# Biflavonoid Profiling of _Juniperus_ Species: The Influence of Plant Part and Growing Location
## 4 Discussion
Amentoflavone is known for its significant biological activities, including strong anticancer, neuroprotective, antiviral, and anti-inflammatory effects [27]. The total biflavonoid content in both analyzed species, _J. communis_ and _J. oxycedrus_, exceeded 5 mg/g, which is comparable to the amount we previously reported for yellow ginkgo leaves [25]. In cones, the total biflavonoid content was lower for both _J. communis_ and _J. oxycedrus_, ranging from 0.13 to 1.3 mg/g dw. A general trend was observed where green cones accumulated more biflavonoids than ripe cones, similar to findings by [PERSON] et al. [3] for some _J. communis_ berries. However, this trend was not consistent across all locations. [PERSON] et al. [3] reported a total biflavonoid content in _J. communis_ berries ranging from 0.14 to 1.38 mg/g of fresh weight. Our results suggest that biflavonoids are important phytochemicals in _Juniperus_ species, particularly in needles, which should be included in phytochemical studies. However, they have often been neglected or omitted in previous studies. To investigate how growing location influences biflavonoid accumulation, we collected _J. communis_ samples from various locations. As shown in Table 1, the growing location significantly affected both individual and total biflavonoid accumulation. In some locations, bilobetin was not detected at all, which may be directly related to environmental factors. However, since our samples were collected from wild plants, where environmental conditions were not controlled, it is challenging to determine which specific factors influence biflavonoid accumulation. According to the literature, biflavonoid accumulation is influenced by tissue type, with plant parts directly exposed to the environment containing higher levels, while inner tissues lack detectable biflavonoids, as observed in _Ginkgo biloba_ L. [26]. [PERSON] et al. [16] investigated how harvesting time and geographic area affect polyphenolic compounds in _J. communis_ cones from Romania. While they did not detect biflavonoids, they found that total polyphenol content was significantly higher in areas with greater altitude and in samples collected in November compared to September and October. Similarly, [PERSON] et al. [28] compared the total flavonoid content in _J. communis_ berries from four locations in India and reported a significant influence of growing location. To the best of our knowledge, our study is the first to report biflavonoid content in _J. communis_ across different locations, and our findings align with expectations. Although the exact role of biflavonoids in plants remains unclear, their localization within plant tissues and biological activity suggest a potential role in plant-environment interactions [11]. However, to identify the precise environmental factors affecting biflavonoid accumulation, further controlled laboratory experiments are needed. As evident from the PCA plots in Figure 10, the plant part--particularly needle tissue--emerges as the dominant factor influencing biflavonoid accumulation, surpassing both species identity and geographic origin in its impact. This clear separation of needle samples underscores the tissue-specific nature of biflavonoid biosynthesis within _Juniperus_ species.