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(A) Accumulation of <t>PvCRT08</t> transcripts in whole roots that were either uninoculated or inoculated with R. tropici CIAT899, at the indicated number of days post-inoculation (dpi). (B) Relative accumulation of PvCRT08 transcripts in nodules and nodule-free roots at the indicated number of days post-inoculation (dpi). For both A and B, total RNA was isolated from each biological sample and used in RT-qPCR analysis. The elongation factor EF1α was used as an endogenous reference gene. Each bar represents the mean ± SEM of two independent biological replicates (n> 6) with three technical repeats. ** p <0.005 and **** p <0.0001 based on Student’s t -test.
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(A) Accumulation of <t>PvCRT08</t> transcripts in whole roots that were either uninoculated or inoculated with R. tropici CIAT899, at the indicated number of days post-inoculation (dpi). (B) Relative accumulation of PvCRT08 transcripts in nodules and nodule-free roots at the indicated number of days post-inoculation (dpi). For both A and B, total RNA was isolated from each biological sample and used in RT-qPCR analysis. The elongation factor EF1α was used as an endogenous reference gene. Each bar represents the mean ± SEM of two independent biological replicates (n> 6) with three technical repeats. ** p <0.005 and **** p <0.0001 based on Student’s t -test.
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(A) Accumulation of <t>PvCRT08</t> transcripts in whole roots that were either uninoculated or inoculated with R. tropici CIAT899, at the indicated number of days post-inoculation (dpi). (B) Relative accumulation of PvCRT08 transcripts in nodules and nodule-free roots at the indicated number of days post-inoculation (dpi). For both A and B, total RNA was isolated from each biological sample and used in RT-qPCR analysis. The elongation factor EF1α was used as an endogenous reference gene. Each bar represents the mean ± SEM of two independent biological replicates (n> 6) with three technical repeats. ** p <0.005 and **** p <0.0001 based on Student’s t -test.
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(A) Accumulation of <t>PvCRT08</t> transcripts in whole roots that were either uninoculated or inoculated with R. tropici CIAT899, at the indicated number of days post-inoculation (dpi). (B) Relative accumulation of PvCRT08 transcripts in nodules and nodule-free roots at the indicated number of days post-inoculation (dpi). For both A and B, total RNA was isolated from each biological sample and used in RT-qPCR analysis. The elongation factor EF1α was used as an endogenous reference gene. Each bar represents the mean ± SEM of two independent biological replicates (n> 6) with three technical repeats. ** p <0.005 and **** p <0.0001 based on Student’s t -test.
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(A) Accumulation of <t>PvCRT08</t> transcripts in whole roots that were either uninoculated or inoculated with R. tropici CIAT899, at the indicated number of days post-inoculation (dpi). (B) Relative accumulation of PvCRT08 transcripts in nodules and nodule-free roots at the indicated number of days post-inoculation (dpi). For both A and B, total RNA was isolated from each biological sample and used in RT-qPCR analysis. The elongation factor EF1α was used as an endogenous reference gene. Each bar represents the mean ± SEM of two independent biological replicates (n> 6) with three technical repeats. ** p <0.005 and **** p <0.0001 based on Student’s t -test.
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(A) Accumulation of <t>PvCRT08</t> transcripts in whole roots that were either uninoculated or inoculated with R. tropici CIAT899, at the indicated number of days post-inoculation (dpi). (B) Relative accumulation of PvCRT08 transcripts in nodules and nodule-free roots at the indicated number of days post-inoculation (dpi). For both A and B, total RNA was isolated from each biological sample and used in RT-qPCR analysis. The elongation factor EF1α was used as an endogenous reference gene. Each bar represents the mean ± SEM of two independent biological replicates (n> 6) with three technical repeats. ** p <0.005 and **** p <0.0001 based on Student’s t -test.
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(A) Accumulation of <t>PvCRT08</t> transcripts in whole roots that were either uninoculated or inoculated with R. tropici CIAT899, at the indicated number of days post-inoculation (dpi). (B) Relative accumulation of PvCRT08 transcripts in nodules and nodule-free roots at the indicated number of days post-inoculation (dpi). For both A and B, total RNA was isolated from each biological sample and used in RT-qPCR analysis. The elongation factor EF1α was used as an endogenous reference gene. Each bar represents the mean ± SEM of two independent biological replicates (n> 6) with three technical repeats. ** p <0.005 and **** p <0.0001 based on Student’s t -test.
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(A) Accumulation of PvCRT08 transcripts in whole roots that were either uninoculated or inoculated with R. tropici CIAT899, at the indicated number of days post-inoculation (dpi). (B) Relative accumulation of PvCRT08 transcripts in nodules and nodule-free roots at the indicated number of days post-inoculation (dpi). For both A and B, total RNA was isolated from each biological sample and used in RT-qPCR analysis. The elongation factor EF1α was used as an endogenous reference gene. Each bar represents the mean ± SEM of two independent biological replicates (n> 6) with three technical repeats. ** p <0.005 and **** p <0.0001 based on Student’s t -test.

Journal: bioRxiv

Article Title: Calreticulin modulates the infection process and nodule organogenesis in the Phaseolus vulgaris-Rhizobium symbiosis

doi: 10.64898/2026.04.09.717402

Figure Lengend Snippet: (A) Accumulation of PvCRT08 transcripts in whole roots that were either uninoculated or inoculated with R. tropici CIAT899, at the indicated number of days post-inoculation (dpi). (B) Relative accumulation of PvCRT08 transcripts in nodules and nodule-free roots at the indicated number of days post-inoculation (dpi). For both A and B, total RNA was isolated from each biological sample and used in RT-qPCR analysis. The elongation factor EF1α was used as an endogenous reference gene. Each bar represents the mean ± SEM of two independent biological replicates (n> 6) with three technical repeats. ** p <0.005 and **** p <0.0001 based on Student’s t -test.

Article Snippet: To develop a p PvCRT08 :GUS-GFP construct, a 2086-bp fragment upstream of the PvCRT08 translation start site was amplified using bean genomic DNA and primers p PvCRT08 -Up and p PvCRT08 -Lw ( Table S6 ) and cloned into vector pENTR/SD/D-TOPO (Invitrogen).

Techniques: Isolation, Quantitative RT-PCR

Promoter activity was detected in the vascular bundles of uninoculated P. vulgaris roots (A), and in emerging lateral root primordia (B, D, E, F). Promoter activity was also detected in root hairs of uninoculated roots (C). The spatiotemporal expression pattern of the PvCRT08 promoter was monitored in transgenic roots after-inoculation with R. tropici DsRed (G–J). (G) Infection site at 5 dpi. (H) Nodule primordium at 7 dpi. (I and J) Mature nodules at 21 dpi. (J) Free-hand sections of nodules at 21 dpi. RH: root hair; IT: infection thread; CCD: cortical cell division; IC: infected cell.

Journal: bioRxiv

Article Title: Calreticulin modulates the infection process and nodule organogenesis in the Phaseolus vulgaris-Rhizobium symbiosis

doi: 10.64898/2026.04.09.717402

Figure Lengend Snippet: Promoter activity was detected in the vascular bundles of uninoculated P. vulgaris roots (A), and in emerging lateral root primordia (B, D, E, F). Promoter activity was also detected in root hairs of uninoculated roots (C). The spatiotemporal expression pattern of the PvCRT08 promoter was monitored in transgenic roots after-inoculation with R. tropici DsRed (G–J). (G) Infection site at 5 dpi. (H) Nodule primordium at 7 dpi. (I and J) Mature nodules at 21 dpi. (J) Free-hand sections of nodules at 21 dpi. RH: root hair; IT: infection thread; CCD: cortical cell division; IC: infected cell.

Article Snippet: To develop a p PvCRT08 :GUS-GFP construct, a 2086-bp fragment upstream of the PvCRT08 translation start site was amplified using bean genomic DNA and primers p PvCRT08 -Up and p PvCRT08 -Lw ( Table S6 ) and cloned into vector pENTR/SD/D-TOPO (Invitrogen).

Techniques: Activity Assay, Expressing, Transgenic Assay, Infection

Promoter activity was monitored in root hairs after inoculation with R. tropici -DsRed at 3 and 5 dpi. Control transgenic roots harboring a vehicle without promoter sequence. Promoter activity was observed by confocal microscopy on transgenic roots expressing 2086 pb of the PvCRT08 promoter region fused with GFP and GUS. dpi: days post-inoculation. Arrows indicate infection threads and the arrowhead points out a root hair.

Journal: bioRxiv

Article Title: Calreticulin modulates the infection process and nodule organogenesis in the Phaseolus vulgaris-Rhizobium symbiosis

doi: 10.64898/2026.04.09.717402

Figure Lengend Snippet: Promoter activity was monitored in root hairs after inoculation with R. tropici -DsRed at 3 and 5 dpi. Control transgenic roots harboring a vehicle without promoter sequence. Promoter activity was observed by confocal microscopy on transgenic roots expressing 2086 pb of the PvCRT08 promoter region fused with GFP and GUS. dpi: days post-inoculation. Arrows indicate infection threads and the arrowhead points out a root hair.

Article Snippet: To develop a p PvCRT08 :GUS-GFP construct, a 2086-bp fragment upstream of the PvCRT08 translation start site was amplified using bean genomic DNA and primers p PvCRT08 -Up and p PvCRT08 -Lw ( Table S6 ) and cloned into vector pENTR/SD/D-TOPO (Invitrogen).

Techniques: Activity Assay, Control, Transgenic Assay, Sequencing, Confocal Microscopy, Expressing, Infection

Number or infection events within root hairs or cortical cells of PvCRT08 -RNAi and control transgenic roots at 7 dpi with R. tropici . Values are mean + SEM with n>9 roots for each condition * p<0.05 according to Student’s t-test.

Journal: bioRxiv

Article Title: Calreticulin modulates the infection process and nodule organogenesis in the Phaseolus vulgaris-Rhizobium symbiosis

doi: 10.64898/2026.04.09.717402

Figure Lengend Snippet: Number or infection events within root hairs or cortical cells of PvCRT08 -RNAi and control transgenic roots at 7 dpi with R. tropici . Values are mean + SEM with n>9 roots for each condition * p<0.05 according to Student’s t-test.

Article Snippet: To develop a p PvCRT08 :GUS-GFP construct, a 2086-bp fragment upstream of the PvCRT08 translation start site was amplified using bean genomic DNA and primers p PvCRT08 -Up and p PvCRT08 -Lw ( Table S6 ) and cloned into vector pENTR/SD/D-TOPO (Invitrogen).

Techniques: Infection, Control, Transgenic Assay

Nodulation and nitrogen fixation capacity of PvCRT08 -RNAi and control transgenic roots. (A) Nodules were collected and counted on PvCRT08 -RNAi and control transgenic roots inoculated with R. tropici CIAT-899 at 21 dpi. (B) Nitrogenase activity was determined by acetylene reduction in nodules collected from PvCRT08 -RNAi and control transgenic roots at 21 dpi. Bars represent the mean ± SEM for three biological replicates with n =10. **** p <0.0001 based on Student’s t- test.

Journal: bioRxiv

Article Title: Calreticulin modulates the infection process and nodule organogenesis in the Phaseolus vulgaris-Rhizobium symbiosis

doi: 10.64898/2026.04.09.717402

Figure Lengend Snippet: Nodulation and nitrogen fixation capacity of PvCRT08 -RNAi and control transgenic roots. (A) Nodules were collected and counted on PvCRT08 -RNAi and control transgenic roots inoculated with R. tropici CIAT-899 at 21 dpi. (B) Nitrogenase activity was determined by acetylene reduction in nodules collected from PvCRT08 -RNAi and control transgenic roots at 21 dpi. Bars represent the mean ± SEM for three biological replicates with n =10. **** p <0.0001 based on Student’s t- test.

Article Snippet: To develop a p PvCRT08 :GUS-GFP construct, a 2086-bp fragment upstream of the PvCRT08 translation start site was amplified using bean genomic DNA and primers p PvCRT08 -Up and p PvCRT08 -Lw ( Table S6 ) and cloned into vector pENTR/SD/D-TOPO (Invitrogen).

Techniques: Control, Transgenic Assay, Activity Assay

Number of infection events within root hairs or cortical cells of transgenic PvCRT08 -OE and control roots at 7 dpi with R. tropici . Values are expressed as mean + SEM with n>9 roots for each condition **** p<0.0001 according to Student’s t-test

Journal: bioRxiv

Article Title: Calreticulin modulates the infection process and nodule organogenesis in the Phaseolus vulgaris-Rhizobium symbiosis

doi: 10.64898/2026.04.09.717402

Figure Lengend Snippet: Number of infection events within root hairs or cortical cells of transgenic PvCRT08 -OE and control roots at 7 dpi with R. tropici . Values are expressed as mean + SEM with n>9 roots for each condition **** p<0.0001 according to Student’s t-test

Article Snippet: To develop a p PvCRT08 :GUS-GFP construct, a 2086-bp fragment upstream of the PvCRT08 translation start site was amplified using bean genomic DNA and primers p PvCRT08 -Up and p PvCRT08 -Lw ( Table S6 ) and cloned into vector pENTR/SD/D-TOPO (Invitrogen).

Techniques: Infection, Transgenic Assay, Control

Nodulation and nitrogen-fixing capacity of PvCRT08 -OE transgenic and control roots. (A) Nodules were collected and counted from control and PvCRT08 -OE transgenic roots inoculated with R. tropici CIAT-899 at 21 dpi. (B) Nitrogenase activity determined by acetylene reduction in nodules collected from PvCRT08 -OE and control transgenic roots at 21 dpi. Bars represent the mean ± SEM of four biological replicates with n =10.**** p <0.0001, *** p <0.001, based on Student’s t- test.

Journal: bioRxiv

Article Title: Calreticulin modulates the infection process and nodule organogenesis in the Phaseolus vulgaris-Rhizobium symbiosis

doi: 10.64898/2026.04.09.717402

Figure Lengend Snippet: Nodulation and nitrogen-fixing capacity of PvCRT08 -OE transgenic and control roots. (A) Nodules were collected and counted from control and PvCRT08 -OE transgenic roots inoculated with R. tropici CIAT-899 at 21 dpi. (B) Nitrogenase activity determined by acetylene reduction in nodules collected from PvCRT08 -OE and control transgenic roots at 21 dpi. Bars represent the mean ± SEM of four biological replicates with n =10.**** p <0.0001, *** p <0.001, based on Student’s t- test.

Article Snippet: To develop a p PvCRT08 :GUS-GFP construct, a 2086-bp fragment upstream of the PvCRT08 translation start site was amplified using bean genomic DNA and primers p PvCRT08 -Up and p PvCRT08 -Lw ( Table S6 ) and cloned into vector pENTR/SD/D-TOPO (Invitrogen).

Techniques: Transgenic Assay, Control, Activity Assay