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Chlorosis and cell death induced by transient expression of the H. annosum small secreted protein (HaSS) encoding genes in Nicotiana benthamiana . HaSSPs were custom synthesised into the pUC57 vector. Each candidate gene was cloned into the <t>pICH86988</t> expression vector by Golden Gate cloning, generating pICH86988-HaSSP. The vector was introduced into Agrobacterium tumefaciens GV3101 followed by agroinfiltration into Nicotiana benthamiana leaves. The plants were incubated in a growth room with 12 h/12 h, night/day photoperiods at 20–24 °C and 60% relative humidity (RH). The infiltrated leaves were monitored for 2 to 6 days post-infiltration (dpi).
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List of strains and DNA sequences used in this study.
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Chlorosis and cell death induced by transient expression of the H. annosum small secreted protein (HaSS) encoding genes in Nicotiana benthamiana . HaSSPs were custom synthesised into the pUC57 vector. Each candidate gene was cloned into the pICH86988 expression vector by Golden Gate cloning, generating pICH86988-HaSSP. The vector was introduced into Agrobacterium tumefaciens GV3101 followed by agroinfiltration into Nicotiana benthamiana leaves. The plants were incubated in a growth room with 12 h/12 h, night/day photoperiods at 20–24 °C and 60% relative humidity (RH). The infiltrated leaves were monitored for 2 to 6 days post-infiltration (dpi).

Journal: Scientific Reports

Article Title: Small secreted proteins from the necrotrophic conifer pathogen Heterobasidion annosum s.l . (HaSSPs) induce cell death in Nicotiana benthamiana

doi: 10.1038/s41598-017-08010-0

Figure Lengend Snippet: Chlorosis and cell death induced by transient expression of the H. annosum small secreted protein (HaSS) encoding genes in Nicotiana benthamiana . HaSSPs were custom synthesised into the pUC57 vector. Each candidate gene was cloned into the pICH86988 expression vector by Golden Gate cloning, generating pICH86988-HaSSP. The vector was introduced into Agrobacterium tumefaciens GV3101 followed by agroinfiltration into Nicotiana benthamiana leaves. The plants were incubated in a growth room with 12 h/12 h, night/day photoperiods at 20–24 °C and 60% relative humidity (RH). The infiltrated leaves were monitored for 2 to 6 days post-infiltration (dpi).

Article Snippet: From Level0 plasmids, the HaSSP coding sequences were cloned into the expression vector, Level2 pICH86988, creating the final vector pICH86988-HaSSP (35 S promoter and octopine synthase terminator, Addgene, UK) using the Bsa I restriction enzyme (NEB, Finland).

Techniques: Expressing, Plasmid Preparation, Clone Assay, Incubation

Gene expression levels of several markers related to plant immunity in Nicotiana benthamiana leaves transiently expressing HaSSP30 for 1, 2, and 3 days. Nicotiana benthamiana leaves were infiltrated with Agrobacterium tumefaciens GV3101 carrying either the pICH86988-empty (Control) or pICH86988-HaSSP30 (HaSSP30) vector. The plants were incubated in a plant growth room with 12 h/12 h, night/day photoperiods at 20–24 °C with 60% relative humidity (RH). The leaves were harvested after 1, 2, and 3 days post-infiltration (dpi). Total RNA was extracted and reverse transcribed into cDNA. The qPCR was performed on the cDNA using specific primers for several genes related to activation of plant immunity. Two reference genes were used to normalize the data, which are presented as normalized relative quantities scaled to control. Error bars indicated the standard deviation (SD) of 3 independent replicates (n = 3). Two-tailed Student t-test was used to compare HaSSP30 versus Control in each day and p-values < 0.05 are indicated above the bars.

Journal: Scientific Reports

Article Title: Small secreted proteins from the necrotrophic conifer pathogen Heterobasidion annosum s.l . (HaSSPs) induce cell death in Nicotiana benthamiana

doi: 10.1038/s41598-017-08010-0

Figure Lengend Snippet: Gene expression levels of several markers related to plant immunity in Nicotiana benthamiana leaves transiently expressing HaSSP30 for 1, 2, and 3 days. Nicotiana benthamiana leaves were infiltrated with Agrobacterium tumefaciens GV3101 carrying either the pICH86988-empty (Control) or pICH86988-HaSSP30 (HaSSP30) vector. The plants were incubated in a plant growth room with 12 h/12 h, night/day photoperiods at 20–24 °C with 60% relative humidity (RH). The leaves were harvested after 1, 2, and 3 days post-infiltration (dpi). Total RNA was extracted and reverse transcribed into cDNA. The qPCR was performed on the cDNA using specific primers for several genes related to activation of plant immunity. Two reference genes were used to normalize the data, which are presented as normalized relative quantities scaled to control. Error bars indicated the standard deviation (SD) of 3 independent replicates (n = 3). Two-tailed Student t-test was used to compare HaSSP30 versus Control in each day and p-values < 0.05 are indicated above the bars.

Article Snippet: From Level0 plasmids, the HaSSP coding sequences were cloned into the expression vector, Level2 pICH86988, creating the final vector pICH86988-HaSSP (35 S promoter and octopine synthase terminator, Addgene, UK) using the Bsa I restriction enzyme (NEB, Finland).

Techniques: Expressing, Plasmid Preparation, Incubation, Activation Assay, Standard Deviation, Two Tailed Test

List of strains and DNA sequences used in this study.

Journal: PLoS ONE

Article Title: Bandoniozyma gen. nov., a Genus of Fermentative and Non-Fermentative Tremellaceous Yeast Species

doi: 10.1371/journal.pone.0046060

Figure Lengend Snippet: List of strains and DNA sequences used in this study.

Article Snippet: In this study we propose the new yeast genus Bandoniozyma , with seven species Bandoniozyma noutii sp. nov. (type species of genus; CBS 8364 T = DBVPG 4489 T ), Bandoniozyma aquatica sp. nov. (UFMG-DH4.20 T = CBS 12527 T = ATCC MYA-4876 T ), Bandoniozyma complexa sp. nov. (CBS 11570 T = ATCC MYA-4603 T = MA28a T ), Bandoniozyma fermentans sp. nov. (CBS 12399 T = NU7M71 T = BCRC 23267 T ), Bandoniozyma glucofermentans sp. nov. (CBS 10381 T = NRRL Y-48076 T = ATCC MYA-4760 T = BG 02-7-15-015A-1-1 T ), Bandoniozyma tunnelae sp. nov. (CBS 8024 T = DBVPG 7000 T ), and Bandoniozyma visegradensis sp. nov. (CBS 12505 T = NRRL Y-48783 T = NCAIM Y.01952 T ).

Techniques: Transmission Assay

Selected physiological/biochemical test responses for differentiation of the newly proposed Bandoniozyma species. The full data can be seen in <xref ref-type= Table S1 ." width="100%" height="100%">

Journal: PLoS ONE

Article Title: Bandoniozyma gen. nov., a Genus of Fermentative and Non-Fermentative Tremellaceous Yeast Species

doi: 10.1371/journal.pone.0046060

Figure Lengend Snippet: Selected physiological/biochemical test responses for differentiation of the newly proposed Bandoniozyma species. The full data can be seen in Table S1 .

Article Snippet: In this study we propose the new yeast genus Bandoniozyma , with seven species Bandoniozyma noutii sp. nov. (type species of genus; CBS 8364 T = DBVPG 4489 T ), Bandoniozyma aquatica sp. nov. (UFMG-DH4.20 T = CBS 12527 T = ATCC MYA-4876 T ), Bandoniozyma complexa sp. nov. (CBS 11570 T = ATCC MYA-4603 T = MA28a T ), Bandoniozyma fermentans sp. nov. (CBS 12399 T = NU7M71 T = BCRC 23267 T ), Bandoniozyma glucofermentans sp. nov. (CBS 10381 T = NRRL Y-48076 T = ATCC MYA-4760 T = BG 02-7-15-015A-1-1 T ), Bandoniozyma tunnelae sp. nov. (CBS 8024 T = DBVPG 7000 T ), and Bandoniozyma visegradensis sp. nov. (CBS 12505 T = NRRL Y-48783 T = NCAIM Y.01952 T ).

Techniques: Starch

Bandoniozyma visegradensis CBS 12505 T a) Growth on 5% malt extract agar, 3 days, 25°C, b) Septate hypha in 5% malt extract broth, 19 days, 25°C, c) Curved buds connecting independent cells (c.1) or mother cell bud pairs (c.2), YM agar, 10 days, 25°C, d) Conjugating cells, YM agar, 10 days, 25°C, e) Conjugation tubes, 2% malt extract agar, 18 days, 6°C. Bar = 10 µm.

Journal: PLoS ONE

Article Title: Bandoniozyma gen. nov., a Genus of Fermentative and Non-Fermentative Tremellaceous Yeast Species

doi: 10.1371/journal.pone.0046060

Figure Lengend Snippet: Bandoniozyma visegradensis CBS 12505 T a) Growth on 5% malt extract agar, 3 days, 25°C, b) Septate hypha in 5% malt extract broth, 19 days, 25°C, c) Curved buds connecting independent cells (c.1) or mother cell bud pairs (c.2), YM agar, 10 days, 25°C, d) Conjugating cells, YM agar, 10 days, 25°C, e) Conjugation tubes, 2% malt extract agar, 18 days, 6°C. Bar = 10 µm.

Article Snippet: In this study we propose the new yeast genus Bandoniozyma , with seven species Bandoniozyma noutii sp. nov. (type species of genus; CBS 8364 T = DBVPG 4489 T ), Bandoniozyma aquatica sp. nov. (UFMG-DH4.20 T = CBS 12527 T = ATCC MYA-4876 T ), Bandoniozyma complexa sp. nov. (CBS 11570 T = ATCC MYA-4603 T = MA28a T ), Bandoniozyma fermentans sp. nov. (CBS 12399 T = NU7M71 T = BCRC 23267 T ), Bandoniozyma glucofermentans sp. nov. (CBS 10381 T = NRRL Y-48076 T = ATCC MYA-4760 T = BG 02-7-15-015A-1-1 T ), Bandoniozyma tunnelae sp. nov. (CBS 8024 T = DBVPG 7000 T ), and Bandoniozyma visegradensis sp. nov. (CBS 12505 T = NRRL Y-48783 T = NCAIM Y.01952 T ).

Techniques: Conjugation Assay

° C. Bar = 10 µm, d) Bandoniozyma glucofermentans CBS 10381 T after 7 days in YM broth at 25°C. Bar = 10 µm, e) Bandoniozyma aquatica CBS 12527 T grown in YEPD broth for 3 days at 25°C. Bar = 10 µm, and f) Bandoniozyma complexa CBS 11570 T grown in YEPD broth for 3 days at 25°C. Bar = 10 µm.

Journal: PLoS ONE

Article Title: Bandoniozyma gen. nov., a Genus of Fermentative and Non-Fermentative Tremellaceous Yeast Species

doi: 10.1371/journal.pone.0046060

Figure Lengend Snippet: ° C. Bar = 10 µm, d) Bandoniozyma glucofermentans CBS 10381 T after 7 days in YM broth at 25°C. Bar = 10 µm, e) Bandoniozyma aquatica CBS 12527 T grown in YEPD broth for 3 days at 25°C. Bar = 10 µm, and f) Bandoniozyma complexa CBS 11570 T grown in YEPD broth for 3 days at 25°C. Bar = 10 µm.

Article Snippet: In this study we propose the new yeast genus Bandoniozyma , with seven species Bandoniozyma noutii sp. nov. (type species of genus; CBS 8364 T = DBVPG 4489 T ), Bandoniozyma aquatica sp. nov. (UFMG-DH4.20 T = CBS 12527 T = ATCC MYA-4876 T ), Bandoniozyma complexa sp. nov. (CBS 11570 T = ATCC MYA-4603 T = MA28a T ), Bandoniozyma fermentans sp. nov. (CBS 12399 T = NU7M71 T = BCRC 23267 T ), Bandoniozyma glucofermentans sp. nov. (CBS 10381 T = NRRL Y-48076 T = ATCC MYA-4760 T = BG 02-7-15-015A-1-1 T ), Bandoniozyma tunnelae sp. nov. (CBS 8024 T = DBVPG 7000 T ), and Bandoniozyma visegradensis sp. nov. (CBS 12505 T = NRRL Y-48783 T = NCAIM Y.01952 T ).

Techniques: