摘要
【目的】为油茶的单倍体育种提供参考。【方法】以普通油茶‘华硕’为试材,对其未授粉子房进行离体培养,诱导愈伤组织,研究消毒处理对油茶子房褐化和染菌的影响,及激素的种类和浓度、子房切割方式、子房发育时期、子房预处理方式等因素对愈伤组织诱导的影响,并使用筛选出的2种激素和蔗糖进行L9(33)正交试验,将正交试验结果进行极差分析和方差分析,进一步筛选油茶子房愈伤组织诱导的最佳培养基。【结果】选择开花当天的油茶未授粉花蕾,热激(35℃)预处理1 d后,剥出子房部分,采用75%乙醇溶液浸泡60 s、0.1%升汞溶液浸泡5 min的方式进行消毒处理,纵切后,接种到MS+60 g/L蔗糖+7 g/L琼脂+0.3 mg/L NAA+2 mg/L TDZ培养基上,愈伤组织诱导率最高,为72.34%。诱导出的愈伤组织呈浅绿色,质地紧密。TDZ和NAA显著影响了油茶子房的愈伤组织诱导率,且TDZ的影响大于NAA,蔗糖对诱导率的影响不显著。【结论】在各影响因素中,对油茶未授粉子房愈伤组织诱导率影响最明显的是激素的种类和浓度,适宜激素为NAA和TDZ,适宜的质量浓度分别为0.3、2.0 mg/L。子房的发育时期、切割方式、预处理方式等均会影响其愈伤组织诱导率,选择开花当天的子房,热激(35℃)预处理1 d并纵切,可以提高愈伤组织的诱导率。
【Objective】It provided reference for haploid breeding of Camellia oleifera.【Method】The unpollinated ovaries of C.oleifera‘Huashuo’were cultured and induced callus in vitro to study the effects of camellia ovary browning and infecting bacterium by disinfection treatment,and the combinations and concentrations of plant growth regulators,cutting methods,development stages,and pre-treatments on ovary callus induction.L9(33)orthogonal test was carried out using the two selected plant growth regulators and sucrose,and the results of orthogonal test were used for range analysis and variance analysis to further screen the optimal medium for callus induction of C.oleifera ovaries.【Result】Unpollinated buds of C.oleifera on the day of flowering were selected.After 1 day of heat shock pretreatment at 35℃,stripping out the ovary part and the ovaries should be sterilized with 75%ethanol for 1 min and 0.1%mercury dichloride solution for 5 min.The highest callus induction rate of ovaries was 72.34%when the ovules were cut lengthwise and inoculated on MS medium supplemented with 60 g/L sucrose,7 g/L AGAR,0.3 mg/L NAA and 2 mg/L TDZ.The induced callus was light green and dense in texture.NAA and TDZ significantly affected the callus induction rate of ovaries,and the effect of TDZ was greater than that of NAA,while the effect of sucrose on the induction rate was not significant.【Conclusion】Among the factors,the types and concentrations of plant growth regulators had the most obvious influence on callus induction rate.The optimal hormones were 0.3 mg/L NAA and 2 mg/L TDZ.In addition,the stage of ovary development,cutting and pretreatment all affected the callus induction rate of ovaries.Selecting ovaries on the day of flowering,pretreating at 35℃for 1 day and cutting ovaries longitudinally can improve the callus induction rate.
作者
靳皓然
鲁建军
余安
何杰
张紫缇
盘鑫海
袁德义
范晓明
JIN Haoran;LU Jianjun;YU An;HE Jie;ZHANG Ziti;PAN Xinhai;YUAN Deyi;FAN Xiaoming(The Key Laboratory of Non-wood Forest Products of National Forestry and Grassland Administration,Central South University of Forestry&Technology,Changsha 410004,Hunan,China;Changsha Forest Seedling Management Station,Changsha 410339,Hunan,China)
出处
《经济林研究》
北大核心
2022年第2期62-70,共9页
Non-wood Forest Research
基金
长沙市“揭榜挂帅”重大科技专项“高产油茶优异种质创制”(Kq2102007-02)
国家重点研发计划项目“木本油料重要性状形成与调控”(2018YFD1000603)。
关键词
普通油茶
未授粉子房
离体培养
单倍体育种
Camellia oleifera
unpollinated ovary
in vitro culture
haploid breeding
