# Genome-sequenced bacterial collection from sorghum aerial root mucilage

Marco Mechan-Llontop, John Mullet, Ashley Shade

► To cite this version:

Marco Mechan-Llontop, John Mullet, Ashley Shade. Genome-sequenced bacterial collection from sorghum aerial root mucilage. Microbiology Resource Announcements, In press, 10.1128/MRA.00484-23. [https://doi.org/10.1128/MRA.00484-23](https://doi.org/10.1128/MRA.00484-23)

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**Article title**

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Genome-sequenced bacterial collection from sorghum aerial root mucilage.

4

**Authors**

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Marco E. Mechan-Llontopª, John Mulletª,b, and Ashley Shadec#

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**Affiliations**

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a. Great Lakes Bioenergy Research Center, Michigan State University, East Lansing, Michigan, USA.

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b. Department of Biochemistry & Biophysics, Texas A&M University, College Station, Texas, USA.

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c. Universite Claude Bernard Lyon 1; CNRS, UMR 5557 Écologie Microbienne, 16 rue Dubois 69622

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Villeurbanne Cedex, FRANCE

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**Running title**

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Bacteria from sorghum aerial root mucilage.

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**Corresponding author's email address**

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\#Address correspondence to: Ashley Shade, [ashley.shade@cnrs.fr](mailto:ashley.shade@cnrs.fr)

15

**Abstract**

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A collection of 47 bacteria isolated from the mucilage of aerial roots of energy sorghum is available at

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the Great Lakes Bioenergy Research Center, Michigan State University, MI, USA. We enriched bacteria

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with putative plant-beneficial phenotypes and include information on phenotypic diversity, taxonomy,

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and whole genome sequences.

20

**Announcement**

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Plant phyllosphere exudates harbor microbial communities that can engage with the host plant, for

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example by supporting plant growth or resilience to various stresses (1). The heat and drought-tolerant

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crop bioenergy sorghum (*Sorghum bicolor* L. Moench) secretes abundant polysaccharide-rich mucilage

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on aerial roots (2) that selects for specialized microbiome members that are expected to have plant-
beneficial functions (3). As a resource for the genomic and functional diversities of bacteria associated
with phyllosphere exudates, we present a collection of 47 bacterial strains isolated from sorghum aerial
root mucilage. These strains represent 17 families and include 21 genera with putative plant-beneficial
capabilities, including nitrogen fixation, phosphate solubilization, resistance to terpenoids, use of
methanol as a carbon source, and tolerance to desiccation, based on the culture media in which the
strains were originally isolated.

Bacterial strains were enriched from the mucilage of aerial roots of bioenergy sorghum cultivar TAM 17651 grown at the Great Lakes Bioenergy Research Center (GLBRC), as part of the Biofuel Cropping System Experiment (BCSE) in Hickory Corners, Michigan (42°23′41.6″ N, 85°22′23.1″ W). Aerial roots were harvested from sorghum plants grown on nitrogen-fertilized plots on September 25, 2020. We used sterile razor blades to collect between 3 to 5 aerial roots per plant into sterile 50 ml Eppendorf tubes. Within 2 hours of collection, the samples were transported to the laboratory on ice and stored at -80°C for 18 days until processing. Sorghum mucilage was first fully hydrated with sterile water, removed from roots, and then collected into sterile 1.5 ml Eppendorf tubes. Mucilage collected from different plants was pooled and serially diluted from 10⁻¹ to 10⁻⁴. To capture a diversity of bacteria from the rehydrated mucilage, we used a variety of cultivation media (Table 1). To select for anaerobic bacteria, plates were placed in anaerobic jars containing three bags of anaerobic gas generator (Mitsubishi AnaeroPack System). All plates were incubated at 25°C and 37°C for up to 14 days. Isolated colonies were transferred on new plates with the same medium as used for isolation to confirm purity. Bacteria were grown overnight on R2A broth at 28°C with shaking at 200rpm and glycerol stock (25% v/v) of pure isolates was stored at -80°C. Prior to performing whole genome sequencing, we performed full-length 16S rRNA gene sequencing, with universal primers 27F (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (5'-TACGGTTACCTTGTTACGACTT-3') (4), of each isolate using the Sanger protocol to link
48 them to corresponding 16S rRNA gene amplicon dynamics in a field microbiome study that was
49 executed at the same field site at Kellogg Biological Station (3).

50 Individual isolates or the full cryopreserved collection is available. Cultures can be regrown
51 using standard microbiological procedures to transfer a sterile inoculation loop of freezer stock onto the
52 original isolation medium and incubation time (Table 1).

53 The high molecular weight genomic DNA of each bacterial isolate was extracted using a
54 phenol/chloroform extraction protocol (5). The genomic DNA of each strain was submitted to the U.S.
55 Department of Energy Joint Genome Institute (DOE-JGI), for library preparation and sequencing with the
56 Pacific Biosciences (PacBio) platform (6). DNA was sheared to 10kb using g-TUBE columns (Covaris) and
57 subjected to library preparation using the SMARTbell Express Template prep 2.0 kit and sequenced on
58 the PacBio Sequel platform. Samples that failed DNA quality control required by the PacBio pipeline
59 were instead sequenced using the NovaSeq S4 Illumina platform (7). High fidelity CCS reads >5kb were
60 assembled with Flye 2.8.3 (8) using default settings. Raw Illumina sequences were quality-filtered using
61 BBTools (9). Artifact filtered and normalized Illumina reads were assembled with SPAdes v3.15.3 (10) (-
62 phred-offset 33 -cov-cutoff auto -t 16 -m 64 -careful -k 25,55,95), and contigs were discarded if the
63 length was <1kb (BBTools reformat.sh: minlength=1000 ow=t). Genome completeness and
64 contamination was estimated with CheckM v1.2.2 (11). The assembly was annotated using Prokka 1.14.6
65 (12) and the phylogenetic tree was inferred using Orthofinder 2.5.5 (13) and edited with iTOLs v.6.5.8
66 (14) (Figure 1).

67 **Data availability statement**

68 Raw sequencing data are available at the Joint Genome Institute's GOLD database under Study ID
69 [Gs0157305](https://www.ncbi.nlm.nih.gov/gs/0157305), as well as with the National Institutes of Health Sequence Read Archive (see Table 1).

70 Genome sequencing assembly data has been deposited in the GenBank with accession numbers listed in
71 Table 1. The bacterial collection is available at The Great Lakes Bioenergy Research Center at Michigan State University, USA (glbrc@msu.edu).

72

73 Acknowledgments

74 This work was supported by the Great Lakes Bioenergy Research Center, U.S. Department of Energy,

75 Office of Science, Office of Biological and Environmental Research under Award Number DE-SC0018409.

76 Support for field research was provided by the Great Lakes Bioenergy Research Center, U.S. Department

77 of Energy, Office of Science, Office of Biological and Environmental Research (Awards DE-SC0018409 and

78 DE-FC02-07ER64494), by the National Science Foundation Long-term Ecological Research Program (DEB

79 1637653) at the Kellogg Biological Station, and by Michigan State University AgBioResearch. The work

80 conducted by the U.S. Department of Energy Joint Genome Institute, a DOE Office of Science User

81 Facility, is supported under Contract No. DE-AC02-05CH11231.

82 This data was generated for JGI Proposal #508590.

83 The authors declare no conflict of interest.

84 References

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132 Tables
<table><caption>TABLE 1.- Taxonomy, colony phenotype, and genome characteristics of 49 bacterial isolates from sorghum aerial root mucilage, as described in this study. SRA is the sequence read archive of the U.S. National Institutes of Health.</caption><thead><tr><th>Isolate ID</th><th>Bacterial species</th><th>Initial isolation media</th><th>Temperature, oxygen availability</th><th>Colony phenotype</th><th>Sequencing Platform</th><th>SRA accession number</th><th>GenBank accession</th><th>Genome length (bp)</th><th>N° contigs</th><th>Coverage</th><th>GC %</th><th>Genome completeness</th></tr></thead><tbody><tr><td>SORGH_AS_0335</td><td>Acidovorax wautersii</td><td>R2A</td><td>25 °C, anaerobic</td><td>Small, white</td><td>PacBio</td><td>SRP437222</td><td>JAVIZX000000000</td><td>4,571,377</td><td>1</td><td>70.4 X</td><td>68.30%</td><td>99.52%</td></tr><tr><td>SORGH_AS_0440</td><td>Agrobacterium sp.</td><td>M9 + 0.4% sucrose</td><td>25 °C, aerobic</td><td>White</td><td>PacBio</td><td>SRP437134</td><td>JAVIYR000000000</td><td>5,277,481</td><td>3</td><td>166.0 X</td><td>59.30%</td><td>99.04%</td></tr><tr><td>SORGH_AS_0745</td><td>Agrobacterium sp.</td><td>M9 nitrogen-free + 1% xylose</td><td>37 °C, aerobic</td><td>White, EPS production</td><td>PacBio</td><td>SRP437174</td><td>JAUTBI000000000</td><td>5,670,289</td><td>4</td><td>177.6 X</td><td>58.60%</td><td>99.88%</td></tr><tr><td>SORGH_AS_0749</td><td>Agrobacterium tumefaciens</td><td>M9 nitrogen-free, Carbon-free</td><td>37 °C, aerobic</td><td>White, EPS production</td><td>PacBio</td><td>SRP437121</td><td>JAURUU000000000</td><td>5,669,583</td><td>4</td><td>151.8 X</td><td>58.40%</td><td>99.93%</td></tr><tr><td>SORGH_AS_0212</td><td>Arthrobacter sp.</td><td>Jensen</td><td>25 °C, aerobic</td><td>White, EPS production</td><td>PacBio</td><td>SRP437201</td><td>JAUTAA000000000</td><td>4,369,871</td><td>1</td><td>117.7 X</td><td>66.10%</td><td>99.71%</td></tr><tr><td>SORGH_AS_0304</td><td>Atlantibacter sp.</td><td>Jensen</td><td>25 °C, anaerobic</td><td>White</td><td>PacBio</td><td>SRP437126</td><td>JAVLWF000000000</td><td>4,876,478</td><td>2</td><td>140.5 X</td><td>56.20%</td><td>99.39%</td></tr><tr><td>SORGH_AS_0510</td><td>Bacillus sp.</td><td>R2A + 0.1% linalool</td><td>37 °C, aerobic</td><td>White</td><td>PacBio</td><td>SRP437241</td><td>JAUTAU000000000</td><td>4,828,420</td><td>1</td><td>171.6 X</td><td>38.90%</td><td>98.52%</td></tr><tr><td>SORGH_AS_0407</td><td>Bacillus thuringiensis</td><td>R2A + PEG6000 - 0.73MPa</td><td>25 °C, aerobic</td><td>White</td><td>PacBio</td><td>SRP437118</td><td>JAURUT000000000</td><td>5,894,929</td><td>5</td><td>106.5 X</td><td>35.40%</td><td>99.34%</td></tr><tr><td>SORGH_AS_0431</td><td>Brevundimonas vesicularis</td><td>M9 + 0.4% sucrose</td><td>25 °C, aerobic</td><td>pink</td><td>PacBio</td><td>SRP437147</td><td>JAUTAQ000000000</td><td>3,191,065</td><td>1</td><td>68.5 X</td><td>66.20%</td><td>99.05%</td></tr><tr><td>SORGH_AS_0447</td><td>Chryseobacterium sp.</td><td>TSA</td><td>25 °C, aerobic</td><td>Yellow</td><td>PacBio</td><td>SRP437232</td><td>JAUTAR000000000</td><td>4,374,394</td><td>1</td><td>132.6 X</td><td>39.90%</td><td>99.51%</td></tr><tr><td>SORGH_AS_0776</td><td>Curtobacterium sp.</td><td>M9 nitrogen-free + 1% xylose</td><td>37 °C, aerobic</td><td>Yellow</td><td>PacBio</td><td>SRP437163</td><td>JAVIYU000000000</td><td>3,877,562</td><td>1</td><td>164.5 X</td><td>71.30%</td><td>98.99%</td></tr><tr><td>SORGH_AS_0287</td><td>Enterobacter sp.</td><td>Jensen</td><td>37 °C, anaerobic</td><td>White, EPS production</td><td>PacBio</td><td>SRP437233</td><td>JAVIZW000000000</td><td>4,947,816</td><td>2</td><td>108.4 X</td><td>54.60%</td><td>99.97%</td></tr><tr><td>SORGH_AS_0622</td><td>Flavobacterium sp.</td><td>MMS methanol</td><td>25 °C, aerobic</td><td>Orange yellow</td><td>PacBio</td><td>SRP437242</td><td>JAUTAX000000000</td><td>5,243,061</td><td>1</td><td>137.1 X</td><td>33.80%</td><td>98.94%</td></tr><tr><td>SORGH_AS_0428</td><td>Microbacterium sp.</td><td>M9 + 0.4% sucrose</td><td>25 °C, aerobic</td><td>Light yellow, EPS production</td><td>PacBio</td><td>SRP437182</td><td>JAVIZT000000000</td><td>3,300,177</td><td>1</td><td>89.6 X</td><td>69.60%</td><td>98.23%</td></tr><tr><td>SORGH_AS_0445</td><td>Microbacterium foliorum</td><td>TSA</td><td>25 °C, aerobic</td><td>Light yellow</td><td>PacBio</td><td>SRP437225</td><td>JAVIZQ000000000</td><td>3,850,168</td><td>1</td><td>187.5 X</td><td>67.90%</td><td>99.14%</td></tr><tr><td>SORGH_AS_0209</td><td>Microbacterium proteolyticum</td><td>Jensen</td><td>25 °C, aerobic</td><td>White, EPS production</td><td>PacBio</td><td>SRP437239</td><td>JAUSZZ000000000</td><td>3,767,688</td><td>1</td><td>122.8 X</td><td>70.20%</td><td>98.48%</td></tr><tr><td>SORGH_AS_0344</td><td>Microbacterium sp.</td><td>R2A</td><td>37 °C, aerobic</td><td>Pink</td><td>PacBio</td><td>SRP437155</td><td>JAUTAD000000000</td><td>3,750,535</td><td>1</td><td>78.7 X</td><td>70.20%</td><td>97.98%</td></tr><tr><td>SORGH_AS_0421</td><td>Microbacterium sp.</td><td>M9 + 0.4% sucrose</td><td>25 °C, aerobic</td><td>Orange</td><td>PacBio</td><td>SRP437226</td><td>JAUTAO000000000</td><td>3,450,105</td><td>1</td><td>228.2 X</td><td>69.50%</td><td>97.72%</td></tr></tbody></table>
<table><tr><td>SORGH_AS_0454</td><td>Microbacterium sp.</td><td>TSA</td><td>25 °C, aerobic</td><td>Orange</td><td>PacBio</td><td>SRP437164</td><td>JAVIYS000000000</td><td>3,569,378</td><td>1</td><td>197.0 X</td><td>69.60%</td><td>97.98%</td></tr><tr><td>SORGH_AS_0505</td><td>Microbacterium sp.</td><td>M9 nitrogen-free +</td><td>25 °C, aerobic</td><td>Orange, EPS production</td><td>PacBio</td><td>SRP437119</td><td>JAUTAT000000000</td><td>2,952,848</td><td>1</td><td>139.1 X</td><td>69.60%</td><td>97.79%</td></tr><tr><td>SORGH_AS_0426</td><td>Microbacterium testaceum</td><td>M9 + 0.4% sucrose</td><td>25 °C, aerobic</td><td>Orange</td><td>PacBio</td><td>SRP437123</td><td>JAUTAP000000000</td><td>3,569,373</td><td>1</td><td>202.9 X</td><td>69.80%</td><td>97.31%</td></tr><tr><td>SORGH_AS_0594</td><td>Microbacterium testaceum</td><td>Jensen</td><td>37 °C, aerobic</td><td>Orange, EPS production</td><td>PacBio</td><td>SRP437144</td><td>JAUTAW000000000</td><td>3,750,737</td><td>1</td><td>192.6 X</td><td>70.20%</td><td>97.98%</td></tr><tr><td>SORGH_AS_0422</td><td>Mucilaginibacter terrae</td><td>M9 + 0.4% sucrose</td><td>25 °C, aerobic</td><td>Pink, EPS production</td><td>PacBio</td><td>SRP437139</td><td>JAVLVU000000000</td><td>5,485,925</td><td>1</td><td>173.4 X</td><td>43.00%</td><td>97.62%</td></tr><tr><td>SORGH_AS_0306</td><td>Paenibacillus sp.</td><td>Jensen</td><td>25 °C, anaerobic</td><td>White, EPS production</td><td>PacBio</td><td>SRP437208</td><td>JAUTBP000000000</td><td>5,288,478</td><td>1</td><td>208.8 X</td><td>39.20%</td><td>99.45%</td></tr><tr><td>SORGH_AS_0338</td><td>Paenibacillus sp.</td><td>R2A</td><td>25 °C, anaerobic</td><td>Pink, EPS production</td><td>PacBio</td><td>SRP437186</td><td>JAVIYQ000000000</td><td>5,288,506</td><td>1</td><td>244.2 X</td><td>39.20%</td><td>99.36%</td></tr><tr><td>SORGH_AS_0197</td><td>Pantoea ananatis</td><td>Pirovskaya</td><td>25 °C, aerobic</td><td>Yellow</td><td>PacBio</td><td>SRP437179</td><td>JAUTBM000000000</td><td>5,000,175</td><td>5</td><td>197.6 X</td><td>53.40%</td><td>99.34%</td></tr><tr><td>SORGH_AS_0213</td><td>Pantoea ananatis</td><td>Jensen</td><td>25 °C, aerobic</td><td>Yellow, EPS production</td><td>Illumina</td><td>SRP437485</td><td>JAVIZK000000000</td><td>4,919,493</td><td>39</td><td>310.9 X</td><td>56.50%</td><td>100%</td></tr><tr><td>SORGH_AS_0797</td><td>Pantoea anthophila</td><td>M9 nitrogen-free + 1% arabinose</td><td>37 °C, aerobic</td><td>White, EPS production</td><td>PacBio</td><td>SRP437125</td><td>JAUTBH000000000</td><td>4,704,313</td><td>5</td><td>127.5 X</td><td>56.60%</td><td>100%</td></tr><tr><td>SORGH_AS_0585</td><td>Pedobacter agri</td><td>Jensen</td><td>25 °C, aerobic</td><td>Pink</td><td>PacBio</td><td>SRP437169</td><td>JAUTAV000000000</td><td>5,055,982</td><td>1</td><td>181.8 X</td><td>38.00%</td><td>97.61%</td></tr><tr><td>SORGH_AS_0191</td><td>Pseudomonas fluorescens</td><td>KB</td><td>25 °C, aerobic</td><td>White, fluorescens, EPS production</td><td>PacBio</td><td>SRP437154</td><td>JAVIZU000000000</td><td>6,080,362</td><td>1</td><td>43.2 X</td><td>60.10%</td><td>99.52%</td></tr><tr><td>SORGH_AS_0201</td><td>Pseudomonas psychrotolerans</td><td>Pirovskaya</td><td>25 °C, aerobic</td><td>Yellow</td><td>PacBio</td><td>SRP437137</td><td>JAVJAF000000000</td><td>5,262,371</td><td>1</td><td>156.0 X</td><td>64.70%</td><td>98.47%</td></tr><tr><td>SORGH_AS_0199</td><td>Pseudomonas sp.</td><td>Pirovskaya</td><td>25 °C, aerobic</td><td>Yellow</td><td>PacBio</td><td>SRP437202</td><td>JAVJAE000000000</td><td>5,020,881</td><td>1</td><td>146.4 X</td><td>65.90%</td><td>99.77%</td></tr><tr><td>SORGH_AS_0211</td><td>Pseudomonas sp.</td><td>Jensen</td><td>25 °C, aerobic</td><td>Yellow</td><td>PacBio</td><td>SRP437120</td><td>JAVIZV000000000</td><td>4,840,922</td><td>1</td><td>62.3 X</td><td>66.10%</td><td>99.87%</td></tr><tr><td>SORGH_AS_0260</td><td>Rhizobium sp.</td><td>M9 + 0.4% sucrose</td><td>37 °C, anaerobic</td><td>White</td><td>PacBio</td><td>SRP437152</td><td>JAURUW000000000</td><td>5,274,802</td><td>3</td><td>126.6 X</td><td>59.30%</td><td>99.62%</td></tr><tr><td>SORGH_AS_0285</td><td>Rhizobium sp.</td><td>Jensen</td><td>25 °C, aerobic</td><td>Yellow</td><td>PacBio</td><td>SRP437128</td><td>JAURUX000000000</td><td>5,277,463</td><td>3</td><td>136.4 X</td><td>59.30%</td><td>99.36%</td></tr><tr><td>SORGH_AS_0755</td><td>Rhizobium sp.</td><td>M9 nitrogen-free, Carbon-free</td><td>37 °C, aerobic</td><td>White, EPS production</td><td>PacBio</td><td>SRP437200</td><td>JAURUV000000000</td><td>5,423,623</td><td>3</td><td>122.8 X</td><td>59.40%</td><td>99.84%</td></tr><tr><td>SORGH_AS_0787</td><td>Rhizobium sp.</td><td>M9 nitrogen-free + 1% arabinose</td><td>25 °C, aerobic</td><td>White, EPS production</td><td>PacBio</td><td>SRP437131</td><td>JAUTBG000000000</td><td>5,400,756</td><td>3</td><td>141.4 X</td><td>58.20%</td><td>98.19%</td></tr><tr><td>SORGH_AS_0301</td><td>Rhodococcus sp.</td><td>Jensen</td><td>25 °C, aerobic</td><td>Pink, EPS production</td><td>PacBio</td><td>SRP437130</td><td>JAUTAC000000000</td><td>4,603,776</td><td>2</td><td>55.1 X</td><td>68.60%</td><td>97.93%</td></tr><tr><td>SORGH_AS_0303</td><td>Rhodococcus sp.</td><td>Jensen</td><td>25 °C, aerobic</td><td>Pink, EPS production</td><td>PacBio</td><td>SRP437177</td><td>JAUTBO000000000</td><td>4,508,481</td><td>1</td><td>162.5 X</td><td>68.50%</td><td>98.78%</td></tr></table>
<table><thead><tr><th></th><th></th><th></th><th></th><th></th><th></th><th></th><th></th><th></th><th></th><th></th><th></th><th></th><th></th><th></th><th></th><th></th><th></th><th></th><th></th></tr></thead><tbody><tr><td>SORGH_AS_0500</td><td>Siphonobacter sp.</td><td>M9 nitrogen-free +</td><td>25 °C, aerobic</td><td>Orange, EPS production</td><td>PacBio</td><td>SRP437204</td><td>JAVIYT000000000</td><td>6,158,200</td><td>9</td><td>119.3 X</td><td>44.10%</td><td>99.70%</td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>SORGH_AS_0438</td><td>Sphingomonas sp.</td><td>M9 + 0.4% sucrose</td><td>25 °C, aerobic</td><td>Orange</td><td>PacBio</td><td>SRP437229</td><td>JAVIZS000000000</td><td>4,040,749</td><td>4</td><td>165.8 X</td><td>67.80%</td><td>98.99%</td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>SORGH_AS_0742</td><td>Sphingomonas sp.</td><td>M9 nitrogen-free + 1% xylose</td><td>37 °C, aerobic</td><td>Yellow</td><td>PacBio</td><td>SRP437158</td><td>JAVIZP000000000</td><td>4,247,105</td><td>2</td><td>30.2 X</td><td>67.10%</td><td>99.25%</td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>SORGH_AS_0789</td><td>Sphingomonas sp.</td><td>M9 nitrogen-free + 1% arabinose</td><td>37 °C, aerobic</td><td>Yellow</td><td>PacBio</td><td>SRP437240</td><td>JAVIYV000000000</td><td>4,247,089</td><td>2</td><td>13.5 X</td><td>67.10%</td><td>99.42%</td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>SORGH_AS_0802</td><td>Sphingomonas sp.</td><td>M9 nitrogen-free + 1% galactose</td><td>25 °C, aerobic</td><td>Orange</td><td>Illumina</td><td>SRP437478</td><td>JAVIZL000000000</td><td>3,943,888</td><td>20</td><td>398.6 X</td><td>67.10%</td><td>99.55%</td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>SORGH_AS_0457</td><td>Stenotrophomonas rhizophila</td><td>TSA</td><td>25 °C, aerobic</td><td>White</td><td>PacBio</td><td>SRP437136</td><td>JAUTAS000000000</td><td>4,160,543</td><td>1</td><td>107.6 X</td><td>66.70%</td><td>99.71%</td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>SORGH_AS_0321</td><td>Stenotrophomonas sp.</td><td>Jensen</td><td>25 °C, anaerobic</td><td>Yellow</td><td>PacBio</td><td>SRP437127</td><td>JAVIZG000000000</td><td>4,110,757</td><td>1</td><td>159.9 X</td><td>66.70%</td><td>99.20%</td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>SORGH_AS_0282</td><td>Stenotrophomonas sp.</td><td>Jensen</td><td>37 °C, aerobic</td><td>Yellow</td><td>PacBio</td><td>SRP437122</td><td>JAUTAB000000000</td><td>4,172,479</td><td>1</td><td>166.1 X</td><td>66.70%</td><td>99.77%</td><td></td><td></td><td></td><td></td><td></td><td></td></tr></tbody></table>

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Figures

Figure 1.- Phylogenetic diversity of the bacterial collection cultivated from sorghum aerial root mucilage. The archaea *Sulfolobus acidocaldarius* DSM 639 (ASM2847236v1) was included as an outgroup. The bacterial reference strains *Pseudomonas aeruginosa* PA01 (ASM676v1), *Microbacterium foliorum* DSM 12966 (ASM95641v1), *Stenotrophomonas lactitubi* M15 (ASM280351v1), *Bacillus subtilis* subsp. *subtilis* NCIB 3610 (ASM608879v1), *Flavobacterium johnsoniae* UW101 (ASM1664v1), *Sphingomonas paucimobilis* FDAARGOS_908 (ASM1602709v1), and *Rhizobium leguminosarum* SM52 (ASM430655v1) were also included. The species Tree was inferred from unrroted orthogroups gene trees using STAG and then rooted using the STRIDE algorithms implemented in OrthoFinder. Species tree was annotated with iTOLs.

<figure><img src="image_4.png" /></figure>

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