双单倍体技术在水稻育种中的应用进展

doi:10.11924/j.issn.1000-6850.casb2025-0594

摘要/Abstract

摘要：

双单倍体(DH)技术能快速实现基因型纯合、缩短选育周期、提高育种效率，该技术在玉米育种中应用广泛。然而，DH技术在水稻品种选育中的研究与报道尚显不足。本文系统梳理了DH技术在水稻中的研究进展，详细阐述了单倍体诱导、染色体加倍、筛选与鉴定的关键流程，分析了其在基础研究中的多元化应用，包括构建遗传图谱、定位数量性状位点(QTLs)、在单倍体阶段进行基因编辑以快速创制纯合突变体，以及在基因组测序等前沿领域的探索。此外，还总结了DH技术在水稻新品种培育中的应用情况。针对当前单倍体技术在水稻育种过程中面临的植株矮小、存活率低、诱导效率不稳定等问题，本文提出了优化该技术的策略，例如通过利用多样化的种质资源进行杂交，在单倍体阶段进行基因编辑，以及整合分子标记辅助选择、全基因组选择或多基因协同编辑等方法。最后，本文对未来水稻DH育种的发展方向进行了前瞻性展望，以期为此项技术在水稻新品种培育中的应用提供理论参考。

关键词: 水稻, 双单倍体, 育种技术, 品种选育, 遗传改良

Abstract:

Doubled Haploid (DH) technology can rapidly achieve genotypic homozygosity., shorten the breeding cycle, and enhance breeding efficiency. It has been widely applied in maize breeding. However, reports on its application in rice variety selection remain relatively limited. This paper reviews the research progress of DH technology in rice, summarizing the processes of haploid induction, chromosome doubling, screening, and identification. It analyzes the applications of this technology in fundamental research areas such as constructing genetic maps, mapping quantitative trait loci (QTLs), editing haploid genes, creating homozygous mutants, and genome sequencing. Additionally, it summarizes the application of DH technology in the development of new rice varieties. To address challenges like plant dwarfism, low survival rates, and unstable induction efficiency encountered during rice breeding using haploid technology, strategies are proposed to optimize the technology, including utilizing diverse germplasm resources for hybridization, gene editing at the haploid stage, and integrating molecular marker-assisted selection, genomic selection, or multi-gene synergistic editing. Finally, this review provides insights into the future development directions of DH breeding in rice, offering a theoretical reference for its application in cultivating new rice varieties.

Key words: rice, double haploid, breeding technology, variety selection, genetic improvement

中图分类号:

S511稻

李堂, 李潜龙, 杜明, 方玉. 双单倍体技术在水稻育种中的应用进展[J]. 中国农学通报, 2026, 42(17): 1-10.

LI Tang, LI Qianlong, DU Ming, FANG Yu. Application Progress of Double Haploid Breeding Technology in Rice Breeding[J]. Chinese Agricultural Science Bulletin, 2026, 42(17): 1-10.

图/表 2

参考文献 64

[1]	GHALAGI C, NAMRATHA M R, KOTYAL K, et al. A novel visual marker to distinguish haploids from doubled haploids in rice (Oryza sativa L.) at early growth stages[J]. Plant methods, 2023, 19(1):137. doi: 10.1186/s13007-023-01085-z
[2]	TOURAEV A, FORSTER B P, JAIN S M. Advances in haploid production in higher plants[M]. Berlin: Springer, 2009.
[3]	GUHA S, MAHESHWARI S. In vitro production of embryos from anthers of Datura[J]. Nature, 1964, 204(4957):497-497.
[4]	黄安平, 谭炎宁, 王伟平. 水稻双单倍体育种技术研究进展[J]. 杂交水稻, 2022, 37(6):1-5.
[5]	张建军, 范昆华. “水稻双单倍体育种技术及应用研究”通过成果鉴定[J]. 上海农业学报, 1998(4):6.
[6]	ZHANG Z, QIU F, LIU Y, et al. Chromosome elimination and in vivo haploid production induced by Stock 6-derived inducer line in maize (Zea mays L.)[J]. Plant cell reports, 2008, 27(12):1851-1860. doi: 10.1007/s00299-008-0601-2 URL
[7]	LIU C, LI X, MENG D, et al. A 4-bp insertion at ZmPLA1 encoding a putative phospholipase A generates haploid induction in maize[J]. Molecular plant, 2017, 10(3):520-522. doi: 10.1016/j.molp.2017.01.011 URL
[8]	ZHONG Y, LIU C, QI X, et al. Mutation of ZmDMP enhances haploid induction in maize[J]. Nature plants, 2019, 5(6):575-580. doi: 10.1038/s41477-019-0443-7
[9]	冯红玉, 姚碧娇, 陈媚, 等. 单倍体育种技术的应用进展[J]. 中国农学通报, 2021, 37(30):1-6. doi: 10.11924/j.issn.1000-6850.casb2021-0110
[10]	陈海强, 刘会云, 王轲, 等. 植物单倍体诱导技术发展与创新[J]. 遗传, 2020, 42(5):466-482.
[11]	JACQUIER N M, GILLES L M, PYOTT D E, et al. Puzzling out plant reproduction by haploid induction for innovations in plant breeding[J]. Nature plants, 2020, 6(6):610-619. doi: 10.1038/s41477-020-0664-9 pmid: 32514145
[12]	王健. 水稻单倍体诱导系的开发与应用[D]. 北京: 中国农业科学院, 2023.
[13]	GUO G, LIU S, ZHANG S, et al. Generic workflow of a highly effective and easy anther culture method for both japonica and indica rice[J]. Plants, 2024, 13(17):2531. doi: 10.3390/plants13172531 URL
[14]	QUIROZ L F, GONDALIA N, BRYCHKOVA G, et al. Haploid rhapsody: the molecular and cellular orchestra of in vivo haploid induction in plants[J]. New phytologist, 2024, 241(5):1936-1949. doi: 10.1111/nph.19523 pmid: 38180262
[15]	文钦. 水稻孤雌生殖单倍体诱导基因OsMATL突变体的创制与分析[D]. 广州: 华南农业大学, 2021.
[16]	肖国樱, 肖友伦, 刘建丰, 等. 多抗、优质、高产杂交水稻新品种的培育[A]//中国作物学会.第二十届中国作物学会学术年会论文摘要集[C].中国科学院亚热带农业生态研究所,湖南省植物保护研究所,中国种子集团有限公司, 2023.
[17]	迟铭, 方兆伟, 李健, 等. 花药培养在水稻育种中的应用研究进展[J]. 江苏农业科学, 2011, 39(6):111-113.
[18]	王敏, 段海燕, 姜恭好, 等. 水稻花药培养技术的研究进展[J]. 中国农学通报, 2022, 38(14):18-22. doi: 10.11924/j.issn.1000-6850.casb2021-0686
[19]	肖婉露, 马培芳, 张伟, 等. 植物小孢子培养技术研究进展[J]. 陕西农业科学, 2021, 67(12):70-73.
[20]	代金英, 郭桂梅, 程新杰, 等. 水稻F₁小孢子培养再生DH系的耐盐性鉴定及稻米品质分析[J]. 大麦与谷类科学, 2022, 39(6):1-5,10.
[21]	王亮, 邢虎成, 揭雨成. 作物孤雌生殖及其应用研究进展[J]. 安徽农业科学, 2012, 40(12):6993-6995,7003.
[22]	姬亚捷, 熊杰, 邱先进, 等. 植物孤雌生殖研究进展:助力无融合生殖走向应用[J]. 遗传, 2025, 47(4):448-455.
[23]	YAO L, ZHANG Y, LIU C, et al. OsMATL mutation induces haploid seed formation in indica rice[J]. Nature plants, 2018, 4(8):530-533. doi: 10.1038/s41477-018-0193-y
[24]	RAVI M, KWONG P N, MENORCA R M, et al. The rapidly evolving centromere-specific histone has stringent functional requirements in Arabidopsis thaliana[J]. Genetics, 2010, 186(2):461-471. doi: 10.1534/genetics.110.120337 URL
[25]	田光蕾. 基于OsCENH3基因突变创制水稻单倍体诱导系的研究[D]. 福州: 福建农林大学, 2020.
[26]	KUPPU S, RON M, MARIMUTHU M P, et al. A variety of changes, including CRISPR/Cas9-mediated deletions, in CENH3 lead to haploid induction on outcrossing[J]. Plant biotechnology journal, 2020, 18(10):2068-2080. doi: 10.1111/pbi.v18.10 URL
[27]	代雨婷. 水稻籼粳杂交DH群体的构建与遗传偏分离位点定位[D]. 南昌: 江西农业大学, 2023.
[28]	董俊杰, 张馨月, 富昊伟, 等. “粳不籼恢”杂交稻主要农艺性状的遗传参数分析[J]. 杂交水稻, 2023, 38(6):14-21.
[29]	井文, 杨世翌, 仇泽宇, 等. 磷脂酶在植物单倍体诱导中的功能研究进展[J]. 植物生理学报, 2023, 59(6):1091-1100.
[30]	沈任佳, 梁思怡, 郭涛, 等. 一种创制可高效固定变异的水稻单倍体诱导系的方法及其应用:CN119020519A[P]. 2024-11-26.
[31]	郭书磊, 魏昕, 魏良明, 等. 玉米单倍体诱导、加倍技术及相关机理探讨[J]. 玉米科学, 2020, 28(3):52-59,65.
[32]	王葆生, 刘湘萍, 廉勇, 等. 单倍体育种技术研究进展[J]. 北方农业学报, 2018, 46(5):44-49.
[33]	CHAIKAM V, NAIR S K, BABU R, et al. Analysis of effectiveness of R1-nj anthocyanin marker for in vivo haploid identification in maize and molecular markers for predicting the inhibition of R1-nj expression[J]. Theoretical and applied genetics, 2015, 128(1):159-171. doi: 10.1007/s00122-014-2419-3 pmid: 25385333
[34]	MELCHINGER A E, SCHIPPRACK W, WÜRSCHUM T, et al. Rapid and accurate identification of in vivo-induced haploid seeds based on oil content in maize[J]. Scientific reports, 2013, 3(1):2129. doi: 10.1038/srep02129
[35]	VIJAY CHAIKAM V C, MARTINEZ L, MELCHINGER A, et al. Development and validation of red root marker-based haploid inducers in maize[J]. Crop science, 2016, 56(4):1678-1688. doi: 10.2135/cropsci2015.10.0653 URL
[36]	CHEN C, LIU X, LI S, et al. Co-expression of transcription factors ZmC1 and ZmR2 establishes an efficient and accurate haploid embryo identification system in maize[J]. The plant journal, 2022, 111(5):1296-1307. doi: 10.1111/tpj.v111.5 URL
[37]	郭涛, 王佳峰, 黄翠红, 等. 一种基于单倍体诱导系的水稻双单倍体育种方法:CN109997683B[P]. 2019-07-12.
[38]	王克剑, 刘朝雷. 一种培育水稻两系不育系的育种方法:CN115537426A[P]. 2022-12-30.
[39]	ZHU L, SHEN Y, DAI Z, et al. Gγ-protein GS3 function in tight genetic relation with OsmiR396/GS2 to regulate grain size in rice[J]. Rice, 2024, 17(1):59. doi: 10.1186/s12284-024-00736-6 pmid: 39249660
[40]	XIAO W, YANG Q, HUANG M, et al. Improvement of rice blast resistance by developing monogenic lines, two-gene pyramids and three-gene pyramid through MAS[J]. Rice, 2019, 12:1-11. doi: 10.1186/s12284-018-0262-x
[41]	RAVI M, CHAN SW. Haploid plants produced by centromere-mediated genome elimination[J]. Nature, 2010, 464(7288):615-618. doi: 10.1038/nature08842
[42]	GILLES L M, KHALED A, LAFFAIRE J B, et al. Loss of pollen-specific phospholipase NOT LIKE DAD triggers gynogenesis in maize[J]. The EMBO journal, 2017, 36(6):707-717. doi: 10.15252/embj.201796603 URL
[43]	WANG J, CAO Y, WANG K, et al. Development of multiple-heading-date mtl haploid inducer lines in rice[J]. Agriculture, 2022, 12(6):806. doi: 10.3390/agriculture12060806 URL
[44]	LIU C, YAN S, MAO F, et al. Large-scale production of rice haploids by combining superior haploid inducer with PTGMS lines[J]. Plant communications, 2024, 5(12):101067. doi: 10.1016/j.xplc.2024.101067 URL
[45]	LIANG S, WEN Q, LU W, et al. The haploid induction ability analysis of various mutation of OsMATL and OsDMPs in rice[J]. BMC biology, 2025, 23(1):30-30. doi: 10.1186/s12915-025-02140-y
[46]	SINGH A, BARANWAL V, SHANKAR A, et al. Rice phospholipase a superfamily: organization, phylogenetic and expression analysis during abiotic stresses and development[J]. Plos one, 2012, 7(2):e30947. doi: 10.1371/journal.pone.0030947 URL
[47]	安保光, 金雄霞, 王健华, 等. 一种提高单倍体诱导效率的突变型OsPLA1m1及其应用.CN116063423A[P]. 2023-05-05.
[48]	LI Y, LIN Z, YUE Y, et al. Loss-of-function alleles of ZmPLD3 cause haploid induction in maize[J]. Nature plants, 2021, 7(12):1579-1588. doi: 10.1038/s41477-021-01037-2
[49]	胡风越, 王健, 王春, 等. 水稻DMP1、DMP2、DMP3基因突变体的创制及其单倍体诱导能力鉴定[J]. 中国水稻科学, 2025, 39(1):55-66. doi: 10.16819/j.1001-7216.2025.231112
[50]	KHANDAY I, SKINNER D, YANG B, et al. A male-expressed rice embryogenic trigger redirected for asexual propagation through seeds[J]. Nature, 2019, 565(7737):91-95. doi: 10.1038/s41586-018-0785-8
[51]	WEI X, LIU C, CHEN X, et al. Synthetic apomixis with normal hybrid rice seed production[J]. Molecular plant, 2023, 16(3):489-492. doi: 10.1016/j.molp.2023.01.005 URL
[52]	LIAN X, ZHONG L, BAI Y, et al. Spatiotemporal transcriptomic atlas of rhizome formation in Oryza longistaminata[J]. Plant biotechnology journal, 2024, 22(6):1652-1668. doi: 10.1111/pbi.14294 pmid: 38345936
[53]	GURUSHIDZE M, HENSEL G, HIEKEL S, et al. True-breeding targeted gene knock-out in barley using designer TALE-nuclease in haploid cells[J]. Plos one, 2014, 9(3):e92046. doi: 10.1371/journal.pone.0092046 URL
[54]	DWIVEDI SL, BRITT AB, TRIPATHI L, et al. Haploids: constraints and opportunities in plant breeding[J]. Biotechnology advances, 2015, 33(6):812-829. doi: 10.1016/j.biotechadv.2015.07.001 URL
[55]	相志国, 海燕, 康明辉, 等. 单倍体的产生途径及其在作物遗传育种中的应用[J]. 河南农业科学, 2011, 40(11):17-21.
[56]	郭锐, 钱前, 高振宇. 水稻生物育种研究进展[J]. 中国基础科学, 2022, 24(4):9-17.
[57]	江杰. 低镉水稻突变系的综合评价与分子标记辅助花培加倍单倍体群体的构建[D]. 杭州: 浙江大学, 2020.
[58]	张馨月, 钱秋, 陈婕, 等. 分子标记辅助选择与花药培养相结合选育携带抗白叶枯病基因Xa39的水稻恢复系[J]. 浙江农业科学, 2023, 64(11):2607-2610. doi: 10.16178/j.issn.0528-9017.20221182
[59]	黄卫峰, 闫影, 王凯, 等. 利用花药培养和分子标记辅助相结合方法选育香软型粳稻新品种[J]. 上海农业学报, 2022, 38(6):41-46.
[60]	徐华山, 周雷, 李三和, 等. 利用花药培养技术快速创制长粒型优质籼稻材料[A].中国作物学会.第十九届中国作物学会学术年会论文摘要集[C].粮食作物种质创新与遗传改良湖北省重点实验室/湖北省农业科学院粮食作物研究所, 2020.
[61]	张承妹, 陆家安, 万常照, 等. 粳型杂交稻双单倍体的遗传及其育种意义[J]. 上海农业学报, 2000(4):24-30.
[62]	王克剑, 熊杰, 姬亚杰. 一种水稻单倍体诱导方法:CN117660525B[P]. 2024-04-26.
[63]	胡兴明, 钱前. 中国杂交粳稻研究的回顾与思考[J]. 中国稻米, 2021, 27(4):9-11. doi: 10.3969/j.issn.1006-8082.2021.04.003
[64]	ZHENG X, PENG Y, QIAO J, et al. Wild rice: unlocking the future of rice breeding[J]. Plant biotechnology journal, 2024, 22(11):3218-3226. doi: 10.1111/pbi.14443 pmid: 39150344

基因名称	诱导过程	诱导方式	诱导产生单倍体的原因	参考文献
OsMTL/PLA1	卵细胞受精依赖性	孤雌生殖	基因突变，诱导父系基因组消除	[7]
OspPLAγ	卵细胞受精依赖性	孤雌生殖	基因突变，诱导父系基因组消除	[46]
OsPLD	卵细胞受精依赖性	孤雌生殖	基因突变，在磷脂酶单倍体诱导途径中具有协同作用	[48]
OsDMP	卵细胞受精依赖性	不具备独立的单倍体诱导能力	与MTL互作诱导产生单倍体	[49]
OsCENH3	卵细胞受精依赖性	着丝粒介导，孤雌生殖	自身基因组消除，CENH3蛋白功能缺失，与cenh3突变体共同诱导	[41]
OsBBM	卵细胞受精非依赖性	孤雌生殖,无融合生殖	卵细胞异位表达，导致亲本基因组消除	[50-51]

基因名称	诱导过程	诱导方式	诱导产生单倍体的原因	参考文献
OsMTL/PLA1	卵细胞受精依赖性	孤雌生殖	基因突变，诱导父系基因组消除	[7]
OspPLAγ	卵细胞受精依赖性	孤雌生殖	基因突变，诱导父系基因组消除	[46]
OsPLD	卵细胞受精依赖性	孤雌生殖	基因突变，在磷脂酶单倍体诱导途径中具有协同作用	[48]
OsDMP	卵细胞受精依赖性	不具备独立的单倍体诱导能力	与MTL互作诱导产生单倍体	[49]
OsCENH3	卵细胞受精依赖性	着丝粒介导，孤雌生殖	自身基因组消除，CENH3蛋白功能缺失，与cenh3突变体共同诱导	[41]
OsBBM	卵细胞受精非依赖性	孤雌生殖,无融合生殖	卵细胞异位表达，导致亲本基因组消除	[50-51]