 683-4 424 830 | ZOS4-04-C2H2锌指蛋白 ZOS4-04-C2H2 zinc finger protein |
LOC_Os04g08300 | 4 433 624-4 433 839 | 反转录转座子蛋白,Ty3-gypsy亚类 Retrotransposon protein, putative, Ty3-gypsy subclass |
LOC_Os04g08310 | 4 436 856-4 441 657 | 表达蛋白 Expressed protein |
LOC_Os04g08320 | 4 444 004-4 441 881 | 假定蛋白YIF1B YIF1B, putative |
LOC_Os04g08330 | 4 446 960-4 446 649 | 表达蛋白 Expressed protein |
LOC_Os04g08340 | 4 452 696-4 455 350 | OsSigP3-假定的Ⅰ型信号肽酶同系物;采用假定的Ser/Lys催化二元体 OsSigP3-putative Type I signal peptidase homologue, employs a putative Ser/Lys catalytic dyad |
LOC_Os04g08350 | 4 461 845-4 457 458 | 半胱氨酸合酶,叶绿体/染色质前体 Cysteine synthase, chloroplast/chromoplast precursor |
LOC_Os04g08360 | 4 463 046-4 463 552 | 表达蛋白 Expressed protein |
LOC_Os04g08370 | 4 473 512-4 470 642 | 富含亮氨酸的重复家族蛋白 Leucine rich repeat family protein |
LOC_Os04g08390 | 4 490 101-4 484 997 | 富含亮氨酸的重复家族蛋白 Leucine rich repeat family protein |
LOC_Os04g08400 | 4 497 789-4 503 741 | 反转录转座子蛋白 Retrotransposon protein |
LOC_Os04g08410 | 4 513 883-4 508 804 | 含有ELM2结构域蛋白 ELM2 domain containing protein |
Table 3 The 12 genes in the candidate interval on chromosome 4.
登录号 | 位置 | 功能注释 |
---|---|---|
Gene accession | Location/bp | Functional annotation |
LOC_Os04g08290 | 4 427 683-4 424 830 | ZOS4-04-C2H2锌指蛋白 ZOS4-04-C2H2 zinc finger protein |
LOC_Os04g08300 | 4 433 624-4 433 839 | 反转录转座子蛋白,Ty3-gypsy亚类 Retrotransposon protein, putative, Ty3-gypsy subclass |
LOC_Os04g08310 | 4 436 856-4 441 657 | 表达蛋白 Expressed protein |
LOC_Os04g08320 | 4 444 004-4 441 881 | 假定蛋白YIF1B YIF1B, putative |
LOC_Os04g08330 | 4 446 960-4 446 649 | 表达蛋白 Expressed protein |
LOC_Os04g08340 | 4 452 696-4 455 350 | OsSigP3-假定的Ⅰ型信号肽酶同系物;采用假定的Ser/Lys催化二元体 OsSigP3-putative Type I signal peptidase homologue, employs a putative Ser/Lys catalytic dyad |
LOC_Os04g08350 | 4 461 845-4 457 458 | 半胱氨酸合酶,叶绿体/染色质前体 Cysteine synthase, chloroplast/chromoplast precursor |
LOC_Os04g08360 | 4 463 046-4 463 552 | 表达蛋白 Expressed protein |
LOC_Os04g08370 | 4 473 512-4 470 642 | 富含亮氨酸的重复家族蛋白 Leucine rich repeat family protein |
LOC_Os04g08390 | 4 490 101-4 484 997 | 富含亮氨酸的重复家族蛋白 Leucine rich repeat family protein |
LOC_Os04g08400 | 4 497 789-4 503 741 | 反转录转座子蛋白 Retrotransposon protein |
LOC_Os04g08410 | 4 513 883-4 508 804 | 含有ELM2结构域蛋白 ELM2 domain containing protein |
Fig. 6. Expression profile of the genes involved in the number and development of flower organs of dps2 mutant and wild type Chunjiang 06. Mean±SD(n=3). * and ** indicate the difference between the wild type Chunjiang 06 and mutant dps2 is significant at 0.05 and 0.01 levels, respectively according to Student’s t test.
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