摘要
为推动银白杨倍性育种进展,利用秋水仙碱溶液注射处理,对银白杨花粉染色体加倍技术进行了研究,并通过常规压片及间接免疫荧光技术观察了秋水仙碱处理对小孢子母细胞染色体行为和微管骨架分布的影响。结果表明:(1)施加秋水仙碱处理可诱导银白杨花粉染色体加倍,在小孢子母细胞减数分裂细线末期-粗线期施加0.5%秋水仙碱溶液注射处理7次,可获得最高达82.23%的2n花粉。由于注射处理5次与7次诱导效果差异不显著,从节约成本和降低毒害考虑,在实际应用中以0.5%秋水仙碱溶液注射处理5次为宜;(2)秋水仙碱处理导致银白杨小孢子母细胞减数分裂过程染色体行为异常,包括双线期细胞核仁数目增多、微核形成,终变期染色体紧贴核仁,中期I染色体发生极度收缩聚集成团;(3)受秋水仙碱影响,银白杨小孢子母细胞细胞质微管骨架分布发生紊乱。粗线期小孢子母细胞内微管骨架不再呈网状均匀分布于细胞质,而是聚集呈短带状,从而失去对染色体行为的控制。
In order to promote polyploid breeding of Populus alba L. , pollen chromosome doubling of P. alba with eolchicine injection and its cytological effect for chromosome behaviors and microtubule distribution Were investigated by squashed technique and indirect immunofluorescence. The results showed that, 1 ） Colchicine-treatment can induce pollen chromosome doubling of P. alba. Injecting 0.5% colchicine solution to male buds under late leptotene to pachytene stages for 7 times produced 82.23% 2n pollen. Because there was no significant difference for 2n pollen induction between 5 and 7 times injection, injecting 5 times with 0.5% colchicine solution is feasible in practice in consideration of cost and toxic action. 2 ） Colchicine-treatment led to abnormal meiotic chromosome behavior of microsporocytes, including increase of nucleolus number and micronuclei formation in diplotene stage, chromosomes clung to the nucleolus in diakinesis stage, and highly condensed chromosomes in metaphase I stage. 3） Distribution of microtubular cytoskeleton in microsporocytes of P. alba was abnormal, owing to colchicine-treatment. Mierotubules were no longer distributed as random networks in cytoplasm, but gathered together to form a short belt, resulting in losing control to chromosome behavior.
出处
《核农学报》
CAS
CSCD
北大核心
2014年第5期749-756,共8页
Journal of Nuclear Agricultural Sciences
基金
国家自然科学基金(31000306)
高等学校全国优秀博士学位论文作者专项资金(201267)
关键词
银白杨
花粉染色体加倍
秋水仙碱
染色体行为
微管骨架
Populus alba L.
Pollen chromosome doubling
Colchicine
Chromosome behavior
Microtubular cytoskeleton
