Biotechnology Department, CSIR-Institute of Himalayan Bioresource Technology, Palampur, Himachal Pradesh 176061, India.
Department of Biotechnology, Guru Nanak Dev University, Amritsar 143005, India.
Int J Mol Sci. 2019 Jan 28;20(3):539. doi: 10.3390/ijms20030539.
This study explicates molecular insights commencing Self-Incompatibility (SI) and CC (cross-compatibility/fertilization) in self (SP) and cross (CP) pollinated pistils of tea. The fluorescence microscopy analysis revealed ceased/deviated pollen tubes in SP, while successful fertilization occurred in CP at 48 HAP. Global transcriptome sequencing of SP and CP pistils generated 109.7 million reads with overall 77.9% mapping rate to draft tea genome. Furthermore, concatenated de novo assembly resulted into 48,163 transcripts. Functional annotations and enrichment analysis (KEGG & GO) resulted into 3793 differentially expressed genes (DEGs). Among these, de novo and reference-based expression analysis identified 195 DEGs involved in pollen-pistil interaction. Interestingly, the presence of 182 genes [PT germination & elongation (67), S-locus (11), fertilization (43), disease resistance protein (30) and abscission (31)] in a major hub of the protein-protein interactome network suggests a complex signaling cascade commencing SI/CC. Furthermore, tissue-specific qRT-PCR analysis affirmed the localized expression of 42 DE putative key candidates in stigma-style and ovary, and suggested that LSI initiated in style and was sustained up to ovary with the active involvement of RNS, SRKs & SKIPs during SP. Nonetheless, COBL10, RALF, FERONIA-rlk, LLG and MAPKs were possibly facilitating fertilization. The current study comprehensively unravels molecular insights of phase-specific pollen-pistil interaction during SI and fertilization, which can be utilized to enhance breeding efficiency and genetic improvement in tea.
本研究阐述了自交不亲和(SI)和 CC(交叉亲和/受精)在茶树自交(SP)和杂交(CP)授粉雌蕊中的分子机制。荧光显微镜分析显示 SP 中的花粉管停止/偏离,而 CP 在 48 HAP 时发生了成功受精。SP 和 CP 雌蕊的全转录组测序产生了 1.097 亿条reads,总体映射率为 77.9%至草案茶树基因组。此外,拼接从头组装得到 48163 个转录本。功能注释和富集分析(KEGG 和 GO)产生了 3793 个差异表达基因(DEGs)。其中,从头和基于参考的表达分析鉴定出 195 个参与花粉-雌蕊相互作用的 DEGs。有趣的是,在蛋白质-蛋白质相互作用网络的一个主要枢纽中存在 182 个基因[花粉萌发和伸长(67)、S 座位(11)、受精(43)、抗病蛋白(30)和离层(31)],这表明 SI/CC 起始了一个复杂的信号级联反应。此外,组织特异性 qRT-PCR 分析证实了 42 个推定关键候选基因在柱头-花柱中的局部表达,并表明 LSI 从花柱开始,并一直持续到子房,在 SP 过程中,RNS、SRKs 和 SKIPs 积极参与。然而,COBL10、RALF、FERONIA-rlk、LLG 和 MAPKs 可能有助于受精。本研究全面揭示了 SI 和受精过程中特定阶段花粉-雌蕊相互作用的分子机制,可用于提高茶树的繁殖效率和遗传改良。