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辣椒花粉发育相关基因CaPME1的表达与功能鉴定研究

发表时间:2016-11-15  浏览量:1351  下载量:451
全部作者: 王 丽,陈国菊,曹必好,陈长明,邹丽芳,朱张生,尹 玲,雷建军
作者单位: 华南农业大学园艺学院
摘 要: 辣椒育种主要采用优势育种,利用植物雄性不育系育种是优势育种的重要手段,弄清花粉发育的分子机理是研究雄性不育的关键所在。研究以辣椒(Capsicum annuum L.)隐性核雄性不育两用系AB114为材料,构建了一个载体即CaPME1基因的烟草脆裂病毒(tobacco rattle virus,TRV)重组质粒,采用病毒诱导的基因沉默(virus induced gene silencing,VIGS)方法对目标基因在辣椒可育植株中的表达进行特异性地抑制。 结果显示,相对于野生型和空载体对照植株(pTRV2),CaPME1基因沉默植株的花药开裂推迟,花药中的花粉减少;沉默植株的花粉萌发率显著降低,且沉默植株的花粉管比野生型和对照植株的花粉管短。扫描电镜(scanning electron microscope,SEM)结果显示,相对于野生型和对照植株,CaPME1沉默植株花粉萌发沟的数目及位置分布不规则,花粉的整体形状也不规则,呈现不同程度的皱缩、卷曲,有部分花粉严重皱缩。对花粉进行统计,结果表明沉默植株的花粉畸形率显著升高,说明抑制CaPME1的表达能够降低花粉的萌发率和导致花粉畸形,进而推测CaPME1在辣椒的花粉发育过程中起着一定的作用。实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)法检测基因的表达水平表明,CaPME1基因在VIGS沉默植株中的表达量比在对照植株(pTRV2)中的表达量明显偏低。
关 键 词: 园艺学;辣椒;花粉发育;CaPME1;病毒诱导的基因沉默;功能鉴定
Title: Expression and functional identification of CaPME1 related to pollen development in pepper (Capsicum annuum L.)
Author: WANG Li, CHEN Guoju, CAO Bihao, CHEN Changming, ZOU Lifang, ZHU Zhangsheng,
Organization: College of Horticultrue, South China Agriculutral University
Abstract: Hybrids is very popular in pepper production. The hybrid seed production is mainly by means of male sterile lines. The molecular mechanism of pollen development is the key to study male sterility. The recessive nuclear male sterility ab line AB114 was used to study. Tobacco rattle virus (TRV) recombinant plasmid, a vector, which included CaPME1 genes, was constructed using the virus induced gene silencing (VIGS) method, and it can silence the target gene expression in fertile pepper plants. Results showed that compared with wild type plants and transgenic plant with empty vector (pTRV2), the pollen became lesser and the anther dehiscence was significantly delayed in CaPME1gene silencing plants. Under the condition in vitro, pollen was placed on agar gel to germinate for us to test the pollen viability. Results indicated that the pollen germination rate of silent plant was significantly reduced and the pollen tube of silent plant was shorter than that of wild type and the pTRV2. We observed the surface of the pollen over the scanning electron microscope (SEM), the results showed that compared with that of wild type and pTRV2, the quantitative and irregular distribution of the CaPME1 silent plant’s pollen germination ditch had an impact on the global shape feature and it showed different levels of shrinking, intertangling and some part of the pollen got serious shrinkage. The results of statistical analysis about pollen showed that the deformity rate of the plant with silenced CaPME1 was significantly increased. This shows that the substance that restrains the CaPME1 can reduce pollen’s germination ratio and cause the pollen deformity, therefore we can assume that the CaPME1 plays a certain role in the pollen development process. The quantitative real-time PCR (qRT-PCR) was used to detect the gene expression level. The results showed that the CaPME1 expression level was obviously lower in the plants with scilenced CaPME1 than that in the control (wild and transformant with pTRV2).
Key words: horticulture; pepper; pollen development; CaPME1; virus induced gene silencing; function identification
发表期数: 2016年11月第21期
引用格式: 王 丽,陈国菊,曹必好,等. 辣椒花粉发育相关基因CaPME1的表达与功能鉴定研究[J]. 中国科技论文在线精品论文,2016,9(21):2226-2238.
 
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