Fig 1.
Fig 2.
Fig 3.
Fig 4.
Enrichment factor (EF) scale, according to Sutherland (2000)_
| EF = < 2 | deficient to low enrichment |
| EF = 2–5 | moderate enrichment |
| EF = 5–20 | considerable enrichment |
| EF = 20–40 | high enrichment |
| E = > 40 | extremely high enrichment |
The content of macroelements in soil collected from Donja Paklenica_
| Locality/Element | Ca (%) | Mg (%) | K (%) | Ca/Mg |
|---|---|---|---|---|
| Donja Paklenica | 0.009 | 1.439 | 0.036 | 0.0063 |
Contamination categories for Igeo values (Huu et al_ 2010)_
| Igeo < 0 | unpolluted |
| 0 ≤ Igeo < 1 | unpolluted to moderately contaminated |
| 1 ≤ Igeo < 2 | moderately contaminated |
| 2 ≤ Igeo < 3 | moderately to strongly contaminated |
| 3 ≤ Igeo < 4 | strongly contaminated |
| 4 ≤ Igeo < 5 | strongly to extremely contaminated |
| Igeo ≥ 5 | extremely contaminated |
All Identified Rb and Eb with heavy metal resistance, nitrogen fixation, and ACC deaminase activity_
| Isolate | PGP properties | Heavy metal resistance | |||||
|---|---|---|---|---|---|---|---|
| Nitrogen fixation | ACC deaminase activity | Growth on TY agar plates supplemented with heavy metals 14 d | |||||
| Blue color, pellicle 7 d | Growth on DF + ACC agar plates | Ni 100 mg/l | Ni 200 mg/l | Cu 100 mg/l | Cu 200 mg/l | Co 100 mg/l | |
| Rb1-Pseudomonas sp. | + | + | + | + | + | − | + |
| Rb2-Pseudomonas sp. | + | + | + | + | + | − | + |
| Rb3-Pseudomonas sp. | + | + | + | + | + | − | + |
| Rb4-Pseudomonas sp. | + | + | + | + | + | − | + |
| Rb5-Pseudomonas sp. | + | + | + | + | + | − | + |
| Rb6-Pseudomonas sp. | + | + | + | + | + | − | + |
| Rb7-Pseudomonas sp. | + | + | + | + | + | − | + |
| Rb8-Pseudomonas sp. | + | + | + | + | + | − | + |
| Rb9-Bacillus sp. | + | + | + | − | − | − | − |
| Rb10-Pseudomonas sp. | + | + | + | + | + | − | + |
| Rb11-Pseudomonas sp. | + | + | + | + | + | − | + |
| Rb12-Pseudomonas sp. | + | + | + | + | + | − | + |
| Rb13-Pseudomonas sp. | + | + | + | + | + | − | + |
| Rb14-Pseudomonas sp. | + | − | + | + | + | − | + |
| Eb1-Bacillus sp. | + | + | − | − | + | − | + |
| Eb2-Staphylococcus sp. | + | + | + | − | + | − | + |
| Eb3-Bacillus sp. | + | + | − | − | + | − | + |
| Eb4-Bacillus sp. | + | − | − | − | + | − | + |
| Eb5-Bacillus sp. | − | − | − | − | − | − | − |
| Eb6-Bacillus sp. | + | + | + | − | − | − | − |
| Eb7-Pseudomonas sp. Eb8-Brevibacterium/ | + | − | − | − | − | − | − |
| Peribacillus/Bacillus sp. | − | + | + | − | − | − | − |
| Eb9-Lysinibacillus sp. | + | − | − | − | − | − | − |
| Eb10-Bacillus sp. | − | − | − | − | − | − | − |
| Eb11-Bacillus sp. | + | + | − | − | − | − | − |
BLAST results after sequence editing of Rb and Eb sequences_
| Isolate | Max score | Total score | Query cover | Query length | E-value | Percentage identity | Accession |
|---|---|---|---|---|---|---|---|
| Rb1-Pseudomonas sp. | 1033 | 1033 | 100% | 559 | 0.0 | 100.00% | MT555396.1 |
| Rb2-Pseudomonas sp. | 1029 | 1029 | 100% | 567 | 0.0 | 99.47% | MT955582.1 |
| Rb3-Pseudomonas sp. | 1007 | 1007 | 98% | 569 | 0.0 | 98.93% | MH018885.1 |
| Rb4-Pseudomonas sp. | 1026 | 1026 | 99% | 560 | 0.0 | 99.64% | KJ598026.1 |
| Rb5-Pseudomonas sp. | 1035 | 1035 | 100% | 560 | 0.0 | 100.00% | MT626826.1 |
| Rb6-Pseudomonas sp. | 1033 | 1033 | 99% | 562 | 0.0 | 100.00% | PP738394.1 |
| Rb7-Pseudomonas sp. | 1024 | 1024 | 100% | 558 | 0.0 | 98.82% | MN006008.1 |
| Rb8-Pseudomonas sp. | 1024 | 1024 | 100% | 558 | 0.0 | 98.82% | MT373558.1 |
| Rb9-Bacillus sp. | 1037 | 1037 | 97% | 573 | 0.0 | 100.00% | MG571235.1 |
| Rb10-Pseudomonas sp. | 1026 | 1026 | 99% | 561 | 0.0 | 99.64% | HM755479.1 |
| Rb11-Pseudomonas sp. | 1026 | 1026 | 99% | 560 | 0.0 | 99.64% | MH717340.1 |
| Rb12-Pseudomonas sp. | 1024 | 1024 | 100% | 558 | 0.0 | 99.82% | MT280204.1 |
| Rb13-Pseudomonas sp. | 957 | 957 | 100% | 518 | 0.0 | 100.00% | KT695830.1 |
| Rb14-Pseudomonas sp. | 1024 | 1024 | 100% | 558 | 0.0 | 99.82% | MT533925.1 |
| Eb1-Bacillus sp. | 1040 | 1040 | 98% | 574 | 0.0 | 100.00% | MT071453.1 |
| Eb2-Staphylococcus sp. | 1038 | 1038 | 100% | 562 | 0.0 | 100.00% | KM021292.1 |
| Eb3-Bacillus sp. | 1033 | 1033 | 100% | 559 | 0.0 | 100.00% | MT586023.1 |
| Eb4-Bacillus sp. | 992 | 992 | 100% | 540 | 0.0 | 99.81% | MT032498.1 |
| Eb5-Bacillus sp. | 933 | 933 | 99% | 506 | 0.0 | 100.00% | MW767008.1 |
| Eb6-Bacillus sp. | 963 | 963 | 100% | 521 | 0.0 | 100.00% | MF375095.1 |
| Eb7-Pseudomonas sp. | 966 | 966 | 100% | 523 | 0.0 | 100.00% | OM527246.1 |
| Eb8-Brevibacterium/Peribacillus/Bacillus sp. | 996 | 996 | 100% | 555 | 0.0 | 98.93% | MT538263.1 |
| Eb9-Lysinibacillus sp. | 942 | 942 | 100% | 510 | 0.0 | 100.00% | OR104997.1 |
| Eb10-Bacillus sp. | 957 | 957 | 100% | 518 | 0.0 | 100.00% | MF111532.1 |
| Eb11-Bacillus sp. | 1033 | 1033 | 100% | 559 | 0.0 | 100.00% | MF111382.1 |
Heavy metal concentration in soil collected from Donja Paklenica_
| Soil type | Cd | Ni | Mn | Pb | Co | Zn | Cr | Cu | Fe | |
|---|---|---|---|---|---|---|---|---|---|---|
| Donja Paklenica | HM (ppm) | 0 | 1,912.45 | 1042.51 | 5.76 | 87.7 | 33.69 | 203.42 | 22.19 | 16,983 |
| EF | − | 56.305 | 1.637 | 0.242 | 8.607 | 0.749 | 5.208 | 2.904 | − | |
| Igeo | − | 5.202 | 0.238 | -2.505 | 2.634 | -0.891 | 1.9178 | 1.0570 | -0.472 | |
| TV | HM (ppm) | 0.8a | 35a | − | 85a | 20a | 50a | 100a | 36a | − |
| IV | HM (ppm) | 12a | 210a | 3,000–40,000c | 530b | 240b | 720b | 380a | 190b | 3,000–500,000d |