2个荔枝品种胚性愈伤组织诱导体胚发生过程的细胞学观察
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摘要
以'新球蜜荔'和'大丁香'2个荔枝品种胚性愈伤组织为材料,利用石蜡切片技术研究了'新球蜜荔'和'大丁香'接种在TD(MS+0.1 mg/L NAA+0.5 mg/L TDZ+0.1 g/L肌醇+0.4 g/L水解乳蛋白(LH)+100 m L/L椰汁+60 g/L蔗糖+10 g/L琼脂)培养基(XQTD和DDXTD)与'新球蜜荔'接种在TX(MS+0.1 mg/L NAA+5 mg/L ZT+0.1 g/L肌醇+0.4 g/L水解乳蛋白(LH)+100 ml/L椰汁+60 g/L蔗糖+10 g/L琼脂)培养基(XQTX)上各培养物体胚发生过程中细胞学变化。结果显示:XQTD从6 d开始变褐,9 d时观察到球状凸起,石蜡切片中观察到球形胚结构,12~21 d时观察到球形胚、心形胚和鱼雷形胚等不同形态体胚;而DDXTD从9 d开始逐渐变褐,15~21 d时石蜡切片中能看到少量的球形胚和心形胚。XQTX在第12 d时观察到有完整原形成层的球形胚,21 d时石蜡切片中观察到子叶形胚。2个荔枝品种的体胚发生过程中都观察到内起源和外起源两种起源方式,体胚发生均为单细胞起源,都经历了球形胚、心形胚、鱼雷形胚和子叶形胚等相似的发育过程。
Comparative analysis of the structural changes in the embryonic callus of litchi cultivar 'Xinqiumili'(XQTD)and 'Dadingxiang'(DDXTD)inoculated on TD medium(MS+0.1 mg/L NAA+0.5 mg/L TDZ+0.1 g/L inositol+0.4 g/L LH+100 ml/L coconut juice+60 g/L sucrose+10 g/L agar), and the embryonic callus of 'Xingqiumili'(XQTX) inoculated on TX medium(MS+0.1 mg/L NAA+5 mg/L ZT+0.1 g/L inositol+0.4 g/L LH+100 ml/L coconut juice+60 g/L sucrose+10 g/L agar), were studied during the somatic embryogenesis induced on different induction medium through paraffin sections. XQTD became brown at 6 d. Globular bulges were observed at 9 d, and globular embryo structures were observed in paraffin sections. Different somatic embryos such as globular embryo, heart-shaped embryo and torpedo-shaped embryo were observed at 12-21 d. DDXTD was brown at 9 d. A small amount of globular embryos and heart-shaped embryos could be seen in paraffin sections at 15-21 d. XQTX were observed globular embryos with intact primary cambium at 12 d, and cotyledon embryos were observed in paraffin sections at 21 d. In the course of somatic embryogenesis, two sources of internal and external origin were observed in the two litchi cultivars. Somatic embryogenesis was a single cell origin, and it had similar developmental processes, such as globular embryo, heart-shaped embryo, torpedo shaped embryo and cotyledon embryo.
Comparative analysis of the structural changes in the embryonic callus of litchi cultivar 'Xinqiumili'(XQTD)and 'Dadingxiang'(DDXTD)inoculated on TD medium(MS+0.1 mg/L NAA+0.5 mg/L TDZ+0.1 g/L inositol+0.4 g/L LH+100 ml/L coconut juice+60 g/L sucrose+10 g/L agar), and the embryonic callus of 'Xingqiumili'(XQTX) inoculated on TX medium(MS+0.1 mg/L NAA+5 mg/L ZT+0.1 g/L inositol+0.4 g/L LH+100 ml/L coconut juice+60 g/L sucrose+10 g/L agar), were studied during the somatic embryogenesis induced on different induction medium through paraffin sections. XQTD became brown at 6 d. Globular bulges were observed at 9 d, and globular embryo structures were observed in paraffin sections. Different somatic embryos such as globular embryo, heart-shaped embryo and torpedo-shaped embryo were observed at 12-21 d. DDXTD was brown at 9 d. A small amount of globular embryos and heart-shaped embryos could be seen in paraffin sections at 15-21 d. XQTX were observed globular embryos with intact primary cambium at 12 d, and cotyledon embryos were observed in paraffin sections at 21 d. In the course of somatic embryogenesis, two sources of internal and external origin were observed in the two litchi cultivars. Somatic embryogenesis was a single cell origin, and it had similar developmental processes, such as globular embryo, heart-shaped embryo, torpedo shaped embryo and cotyledon embryo.
引文
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[31]张宝红,刘方,刘志红,等.外源激素对棉花体细胞胚胎发生及发育的调控作用[J].棉花学报,2000,12(1):17-21.
[32]杨凤美.葡萄风信子体细胞胚发生与植株再生[D].西安:西北农林科技大学,2013.
[33]方少秋.菠萝体细胞胚发生及遗传转化的研究[D].广州:华南农
[2]Yu C H,Chen Z G,Lu L X,et al.Somatic embryogenesis and plant regeneration from litchi protoplasts isolated from embryogenic suspensions[J].Plant Cell,Tissue and Organ Culture,2000,61(1):51-58.
[3]Wang G,Li H L,Wang J B.In vitro regeneration of‘Feizixiao’litchi(Litchi chinensis Sonn.)[J].African Journal of Biotechnology,2016,15(22):1 026-1 034.
[4]俞长河,陈振光.荔枝胚性悬浮培养物的建立、保持和优化原生质体分离的研究[J].热带作物学报,1998,19(3):16-20.
[5]邝哲师,周丽侬,马雪筠,等.荔枝体胚发生及成苗研究[J].广东农业科学,1997,24(1):16-18.
[6]吕柳新,余小玲.荔枝幼胚的离体培养[J].福建农学院学报,1993(4):410-413.
[7]王果,李焕苓,王家保.2个海南荔枝稀优品种花药培养诱导胚性愈伤组织的研究[J].热带作物学报,2013,34(4):669-674.
[8]邝哲师,周丽依,马雪筠,等.荔枝组织培养中胚状体产生的类型及分析[J].果树科学,1996(1):25-28.
[9]邓朝军,易干军,曾继吾,等.荔枝体胚发生过程中的细胞学观察[J].果树学报,2006,23(5):760-762.
[10]王果.几个荔枝品种离体再生技术体系的研究[D].海口:海南大学,2013.
[11]陈春玲,赖钟雄.龙眼胚性愈伤组织体胚发生同步化调控及组织细胞学观察[J].福建农林大学学报(自然科学版),2002,31(2):192-194.
[12]达克东,张松,臧运祥,等.苹果离体叶片培养直接体细胞胚胎发生的形态学研究[J].核农学报,2004,18(2):118-120.
[13]肖洁凝,吴永杰,陈云凤,等.芒果植株再生及其在生物技术中的应用[J].植物生理学通讯,2004(5):638-642.
[14]桑庆亮.荔枝体细胞胚胎发生的研究进展[J].福建农业大学学报,2000,29(3):311-315.
[15]关亚丽,潘琴,丁朝.影响植物体细胞胚发生的几个因素[J].海南广播电视大学学报,2009(2):114-116.
[16]王非,薛白鹭,王一然,等.园林植物体胚发生的研究进展[J].安徽农业科学,2011,39(1):18-20.
[17]王萍,钟影,王恩旭,等.诱导不同基因型大豆未成熟子叶体细胞胚胎发生的研究[J].作物杂志,2012(2):65-68.
[18]刘勇刚,徐子勤,郝建国.温度和水分对不同基因型小麦未成熟胚体细胞胚胎发生以及分化能力的影响[J].西北植物学报,2001,21(3):425-431,612.
[19]朱彦涛,李殿荣,杨淑慎.低温预处理和基因型对甘蓝型油菜小孢子胚胎发生的影响[J].西北农林科技大学学报(自然科学版),2005,33(5):88-94.
[20]范国强,翟晓巧,马新业,等.两种基因型泡桐体细胞胚胎发生及植株再生[J].核农学报,2005,19(4):274-278,254.
[21]冯大领,孟祥书,王艳辉,等.植物生长调节剂在植物体细胞胚发生中的应用[J].核农学报,2007,21(3):256-260.
[22]陈云凤,张春荣,黄霞,等.TDZ对植物体细胞胚胎发生的作用[J].植物生理学通讯,2006,42(1):127-133.
[23]崔凯荣,邢更生,周功克,等.植物激素对体细胞胚胎发生的诱导与调节[J].遗传,2000,22(5):349-354.
[24]陈金慧,施季森,诸葛强,等.植物体细胞胚胎发生机理的研究进展[J].南京林业大学学报(自然科学版),2003,27(1):75-80.
[25]郭文义,白永富,赖钟雄,等.烟草体细胞胚胎发生再生植株[J].福建农业大学学报,2004(3):369-372
[26]林荣双,王庆华,梁丽琨,等.TDZ诱导花生幼叶的不定芽和体细胞胚发生的组织学观察[J].植物研究,2003,23(2):169-171,263.
[27]Kumar V,Moyo M,Staden J V.Somatic embryogenesis of Pelargonium sidoides DC[J].Plant Cell Tissue&Organ Culture,2015,121(3):571-577.
[28]李红.防城金花茶体细胞胚状体发生的研究[J].广西园艺,1999(3):2-3.
[29]刘宝光.红皮云杉体细胞胚形成及不定芽诱导[D].东北林业大学,2008.
[30]陈金慧,施季森,诸葛强,等.杂交鹅掌楸体细胞胚胎发生研究[J].林业科学,2003,39(4):49-53,177.
[31]张宝红,刘方,刘志红,等.外源激素对棉花体细胞胚胎发生及发育的调控作用[J].棉花学报,2000,12(1):17-21.
[32]杨凤美.葡萄风信子体细胞胚发生与植株再生[D].西安:西北农林科技大学,2013.
[33]方少秋.菠萝体细胞胚发生及遗传转化的研究[D].广州:华南农