摘要
以‘突尼斯软籽’石榴为试材,对雌蕊败育的原因及关键时期、PgAGL11序列特征及功能进行研究。石蜡切片观察结果表明,雌蕊败育的原因为胚珠内珠被原基形成后停止发育,胚珠败育的关键时期为花蕾纵径8.1~15.0 mm时;采用TA克隆得到PgAGL11全长CDS序列696 bp,系统进化分析发现其与拟南芥的AGL11序列相似性最高;实时荧光定量PCR结果显示,PgAGL11在雌蕊中表达量显著高于其他花器官,且在胚珠败育关键时期(花蕾纵径8.1~15.0 mm),可育花雌蕊中表达量极显著高于败育花雌蕊;构建PgAGL11过量表达载体,利用农杆菌介导法侵染野生型拟南芥,转基因拟南芥花表现为雄蕊变短,花瓣变小,花柱变长,柱头表面乳突状物质变长。本研究确定了石榴雌蕊败育的关键原因及时期,并初步证明了PgAGL11可能在石榴雌蕊败育过程中发挥重要作用。
To confirm the key reason and stage of pomegranate female sterility and explore the role of PgAGL11 for pomegranate female sterility,we investigated the key stage of pomegranate female sterility as well as sequence characteristic and function of PgAGL11 using‘Tunisiruanzi＇. The results of paraffin section revealed that the key reason of pomegranate female sterility was ovule termination after the formation of inner integument primordial. The key stage was when the flower bud vertical diameter developed into 8.1–15.0 mm. The 696 bp coding sequence（CDS）of PgAGL11 was obtained by TA cloning,which showed a high similarity with AGL11 in Arabidopsis thaliana. Transcription analysis indicated the expression level of PgAGL11 in bisexual flowers was significantly higher than that in functional male flowers during the flower bud vertical diameter was 8.1–15.0 mm（P 0.05）,the expression level of PgAGL11 in pistil was significantly higher than that in petal,calyx,seed,and pericarp（P 0.01）,which indicated it＇s crucial roles in pistil development. Furthermore,we induced the PgAGL11 gene into Arabidopsis thaliana using a Agrobacterium-mediated transgenic method and produced shortened stamen,smaller petal,thicker style,and elongated mastoid cells on the surface of stigma phenotypes. The results confirmed the key reason and stage of pomegranate female sterility and the important roles of PgAGL11 in pomegranate female sterility.
出处
《园艺学报》
CAS
CSCD
北大核心
2017年第11期2089-2098,共10页
Acta Horticulturae Sinica
基金
中国农业科学院科技创新工程专项经费项目(CAAS-ASTIP-2017-ZFRI-03)
国家科技基础性工作专项重点项目(2012 FY110100)
关键词
石榴
雌蕊
败育
胚珠
表达分析
转基因
pomegranate
female
sterility
ovule
expression analysis
transgenic
