郁金香小孢子发育与花粉萌发特性研究  

Characteristics of microspore development and pollen germination of Tulipa gesneriana in vitro

作　　者：杨延红 宋桂全[1] 李孔栋 姜倩倩[1] 李光耀 孟迎迎 YANG Yanhong;SONG Guiquan;LI Kongdong;JIANG Qianqian;LI Guangyao;MENG Yingying(Key Laboratory of Biochemistry&Molecular Biology in Universities of Shandong,Weifang University,Weifang,Shandong 261000,China)

机构地区：[1]潍坊学院山东省高校生物化学与分子生物学重点实验室,山东潍坊261000

出　　处：《西北农林科技大学学报（自然科学版）》2022年第6期93-100,共8页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家自然科学基金项目(31301733);山东省高等学校科技发展计划项目(J18KA157);潍坊学院博士基金资助项目(2012BS19)。

摘　　要：【目的】探究郁金香(Tulipa gesneriana)小孢子发育过程及液体培养基组分对花粉萌发和生长的影响,明确花粉发育过程中的细胞学变化及花粉萌发的适宜培养基组分,为郁金香杂交育种及胚胎学研究奠定基础。【方法】以郁金香品种“阿帕尔顿”种球为材料,用体积分数0.05%高锰酸钾溶液消毒处理后,剥出种球内大小不同的花芽,利用4′,6-二脒基-2-苯基吲哚(DAPI)进行荧光染色,确定小孢子发育时期;将郁金香种球定植于花盆内,按常规方法培养8周后取新鲜花粉进行离体培养,以200 mg/L磷酸二氢钾+200 mg/L硫酸镁为基础培养基,分别添加蔗糖(0,50,100,150,200,250 g/L)、硼酸(0,60,80,100,120,140 mg/L)、氯化钙(0,100,300,500,700,900 mg/L),在1,3,5,7 h取花粉镜检确定各组分最合适的质量浓度;再在筛选的最佳培养基组分下培养1,3,5,7 h,确定花粉萌发的最佳培养时间。【结果】郁金香小孢子发育历经四分体时期、单核早中期、单核靠边期和二胞花粉时期,其中以四面体型四分体居多,偶见左右对称型;花粉发育过程中存在发育不同步、异常花粉等现象;蔗糖、硼酸和钙对郁金香花粉萌发和花粉管生长影响显著,最适液体培养基组分为基础培养基(200 mg/L磷酸二氢钾+200 mg/L硫酸镁)+150 g/L蔗糖+100 mg/L硼酸+500 mg/L氯化钙,在此条件下培养5 h,花粉萌发率高达69.60%,花粉管长度为439.12μm。【结论】郁金香小孢子发育过程中以四面体型四分体居多,偶见左右对称型。在最佳培养条件下,郁金香花粉萌发情况及花粉管生长状态最好。【Objective】This study explored the microspore development process and the effects of liquid medium components on pollen germination and pollen tube growth of Tulipa gesneriana,and clarified the cytological changes of pollen development process and the suitable medium components for pollen germination to lay foundation for hybrid breeding and embryology research of Tulipa gesneriana.【Method】The bulbs of Tulipa gesneriana‘Apeldoorn’were disinfected with potassium permanganate solution at volume fraction of 0.05%and flower buds with different sizes were peeled out.The microspore development period was determined by fluorescence staining with 4′,6-diamidino-2-phenylindole(DAPI).After 8 weeks of culture according to the conventional method,fresh pollen was taken for in vitro culture.Basic medium was 200 mg/L potassium dihydrogen phosphate and 200 mg/L magnesium sulfate,and sucrose(0,50,100,150,200 and 250 g/L),boric acid(0,60,80,100,120 and 140 mg/L)and calcium chloride(0,100,300,500,700 and 900 mg/L)were added into the basic medium.Pollen was taken for microscopic examination at 1,3,5 and 7 hours to determine the most appropriate concentration of each component.Then pollen cultured for 1,3,5 and 7 h under optimal medium components was analyzed to determine appropriate culture time for pollen germination.【Result】The microspore of Tulipa gesneriana‘Apeldoorn’developed in four periods of tetrad,early-or mid-uninucleate,late uninucleate and bicellular pollen with most were tetrahedral and occasionally some were isobilateral.In the process of pollen development,there were phenomena such as development asynchrony and abnormal pollen.Sucrose,boric acid and calcium in medium had significant influence on pollen germination and pollen tube growth.The optimum liquid medium components included basic medium(200 mg/L potassium dihydrogen phosphate+200 mg/L magnesium sulfate),150 g/L sucrose,100 mg/L H_(3)BO_(3) and 500 mg/L CaCl_(2).Under these conditions,the pollen germination rate reached 69.60%and the poll

关 键 词：郁金香 小孢子发育 花粉萌发 培养基组分 

分 类 号：S682.263[农业科学—观赏园艺]