油茶胚珠败育的组织学特性及相关基因表达分析  

Histological characteristics and related gene expression analysis of ovule abortion in Camellia oleifera

作　　者：陈雨晴 杜兵帅 王胜楠 曹一博 袁德义[2] 张凌云[1] Chen Yuqing;Du Bingshuai;Wang Shengnan;Cao Yibo;Yuan Deyi;Zhang Lingyun(School of Forestry,Key Laboratory of Forest Silviculture and Conservation of Ministry of Education,Beijing Forestry University,Beijing 100083,China;Key Laboratory of Non-Wood Forest Product of State Forestry Administration,Key Laboratory of Cultivation and Protection for Non-Wood Forest Trees of Ministry of Education,Central South University of Forestry and Technology,Changsha 410004,Hunan,China)

机构地区：[1]森林培育与保护教育部重点实验室,北京林业大学林学院,北京100083 [2]经济林育种与栽培国家林业局重点实验室,经济林培育与保护教育部重点实验室,中南林业科技大学,湖南长沙410004

出　　处：《北京林业大学学报》2023年第9期9-20,共12页Journal of Beijing Forestry University

基　　金：国家自然科学基金项目(32071798)。

摘　　要：【目的】油茶胚珠败育现象严重,只有少数胚珠能发育为成熟种子,但其败育机制尚不清楚。本研究对油茶胚珠败育时期、组织结构变化和败育原因展开探究,旨在明确油茶胚珠的败育过程,为提高油茶产量提供一定的理论基础和实践意义。【方法】本研究以‘华硕’品种果实为试验材料,在体视显微镜下观察油茶果实内胚珠的形态,统计败育率,采用荧光素二乙酸酯(FDA)染色观察败育胚珠失去活性的时间;通过石蜡切片和显微镜观察明确可育胚珠和败育胚珠的组织结构变化,通过碘-碘化钾染色和PAS反应标记可育胚珠与败育胚珠中淀粉粒的分布。利用CFDA荧光示踪和激光共聚焦成像技术揭示同化物在可育胚珠与败育胚珠中的运输路径,通过实时荧光定量PCR试验分析与糖和能量代谢、活性氧代谢、细胞凋亡等过程相关的基因在可育胚珠与败育胚珠中的表达情况。【结果】(1)体式显微镜的观察结果显示26 WAA(花后周数)后油茶可育胚珠与败育胚珠的大小产生差异;FDA标记结果说明,败育胚珠在果实发育过程中逐步失去活性。(2)37 WAA时,胚珠败育率达到64.08%。(3)显微观察显示:可育胚珠的胚和胚乳均正常发育,内外珠被结合紧密;败育胚珠无胚乳细胞,内珠被与外珠被之间的空隙较大;可育胚珠的胚柄和胚乳中均有淀粉粒存在,败育胚珠仅在萎缩的内珠被上观察到少量淀粉粒。可育胚珠的内珠被上无胼胝质沉积,败育胚珠的内珠被上可见胼胝质沉积。(4)CFDA荧光示踪结果发现,败育胚珠与可育胚珠的同化物运输方式存在差异。(5)与糖和能量代谢、活性氧代谢、细胞凋亡等过程相关基因在败育胚珠和可育胚珠中存在差异性表达。【结论】败育胚珠的结构异常,胚珠内缺乏淀粉的积累,内珠被上有胼胝质沉积,同化物的运输方式与可育胚珠不同,参与胚珠物质和能量代谢、抗氧[Objective] The ovule abortion of Camellia oleifera is serious.Only a few ovules could develop into mature seeds,but its abortion mechanism is not clear.The research investigated abortion period,microstructure changes and abortion reasons of ovules to clarify the abortion process and to provide certain theoretical basis and practical significance for increasing the yield of Camellia oleifera.[Method] The fruits of C.oleifera cv.‘Huashuo' were chosen as the experimental material.The ovule morphology in Camellia oleifera fruit was observed under stereomicroscope.We counted the proportion of abortive ovules.The stage of losing activity of abortive ovules was observed by FDA staining technique and stereomicroscope.The histological changes of fertile and abortive ovules were clarified by paraffin technique and microscopic observation,and the starch grain distribution in fertile and abortive ovules was marked by K-KI2 staining and PAS reaction.The CFDA fluorescence tracing experiment and laser confocal microscopy were used to reveal assimilate transport pathways in fertile and abortive ovules.The qRT-PCR was used to analyze the expression of genes related to sugar and energy metabolism,reactive oxygen species metabolism,and apoptosis processes in fertile and abortive ovules.[Result](1) The stereomicroscope observation showed that the size of fertile and abortive ovules differed after 26 WAA(weeks after anthesis).The results of FDA staining indicated that the abortive ovules gradually lost their activity during fruit development.(2) At 37 WAA,the proportion of abortive ovules reached 64.08%.(3) Microscopic observation showed that the embryo and endosperm of fertile ovules developed normally,and the inner and outer integuments were tightly united;the abortive ovules had no endosperm cells,and the space between the inner and outer integument was wild.Starch grains were observed in both the suspensor and endosperm of fertile ovules,and only a few starch grains were observed on the shrunken inner integuments of abortive o

关 键 词：油茶 胚珠 败育 组织学特性 

分 类 号：S792.11[农业科学—林木遗传育种]