Pollination is composed of cell-cell communication and complicated signaling cascades that regulate pollen tube growth and guidance toward the ovules for double fertilization, and is critical for successful sexual reproduction. Exploring expression profiles of in vivo grown pollen tubes is important. Nevertheless, it is difficult to obtain accessible pollen tubes for profiling studies in most model plants. By taking advantage of the hollow styles of lily (Lilium longiflorum), in vivo pollen tubes harvested from pollinated styles which had been cut open were used here to study their protein and transcript profiles. Pollination quantitatively and qualitatively altered the total protein composition of elongating pollen tubes. cDNAs generated and amplified from total RNAs of 24 h in vivo grown and 12 h in vitro cultured pollen tubes were used for suppression subtractive hybridization analyses and preparation of home-made array chips. Microarray analyses conducted with different probe sets revealed 16 transcripts specifically present and/or enriched in in vivo pollen tubes. Reverse transcription-PCR (RT-PCR), in situ hybridization and Northern blotting were applied to validate their unique pollination-induced expression features. Interestingly, several transcripts were simultaneously detected on the stylar transmitting tract epidermis, where in vivo pollen tubes tightly adhered during pollination. Their deduced amino acid sequences showed that most of them encoded small proteins and could be classified into several families. Transient assay revealed filament-like structures decorated by these proteins and one probably localized in the generative cell. These small peptides might be critical for pollen tube growth during pollination, and further exploration of their biological functions and mechanisms of action are of great interest.

Background: Yinqiao Powder (YQP) is administered in Traditional Chinese Medicine (TCM) for febrile infectious diseases and has been demonstrated to fight the prevalence of influenza. Objective: The present study aimed to identify the optimal decoction time of YQP on inhibit influenza A virus FM1 and investigate the molecular mechanisms of different decocting time YQP. Methods: The male BALB/c mice were made into pneumonia model of influenza A virus FM1. They were randomly assigned to twelve groups: blank control group, model control group, tamiflu (27.5 mg/Kg) group, 3 minutes YQP (High-dose group 30 g/Kg•d, Middle-dose group 15 g/Kg•d, and Low-dose group 7.5 g/Kg•d) groups, 6 minutes YQP (High-dose group 30 g/Kg•d, Middle-dose group 15 g/Kg•d, and Low-dose group 7.5 g/Kg•d) groups, and 12 minutes YQP (High-dose group 30 g/Kg•d, Middle-dose group 15 g/Kg•d, and Low-dose group 7.5 g/Kg•d) groups. All groups were administrated for 7 days. The mice were killed after the third and last days. Viral load of lung tissues were observed by Real-time PCR. Pulmonary index and inhibition rate were calculated by formula. Furthermore, the mice macrophage Ana-1 was infected by FM1, treated with different decocting time YQP drug-containing serum for 24 h. TLR3/4, MyD88, TRAF-6, TRAM and TRIF mRNA were detected by Real-time PCR. TLR3/4 proteins were observed by western blot. Results: All YQP intervention groups induced a reduction in the viral load and pulmonary index on the third and last days of infection, especially in 3 minutes YQP H/M/L groups and 6 minutes High-dose group. Meanwhile, YQP intervention groups led to significant decrease in TLR3/4, MyD88, TRAF-6, TRAM and TRIF mRNA levels and TLR3/4 proteins, in particular with 3 and 6 minutes YQP groups. Conclusion: YQP could protect the lung tissue and alleviate the inflammation caused by influenza A virus FM1. The optimal decoction time of YQP was 3to 6minutes on inhibit influenza A virus FM1 in mice.