大花红山茶花粉形态特征和培养条件及其储藏过程的生理动态分析

Pollen Morphology,Storage Condition and Physiologically Dynamic Change during Storage of Camellia magniflora

以大花红山茶花粉为材料,用扫描电子显微镜观察花粉表观形态,采用离体培养法研究花粉的萌发特性,并就不同贮藏温度与时间对花粉萌发率、超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性的影响进行了分析。结果表明:(1)大花红山茶花粉极面观呈正三角形,沟界极区较宽;花粉粒纹饰为穴状、小沟状雕饰纹;花粉粒平均极轴长为71.05μm;说明大花红山茶在山茶属中具有较为特殊的分类地位和系统进化地位。(2)大花红山茶花粉萌发的最适宜培养基为:150g·L-1蔗糖+20mg·L-1硼酸+100mg·L-1 KNO3+50mg·L-1MgSO4+300mg·L-1 Ca(NO3)2,花粉萌发率可达68.10%。(3)大花红山茶花粉最佳贮藏条件为-80℃;-80℃下贮藏360d后,萌发率仍可达到39.40%。(4)-20℃、-80℃贮藏条件下,大花红山茶花粉3种保护酶(SOD、POD、CAT)的活性较高,花粉抗氧化、抗衰老的能力较强;且在花粉贮藏过程中,3种酶活性高峰值前后花粉萌发率出现大幅度下降。(5)不同贮藏温度下,3种保护酶的作用不同,常温下,SOD为敏感性保护酶;4℃、-20℃下,POD为敏感性保护酶;-80℃下CAT为敏感性保护酶;3种酶活性对花粉萌发率的影响依次为:过氧化氢酶>超氧化物歧化酶>过氧化物酶。

The pollen morphology of Camellia magniflora was investigated under scanning electronic microscope（SEM）,Fresh pollen of C.magniflora was took as materials to test the pollen viability by in vitro.Effects of different store methods on the pollen viability,SOD,POD and CAT were also investigated.The results show that（1）The polar face is a equilateral triangle,the germinal aperture is wider on the polar.The exine ornamentation were mainly perforate,sulciform.The average length of polar axis is 71.05μm,which show C.magniflorashould occupy special taxonomic status in the genus Camellia.（2）The best combination for pollen germination is 150g·L^-1 sucrose,20mg·L^-1 boric acid,100mg·L^-1 KNO3,50mg·L^-1 MgSO4,300mg·L^-1 Ca（NO3）2.The germination rate reached 68.10%.（3）-80℃sealed storage is the best storage condition to keep the vitality of pollen,the germination rate of pollen reached 39.40% after 360 d storage.（4）The three kinds of enzyme activities were higher,Antioxidant,aging ability were also higher in-20℃ and -80℃ storage.The germination proportion of pollen decreased significantly when the activities of SOD,POD and CAT were around the highest levels;（5）The three enzymes had different responses to the storage temperature.SOD was a responding protection enzymes at normal temperature,POD was a responding protection enzymes at 4℃ and -20℃,while CAT was a responding protection enzymes at-80℃.The effect of three kinds of enzymes on pollen germination is CATSODPOD.