Accès libre

Reproductive Span of Caenorhabditis elegans Is Extended by Microbacterium sp.

Tho Son Le
Tho Son Le
, 
Thi Hong Gam Nguyen
Thi Hong Gam Nguyen
, 
Bich Hong Ha
Bich Hong Ha
, 
Bui Thi Mai Huong
Bui Thi Mai Huong
, 
Thi Thu Hang Nguyen
Thi Thu Hang Nguyen
, 
Kim Dung Vu
Kim Dung Vu
, 
Tu Cuong Ho
Tu Cuong Ho
 et 
John Wang
John Wang
   | 10 mai 2022
Journal of Nematology's Cover Image
À propos de cet article
Article précédent
Article suivant
Citez
Article Category: Research Paper
Publié en ligne: 10 mai 2022
Pages: -
Reçu: 29 juin 2021
DOI: https://doi.org/10.2478/jofnem-2022-0010
Mots clés
© 2022 Tho Son Le et al., published by Sciendo
This work is licensed under the Creative Commons Attribution 4.0 International License.

Figure 1

RS of individual nematodes on Microbacterium sp. CFBb37 and the E. coli OP50 control. (A) C. elegans, (B) C. briggsae, (C) C. tropicalis, and (D) Protorhabditis sp. on CFBb37 had longer RS than on OP50 (P < 0.001, log-rank test). Full RS data in Table 2. RS, reproductive span.
RS of individual nematodes on Microbacterium sp. CFBb37 and the E. coli OP50 control. (A) C. elegans, (B) C. briggsae, (C) C. tropicalis, and (D) Protorhabditis sp. on CFBb37 had longer RS than on OP50 (P < 0.001, log-rank test). Full RS data in Table 2. RS, reproductive span.

Figure 2

Lifespan assays of C. elegans on Microbacterium sp. CFBb37 and the OP50 control. C. elegans grown on CFBb37 survived longer than OP50 (P < 0.001, log-rank test). Detailed lifespan data are in Tables S4 and S5.
Lifespan assays of C. elegans on Microbacterium sp. CFBb37 and the OP50 control. C. elegans grown on CFBb37 survived longer than OP50 (P < 0.001, log-rank test). Detailed lifespan data are in Tables S4 and S5.

Figure 3

Summary of RS extension in the Caenorhabditis species (orange) and the outgroup Protorhabditis sp. (light blue). Red lines indicate inferred ancestral evolutionary lineages with extended RS. We suggest that other species within the genus Caenorhabditis are likely to have extended RS but this is untested. For simplification, some species are not listed and are grouped together in triangles, with the numbers of known species indicated. Phylogeny is adapted from Kiontke and Fitch (2005) and Felix et al. (2014) with the position of Protorhabditis sp. CFB231 placed next to P. sp. 1 species based on the 18S ribosomal RNA gene sequence (Zhang et al., 2000; Morgulis et al., 2008). Protorhabditis sp. CFB231 is self-fertile but the sexual system, hermaphroditic or parthenogenetic, is unknown (indicated by a question mark). RS, reproductive span.
Summary of RS extension in the Caenorhabditis species (orange) and the outgroup Protorhabditis sp. (light blue). Red lines indicate inferred ancestral evolutionary lineages with extended RS. We suggest that other species within the genus Caenorhabditis are likely to have extended RS but this is untested. For simplification, some species are not listed and are grouped together in triangles, with the numbers of known species indicated. Phylogeny is adapted from Kiontke and Fitch (2005) and Felix et al. (2014) with the position of Protorhabditis sp. CFB231 placed next to P. sp. 1 species based on the 18S ribosomal RNA gene sequence (Zhang et al., 2000; Morgulis et al., 2008). Protorhabditis sp. CFB231 is self-fertile but the sexual system, hermaphroditic or parthenogenetic, is unknown (indicated by a question mark). RS, reproductive span.

Figure S1

Size comparison of the four tested nematode species grown on Microbacterium sp. CFBb37 and Escherichia coli OP50. Images are representative samples.
Size comparison of the four tested nematode species grown on Microbacterium sp. CFBb37 and Escherichia coli OP50. Images are representative samples.

Figure S2

Protorhabditis sp. CFB231 make sperm. (A) DIC image of spermathecal region (outlined). One oocyte is apparently transiting the spermatheca. (B) DAPI staining for nuclei. (C) Merged images of DIC and DAPI. DIC, differential interference contrast.
Protorhabditis sp. CFB231 make sperm. (A) DIC image of spermathecal region (outlined). One oocyte is apparently transiting the spermatheca. (B) DAPI staining for nuclei. (C) Merged images of DIC and DAPI. DIC, differential interference contrast.

Total average brood and RS of tested nematode species.

Reproductive day number OP50 CFBb37 Different between the strains P (paired t-test)
Eggs laid per day (mean ± SE) Greater than zero P (one-sample t-test) Eggs laid per day (mean ± SE) Greater than zero P (one-sample t-test)
C. elegans
Day 1 32.68 ± 5.04 <0.001 3.76 ± 1.74 <0.001 <0.01
Day 2 149.75 ± 4.71 <0.001 24.64 ± 2.53 <0.001 <0.001
Day 3 78.55 ± 6.46 <0.001 23.49 ± 2.12 <0.001 <0.001
Day 4 3.25 ± 0.81 <0.001 22.05 ± 2.31 <0.001 <0.001
Day 5 1.40 ± 0.77 >0.05 15.10 ± 1.75 <0.001 <0.001
Day 6 0.58 ± 0.55 >0.05 7.93 ± 1.14 <0.001 <0.001
Day 7 0.05 ± 0.03 >0.05 4.39 ± 0.79 <0.001 <0.001
Day 8 0.03 ± 0.03 >0.05 1.88 ± 0.51 <0.001 <0.01
Day 9 0 0.56 ± 0.23 <0.05 N.A.
Day 10 0.07 ± 0.04 >0.05 N.A.
Day 11 0.03 ± 0.03 >0.05 N.A.
Day 12 0
na 40 41
Average brood size per wormb 266.28 ± 6.42 137.63 ± 5.74 <0.001
RS (day)c 4.08 ± 0.16 7.29 ± 0.25 <0.001 (log-rank test)
C. briggsae
Day 1 11.59 ± 1.33 <0.001 2.11 ± 0.34 <0.001 <0.001
Day 2 18.21 ± 1.89 <0.001 3.80 ± 0.57 <0.001 <0.001
Day 3 22.56 ± 3.46 <0.001 3.17 ± 0.56 <0.001 <0.001
Day 4 12.00 ± 2.52 <0.001 2.03 ± 0.50 <0.001 <0.001
Day 5 0.88 ± 0.28 <0.01 2.30 ± 0.50 <0.001 0.0109
Day 6 0.16 ± 0.10 >0.05 1.69 ± 0.44 <0.001 <0.01
Day 7 0 0.50 ± 0.17 <0.001 N.A.
Day 8 0.22 ± 0.07 <0.01 N.A.
Day 9 0.08 ± 0.08 >0.05 N.A.
Day 10 0.08 ± 0.08 >0.05 N.A.
Day 11 0
na 32 36
Average brood size per wormb 65.40 ± 7.09 16.00 ± 1.88 <0.001
RS (day)c 3.97 ± 0.25 5.61 ± 0.39 <0.001 (log-rank test)
C. tropicalis
Day 1 4.37 ± 0.78 <0.001 4.75 ± 0.65 <0.001 1.0
Day 2 4.81 ± 1.37 <0.001 4.79 ± 0.92 <0.001 0.8652
Day 3 1.22 ± 0.39 <0.01 7.20 ± 1.71 <0.001 <0.01
Day 4 0.37 ± 0.24 >0.05 3.88 ± 1.05 <0.01 <0.01
Day 5 0 3.29 ± 0.92 <0.01 N.A.
Day 6 0.95 ± 0.29 <0.01 N.A.
Day 7 0.33 ± 0.14 <0.05 N.A.
Day 8 0.04 ± 0.04 >0.05 N.A.
Day 9 0 N.A.
na 27 24
Average brood size per wormb 10.78 ± 1.90 25.25 ± 3.59 <0.01
RS (day)c 2.37 ± 0.20 4.96 ± 0.36 <0.001 (log-rank test)
Protorhabditis sp.
Day 1 45.00 ± 5.02 <0.001 2.70 ± 0.30 <0.001 <0.001
Day 2 95.15 ± 7.18 <0.001 4.21 ± 0.49 <0.001 <0.001
Day 3 18.06 ± 4.95 <0.001 3.23 ± 0.52 <0.001 <0.01
Day 4 2.00 ± 0.69 <0.01 3.12 ± 0.56 <0.001 >0.05
Day 5 0.88 ± 0.70 >0.05 2.38 ± 0.43 <0.001 N.A.
Day 6 0.24 ± 0.18 >0.05 1.36 ± 0.28 <0.001 N.A.
Day 7 0.06 ± 0.06 >0.05 1.09 ± 0.24 <0.001 N.A.
Day 8 0 1.00 ± 0.24 <0.001 N.A.
Day 9 1.17 ± 0.33 <0.01 N.A.
Day 10 0.53 ± 0.18 <0.01 N.A.
Day 11 0.53 ± 0.19 <0.01 N.A.
Day 12 0.42 ± 0.14 <0.01 N.A.
Day 13 0.14 ± 0.06 <0.05 N.A.
Day 14 0.10 ± 0.07 >0.05 N.A.
Day 15 0.06 ± 0.04 >0.05 N.A.
0
na 34 47
Average brood size per wormb 161.38 ± 10.05 22.08 ± 2.26 <0.01
RS (day)c 3.59 ± 0.20 6.68 ± 0.61 <0.001 (log-rank test)

Bacterial isolates with description of colonial morphology, and geography of sample sites.

Cat Tien National Park Cuc Phuong National Park
Strain Morphology of colonya North East Strain Morphology of colonya North East
CFBb1 Small 11°23′57.5″ 107°20′21.4″ CFBb65 Big 20°17′40.4″ 105°39′46.6″
CFBb2 Big 11°23′54.4″ 107°20′21.4″ CFBb66 Big – –
CFBb3 Small – – CFBb67 Small – –
CFBb4 Irregular 11°24′22.8″ 107°24′22″ CFBb68 Small – –
CFBb5 Irregular – – CFBb69 Big 20°17′40″ 105°39′57.5″
CFBb6 Big – – CFBb70 Small – –
CFBb7 Big – – CFBb71 Small 20°17′41.3″ 105°39′59.6″
CFBb8 Small – – CFBb72 Small – –
CFBb9 Big 11°24′20.9″ 107°24′22.3″ CFBb73 Big – –
CFBb10 Irregular 11°24′21.4″ 107°24′25.6″ CFBb74 Big – –
CFBb11 Big 11°24′20.9″ 107°24′22.3″ CFBb75 Small – –
CFBb12 Small 11°24′20.9″ 107°24′22.1″ CFBb76 Irregular 20°20′56.7″ 105°35′48.1″
CFBb13 Big 11°24′19.6″ 107°24′22.1″ CFBb77 Big – –
CFBb14 Small – – CFBb78 Small 20°15′51.2″ 105°41′57.6″
CFBb15 Big 11°24′19″ 107°24′22.2″ CFBb79 Big 20°20′56.6″ 105°35′47.4″
CFBb16 Big 11°24′18.6″ 107°24′22.3″ CFBb80 Small – –
CFBb17 Irregular 11°24′21.4″ 107°24′21.4″ CFBb81 Irregular – –
CFBb18 Big – – CFBb82 Big 20°15′7.6″ 105°42′45″
CFBb19 Big 11°28′46.4″ 107°22′52.9″ CFBb83 Small – –
CFBb20 Small – – CFBb84 Big 20°14′53.5″ 105°42′34.3″
CFBb21 Medium 11°23′18.7″ 107°21′3.4″ CFBb85 Small – –
CFBb22 Medium – – CFBb86 Small – –
CFBb23 Big 11°29′7.2″ 107°22′58″ CFBb87 Medium 20°14′59.8″ 105°42′28.1″
CFBb24 Small – – CFBb88 Medium – –
CFBb25 Medium 11°27′56.1″ 107°22′43.5″ CFBb89 Big – –
CFBb26 Big, yellow 11°27′0.9″ 107°21′27.1″ CFBb90 Small – –
CFBb27 Small – – CFBb91 Big 20°14′56.7″ 105°42′21.1″
CFBb28 Big 11°27′1.5″ 107°21′27.5″ CFBb92 Big – –
CFBb29 Small – – CFBb93 Big 20°14′39.1″ 105°42′25.3″
CFBb30 Big, red 11°26′56.6″ 107°21′35.9″ CFBb94 Small – –
CFBb31 Big – – CFBb95 Big 20°20′56.7″ 105°35′46.5″
CFBb32 Big 11°23′18.8″ 107°21′3.6″ CFBb96 Small – –
CFBb33 Small – – CFBb97 Medium 20°20′58.4″ 105°35′36.5″
CFBb34 Big 11°27′31.9″ 107°20′43.1″ CFBb98 Medium – –
CFBb35 Big 11°27′5″ 107°27′25.6″ CFBb99 Big – –
CFBb36 Irregular 11°27′3″ 107°21′51.7″ CFBb100 Big – –
CFBb37 Small 11°27′5″ 107°27′25.6″ CFBb101 Small – –
CFBb38 Small – – CFBb102 Big 20°21′0.5″ 105°35′36.3″
CFBb39 Big 11°27′17.9″ 107°22′5.6″ CFBb103 Big – –
CFBb40 Big 11°27′17.9″ 107°22′5.6″ CFBb104 Small – –
CFBb41 Small 11°27′17.9″ 107°22′5.6″ CFBb105 Small – –
CFBb42 Big 11°26′47.8″ 107°26′19.8″ CFBb106 Big – –
CFBb43 Small 11°26′47.8″ 107°26″19.8″ CFBb107 Small – –
CFBb44 Big 11°27′50.4″ 107°27′43.5″ CFBb108 Big 20°20′54.1″ 105°35′48.5″
CFBb45 Irregular 11°27′51.8″ 107°27′42.1″ CFBb109 Small – –
CFBb46 Big – –
CFBb47 Small – –
CFBb48 Big 11°23′19.3″ 107°21′3.7″
CFBb49 Small – –
CFBb50 Big, pink 11°26′14.5″ 107°25′24.2″
CFBb51 Small – –
CFBb52 Big 11°26′12.1″ 107°25′25.7″
CFBb53 Small 11°26′12.1″ 107°25′25.7″
CFBb54 Small 11°26′12.1″ 107°25′25.7″
CFBb55 Big 11°25′23″ 107°25′41.4″
CFBb56 Small – –
CFBb57 Medium 11°23′26″ 107°21′1.8″
CFBb58 Big 11°24′49″ 107°25′27.7″
CFBb59 Big – –
CFBb60 Small – –
CFBb61 Small – –
CFBb62 Big – –
CFBb63 Medium 11°24′48.2″ 107°25′25.9″
CFBb64 Small 11°23′39.7″ 107°21′47.9″

Counts of surviving and lifespan of C. elegans on Microbacterium sp. CFBb37 and the OP50 control.

Day of lifespan Number of surviving worms
OP50 CFBb37
1 73 60
2 73 60
3 73 60
4 73 60
5 73 60
6 73 60
7 73 60
8 73 60
9 72 59
10 72 59
11 69 59
12 66 59
13 63 58
14 46 54
15 39 53
16 36 52
17 25 52
18 18 49
19 16 48
20 14 48
21 13 47
22 12 47
23 11 45
24 7 43
25 6 43
26 6 42
27 4 36
28 3 30
29 3 25
30 1 24
31 1 16
32 0 11
33 6
34 1
35 0
Censored 7 0
Lifespan (days) (mean ± 1SE) 17.26 ± 0.57 26.77 ± 0.86
P (log-rank test) <0.001

Average daily individual brood countsa for C. eleganson three environmental bacterial species and the OP50 control.

Reproductive day number Acinetobacter sp. CFBb9 (N = 4, n = 29)b Serratia sp. CFBb17 (N = 4, n = 39)b Microbacterium sp. CFBb37 (N = 3, n = 37)b Escherichia coli OP50 (N = 5, n = 33)b
Eggs laid per individual Greater than zero P (one-sample t-test) Eggs laid per individual Greater than zero P (one-sample t-test) Eggs laid per individual Greater than zero P (one-sample t-test) Eggs laid per individual Greater than zero P (one-sample t-test)
Day 1 41.24 <0.001 69.30 <0.001 35.16 <0.001 151.69 <0.001
Day 2 162.34 <0.001 165.74 <0.001 47.00 <0.001 95.39 <0.001
Day 3 44.51 <0.001 39.23 <0.001 24.08 <0.001 2.96 <0.001
Day 4 2.07 <0.001 0.95 <0.001 13.59 <0.001 1.12 <0.001
Day 5 0 0 8.86 <0.001 0
Day 6 0 0 6.16 <0.001 0
Day 7 3.51 <0.001
Day 8 1.49 <0.001
Day 9 1.02 <0.001
Day 10 0.24 <0.001
Day 11 0.51 <0.001
Day 12 0
Day 13 0
Average brood 250.16 275.22 141.62 251.16
size per wormc
RS (days)d 4 4 11 3.8

Wild-type isolates of nematodes in Cat Tien National Park.

Species name Strain number GenBank accession No. Best BLAST hita Query coverage Identity (%) Bit score (max/total) E value
Caenorhabditis briggsae CFB233 MZ457559 Caenorhabditis briggsae AF16 98% 100% 1,544/1,544 0.0
Protorhabditis sp. CFB231 MZ474671 Protorhabditis sp. 1 GVDU-2019 90% 95.35 1,229/1,229 0.0

Bacterial strains isolated from rotting vegetation samples collected in Cat Tien National Park and their effects on nematodes.

Straina Species Isolation site GenBank accession No. Phenotype
CFBb3 Pseudomonas sp. 11°23′54.4″N107°20′21.4″E OM456346 Late laying egg
CFBb9b Acinetobacter sp. 11°24′20.9″N107°24′22.3″E MZ467312 Normal
CFBb11 Acinetobacter sp. 11°24′20.9″N107°24′22.3″E OM456348 Small body
CFBb17 Serratia sp. 11°24′21.4″N107°24′21.4″E MZ457333 Small body
CFBb35 Serratia sp. 11°27′5″N107°27′25.6″E OM629173 Normal
CFBb37 Microbacterium sp 11°27′5″N107°27′25.6″E MZ453404 RS; small body
CFBb39 Sphingobacterium sp. 11°27′17.9″N107°22′5.6″E OM892000 Normal
CFBb40 Bacillus sp. 11°27′17.9″N107°22′5.6″E OM456347 Normal
CFBb42 Serratia sp. 11°26′47.8″N107°26′19.8″E OM456350 Small body
CFBb53 Bacillus cereus 11°26′12.1″N107°25′25.7″E OM883917 Small body
CFBb54 Enterobacter sp. 11°26′12.1″N107°25′25.7″E OM456349 Small body
CFBb57 Serratia sp. 11°23′26″N107°21′1.8″E OM892003 Small body

Counts of daily uterus eggs of close nematode species.

Nematode Reproductive day number OP50 CFBb37 Different between the strains P (paired t-test)
Number of tested hermaphrodites Number of eggs in uterus/individual (mean ± SE) Greater than zero P (one-sample t-test) Number of tested hermaphrodites Number of eggs in uterus/individual (mean ± SE) Greater than zero P (one-sample t-test)
C. elegans Day 1 32 3.94 ± 0.68 <0.001 35 1.57 ± 0.34 <0.001 0.0051
Day 2 32 25.25 ± 1.12 <0.001 31 3.13 ± 0.32 <0.001 <0.001
Day 3 29 5.38 ± 0.98 <0.001 42 4.02 ± 0.33 <0.001 0.06615
Day 4 30 1.13 ± 0.34 <0.01 32 3.69 ± 0.33 <0.001 <0.001
Day 5 37 0.03 ± 0.03 >0.05 34 3.5 ± 0.46 <0.001 <0.001
Day 6 37 3.22 ± 0.51 <0.001
Day 7 34 2.29 ± 0.36 <0.001
Day 8 35 0.43 ± 0.16 <0.01
C. briggsae Day 1 39 3.30 ± 0.73 <0.001 39 0.51 ± 0.09 <0.001 <0.001
Day 2
Day 3 34 4.09 ± 0.39 <0.001 35 0.6 ± 0.15 <0.001 <0.001
Day 4 39 2.38 ± 0.24 <0.001 32 0.81 ± 0.14 <0.001 <0.001
Day 5 35 0.2 ± 0.07 <0.01 35 1.46 ± 0.15 <0.001 <0.001
Day 6 34 0.09 ± 0.05 >0.05 31 1.45 ± 0.19 <0.001 <0.001
Day 7 32 0.63 ± 0.15 <0.001
C. tropicalis Day 1 33 2.09 ± 0.22 <0.001 32 0.28 ± 0.09 <0.001 <0.001
Day 2 32 3.47 ± 0.23 <0.001 37 2.76 ± 0.32 <0.001 0.2149
Day 3 38 1.87 ± 0.23 <0.001 36 3.19 ± 0.4 <0.001 0.0044
Day 4 28 0.64 ± 0.23 <0.05 38 2.6 ± 0.24 <0.001 <0.001
Day 5 36 0.19 ± 0.13 >0.05 34 2.94 ± 0.36 <0.001 <0.001
Day 6 36 1.53 ± 0.2 <0.001
Day 7 27 1.63 ± 0.22 <0.001
Day 8 17 0.82 ± 0.16 <0.01
P. sp. Day 1 34 3.23 ± 0.55 <0.001 32 3.28 ± 0.26 <0.001 0.595
Day 2 36 3.67 ± 0.63 <0.001 38 1.61 ± 0.18 <0.001 <0.001
Day 3 39 1.38 ± 0.24 <0.001 36 1.58 ± 0.12 <0.001 0.629
Day 4 36 0.19 ± 0.03 <0.05 36 1.56 ± 0.12 <0.001 <0.001
Day 5 34 0.15 ± 0.03 >0.05 40 1.80 ± 0.16 <0.001 <0.001
Day 6 35 1.06 ± 0.13 <0.001 <0.001
Day 7 53 0.81 ± 0.10 <0.001 <0.001
eISSN:
2640-396X
Langue:
Anglais
Périodicité:
Volume Open
Sujets de la revue:
Life Sciences, other
RSS Feed de la revue