木薯同源四倍体的离体诱导及鉴定
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摘要
为获得木薯新品种四倍体种质资源,以木薯主栽品种南植199组培苗单芽茎段为试验材料,利用秋水仙素诱导同源四倍体,统计不同浓度秋水仙素对外植体成活率和诱导效果的影响,并对二倍体与同源四倍体进行形态学、解剖学和生理生化方面的比较。结果表明,外植体成活率随秋水仙素浓度升高而降低,0. 05～0. 10 g·L~(-1)是秋水仙素诱导木薯同源四倍体的适宜浓度范围。本研究获得了10个同源四倍体株系和1个嵌合体株系。与二倍体木薯相比,四倍体木薯叶片增厚,叶型变宽,保卫细胞增大,叶绿素含量增加。本研究拓宽了木薯种质资源,并为诱导鉴定木薯四倍体提供了一套高效的技术方法。
To obtain the tetraploid germplasm resources of cassava new variety,in vitro single-node axillary buds of cassava( cv. Nanzhi 199) were taken as explants to induced the autotetraploid. The effects of various concentration of colchicine on the survival rate and variation rate were investigated and the morphological characters,anatomical characters and chlorophyll content of normal and variant plants were also analyzed. The results showed that the survival rate of explants decreased with the increase of colchicine concentrations. Concentration range of 0. 05 ～ 0. 1 g·L~(-1) was suitable for inducing autotetraploid cassava. Identification of ploidy levels by flow cytometry( FCM) showed that 10 autotetraploid and one chimeras were obtained. Compared to the original diploid plants,the tetraploid plants showed significantly difference in color,thickness and surface of leaves. Moreover,the tetraploid plants had bigger guard cells and higher chlorophyll content.
To obtain the tetraploid germplasm resources of cassava new variety,in vitro single-node axillary buds of cassava( cv. Nanzhi 199) were taken as explants to induced the autotetraploid. The effects of various concentration of colchicine on the survival rate and variation rate were investigated and the morphological characters,anatomical characters and chlorophyll content of normal and variant plants were also analyzed. The results showed that the survival rate of explants decreased with the increase of colchicine concentrations. Concentration range of 0. 05 ～ 0. 1 g·L~(-1) was suitable for inducing autotetraploid cassava. Identification of ploidy levels by flow cytometry( FCM) showed that 10 autotetraploid and one chimeras were obtained. Compared to the original diploid plants,the tetraploid plants showed significantly difference in color,thickness and surface of leaves. Moreover,the tetraploid plants had bigger guard cells and higher chlorophyll content.
引文
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[23] Wang J,Shi L,Song S Y,Tian J,Kang X Y. Tetraploid production through zygotic chromosome doubling in Populus[J]. Silva Fennica,2013,47(2):1-12
[24]姚焱,李岩松,王艺霖,黎雨芯,赵树进,Yan Cui-jun.利用何首乌叶片气孔保卫细胞叶绿体数目鉴定植株倍性[J].中药材,2014,37(6):939-942
[25]闫艳华,杜京旗,高晓丽,李毛毛.燕麦不同品种染色体倍性的检测[J].种子,2017,36(6):42-44
[26] Wang L J,Sheng M Y,Wen P C,Du J Y. Morphological,physiological,cytological and phytochemical studies in diploid and colchicine induced tetraploid plants of Fagopyrumtataricum(L.)[J]. Botanical Studies,2017,58(2):1-12
[27] Xu C P,Huang Z,Liao T,Li Y,Kang X Y. In vitro tetraploid plants regeneration from leaf explants of multiple genotypes in Populus[J]. Plant Cell and Organ Culture,2016,125(1):1-9
[28]刘玲,葛春峰,王涛,陈孟龙,乔玉山.黄毛草莓叶片离体再生及其同源四倍体的诱导[J].核农学报,2017,31(1):51-58
[29]邱芬,辛亚龙,唐军荣,李斌,何承忠,许程,辛培尧.不同倍性滇杨叶片解剖结构差异分析[J].中南林业科技大学学报,2017,37(6):56-59,69
[30] Javadian N,Karimzadeh G,Sharifi M,Moieni A,Behmanesh M. In vitro polyploidy induction:changes in morphology,podophyllotoxin biosynthesis,and expression of the related genes in Linum album(Linaceae)[J]. Planta,2017,245(6):1165-1178
[31] Chen R,Jiang W Z,Li Q L,Li X L,Chen X L,Yang Y S,Wu H.Comparison of seven colchicine-induced tetraploid clones with their original diploid clones in purple coneflower(Echinacea purpurea L.)[J]. Euphytica,2016,207(2):387-399
[32]安飞飞,凡杰,李庚虎,简纯平,李开绵.华南8号木薯及其四倍体诱导株系叶片蛋白质组及叶绿素荧光差异分析[J].中国农业科学,2013,46(19):3978-3987
[33]张虹,龙宏周,路国栋,陈任.黑果枸杞多倍体诱导及鉴定[J].核农学报,2017,31(1):59-65
[2]严华兵,叶剑秋,李开绵.中国木薯育种研究进展[J].中国农学通报,2015,31(15):63-70
[3]凡杰.木薯多倍体的倍性鉴定及蛋白组学研究[D].海口:海南大学,2012
[4] Bagheri M, Mansouri H. Effect of induced polyploidy on somebiochemical parameters in Cannabis sativa L.[J]. Applied Biochemistry&Biotechnology,2015,175(5):2366-237
[5]周慧文,冯斗,严华兵.秋水仙素离体诱导多倍体研究进展[J].核农学报,2015,29(7):1307-1315
[6]石庆华,刘平,刘孟军.果树倍性育种研究进展[J].园艺学报,2012,39(9):1639-1654
[7] Sreekumari M T,Jos J S,Nair S G.‘Sree Harsha’:a superior triploid hybrid in cassava[J]. Euphytica,1999,106(1):1-6
[8]黄慧德.木薯多倍突变体新品种[J].中国农学通报,1985(5):20
[9] Zhou H W,Zeng W D,Yan H B. In vitro induction of tetraploids in cassavavariety‘Xinxuan 048’using colchicine[J]. Plant Cell and Organ Culture,2017,128(3):723-729
[10]张健.木薯离体培养诱导多倍体育种技术研究[D].海口:海南大学,2012
[11]聂扬眉,文峰,郭文武.秋水仙素诱导获得5个木薯品种的同源四倍体植株[J].华中农业大学学报,2013,32(6):8-12
[12]张俊娥,刘继红,邓秀新.采用倍性分析仪鉴定柑橘愈伤组织的遗传变异[J].遗传学报,2003,30(2):169-174
[13]周慧文.木薯四倍体离体诱导与诱变材料评价研究[D].南宁:广西大学,2016
[14]孔祥生.植物生理学实验技术[M].北京:中国农业出版社.2008
[15] Guo L Q,Xu W T,Zhang Y,Zhang J F,Wei Z H. Inducing triploids and tetraploids with high temperatures in Populus sect.Tacamahaca[J]. Plant Cell Reports,2017,36(2):313-326
[16] Leus L,Van Laere K,Dewitte A,Van Huylenbroeck J. Flow cytometry for plant breeding[J]. Acta Horticulturae,2009,836(836):221-226
[17]邵冰洁,万思琦,刘江淼,许建新,赵惠恩.黑果枸杞和宁夏枸杞的多倍体诱导和鉴定[J].分子植物育种,2018,16(8):2593-2599
[18]张源源,郭涵,袁红章,王祥军,黄肖,李媛,李维国.橡胶树不同倍性的气孔性状差异研究[J].热带作物学报,2017,38(3):389-394
[19]熊梅,伊风艳,石凤翎,高霞,高翠萍.苜蓿花培植株倍性的初步鉴定[J].种子,2015,34(5):13-16
[20]谢利,刘芳,易懋升,曾瑞珍,夏晴,黎杨辉,张志胜.蝴蝶兰倍性与叶部和花部性状的相关性[J].华南农业大学学报,2014,35(5):82-87
[21]马艳芝,客绍英.菘蓝四倍体株系细胞形态学和流式细胞术鉴定[J].中药材,2014,37(5):736-740
[22]卢璇.非洲菊组织培养及未受精胚珠苗的倍性鉴定[D].广州:华南农业大学,2016
[23] Wang J,Shi L,Song S Y,Tian J,Kang X Y. Tetraploid production through zygotic chromosome doubling in Populus[J]. Silva Fennica,2013,47(2):1-12
[24]姚焱,李岩松,王艺霖,黎雨芯,赵树进,Yan Cui-jun.利用何首乌叶片气孔保卫细胞叶绿体数目鉴定植株倍性[J].中药材,2014,37(6):939-942
[25]闫艳华,杜京旗,高晓丽,李毛毛.燕麦不同品种染色体倍性的检测[J].种子,2017,36(6):42-44
[26] Wang L J,Sheng M Y,Wen P C,Du J Y. Morphological,physiological,cytological and phytochemical studies in diploid and colchicine induced tetraploid plants of Fagopyrumtataricum(L.)[J]. Botanical Studies,2017,58(2):1-12
[27] Xu C P,Huang Z,Liao T,Li Y,Kang X Y. In vitro tetraploid plants regeneration from leaf explants of multiple genotypes in Populus[J]. Plant Cell and Organ Culture,2016,125(1):1-9
[28]刘玲,葛春峰,王涛,陈孟龙,乔玉山.黄毛草莓叶片离体再生及其同源四倍体的诱导[J].核农学报,2017,31(1):51-58
[29]邱芬,辛亚龙,唐军荣,李斌,何承忠,许程,辛培尧.不同倍性滇杨叶片解剖结构差异分析[J].中南林业科技大学学报,2017,37(6):56-59,69
[30] Javadian N,Karimzadeh G,Sharifi M,Moieni A,Behmanesh M. In vitro polyploidy induction:changes in morphology,podophyllotoxin biosynthesis,and expression of the related genes in Linum album(Linaceae)[J]. Planta,2017,245(6):1165-1178
[31] Chen R,Jiang W Z,Li Q L,Li X L,Chen X L,Yang Y S,Wu H.Comparison of seven colchicine-induced tetraploid clones with their original diploid clones in purple coneflower(Echinacea purpurea L.)[J]. Euphytica,2016,207(2):387-399
[32]安飞飞,凡杰,李庚虎,简纯平,李开绵.华南8号木薯及其四倍体诱导株系叶片蛋白质组及叶绿素荧光差异分析[J].中国农业科学,2013,46(19):3978-3987
[33]张虹,龙宏周,路国栋,陈任.黑果枸杞多倍体诱导及鉴定[J].核农学报,2017,31(1):59-65
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