Mustafa Roma, Hamza Muhammad, Kamal Hira, Mansoor Shahid, Scheffler Jodi, Amin Imran
Agricultural Biotechnology Division, National Institute for Biotechnology and Genetic Engineering, Faisalabad, Pakistan.
Pakistan Institute of Engineering and Applied Sciences (PIEAS), Islamabad, Pakistan.
Mol Biotechnol. 2017 Jul;59(7):241-250. doi: 10.1007/s12033-017-0014-y.
A Tobacco rattle virus (TRV)-based virus-induced gene silencing assay was employed as a reverse genetic approach to study gene function in cotton (Gossypium hirsutum). This approach was used to investigate the function of the Enoyl-CoA reductase (GhECR) gene in pathogen defense. Amino acid sequence alignment of Arabidopsis ECR with homologous sequence from G. hirsutum, G. arboreum, G. herbaceum and G. barbadense showed that ECRs are highly conserved among these species. TRV-based silencing of GhECR gene in G. hirsutum induced a cell death/necrotic lesion-like phenotype. Reverse transcription polymerase chain reaction (RT-PCR) and real-time quantitative PCR showed reduced GhECR mRNA levels in TRV inoculated plants. Three isolates of Verticillium dahliae (V. dahliae) and Fusarium oxysporum f. sp. vasinfectum (FOV) were used to infect GhECR-silenced plants. Out of 6 races of 2 pathogens, down regulation of GhECR gene resulted in reduced resistance. This is the first report showing that cotton GhECR gene is involved in resistance to different strains of V. dahliae and FOV.
采用基于烟草脆裂病毒（TRV）的病毒诱导基因沉默试验作为反向遗传学方法，研究棉花（陆地棉）中的基因功能。该方法用于研究烯酰辅酶A还原酶（GhECR）基因在病原体防御中的功能。拟南芥ECR与陆地棉、亚洲棉、草棉和海岛棉同源序列的氨基酸序列比对表明，这些物种中的ECR高度保守。基于TRV的陆地棉GhECR基因沉默诱导了细胞死亡/坏死样病变表型。逆转录聚合酶链反应（RT-PCR）和实时定量PCR显示，接种TRV的植株中GhECR mRNA水平降低。使用三种大丽轮枝菌（V. dahliae）分离株和尖孢镰刀菌枯萎专化型（FOV）感染GhECR沉默植株。在两种病原体的6个小种中，GhECR基因的下调导致抗性降低。这是首次报道表明棉花GhECR基因参与对不同大丽轮枝菌菌株和FOV的抗性。
