CAO Bingdong,CAI Xia,ZHANG Jie,et al.Seedling Regeneration From Isolated Microspore-derived Embryo in Cabbage[J].Northern Horticulture,2022,(23):29-37.[doi:10.11937/bfyy.20221650]
结球甘蓝游离小孢子培养及胚发育成苗研究
- Title:
- Seedling Regeneration From Isolated Microspore-derived Embryo in Cabbage
- Keywords:
- cabbage; isolated microspore; cotyledon embryos; TDZ
- 文献标志码:
- A
- 摘要:
- 以20种不同基因型结球甘蓝为试材,采用游离小孢子离体培养方法,研究了结球甘蓝小孢子胚发生和植株再生的影响因素,并对再生植株倍性进行鉴定,以期建立高效的游离小孢子培养体系,为后期快速创新结球甘蓝种质材料提供参考依据。结果表明:基因型‘D3’小孢子出胚率最高,为42.33胚·蕾-1,是最低出胚材料‘D9’的17倍,表明基因型是影响小孢子胚发生的重要因素。花瓣与花药长度比值为0.51~0.75时小孢子出胚率最高。0.10~1.00 mg·L-1 TDZ能显著促进小孢子胚发生,获得的子叶胚成苗能力强且周期短。采用流式细胞仪对303株再生植株进行倍性鉴定,单倍体占49.18%,双单倍体占43.89%,嵌合体占6.93%。双单倍体和嵌合体植株均能正常结荚,而单倍体植株蕾期授粉后不能结荚。
- Abstract:
- Twenty different genotypes of cabbage were used as test materials for isolated microspore culture to study the influencing factors of microspore embryogenesis and plant regeneration,and to identify the ploidy and seed setting of regenerated plants,in order to establish an efficient isolated microspore culture system and provide reference for rapid innovation of germplasm materials in the later stage.The results showed that the genotype had a significant impact on the embryoid induction.The highest embryo yield was 42.33 embryo·bud-1 in inbred line ‘D3’.Additionally,the embryo yield of this variety was 17 folds that the lowest embryo yield of variety ‘D9’.The yield of microspore embryo was the highest when the ratio of petal length and anther length ranged from 0.51 to 0.75.The addition of 0.10-1.00 mg·L-1 TDZ significantly increased the yields of all type embryo,and the obtained cotyledon embryos had strong seedling capacity.Flow cytometry detection of 303 regenerated plant revealed that 49.18% were haploid,43.89% were doubled haploid,and chimera only accounted for the remaining 6.93%.Double haploid and chimeric plants could set pods normally after pollination at the bud stage,while the haploid plants could not produce pods.
参考文献/References:
[1]胡永霞,李晓峰,轩淑欣,等.利用小孢子培养创建大白菜-结球甘蓝易位系[J].园艺学报,2014,41(7):1361-1368.[2]LICHTER R.Induction of haploid plants from isolated pollen of Brassica napus[J].Zeitschrift Für Pflanzenphysiologie,1982,105(5):427-434.[3]吴江生,石淑稳,周永明,等.甘蓝型双低油菜品种华双3号的选育和研究[J].华中农业大学学报,1999,18(1):6-9.[4]耿建峰,原玉香,张晓伟,等.利用游离小孢子培养技术育成抗热大白菜新品种‘豫园50’[J].园艺学报,2002,29(1):89-97.[5]曾爱松,高兵,宋立晓,等.秋季专用牛心甘蓝新品种锦秋55的选育[J].江苏农业科学,2015,43(11):230-231.[6]苏贺楠,韩风庆,杨丽梅,等.结球甘蓝小孢子培养条件优化及高代自交系胚状体诱导研究[J].中国蔬菜,2018(4):30-36.[7]于文佳,侯喜林,赵晓嫚,等.小菘菜游离小孢子培养和植株再生[J].南京农业大学学报,2013,36(2):7-12.[8]詹艳,陈劲枫,AHMED A M.黄瓜游离小孢子培养诱导成胚和植株再生[J].园艺学报,2009,36(2):221-226.[9]JIA J X,ZHANG Y,CUI L W,et al.Effect of thidiazuron on microspore embryogenesis and plantlet regeneration in Chinese flowering cabbage(Brassica rapa.var.parachinenis)[J].Plant Breeding,2019,138(6):916-824.[10]黄天虹,张娅,梁超凡,等.不结球白菜游离小孢子培养及植株再生研究[J].核农学报,2019,33(2):240-247.[11]韩阳,叶雪凌,冯辉.大白菜小孢子植株的倍性变异及倍性鉴定方法的研究[J].中国蔬菜,2006(11):9-11.[12]袁素霞,刘玉梅,方智远,等.结球甘蓝和青花菜小孢子胚植株再生[J].植物学报,2010,45(2):226-232.[13]贾凯,吴慧,许建,等.不同处理方法对芜菁游离小孢子出胚率的影响[J].江苏农业科学,2018,46(8):39-42.[14]邓英,陶莲,李正丽,等.大白菜游离小孢子的培养及植株再生[J].贵州农业科学,2012,40(6):19-21.[15]DENG Y,TANG B,ZHOU X,et al.Direct regeneration of haploid or doubled haploid plantlets in cucumber (Cucumis sativus L.) through ovary culture[J].Plant Cell,Tissue and Organ Culture,2020,142(2):253-268.[16]ZHANG W,FU Q,DAI X G,et al.The culture of isolated microspores of ornamental kale (Brassica oleracea var.acephala) and the importance of genotype to embryo regeneration[J].Scientia Horticulturae,2008,117(1):69-72.[17]桑玉芳,张恩慧,杨安平,等.甘蓝游离小孢子培养中影响胚状体形成的主要因素[J].西北农业学报,2007,16(2):125-129.[18]张恩慧,马英夏,杨安平,等.甘蓝小孢子培养中花蕾长度与细胞单核期的关系[J].西北农业学报,2012,21(6):124-128.[19]曾爱松,高兵,宋立晓,等.耐寒结球甘蓝小孢子培养及其发育过程[J].中国农业大学学报,2015,20(2):86-92.[20]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.[21]姜凤英,冯辉,王超楠.小白菜小孢子培养胚状体的诱导[J].沈阳农业大学学报,2006,37(5):763-765.[22]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.[23]李岩,刘凡,曹鸣庆.通过游离小孢子培养方法获得小白菜三个变种的胚胎及植株[J].华北农学报,1993,8(3):92-97.[24]余凤群,刘后利.供体材料和培养基成份对甘蓝型油菜小孢子胚状体产量的影响[J].华中农业大学学报,1995,14(4):327-331.[25]田保明,蒋武生,张晓伟,等.提高油菜游离小孢子胚诱导频率的研究[J].华北农学报,2007,22(1):116-119.[26]SATO T,NISHIO T,HIRAI M.Plant regeneration from isolated microspore cultures of Chinese cabbage (Brassica campestris spp.pekinensis) [J].Plant Cell Reports,1989,8(8):486-488.[27]程利华.甘蓝型油菜小孢子培养技术体系研究[D].重庆:西南大学,2006.[28]LI H C,DEVAUX P.High frequency regeneration of barley doubled haploid plants from isolated microspore culture[J].Plant Science,2003,164(3):379-386.[29]李智军,曾晶,谭铭喜,等.芥蓝游离小孢子胚胎发生途径及2,4-D对成胚的影响[J].园艺学报,2021,48(8):1605-1618.
相似文献/References:
[1]祝海燕.结球甘蓝不同初级三体的花粉整齐度及生活力的研究[J].北方园艺,2013,37(12):90.
ZHU Hai-yan.Comparison of Pollen Regularity and Activity of One Anther of Primary Trisomic Lines[J].Northern Horticulture,2013,37(23):90.
[2]祝朋芳,王卫珍,李珺,等.羽衣甘蓝游离小孢子胚胎发生、植株再生与增殖的研究[J].北方园艺,2015,39(13):111.[doi:10.11937/bfyy.201513032]
ZHU Pengfang,WANG Weizhen,LI Jun,et al.Study on Microspore Embryogenesis,Regeneration and Propagation in vitro on Ornamental Kale (Brassica oleracea var.acephala)[J].Northern Horticulture,2015,39(23):111.[doi:10.11937/bfyy.201513032]
[3]张东昱,李建华,周晓丽,等.土壤水分对结球甘蓝农艺性状及产量品质的影响[J].北方园艺,2011,35(04):0.[doi:10.11937/bfyy.2011040010]
ZHANG Dong-yuI,LI Jian-hua,ZHOU Xiao-li,et al.Influence of Soil Moisture on Agronomic ~aits, Yield and Quality of Cabbage[J].Northern Horticulture,2011,35(23):0.[doi:10.11937/bfyy.2011040010]
[4]申 娟,梁 秋 霞,曹 刚 强,等.蔬菜类作物游离小孢子培养中的影响因素[J].北方园艺,2008,32(06):0.[doi:10.11937/bfyy.200806024]
[5]周 传 余,郎 英.木醋液在结球甘蓝上的应用效果研究[J].北方园艺,2008,32(12):0.[doi:10.11937/bfyy.2008012023]
ZHOU Chuan - yu,LANG Ying.Study on Applying Effects of Wood Vinegar[J].Northern Horticulture,2008,32(23):0.[doi:10.11937/bfyy.2008012023]
[6]张 勇,程 智 慧,张 恩 慧,等.栽培密度和肥料对甘蓝产量的影响[J].北方园艺,2004,28(05):0.[doi:10.11937/bfyy.200405035]
[J].Northern Horticulture,2004,28(23):0.[doi:10.11937/bfyy.200405035]
[7]徐 艳 辉,冯 辉,张 凯.大白菜游离小孢子培养中若干因素对胚状体诱导和植株再生影响[J].北方园艺,2001,25(03):0.[doi:10.11937/bfyy.200103004]
[J].Northern Horticulture,2001,25(23):0.[doi:10.11937/bfyy.200103004]
[8]李江丽,李成凤,王超,等.结球甘蓝抗黑腐病基因SSR标记转CAPS标记[J].北方园艺,2019,43(21):7.[doi:10.11937/bfyy.20183780]
LI Jiangli,LI Chengfeng,WANG Chao,et al.SSR Marker of Black Rot (Xanthommonas campestris pv.campestris) Resistance Gene of Cabbage (Brassica oleracea var.capitata)Turn CAPS Marker[J].Northern Horticulture,2019,43(23):7.[doi:10.11937/bfyy.20183780]
[9]裴云霞,倪天虹,张文君,等.结球甘蓝种子萌发及幼苗生长对不同浓度Cd的适应性[J].北方园艺,2024,(19):9.[doi:10.11937/bfyy.20240877]
PEI Yunxia,NI Tianhong,ZHANG Wenjun,et al.Adaptation of Seed Germination and Seedling Growth of Brassica oleracea L.var.capitata L. to Different Concentrations of Cd[J].Northern Horticulture,2024,(23):9.[doi:10.11937/bfyy.20240877]
备注/Memo
.第一作者简介:曹冰东(1995-),女,硕士研究生,研究方向为蔬菜遗传育种。E-mail:1763701494@qq.com.责任作者:孟平红(1966-),女,博士,研究员,现主要从事蔬菜遗传育种与生物技术等研究工作。E-mail:164931596@qq.com.基金项目:贵州省科技厅资助项目(黔科合支撑[2022]一般086);贵州省农业科学院资源专项资助项目(黔农科院种质资源[2022]03号);贵州省蔬菜现代农业产业技术体系资助项目(GZCYTX2022-01);贵阳市蔬菜种质资源研究中心建设资助项目(筑科合同[2021]5-1号);贵阳市蔬菜新品种新技术试验示范资助项目(筑科合同[2021]3-6号);贵州省教育厅青年科技人才成长资助项目(黔教科合KY字[2017]284号)。收稿日期:2022-04-22