4种冬青属植物花粉离体培养基筛选和贮藏条件的研究

Research on Medium Screening for in Vitro Germination and Storage Conditions of 4 Ilex Species

以4种冬青属植物花粉为试验材料,采用正交试验设计,分析了蔗糖、H3BO3和CaCl2组成的液体培养基对冬青花粉萌发率和花粉管生长的影响,筛选出适合4种冬青花粉离体培养的培养基,同时探讨了4℃和-20℃贮藏条件对冬青花粉生活力的影响。结果表明:1)短梗冬青、全缘冬青和大叶冬青适宜的蔗糖浓度为10%,大别山冬青为5%,短梗冬青、全缘冬青适宜的H3BO3浓度为0.002%,大别山冬青和大叶冬青为0.006%,CaCl2促进大叶冬青的花粉萌发和花粉管的生长,但抑制其他3种冬青。2)在4℃和-20℃贮藏条件下,4种冬青花粉随时间的增加萌发率均表现为先升后降,其中短梗冬青和大别山冬青在-20℃贮藏条件下花粉萌发率高于4℃;全缘冬青和大叶冬青花粉贮藏前期萌发率在4℃条件下高于-20℃,贮藏后期在-20℃条件下随着时间增加花粉萌发率下降趋势较4℃缓慢。3)花粉管长度在2种贮藏条件下随时间增加均表现为先增后减,短梗冬青花粉在4℃条件下花粉管长度增减幅度明显,但在-20℃条件下花粉管长度在贮藏56d无差异变化;全缘冬青、大叶冬青和大别山冬青花粉管在2种贮藏条件下先增后减趋势一致,但-20℃条件下随着时间增加花粉管长度减短趋势较4℃缓慢。4)4种冬青花粉萌发率与花粉管长度均存在一定的正相关性。综合得出,冬青花粉活力个体差异显著;4℃适合冬青花粉短时间贮藏,-20℃更适于冬青花粉的长时间贮藏;根据花期和花粉生活力丧失速度的差异,短梗冬青和全缘冬青适合作为冬青属种间杂交的父本,大叶冬青和大别山冬青适合做母本。

Pollens of 4 Ilex species were used as materials to screen out suitable media for in vitro culture.An orthogonal design was adopted to optimize the culture media and to examine the influences of the components （such as sucrose,boric acid,and CaCl 2） on the germination and the growth of pollen tubes.Impacts of two storage temperatures,i.e.,4℃and -20℃ on the growth vigor of the pollens were investigated.The results showed that 1） the optimum sucrose concentration for culturing I.buergeri,I.integra ,and I.latifolia was 10%,and 5% for I.dabieshanensis .The optimum boric acid concentration for I.buergeri and I.integra was 0.002%,and 0.006% for I.dabieshanensis and I.latifolia .CaCl 2 may promote pollen germination rate and tube growth of I.latifolia ,while inhibit the other 3 species.2） Under the temperatures of 4℃and -20℃,pollen germination rates of all 4 species increased in the early days then decreased.Pollen germination rates of I.buergeri and I.dabieshanensis were higher under -20℃ than 4℃.Pollen germination rates of I.integra and I.latifolia were higher under 4℃ than -20℃in the early days,but in the later days,reduced more slowly than that under 4℃.3） The growth of pollen tube length of 4 Ilex species all increased and then decreased with time,in which the increase and decrease for I.buergeri were significant under 4℃.Under -20℃,the pollen tube length almost kept consistent during 56-day storage,and the length had no significant difference.For the other three species,they all showed the same trends under two temperatures：the tube growth increased in the early days then decreased,while the rate of decrease under -20℃ was slower than under 4℃.4） There was a positive correlation between the germination rate and pollen tube length among 4 Ilex species.In conclusion,there were significant difference among the pollen viabilities of different species of Ilex ; 4℃was suitable for short storage of Ilex pollen ,while -20℃was better for long storage; according to florescence and viabilities of different Ilex species, I.buergeri and I.integra were suitable for male parent in cross breeding, I.latifolia and I.dabieshanensis were suitable for femal parents in cross breeding.