首页 > 过刊浏览>2024年第43卷第2期 >144-153. DOI:10.13300/j.cnki.hnlkxb.2024.02.017
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芥菜小孢子培养及染色体加倍技术体系的优化
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作者:
作者单位:

1.华中农业大学园艺林学学院/果蔬园艺作物种质创新与利用全国重点实验室,武汉 430070;2.贵州省农业科学院园艺研究所,贵阳 550006

作者简介:

储玉凡，E-mail：chuyfan@webmail.hzau.edu.cn

通讯作者:

万正杰，E-mail：wanzj@mail.hzau.edu.cn

中图分类号:

S637

基金项目:

中央高校基本科研业务费专项（2662021YLQD004）；国家自然科学基金项目（32072573；32172601）；芥菜薹优质多抗新品种培育与推广项目（HBZY2023B004-4-4）；贵州省科技支撑计划项目（黔科合支撑［2022］一般086）


Optimization of microspore culture system and chromosome doubling technology in mustard
Author:
Affiliation:

1.National Key Laboratory for Germplasm Innovation & Utilization of Horticultural Crops， College of Horticulture and Forestry Sciences，Huazhong Agricultural University， Wuhan 430070，China;2.Institute of Horticulture， Guizhou Academy of Agricultural Sciences， Guiyang 550006，China

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    摘要:

    为探究不同芥菜基因型和培养条件对小孢子培养效果的影响，建立适合芥菜小孢子培养及染色体加倍的最佳方案，对芥菜8个不同变种的34份自交系进行小孢子培养，比较不同热激时间、活性炭浓度、小孢子密度对出胚率的影响。结果显示，基因型对芥菜小孢子培养成功与否影响较大，34份材料中11份材料成功培养出胚状体；不同材料间出胚效果差异明显，其中，大头菜出胚率最高，可达23.85胚/蕾；芥菜小孢子出胚最佳条件为32 ℃热激1~2 d，培养基含活性炭3~5 g/L，小孢子密度为1.5×105~2.0×105 个/mL。染色体加倍试验结果显示，1 g/L秋水仙素溶液浸泡茎尖1 h的处理加倍效率高、嵌合体少，可应用于芥菜染色体的加倍。

    Abstract:

    The effects of genotypes and culture conditions on the microspore culture in mustard （Brassica juncea） were studied to establish an optimal system of the microspore culture and chromosome doubling in mustard.34 inbred lines of 8 different varieties of mustard were used to conduct the microspore culture.The effects of heat shock times，concentrations of activated carbon，and densities of microspore on the rate of microspore embryogenesis were analyzed.The results showed that genotypes had a significant impact on the success of microspore culture in mustard，with 11 out of 34 inbred lines successfully producing embryoids.There were significant differences in rates of producing embryo among different inbred lines，among which Brassica juncea var. megarrhiza Tsen et Lee had the highest rate of producing embryo，reaching 23.85 embryos/anther.The optimal conditions for producing microspores in mustard were 32 ℃ heat shock for 1-2 days，culture medium containing 3-5 g/L activated carbon，and the density being 1.5×105 to 2.0×105 microspores/mL.The results of chromosome doubling experiment showed that immersing the stem tips in 1 g/L colchicine solution for 1 hour had a high efficiency of chromosome doubling and low chimerism，which could be used to double the chromosomes in mustard.The results of chromosome doubling test showed that 1 g/L colchicine solution soaked in stem tip for 1 hour had high doubling efficiency and less mixed-ploidy formation，which could be used to double the chromosomes in mustard.

    表 3 不同热激处理时间下不同基因型芥菜小孢子培养效果Table 3 Microspore culture effect of different genotypes of mustard under different heat shock treatment times
    表 4 不同质量浓度活性炭处理下不同基因型芥菜小孢子培养效果Table 4 Microspore culture effect of different genotypes of mustard under different concentrations of activated carbon
    表 1 小孢子培养所用34份芥菜自交系Table 1 34 inbred lines of mustard used for microspore culture
    表 6 秋水仙素处理染色体加倍效果Table 6 Chromosome doubling effect of colchicine treatment
    表 2 不同基因型芥菜小孢子培养效果Table 2 Microspore culture effect of different genotypes of mustard
    图1 小孢子发育的过程Fig.1 The process of microspore development
    图2 胚状体发育及植株再生过程Fig.2 Embryoid body development and plant regeneration processes
    图3 倍性鉴定和花粉染色观察Fig.3 Ploidy identification and pollen viability observation
    表 5 不同小孢子密度下不同基因型芥菜小孢子培养效果Table 5 Microspore culture effect of different genotypes of mustard under different microspore density
    参考文献
    [1] WANG N,WANG H,ZHANG A,et al.Genomic prediction across years in a maize doubled haploid breeding program to accelerate early-stage testcross testing[J].Theoretical and applied genetics,2020,133(10):2869-2879.
    [2] ZHAO X,XU X W,XIE H X,et al.Fertilization and uniparental chromosome elimination during crosses with maize haploid inducers[J].Plant physiology,2013,163(2):721-731.
    [3] 万丽丽,王转茸,徐义,等.甘蓝型油菜小孢子培养子叶形胚发育成苗研究[J].湖北农业科学,2021,60(16):15-20.WAN L L,WANG Z R,XU Y,et al.Study on seedling development of cotyledon embryo in microspore culture of Brassica napus L.[J].Hubei agricultural sciences,2021,60(16):15-20 (in Chinese with English abstract).
    [4] 贾俊香.基于小孢子培养的白菜类蔬菜种质创新[D].沈阳:沈阳农业大学,2019.JIA J X.Creation of the germplasm in Brassica rapa via microspore culture[D].Shenyang:Shenyang Agricultural University,2019 (in Chinese with English abstract).
    [5] 万正杰,范永红,孟秋峰,等.中国芥菜种业发展与展望[J].中国蔬菜,2020(12):1-6.WAN Z J,FAN Y H,MENG Q F,et al.Development and prospect of Chinese mustard seed industry[J].China vegetables,2020(12):1-6 (in Chinese).
    [6] 杨美翠,赵诗慧,高源,等.油菜双单倍体诱导育种技术研究进展[J].生物技术进展,2022,12(5):655-663.YANG M C,ZHAO S H,GAO Y,et al.Advances on double haploid induction breeding technology in rapeseed[J].Current biotechnology,2022,12(5):655-663 (in Chinese with English abstract).
    [7] 韩笑,刘希玲,文正华.热激处理对花椰菜小孢子培养的影响[J].天津农林科技,2012(2):9-11.HAN X,LIU X L,WEN Z H.Effect of heat shock treatment on microspore culture of cauliflower[J].Science and technology of Tianjin agriculture and forestry,2012(2):9-11 (in Chinese).
    [8] SHUMILINA D,KORNYUKHIN D,DOMBLIDES E,et al.Effects of genotype and culture conditions on microspore embryogenesis and plant regeneration in Brassica rapa ssp.rapa L.[J/OL].Plants,2020,9(2):278[2023-09-19].https://doi.org/10.3390/plants9020278.
    [9] 邓英,唐兵,付文苑,等.叶用芥菜小孢子培养技术体系的完善及DH系创制[J].中国农业大学学报,2018,23(9):60-67.DENG Y,TANG B,FU W Y,et al.Opertimization of the isolated microspore culture system of leaf mustard and double haploid line producing[J].Journal of China Agricultural University,2018,23(9):60-67 (in Chinese with English abstract).
    [10] 于锡宏,张艳梅,蒋欣梅,等.秋水仙素诱导榨菜四倍体的效应[J].东北农业大学学报,2013,44(10):88-92.YU X H,ZHANG Y M,JIANG X M,et al.Study on effects of colchicines on tetraploid inducement of tuber mustard[J].Journal of Northeast Agricultural University,2013,44(10):88-92 (in Chinese with English abstract).
    [11] 杨以耕,刘念慈,陈学群,等.芥菜分类研究[J].园艺学报,1989,16(2):114-121.YANG Y G,LIU N C,CHEN X Q,et al.A study on classification of mustard[J].Acta horticulturae sinica,1989,16(2):114-121 (in Chinese with English abstract).
    [12] 赵娟,沈佳,李海梅,等.MTG-DAPI双染色法观察黄瓜花粉细胞半薄切片中线粒体DNA的研究[J].南京农业大学学报,2015,38(1):27-32.ZHAO J,SHEN J,LI H M,et al.The research of detecting the mitochondrial DNA in semi-thin sliced pollen grains of Cucumis sativus L. by MTG-DAPI double-staining method[J].Journal of Nanjing Agricultural University,2015,38(1):27-32 (in Chinese with English abstract).
    [13] 唐兵,陶莲,卢松,等.白菜游离小孢子培养高频胚诱导技术体系优化[J].热带作物学报,2017,38(10):1913-1920.TANG B,TAO L,LU S,et al.Optimization of high frequency embryo induction system for isolated microspore culture in Chinese cabbage[J].Chinese journal of tropical crops,2017,38(10):1913-1920 (in Chinese with English abstract).
    [14] 刘晓东,孟川,王明秋,等.一种利用流式细胞仪快速检测大白菜倍性的方法: CN110849795B [P]. 2022-04-26. LIU X D,MENG C,WANG M Q,et al.Method for rapidly detecting ploidy of Chinese cabbages by using flow cytometry:CN110849795B[P].2022-04-26 (in Chinese).
    [15] 石淑稳,周永明,吴江生.秋水仙碱处理油菜离体小孢子的染色体加倍效应[J].华中农业大学学报,2002,21(4):329-333.SHI S W,ZHOU Y M,WU J S.Chromosome doubling of Brassica napus L. haploid by treating in vitro microspores with colchicine[J].Journal of Huazhong Agricultural University,2002,21(4):329-333 (in Chinese with English abstract).
    [16] LI Q F,CHEN Y G,YUE F,et al.Microspore culture reveals high fitness of B.napus-like gametes in an interspecific hybrid between Brassica napus and B.oleracea[J/OL].PloS One,2018,13(3):e0193548[2023-09-19].https://doi.org/10.1371/journal.pone.0193548.
    [17] ZHAO Y,ZHENG W F,LI J,et al.Effects of genotype and sodium p-nitrophenolate on microspore embryogenesis and plant regeneration in broccoli (Brassica oleracea L. var. italica)[J/OL].Scientia horticulturae,2022,293:110711[2023-09-19].https://doi.org/10.1016/j.scienta.2021.110711.
    [18] TAKAHASHI Y,YOKOI S,TAKAHATA Y.Effects of genotypes and culture conditions on microspore embryogenesis and plant regeneration in several subspecies of Brassica rapa L.[J].Plant biotechnology reports,2012,6(4):297-304.
    [19] WANG T T,LI H X,ZHANG J H,et al.Initiation and development of microspore embryogenesis in recalcitrant purple flowering stalk (Brassica campestris ssp. chinensis var. purpurea Hort.) genotypes[J]. Scientia horticulturae,2009,121(4):419-424.
    [20] 顾祥昆.芥菜游离小孢子培养研究[D].北京:中国农业科学院,2013.GU X K.Investigation of microspore culture in Brassica juncea [D].Beijing:Chinese Academy of Agricultural Sciences Dissertation,2013 (in Chinese with English abstract).
    [21] 王萌.根用芥菜(Brassica juncea var. megarrhiza Tsen et Lee)花药培养与游离小孢子培养技术体系研究[D].武汉:华中农业大学,2011. WANG M. Studies on the technical system of anther and isolated microspore culture in root mustard (Brassica juncea var. megarrhiza Tsen et Lee) [D].Wuhan:Huazhong Agricultural University, 2011 (in Chinese with English abstract).
    [22] CUSTERS J B M,CORDEWENER J H G,N?LLEN Y,et al.Temperature controls both gametophytic and sporophytic development in microspore cultures of Brassica napus[J].Plant cell reports,1994,13(5):267-271.
    [23] WEATHERHEAD M A,BURDON J,HENSHAW G G.Some effects of activated charcoal as an additive to plant tissue culture media[J].Zeitschrift für pflanzenphysiologie,1978,89(2):141-147.
    [24] JOHANSSON L.Effects of activated charcoal in anther cultures[J].Physiologia plantarum,1983,59(3):397-403.
    [25] 邓英.芥菜(Brassica juncea Coss.)游离小孢子培养诱导胚状体发生因素研究[D].重庆:西南大学,2010.DENG Y. Study on induction factors of embryogenesis of isolated microspore culture of mustard[D].Chongqing:Southwest University,2010 (in Chinese with English abstract).
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储玉凡,陈野,付文苑,姚培杰,张志琪,兰凯,余长春,万正杰.芥菜小孢子培养及染色体加倍技术体系的优化[J].华中农业大学学报,2024,43(2):144-153

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  • 收稿日期:2023-09-19
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