杂种白杨717-1B_4组培技术研究
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摘要
为保持杂种白杨717-1B4的优良性状,解决生根困难瓶颈,建立了生理年龄一致性和容器化生根炼苗的组织培养繁育技术。结果表明:外植体为母株根蘖的根际萌芽,以有效氯1.0%的乙氰脲酸钠溶液灭菌25 min,接种在MS培养基上效果最好,再生不定芽达75.91%;以再生不定芽3个以上的外植体进行继代培养,继代周期为30 d,平均再生不定芽12.2个;外植体再生的不定芽扩繁1次转移至营养钵基质上生根和练苗,增殖系数达3.73,增殖培养基为MS+6-BA0.5 mg/L+NAA 0.2 mg/L;生根培养基中IBA为2.0 mg/L生根最好,生根率达98.89%;揭开培养瓶盖驯化小植株容器苗,使之适应自然环境,驯化成活率达92.5%。
In order to maintain the maternal excellent features in offspring and refrain from difficult rooting in vegetative propagation of hybrid aspen,established a juvenility and container rooting-hardening micropropagation system in the present work. The result showed that explants taken from the sprouting between root-shoot junctions of maternal plants,sterilized by soaked 1. 0% effective chlorine of sodium dichloroisocyanurate( Na DCC) solution for25 min,cultured on MS medium,75. 91% of them regenerated juvenile shoots. The explants regenerated more than3 juvenile shoots were selected and sub-cultured on the same fresh medium in each 30 days,each explant produced12. 2 shoots on average. The shoots from explants multiplicated one time before did container rooting and hardening.The shoots increased 3. 73 times on MS medium containing 6-BA 0. 5 mg/L and NAA 0. 2 mg/L,respectively. The rooting rate reached the highest at 98. 89% when IBA was 2. 0 mg/L in rooting medium. The container plantlets hardened in reversion and acclimatization by shifting and opening gradually the tissue culture bottles. The container rooted aspen plants survived at 92. 5% in the acclimatization stage.
In order to maintain the maternal excellent features in offspring and refrain from difficult rooting in vegetative propagation of hybrid aspen,established a juvenility and container rooting-hardening micropropagation system in the present work. The result showed that explants taken from the sprouting between root-shoot junctions of maternal plants,sterilized by soaked 1. 0% effective chlorine of sodium dichloroisocyanurate( Na DCC) solution for25 min,cultured on MS medium,75. 91% of them regenerated juvenile shoots. The explants regenerated more than3 juvenile shoots were selected and sub-cultured on the same fresh medium in each 30 days,each explant produced12. 2 shoots on average. The shoots from explants multiplicated one time before did container rooting and hardening.The shoots increased 3. 73 times on MS medium containing 6-BA 0. 5 mg/L and NAA 0. 2 mg/L,respectively. The rooting rate reached the highest at 98. 89% when IBA was 2. 0 mg/L in rooting medium. The container plantlets hardened in reversion and acclimatization by shifting and opening gradually the tissue culture bottles. The container rooted aspen plants survived at 92. 5% in the acclimatization stage.
引文
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[12]卢小三.两个杨树无性系再生体系的建立及抗虫基因Cry1C+9C转化的初步研究[D].武汉:华中农业大学,2010.
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[20]周小红,周艳威,张元莉,等.杉木成年优良无性系的不定芽增殖研究[J].林业科学研究,2013,26(3):299-304.
[21]李金雨,洪丽萍.蝴蝶兰不定芽途径的组织培养技术[J].热带作物学报,2010,31(4):610-613.
[22]刘晓梅.白杨新杂种茎段组培技术的研究[D].杨凌:西北农林科技大学,2010.
[23]Oliveira A B D,Alencar N L M,Prisco J T,et al.Accumulation of organic and inorganic solutes in Na Clstressed sorghum seedlings from aged and primed seeds[J].Scientia Agricola,2011,68(6):632-637.
[24]杜美娥,张伟,刘洁.不同浓度IBA对大叶黄杨扦插生根效果的影响[J].湖北农业科学,2015(24):6268-6271.
[25]朱之悌.毛白杨多圃配套系列育苗新技术研究[J].北京林业大学学报,2002,24(S1):4-44.
[2]麻德业,李丕全,汪清锐.白杨派难生根树种快速繁殖技术研究[J].山西农业大学学报(自然科学版),2003,23(2):161-163.
[3]杨青,王言,申菲.不飞絮741杨转生根基因的研究[J].中国科技教育,2008(4):29-30.
[4]Yan-Min L I,Meng Y E,Zhao X S.Tissue culture and rapid propagation of Prunus serrulata Lindl.‘Royal Burgundy’[J].Plant Physiology Communications,2008,44(6):1163-1164.
[5]宁涛,宁军,韩成明.银中杨茎的组织培养及快速无性繁殖[J].辽宁大学学报(自然科学版),2002,29(4):382-384.
[6]张冰玉,苏晓华,郑书星.山新杨叶片高频率植株再生体系建立及其遗传稳定性[J].北京林业大学学报,2008,30(3):68-73.
[7]Liang C F,Zuo J B,Cheng Y I.Tissue culture and rapid propagation of triploid Populus tomentosa Carr.[J].Research of Agricultural Modernization,2007,28(4):507-509.
[8]郑洪,黄国伟,陈慧玲,等.美洲黑杨杂种优良无性系扦插生根对比研究[J].中国农学通报,2016(4):9-13.
[9]周洲,李永丽,源春彦,等.欧美杨POR组培再生系统研究[J].河南农业大学学报,2015(1):52-56.
[10]Yi L I,Li D I,Hong T.Induction preservation and differentiation of calli of Populus×Jianhumao[J].Acta Botanica Boreali-Occidentalia Sinica,2002,22(3):656-660.
[11]樊军锋,李玲,韩一凡.84 K杨叶片外植体再生系统的建立[J].西北林学院学报,2002,17(2):33-36.
[12]卢小三.两个杨树无性系再生体系的建立及抗虫基因Cry1C+9C转化的初步研究[D].武汉:华中农业大学,2010.
[13]张冰玉,苏晓华,郑书星.山新杨叶片高频率植株再生体系建立及其遗传稳定性[J].北京林业大学报,2008,30(3):68-73.
[14]李世承,佟凤琴.山杨,大青杨,毛白杨组织培养的研究[J].辽宁大学学报(自然科学版),1995(2):59-62.
[15]王斌,李百炼,张金凤,等.杂种白杨离体再生体系的建立[J].西北植物学报,2009,29(4):704-710.
[16]郭长花.白杨年龄与位置效应的生理生化机制研究[D].北京:北京林业大学,2008.
[17]Razdan M K.An introduction to plant tissue culture[M].2nd.Nova Science Publishers,Inc.,2003.
[18]张倩,王华芳.牡丹试管苗生根与移栽技术研究进展[J].园艺学报,2012,39(9):1819-1828.
[19]吕化荣.柑橘多倍体愈伤组织长期继代保存过程中遗传变异的研究[D].武汉:华中农业大学,2014.
[20]周小红,周艳威,张元莉,等.杉木成年优良无性系的不定芽增殖研究[J].林业科学研究,2013,26(3):299-304.
[21]李金雨,洪丽萍.蝴蝶兰不定芽途径的组织培养技术[J].热带作物学报,2010,31(4):610-613.
[22]刘晓梅.白杨新杂种茎段组培技术的研究[D].杨凌:西北农林科技大学,2010.
[23]Oliveira A B D,Alencar N L M,Prisco J T,et al.Accumulation of organic and inorganic solutes in Na Clstressed sorghum seedlings from aged and primed seeds[J].Scientia Agricola,2011,68(6):632-637.
[24]杜美娥,张伟,刘洁.不同浓度IBA对大叶黄杨扦插生根效果的影响[J].湖北农业科学,2015(24):6268-6271.
[25]朱之悌.毛白杨多圃配套系列育苗新技术研究[J].北京林业大学学报,2002,24(S1):4-44.