摘要
【目的】筛选并优化薄壳山核桃花粉活力的测定方法,为薄壳山核桃的杂交育种提供参考。【方法】以韦科、苏普锐尔、基奥瓦、奥多姆4个薄壳山核桃品种的花粉为试验材料,分别采用MTT染色法、TTC染色法、过氧化物酶染色法、醋酸洋红染色法、I2-KI染色法和蓝墨水染色法对花粉进行染色,比较不同染色方法及不同染色溶液浓度处理间花粉活力的差异。采用正交试验对薄壳山核桃花粉萌发培养基配方进行优化,筛选适合薄壳山核桃花粉萌发的培养基。通过回归分析,探讨花粉染色率和离体萌发率之间的相关性,寻找最适合测定薄壳山核桃花粉活力的染色方法。【结果】除TTC染色法外,其余方法均能使薄壳山核桃花粉染色。适宜4个薄壳山核桃品种花粉萌发的培养基并不完全相同,适宜韦科、基奥瓦和奥多姆花粉萌发的培养基为5%蔗糖+0.01%硼酸+0.05%硝酸钙+25%PEG-4000,萌发率分别可达36.80%、46.32%、59.51%。适宜苏普锐尔花粉萌发的培养基为8%蔗糖+0.025%硼酸+0.025%硝酸钙+25%PEG-4000,萌发率为24.19%。采用MTT染色法所染深色花粉比例接近花粉萌发率,且所测花粉活力与花粉萌发率呈线性正相关(R2>0.686,P<0.001)。【结论】薄壳山核桃不同品种的最适花粉萌发培养基组分有差异,与其他染色法相比,MTT法更适用于薄壳山核桃花粉活力的快速检测。
【Objective】In order to screen and optimize the determination method of the pollen viability of pecan,provide a reference for the cross breeding of pecan.【Method】The pollen of four pecan cultivars,Waco,Superior,Kiowa,and Odom,were used as test materials,the pollen was stained with 2,5-diphenylmonotetrazolium bromide(MTT),2,3,5-triphenyl tetrazolium chloride(TTC),peroxide,acetic carmine,I2-KI and blue ink dyeing methods,respectively,the differences in pollen viability between different staining methods and different stain concentrations were compared.The orthogonal test was used to optimize the pecan pollen germination medium formulation and to screen the medium suitable for pecan pollen germination.Through regression analysis,the correlation between pollen staining rate and in vitro germination rate was explored,so as to find the most suitable staining method for measuring the pollen viability of pecan.【Result】Except for TTC dyeing method,other methods could dye pecan pollen.The media suitable for pollen germination of the four pecan cultivars were not identical.The suitable media for pollen of Waco,Kiowa and Odom were:5%sucrose+0.01%boric acid+0.05%calcium nitrate+25%PEG-4000 and the germination rates were 36.80%,46.32%and 59.51%,respectively.The suitable medium for Superior pollen was 8%sucrose+0.025%boric acid+0.025%calcium nitrate+25%PEG-4000,and the germination rate was 24.19%.The proportion of dark pollen stained by MTT staining was close to the pollen germination rate,and the measured pollen viability was linearly positively correlated with the pollen germination rate(R2>0.686,P<0.001).【Conclusion】The most suitable pollen germination medium for different varieties of pecans is different,MTT is more suitable for rapid detection of pollen viability of pecan compared with other methods.
作者
刘帅
彭方仁
LIU Shuai;PENG Fangren(Co-Innovation Center for Sustainable Forestry in Southern China,Nanjing Forestry University,Nanjing 210037,Jiangsu,China;College of Forestry,Nanjing Forestry University,Nanjing 210037,Jiangsu,China)
出处
《经济林研究》
北大核心
2023年第2期110-119,共10页
Non-wood Forest Research
基金
中央财政林业科技推广示范资金项目(苏[2022]TG04)
江苏省高校优势学科建设工程资助项目(PAPD)。
关键词
薄壳山核桃
花粉活力
萌发率
MTT
Carya illinoinensis
pollen viability
germination rate
MTT
