Department of Biology, Tufts University, Medford, Massachusetts, USA.
Department of Biology, Tufts University, Medford, Massachusetts, USA
Appl Environ Microbiol. 2019 Jun 17;85(13). doi: 10.1128/AEM.00269-19. Print 2019 Jul 1.
Patterns of phyllosphere diversity have become increasingly clear with high-throughput sequencing surveys, but the processes that control phyllosphere diversity are still emerging. Through a combination of lab and field experiments using Napa cabbage and lactic acid bacteria (LAB), we examined how dispersal and establishment processes shape the ecological distributions of phyllosphere bacteria. We first determined the abundance and diversity of LAB on Napa cabbage grown at three sites using both culture-based approaches and 16S rRNA gene amplicon sequencing. Across all sites, LAB made up less than 0.9% of the total bacterial community abundance. To assess whether LAB were low in abundance in the Napa cabbage phyllosphere due to a limited abundance in local species pools (source limitation), we quantified LAB in leaf and soil samples across 51 vegetable farms and gardens throughout the northeastern United States. Across all sites, LAB comprised less than 3.2% of the soil bacterial communities and less than 1.6% of phyllosphere bacterial communities. To assess whether LAB are unable to grow in the phyllosphere even if they dispersed at high rates (establishment limitation), we used a gnotobiotic Napa cabbage system in the lab with experimental communities mimicking various dispersal rates of LAB. Even at high dispersal rates, LAB became rare or completely undetectable in experimental communities, suggesting that they are also establishment limited. Collectively, our data demonstrate that the low abundance of LAB in phyllosphere communities may be explained by establishment limitation. The quality and safety of vegetable fermentations are dependent on the activities of LAB naturally present in the phyllosphere. Despite their critical role in determining the success of fermentation, the processes that determine the abundance and diversity of LAB in vegetables used for fermentation are poorly characterized. Our work demonstrates that the limited ability of LAB to grow in the cabbage phyllosphere environment may constrain their abundance on cabbage leaves. These results suggest that commercial fermentation of Napa cabbage proceeds despite low and variable abundances of LAB across different growing regions. Propagule limitation may also explain ecological distributions of other rare members of phyllosphere microbes.
随着高通量测序调查的开展,叶际多样性模式变得越来越清晰,但控制叶际多样性的过程仍在不断出现。通过使用纳帕白菜和乳酸菌(LAB)进行实验室和野外实验的组合,我们研究了扩散和建立过程如何塑造叶际细菌的生态分布。我们首先使用基于培养的方法和 16S rRNA 基因扩增子测序来确定在三个地点生长的纳帕白菜上 LAB 的丰度和多样性。在所有地点,LAB 占总细菌群落丰度的不到 0.9%。为了评估 LAB 是否由于当地物种库(源限制)中数量有限而在纳帕白菜叶际中数量较少,我们在美国东北部的 51 个蔬菜农场和花园中量化了叶片和土壤样本中的 LAB。在所有地点,LAB 占土壤细菌群落的不到 3.2%,占叶际细菌群落的不到 1.6%。为了评估 LAB 是否即使以高速度扩散也无法在叶际中生长(建立限制),我们在实验室中使用了无菌纳帕白菜系统,使用模拟 LAB 各种扩散率的实验群落。即使在高扩散率下,LAB 在实验群落中也变得稀有或完全无法检测到,这表明它们也受到建立限制。总的来说,我们的数据表明,叶际群落中 LAB 的低丰度可能是由建立限制引起的。蔬菜发酵的质量和安全性取决于叶际中自然存在的 LAB 的活动。尽管它们在确定发酵成功方面起着至关重要的作用,但决定用于发酵的蔬菜中 LAB 的丰度和多样性的过程仍未得到很好的描述。我们的工作表明,LAB 在白菜叶际环境中生长的能力有限可能会限制它们在白菜叶片上的丰度。这些结果表明,尽管在不同的种植地区 LAB 的丰度低且变化不定,但商业纳帕白菜发酵仍在进行。传播限制也可能解释叶际微生物其他稀有成员的生态分布。