摘要
以‘月光’枣为试材,研究除草剂二甲戊乐灵在枣树上的多倍体诱变效果,包括二甲戊乐灵浓度、作用时间、施用时期对愈伤组织形成、芽再生及多倍体诱变的影响等。结果表明:二甲戊乐灵明显抑制了枣愈伤组织生长和芽再生,且随着浓度(0.01%～0.1%)的升高,出愈率和单枝截面出芽数均呈逐渐下降趋势;作用时间(12～48 h)对愈伤组织生长及芽再生能力影响的差异不明显;随着施用时期(6～10 d)的推移,单枝截面出芽数增多,截枝处理10 d后施用二甲戊乐灵的出愈率、单枝截面出芽数均最高;优化后的条件下,即二甲戊乐灵浓度0.025%～0.05%,作用时间24～48 h,施用时期为愈伤组织诱导(施用4.0 mg/L TDZ+2.0 mg/L AgNO_3,12～14 h)6～10 d后,获得了80个再生芽,成功诱变出3个四倍体再生芽;与秋水仙素相比,二甲戊乐灵诱导的出芽时期晚10 d左右,出芽总数较低,但其诱导的芽变异率为3.57%,较秋水仙素诱导的高659.57%。
The application of pendimethalin, a novel polyploid agent for Chinese jujube, on ‘Yueguang' jujube was explored to establish a suitable polyploid induction method. The prior concentration, duration and application time of pendimethalin were settled and we evaluated pendimethalin induction efficiency. Pendimethalin evidently inhibited the callus growth and shoot regeneration. The regeneration of callus and bud decreased gradually with the increase of pendimethalin concentration(0.01%～0.1%), and extended when polyploid induction commencement on 6 d to 10 d after TDZ treatment, approaching the top on 10 d. No significant effect on callus growth and shoot regeneration was observed when inducted 12 h to 48 h. The suitable induction protocol was applying pendimethalin(0.025%～0.05%) on callus(for about24～48 h) on 6～10 d after callus induction with thidiazuro(4.0 mg/L TDZ+2.0 mg/L AgNO_3 for 12～14 h), especially 10 d after induction. The modified protocol successfully caused three tetraploid regenerated shoots emerging out of all the 80 branches in this test. When compared with traditional mutagenic agent colchicine, pendimethalin treatment produced fewer regeneration buds, emerging 10 d later as well, yet higher mutation ratio(3.57%), which was 659.57% higher than colchicine induced.
引文
[1]陶抵辉,刘明月,肖君泽,等.生物多倍体诱导方法研究进展.生命科学研究,2007,11(4):6-13.
[2]Barandalla L,Ritter E,Galarreta J I R D.Oryzalin treatment of potato diploids yields tetraploid and chimeric plants from which euploids could be derived by callus induction.Potato Resarch,2006,49(2):143-154.
[3]Allum J F,Bringloe D H,Roberts A V.Chromosome doubling in a Rosa rugosa Thunb.hybrid by exposure of in vitro nodes to oryzalin:the effects of node length,oryzalin concentration and exposure time.Plant Cell Reports,2007,26:1977-1984.
[4]Nadler J D,Pooler M,Olsen R T,et al.In vitro induction of polyploidy in Cercis glabra Pamp.Scientia Horticulturae,2012,148:126-130.
[5]简佳,方李琴,谭欣,等.盆栻亚洲百合的杂家和染色体加倍.农业生物技术学报,2013,21(5):627-630.
[6]Chalak L,Legave J M.Oryzalin combined with adventitious regeneration for an efficient chromosomoe doubling of trihaploid kiwifruit.Plant Cell Reports,1996,16(1):97.
[7]黄新文.氟乐灵对辣椒染色体加倍的诱导效果.贵州农业科学,2011,39(7):13-15.
[8]魏育国,蒋菊芳.氟乐灵诱导甜瓜四倍体研究初探.华北农学报,2006(增刊):73-76.
[9]邱长玉,林强,朱方容,等.氟乐灵在桑树染色体加倍试验上应用的初步研究.广西蚕叶,2017,54(4):1-5.
[10]Li Y,Whitesides J F,Rhosdes B.Preparation of nuclei from in vitro watermelon shoot tissue for cell flow cyto metry.Cueurbit Genetics Coop RPT,1999,22:43-46.
[11]Le C T,姜二花,王康,等.利用氟乐灵进行同源四倍体萝卜种质创新研究.基因组学与应用生物学,2012,31(3):276-281.
[12]赖红英,赵秀萍,林婉玉,等.二甲戊乐灵诱导金线莲多倍体.分子植物育种,2015,13(10):2355-2361.
[13]刘欢,高素萍,姜福星,等.二甲戊灵与秋水仙素离体诱导虎眼万年青多倍体发生的比较研究.核农学报,2014,28(11):1985-1992.
[14]阎志红,刘文革,赵胜杰,等.利用二硝基苯胺类除草剂离体诱导西瓜四倍体.园艺学报,2008,35(11):1621-1626.
[15]王康,何林池,魏小云,等.利用二甲戊乐灵创制薄皮甜瓜同源四倍体.西北农业学报,2015,24(4):114-119.
[16]路芳,代丼,刘孟军,等.二甲戊乐灵离体诱导酸枣八倍体的研究.河北农业大学学报,2010,33(3):42-45.
[17]蒋洪恩.枣(Ziziphus jujuba Mill.)多倍体诱导研究.保定:河北农业大学,2003.
[18]吴改娥.二倍体和四倍体枣的比较研究.保定:河北农业大学,2010.
[19]Shi Q H,Liu P,Wang J R,et al.A novel in vivo shoot regeneration system via callus in woody fruit tree Chinese jujube(Ziziphus jujuba Mill.).Scientia Horticulturae,2015,188:30-35.
[20]周香君,程智慧,孟焕文.二甲戊灵对离体大蒜染色体的加倍研究.西北植物学报,2009,29(12):2571-2575.
