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当前位置: 首页 > 文章 > 利用共聚焦扫描显微镜观察甜樱桃雌配子体的发育 果树学报 2014 (1) 36-40
Position: Home > Articles > Observation of sweet cherry female gametophyte development with Con-focal Scanning Microscope Journal of Fruit Science 2014 (1) 36-40

利用共聚焦扫描显微镜观察甜樱桃雌配子体的发育

作  者：
朱东姿;王甲威;宗晓娟;魏海蓉;刘庆忠
单  位：
山东省果树研究所·山东省果树生物技术育种重点实验室
关键词：
甜樱桃;雌配子体;激光共聚焦扫描显微镜;延迟发育
摘  要：
【目的】快速准确的观察甜樱桃雌配子体的发育过程。【方法】应用激光共聚焦扫描显微镜法(LSCM),对甜樱桃品种‘美早’和‘布鲁克斯’的雌配子体发育过程进行了观察。将甜樱桃胚囊用戊二醛固定,脱水透明后直接用激光扫描共聚焦显微镜观察,用488 nm激光激发后,细胞核和核仁有很强的自发荧光,细胞质液泡则没有自发荧光,因此根据胚囊内核的数量、位置、大小,液泡的有无、位置等可以很清楚地观察雌配子体的发育时期。【结果】利用这种新方法观察到的雌配子体发育过程与用石蜡切片法观察的一致,但是LSCM法更简便、省时、准确;依照拟南芥中雌配子体发生的划分方法,我们把甜樱桃的雌配子体发生过程划分为6个时期。利用激光共聚焦扫描显微镜法通过延迟胚囊的发育,对‘美早’和‘布鲁克斯’甜樱桃胚囊进行观察统计,结果发现开花当天的‘美早’中没有胚囊的胚珠和胚囊发生解体的胚珠占41%,而‘布鲁克斯’中仅为21%。当胚囊延迟发育3 d后,‘美早’中胚囊的解体率达45%,败育率高达66%;‘布鲁克斯’中有35.6%的胚囊发生解体,败育率是46.6%。【结论】由此推测,‘美早’胚囊容易败育且发育成熟后解体快,而‘布鲁克斯’的胚囊败育率低,活力高。
译  名：
Observation of sweet cherry female gametophyte development with Con-focal Scanning Microscope
作  者：
ZHU Dong-zi;WANG Jia-wei;ZONG Xiao-juan;WEI Hai-rong;LIU Qing-zhong;Shandong Institute of Pomology·Key Laboratory for Fruit Biotechnology Breeding of Shandong Province;
关键词：
Sweet cherry;;Female gametophyte;;Confocal laser scanning microscopy;;Delayed development
摘  要：
【Objective】 In order to quickly and accurately observe the development of the sweet cherry female gametophyte, we observed the female gametophyte development of sweet cherry varieties ‘Tieton'and ‘Brooks' s' by laser scanning confocal microscopy(LSCM). 【Method】 Sweet cherry pistils were fixed in glutaraldehyde, then dehydrated and cleared. Then embryo sacs were dissected and observed with a laser scanning microscope with a 488 nm argon laser. When illuminated by 488 nm laser lines, cytoplasm,nucleoplasm and nucleoli exhibited different autofluorescence, nucleoli were distinguished by their extremely bright autofluorescence, and vacuoles were distinguished without fluorescence. Therefore, according to nucleoli's number, size, location, and vacuole's presence or absence, location etc., the developmental stages of female gametophyte could be clearly distinguished. 【Result】 Results from LSCM method were consistent with the paraffin section method's. However, the LSCM method was simple, time-saving and accurate. According to Arabidopsis female gametophyte development stages, we divided sweet cherry female gametophyte development into six stages. Using this method to observe delayed embryo sacs, ‘Tieton' and ‘Brooks' embryo sac development periods were observed. The results showed that the aborted ovule rate was 41% in ‘Tieton', while only 21% in ‘Brooks', on day of flowering. When the embryo sacs were delayed by 3 d, the embryo sac degradation rate of ‘Tieton' was 45%, abortion rate was 66%; however,the degradation rate of ‘Brooks' was 35.6%, abortion rate was 46.6%. 【Conclusion】 It was suggestedthat the embryo sac development in ‘Tieton' was easyly aborted and rapidly degraded after maturation,while embryo sac in ‘Brooks' had higher vitality, lower aborted rate and better adapted to the environment.

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