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, Available online , doi: 10.15886/j.cnki.rdswxb.20250007
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To further explore the key genes regulating drought resistance in soybean, the peroxidase gene GmPOD123, which is significantly induced under drought stress, was screened based on previous transcriptomic analyses (RNA-Seq) of soybean leaves under drought stress. An expression analysis of the soybean GmPOD123 gene under drought stress was performed to identify whether GmPOD123 is involved in response to drought stress, and the results showed that GmPOD123 was significantly up-regulated after 3 h stress treatment. Moreover, phenotype identification of transgenic Arabidopsis plants of overexpressing GmPOD123 showed that overexpression of GmPOD123 in Arabidopsis could increase the drought tolerance, which was compared to wild type Arabidopsis. To further explore the drought-resistant molecular mechanism of GmPOD123, fructose-1,6-bisphosphate aldolase (GmFBA2), an interacting protein of GmPOD123, was screened from the soybean cDNA yeast library by Mating, and its interaction with GmPOD123 was identified by yeast two-hybrid system and luciferase complementation assay (LCA). In summary, GmFBA2 may influence ROS levels by interacting with GmPOD123, thereby regulating soybean drought resistance.
To further explore the key genes regulating drought resistance in soybean, the peroxidase gene GmPOD123, which is significantly induced under drought stress, was screened based on previous transcriptomic analyses (RNA-Seq) of soybean leaves under drought stress. An expression analysis of the soybean GmPOD123 gene under drought stress was performed to identify whether GmPOD123 is involved in response to drought stress, and the results showed that GmPOD123 was significantly up-regulated after 3 h stress treatment. Moreover, phenotype identification of transgenic Arabidopsis plants of overexpressing GmPOD123 showed that overexpression of GmPOD123 in Arabidopsis could increase the drought tolerance, which was compared to wild type Arabidopsis. To further explore the drought-resistant molecular mechanism of GmPOD123, fructose-1,6-bisphosphate aldolase (GmFBA2), an interacting protein of GmPOD123, was screened from the soybean cDNA yeast library by Mating, and its interaction with GmPOD123 was identified by yeast two-hybrid system and luciferase complementation assay (LCA). In summary, GmFBA2 may influence ROS levels by interacting with GmPOD123, thereby regulating soybean drought resistance.
, Available online , doi: 10.15886/j.cnki.rdswxb.20240161
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Torulaspora delbrueckii is one of non-Saccharomyces yeasts mostly used for research, and is widely used for brewing of various fruit wines. It is co-fermented with Saccharomyces cerevisiae for improving fruit wine flavors. In this experiment, a wild-type yeast strain was selected from the peel of cantaloupe in Fulo Town, Ledong County, Hainan Province. Through phenotypic observation of the strain and constructing a phylogenetic tree based on 18S rDNA sequencing, the strain was identified as T. delbrueckii. Under the fermentation condition of fruit wine, T. delbrueckii grew normally in the range of alcohol content 4−20%, pH value 2.8−3.6 and sulfur dioxide concentration 100−500 mg·L−1, and the growth rate was the highest under the condition of alcohol content 4%, pH value 3.6 and sulfur dioxide concentration 300 mg·L−1. The results show that T. delbrueckii did not produce hydrogen sulfide, had no killing effect on S. cerevisiae, and improved the flavor of cantaloupe fruit wine, which can be used in the production of cantaloupe fruit wine.
Torulaspora delbrueckii is one of non-Saccharomyces yeasts mostly used for research, and is widely used for brewing of various fruit wines. It is co-fermented with Saccharomyces cerevisiae for improving fruit wine flavors. In this experiment, a wild-type yeast strain was selected from the peel of cantaloupe in Fulo Town, Ledong County, Hainan Province. Through phenotypic observation of the strain and constructing a phylogenetic tree based on 18S rDNA sequencing, the strain was identified as T. delbrueckii. Under the fermentation condition of fruit wine, T. delbrueckii grew normally in the range of alcohol content 4−20%, pH value 2.8−3.6 and sulfur dioxide concentration 100−500 mg·L−1, and the growth rate was the highest under the condition of alcohol content 4%, pH value 3.6 and sulfur dioxide concentration 300 mg·L−1. The results show that T. delbrueckii did not produce hydrogen sulfide, had no killing effect on S. cerevisiae, and improved the flavor of cantaloupe fruit wine, which can be used in the production of cantaloupe fruit wine.
, Available online , doi: 10.15886/j.cnki.rdswxb.20240152
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The fitness cost of resistance to dimefluthrin in Aedes albopictus was analyzed by population life table comparison method using the resistant line (RR) of Ae. albopictus for experiment with the susceptible line SS as control. The results showed that the net reproduction rate (R0), mean generation time (T), intrinsic rate of increase (rm), finite rate of increase (λ), and population doubling time (D) were 41.13, 11.77 d, 0.32 d−1, 1.38 d−1, and 2.17 d, respectively, for the susceptible line population, and. 26.48, 14.27 d, 0.23 d−1, 1.26 d−1, and 3.01 d, respectively, for the resistant line. The resistant line of Ae. albopictus was longer in population doubling time than the susceptible line, indicating that its reproductive rate has slowed down. The pupation rate and emergence rate of the susceptible and resistant lines were all 100%, indicating there was no significant difference between them. The average development time from first instar larvae to pupae was 9.66±0.17 d for the susceptible line and 11.31±0.17 d for the resistant line; the average emergence time of pupae for both of the lines was 2.15±0.04 d and 2.90±0.03 d, respectively; the average survival time of female mosquitoes was 30.03±1.60 d and 21.83±1.12 d, respectively, for both of the lines, and there were all significant differences. The development time of larval and pupal stages and the life span of adult mosquitoes were prolonged, indicating that there is a certain fitness cost of resistance to dimefluthrin in Ae. albopictus.
The fitness cost of resistance to dimefluthrin in Aedes albopictus was analyzed by population life table comparison method using the resistant line (RR) of Ae. albopictus for experiment with the susceptible line SS as control. The results showed that the net reproduction rate (R0), mean generation time (T), intrinsic rate of increase (rm), finite rate of increase (λ), and population doubling time (D) were 41.13, 11.77 d, 0.32 d−1, 1.38 d−1, and 2.17 d, respectively, for the susceptible line population, and. 26.48, 14.27 d, 0.23 d−1, 1.26 d−1, and 3.01 d, respectively, for the resistant line. The resistant line of Ae. albopictus was longer in population doubling time than the susceptible line, indicating that its reproductive rate has slowed down. The pupation rate and emergence rate of the susceptible and resistant lines were all 100%, indicating there was no significant difference between them. The average development time from first instar larvae to pupae was 9.66±0.17 d for the susceptible line and 11.31±0.17 d for the resistant line; the average emergence time of pupae for both of the lines was 2.15±0.04 d and 2.90±0.03 d, respectively; the average survival time of female mosquitoes was 30.03±1.60 d and 21.83±1.12 d, respectively, for both of the lines, and there were all significant differences. The development time of larval and pupal stages and the life span of adult mosquitoes were prolonged, indicating that there is a certain fitness cost of resistance to dimefluthrin in Ae. albopictus.
, Available online , doi: 10.15886/j.cnki.rdswxb.20240173
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An attempt was made to make full use of the advantages of plant extracts and chemical pesticides to reduce the dosage of chemical pesticides. Syzygium infrarubiginosum was extracted with methanol, and mixed with four chemical fungicides, including ethirimol, pyraclostrobin, thifluzamide and difenoconazole to screen the synergistic ratio of their mixtures against Rhizoctonia solani, the pathogen of rice sheath blight by indoor activity determination of mycelial growth rate of R. solani. The control effects of the synergistic mixtures on rice sheath blight and cowpea powdery mildew were determined by indoor pot experiment. The indoor activity determination showed that the mixtures at 11 ratios were found to have significant synergistic effects in the indoor activity determination of rice sheath blight, including the mixture of 5% (w/w) methanol extract microemulsion with 5% (w/w) pyraclostrobin suspension at the ratios of 4:1 and 5:1 ratio (the synergistic coefficient was 2.349 and 2.987, respectively), with 5% thifluzamide suspension at the respective ratios of 1:3, 1:4 and 1:5 (the synergistic coefficient values were 2.714, 2.455 and 2.167, respectively). The pot experiment showed that the mixtures screened could effectively control the occurrence of rice sheath blight and cowpea powdery mildew.
An attempt was made to make full use of the advantages of plant extracts and chemical pesticides to reduce the dosage of chemical pesticides. Syzygium infrarubiginosum was extracted with methanol, and mixed with four chemical fungicides, including ethirimol, pyraclostrobin, thifluzamide and difenoconazole to screen the synergistic ratio of their mixtures against Rhizoctonia solani, the pathogen of rice sheath blight by indoor activity determination of mycelial growth rate of R. solani. The control effects of the synergistic mixtures on rice sheath blight and cowpea powdery mildew were determined by indoor pot experiment. The indoor activity determination showed that the mixtures at 11 ratios were found to have significant synergistic effects in the indoor activity determination of rice sheath blight, including the mixture of 5% (w/w) methanol extract microemulsion with 5% (w/w) pyraclostrobin suspension at the ratios of 4:1 and 5:1 ratio (the synergistic coefficient was 2.349 and 2.987, respectively), with 5% thifluzamide suspension at the respective ratios of 1:3, 1:4 and 1:5 (the synergistic coefficient values were 2.714, 2.455 and 2.167, respectively). The pot experiment showed that the mixtures screened could effectively control the occurrence of rice sheath blight and cowpea powdery mildew.
, Available online , doi: 10.15886/j.cnki.rdswxb.20240175
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To explore the evolutionary history of the Bradyrhizobium and its plastic degradation-related gene the hydroxylase gene CYP450, and to understand the mechanisms by which the genus acquires its plastic degradation potential. In this study, 28 fully assembled strains of Bradyrhizobium were collected to investigate the evolution and dissemination of Bradyrhizobium and their hydroxylase genes CYP450 through comparative genomic and phylogenetic analyses. The results revealed that, compared to the core genes, the unique and accessory genes of Bradyrhizobium exhibited an increase in functions related to plastic degradation, such as membrane transport, xenobiotic biodegradation and metabolism, lipid metabolism, and signal transduction. Bradyrhizobium has an open pan-genome, enabling it to acquire genes from other species, adapt to new environments, and evolve new functions. Analysis of the flow of the hydroxylase gene CYP450 in bacteria and archaea revealed that Bradyrhizobium likely acquired hydroxylase gene CYP450 from the Actinobacteria phylum through horizontal gene transfer, thus gaining the potential for plastic degradation.
To explore the evolutionary history of the Bradyrhizobium and its plastic degradation-related gene the hydroxylase gene CYP450, and to understand the mechanisms by which the genus acquires its plastic degradation potential. In this study, 28 fully assembled strains of Bradyrhizobium were collected to investigate the evolution and dissemination of Bradyrhizobium and their hydroxylase genes CYP450 through comparative genomic and phylogenetic analyses. The results revealed that, compared to the core genes, the unique and accessory genes of Bradyrhizobium exhibited an increase in functions related to plastic degradation, such as membrane transport, xenobiotic biodegradation and metabolism, lipid metabolism, and signal transduction. Bradyrhizobium has an open pan-genome, enabling it to acquire genes from other species, adapt to new environments, and evolve new functions. Analysis of the flow of the hydroxylase gene CYP450 in bacteria and archaea revealed that Bradyrhizobium likely acquired hydroxylase gene CYP450 from the Actinobacteria phylum through horizontal gene transfer, thus gaining the potential for plastic degradation.
, Available online , doi: 10.15886/j.cnki.rdswxb.20240176
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An attempt was made to investigate the expression of lactate dehydrogenase (ptLDHA), a key enzyme in lactate metabolism, and to explore the acetylation modification sites on ptLDHA and their potential role in the interplay between lactylation and acetylation modifications in Phaeodactylum tricornutum Bohlin. The ptLDHA gene sequence was first cloned from P. tricornutum Bohlin cDNA, and then a prokaryotic expression vector, pMBP-C-LDHA, was constructed. The recombinant plasmid was then transferred into Escherichia coli BL21 (DE3) for induced expression. Under optimized conditions (16℃, 0.2 mmol·L−1 IPTG for 24 hours), ptLDHA protein was successfully expressed, predominantly in a soluble form. The fusion protein was purified using His-tag affinity chromatography and identified by Western blot with ptLDHA polyclonal antibodies. A single protein band at approximately 78 kDa was observed, confirming that the purified protein was ptLDHA. The expression and purification of ptLDHA in prokaryotic cells was successfully established, laying foundations for subsequent site-directed mutagenesis of acetylation modification sites and for investigation of the effects of modification and demodification on enzyme activity.
An attempt was made to investigate the expression of lactate dehydrogenase (ptLDHA), a key enzyme in lactate metabolism, and to explore the acetylation modification sites on ptLDHA and their potential role in the interplay between lactylation and acetylation modifications in Phaeodactylum tricornutum Bohlin. The ptLDHA gene sequence was first cloned from P. tricornutum Bohlin cDNA, and then a prokaryotic expression vector, pMBP-C-LDHA, was constructed. The recombinant plasmid was then transferred into Escherichia coli BL21 (DE3) for induced expression. Under optimized conditions (16℃, 0.2 mmol·L−1 IPTG for 24 hours), ptLDHA protein was successfully expressed, predominantly in a soluble form. The fusion protein was purified using His-tag affinity chromatography and identified by Western blot with ptLDHA polyclonal antibodies. A single protein band at approximately 78 kDa was observed, confirming that the purified protein was ptLDHA. The expression and purification of ptLDHA in prokaryotic cells was successfully established, laying foundations for subsequent site-directed mutagenesis of acetylation modification sites and for investigation of the effects of modification and demodification on enzyme activity.
, Available online , doi: 10.15886/j.cnki.rdswxb.20240138
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In order to fully understand the reasonable nutrients of Yunnan rubber (Hevea brasiliensis) forest, soil ecological restoration of rubber forest provides data support. Using the soil of Yunnan (Jinghong, Hekou, Ruili), the typical samples of 20 m 20 m in different rubber forests, the soil samples were stratified (0−20, 20−40, 40−60, 60−80, 80−100 cm), and the spatial distribution characteristics of the soil of the rubber forest soil with the forest age and the dynamic changes were analyzed. The result show that the soil TC, TN, and TP contents in the rubber plantations generally increased and then decreased with the plantation age and soil depth. Significant differences were observed in the soil TC, TN, and TP contents among different ages of rubber plantations in Ruili and Jinghong, while the differences in Hekou were not significant. In Jinghong, the soil C∶N ratio increased and then decreased with soil depth, reaching its highest value at (40−60 cm). The soil C∶P ratio decreased with increasing soil depth, peaking at the surface layer (0−20 cm). The N∶P ratio initially decreased and then increased with soil depth. The C∶P and N∶P ratios showed a trend of decreasing and then increasing, with the highest values at 40−60 cm. In Ruili, the C∶N and C∶P ratios decreased and then increased with soil depth, also peaking at 40−60 cm, while the N∶P ratio continuously decreased. In Hekou, the C∶P ratio decreased and then increased with soil depth, while both C∶P and N∶P ratios showed a general decreasing trend. The contents of soil nutrients (C, N, P) and their ecological stoichiometric ratios in rubber plantations generally increased and then decreased with plantation age. Significant differences were found in the soil C, N, P contents and their stoichiometric ratios among different ages and soil layers in the rubber plantations (P<0.05), with highly significant differences observed between areas (P<0.01). It is suggested that during the rapid growth stage of rubber plantations, appropriate application of nitrogen and phosphorus fertilizers should be considered, as the soil N and P affect the growth of mature rubber plantations.
In order to fully understand the reasonable nutrients of Yunnan rubber (Hevea brasiliensis) forest, soil ecological restoration of rubber forest provides data support. Using the soil of Yunnan (Jinghong, Hekou, Ruili), the typical samples of 20 m 20 m in different rubber forests, the soil samples were stratified (0−20, 20−40, 40−60, 60−80, 80−100 cm), and the spatial distribution characteristics of the soil of the rubber forest soil with the forest age and the dynamic changes were analyzed. The result show that the soil TC, TN, and TP contents in the rubber plantations generally increased and then decreased with the plantation age and soil depth. Significant differences were observed in the soil TC, TN, and TP contents among different ages of rubber plantations in Ruili and Jinghong, while the differences in Hekou were not significant. In Jinghong, the soil C∶N ratio increased and then decreased with soil depth, reaching its highest value at (40−60 cm). The soil C∶P ratio decreased with increasing soil depth, peaking at the surface layer (0−20 cm). The N∶P ratio initially decreased and then increased with soil depth. The C∶P and N∶P ratios showed a trend of decreasing and then increasing, with the highest values at 40−60 cm. In Ruili, the C∶N and C∶P ratios decreased and then increased with soil depth, also peaking at 40−60 cm, while the N∶P ratio continuously decreased. In Hekou, the C∶P ratio decreased and then increased with soil depth, while both C∶P and N∶P ratios showed a general decreasing trend. The contents of soil nutrients (C, N, P) and their ecological stoichiometric ratios in rubber plantations generally increased and then decreased with plantation age. Significant differences were found in the soil C, N, P contents and their stoichiometric ratios among different ages and soil layers in the rubber plantations (P<0.05), with highly significant differences observed between areas (P<0.01). It is suggested that during the rapid growth stage of rubber plantations, appropriate application of nitrogen and phosphorus fertilizers should be considered, as the soil N and P affect the growth of mature rubber plantations.
, Available online , doi: 10.15886/j.cnki.rdswxb.20240199
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To explore the role of jasmonate-induced oxygenases JOX family genes in the growth, development, and stress resistance of cassava (Manihot esculenta), MeJOXs family members were identified in cassava genome by bioinformatics methods, and their gene structure, promoter cis-acting elements, evolutionary relationships and expression patterns were analyzed. The results showed that there were four MeJOXs family genes in cassava genome, and each member had similar gene structure, conserved motifs, and protein domains. Each gene member exhibited at least 68% protein homology, with the highest similarity observed between MeJOX1 and MeJOX4, as well as between MeJOX2 and MeJOX3. Promoter cis-acting elements analysis showed that MeJOXs contained numerous light-responsive elements. Furthermore, each gene member possessed a varying number of hormone-responsive elements. Phylogenetic analysis showed that MeJOXs were more closely related to JOX genes in dicotyledonous plants. Transcriptome analysis revealed that all the genes were differentially expressed in stems, leaves, midveins, and fibrous roots, with the exception of MeJOX2 that was scarcely expressed in various cassava tissues. MeJOXs were induced by MeJA in different cassava germplasm, with MeJOX3 demonstrating the most significant expression by inducing. Upon infection by pathogen Xpm, MeJOX1/3/4 responded promptly, but their response patterns were distinctly different. MeJOX1/3 were upregulated, whereas MeJOX4 exhibited a trend of downregulation, and MeJOX2 exhibited negligible response. This study provides a theoretical foundation for further elucidating the functions of the MeJOXs gene family in cassava.
To explore the role of jasmonate-induced oxygenases JOX family genes in the growth, development, and stress resistance of cassava (Manihot esculenta), MeJOXs family members were identified in cassava genome by bioinformatics methods, and their gene structure, promoter cis-acting elements, evolutionary relationships and expression patterns were analyzed. The results showed that there were four MeJOXs family genes in cassava genome, and each member had similar gene structure, conserved motifs, and protein domains. Each gene member exhibited at least 68% protein homology, with the highest similarity observed between MeJOX1 and MeJOX4, as well as between MeJOX2 and MeJOX3. Promoter cis-acting elements analysis showed that MeJOXs contained numerous light-responsive elements. Furthermore, each gene member possessed a varying number of hormone-responsive elements. Phylogenetic analysis showed that MeJOXs were more closely related to JOX genes in dicotyledonous plants. Transcriptome analysis revealed that all the genes were differentially expressed in stems, leaves, midveins, and fibrous roots, with the exception of MeJOX2 that was scarcely expressed in various cassava tissues. MeJOXs were induced by MeJA in different cassava germplasm, with MeJOX3 demonstrating the most significant expression by inducing. Upon infection by pathogen Xpm, MeJOX1/3/4 responded promptly, but their response patterns were distinctly different. MeJOX1/3 were upregulated, whereas MeJOX4 exhibited a trend of downregulation, and MeJOX2 exhibited negligible response. This study provides a theoretical foundation for further elucidating the functions of the MeJOXs gene family in cassava.
, Available online , doi: 10.15886/j.cnki.rdswxb.20240177
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In order to understand the anti-actinomycetes with biocontrol value isolated and screened from the soil in Hainan Island, actinomycetes were isolated from six soil samples from different regions of Hainan by using a gradient dilution method, and the actinomycete strains were screened by using the confrontation culture method and identified based on their morphology, physiological and biochemical characteristics and molecular biology methods. The potential application value of the active strains was determined by fruit soaking method and pot experiment. A total of 285 actinomycete strains were isolated from the soil samples, and the strain Q2-02 had the highest antibacterial activity. Its inhibition rates of Botryodiplodia theobromae, Colletotrichum gloeosporioides and Rhizoctonia solani were 92.8%, 90.0% and 88.2%, respectively, and the inhibition rates of the other six pathogens were also more than 60.0%. The identification of this strain showed that the strain Q2-02 was Streptomyces lunalinharesii. The pot experiments showed that the control effect of the 10-fold dilution of the fermented supernatant of the strain on mango stemend rot and rice sheath blight was significantly higher than that of the positive control 45% thiophandazim suspension diluted 800 times and 10% validacin aqueous solution diluted1000 times, indicating that the strain Q2-02 has the value of further development and application.
In order to understand the anti-actinomycetes with biocontrol value isolated and screened from the soil in Hainan Island, actinomycetes were isolated from six soil samples from different regions of Hainan by using a gradient dilution method, and the actinomycete strains were screened by using the confrontation culture method and identified based on their morphology, physiological and biochemical characteristics and molecular biology methods. The potential application value of the active strains was determined by fruit soaking method and pot experiment. A total of 285 actinomycete strains were isolated from the soil samples, and the strain Q2-02 had the highest antibacterial activity. Its inhibition rates of Botryodiplodia theobromae, Colletotrichum gloeosporioides and Rhizoctonia solani were 92.8%, 90.0% and 88.2%, respectively, and the inhibition rates of the other six pathogens were also more than 60.0%. The identification of this strain showed that the strain Q2-02 was Streptomyces lunalinharesii. The pot experiments showed that the control effect of the 10-fold dilution of the fermented supernatant of the strain on mango stemend rot and rice sheath blight was significantly higher than that of the positive control 45% thiophandazim suspension diluted 800 times and 10% validacin aqueous solution diluted
, Available online , doi: 10.15886/j.cnki.rdswxb.20240169
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Cassava is an important food crop in tropical regions, but the yield of cassava is affected by salt stress, which endangers food security. JAZ (jasmonate ZIM-domain, JAZ) proteins, as essential components in the jasmonate signaling pathway, are involved in regulating the tolerance to salt stress in a variety of crops. In order to investigate response of JAZ proteins in cassava to salt stress, as well as the underlying regulatory mechanisms two homologous genes MeJAZ2.1 and MeJAZ2.2 were identified from cassava variety SC124 through bioinformatics. The evolutionary tree and conserved domain analysis indicated that they both contain two conserved domains, ZIM and Jas, which belong to the JAZ gene family. Further research discovered that the expression level of MeJAZ2.2 changed more significantly in response to salt stress in cassava. MeJAZ2.2-silenced cassava plants were more susceptibility to salt stress compared to the wild type, indicating that MeJAZ2.2 may positively regulate cassava resistance to salt stress. The pGADT7-MeJAZ2.2 bait vector was constructed and no self-activating activity was found by yeast two-hybrid experiment. Moreover, three candidate interacting proteins of MeJAZ2.2 were screened, including glutamine synthetase (GS), ubiquitin 3 (Ub3), and FRIGIDA-LIKE PROTEIN (FRI-L), which provides a preliminary framework for analyzing the function and molecular mechanism of JAZ proteins to salt stress in cassava.
Cassava is an important food crop in tropical regions, but the yield of cassava is affected by salt stress, which endangers food security. JAZ (jasmonate ZIM-domain, JAZ) proteins, as essential components in the jasmonate signaling pathway, are involved in regulating the tolerance to salt stress in a variety of crops. In order to investigate response of JAZ proteins in cassava to salt stress, as well as the underlying regulatory mechanisms two homologous genes MeJAZ2.1 and MeJAZ2.2 were identified from cassava variety SC124 through bioinformatics. The evolutionary tree and conserved domain analysis indicated that they both contain two conserved domains, ZIM and Jas, which belong to the JAZ gene family. Further research discovered that the expression level of MeJAZ2.2 changed more significantly in response to salt stress in cassava. MeJAZ2.2-silenced cassava plants were more susceptibility to salt stress compared to the wild type, indicating that MeJAZ2.2 may positively regulate cassava resistance to salt stress. The pGADT7-MeJAZ2.2 bait vector was constructed and no self-activating activity was found by yeast two-hybrid experiment. Moreover, three candidate interacting proteins of MeJAZ2.2 were screened, including glutamine synthetase (GS), ubiquitin 3 (Ub3), and FRIGIDA-LIKE PROTEIN (FRI-L), which provides a preliminary framework for analyzing the function and molecular mechanism of JAZ proteins to salt stress in cassava.
, Available online , doi: 10.15886/j.cnki.rdswxb.20240116
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To explore the volatile components and aroma composition of the flowers of longan (Dimocarpus longan), and to investigate the antioxidant activity of their extracts, the flowers of longan were collected from Fujian and extracted with different solvents, and their volatile components and the aroma of the components were analyzed by using the GC-MS technology. Meanwhile, DPPH and ABTS free radical scavenging assays were used to detect the antioxidant activity of the longan flower extracts. The results showed that 59 components were identified from the essential oil extracted from the longan flowers, accounting for approximately 82.2% of the total volatile components. In the essential oil 4 key aroma compounds, 3 potential aroma compounds and 1 modified aroma compound were found. The DPPH and ABTS free radical scavenging assay showed that different solvent extracts of longan flowers had a certain degree of antioxidant activity. Water extract, methanol extract and ethanol extract had significant antioxidant activity, while n-butyl alcohol extract and ethyl acetate extract had low antioxidant activity. All the results show that longan flowers have a great development potential in the fields of medicine, cosmetics, health products, functional foods and so on, which provides a scientific basis for the development and utilization of longan flowers.
To explore the volatile components and aroma composition of the flowers of longan (Dimocarpus longan), and to investigate the antioxidant activity of their extracts, the flowers of longan were collected from Fujian and extracted with different solvents, and their volatile components and the aroma of the components were analyzed by using the GC-MS technology. Meanwhile, DPPH and ABTS free radical scavenging assays were used to detect the antioxidant activity of the longan flower extracts. The results showed that 59 components were identified from the essential oil extracted from the longan flowers, accounting for approximately 82.2% of the total volatile components. In the essential oil 4 key aroma compounds, 3 potential aroma compounds and 1 modified aroma compound were found. The DPPH and ABTS free radical scavenging assay showed that different solvent extracts of longan flowers had a certain degree of antioxidant activity. Water extract, methanol extract and ethanol extract had significant antioxidant activity, while n-butyl alcohol extract and ethyl acetate extract had low antioxidant activity. All the results show that longan flowers have a great development potential in the fields of medicine, cosmetics, health products, functional foods and so on, which provides a scientific basis for the development and utilization of longan flowers.
, Available online , doi: 10.15886/j.cnki.rdswxb.20240147
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Cassava (Manihot esculenta Crantz) has important economic value. Exploring the key genes of cassava is of great significance for improving cassava quality. To mine disease-resistant genes of cassava, a specific immune receptor-related gene resistant to Xam (MeRXam2) was cloned, and its protein structure was analyzed. The analysis was found that the coding sequence (CDS) region of this gene is3561 bp, with the protein size of about 132.847 kD, Real-time fluorescent quantitative (Real-time quantitative reverse transcription PCR, qRT-PCR) analysis showed that the expression level of MeRXam2 increased with time under pathogen infection and reached the maximum at 24 h, indicating that MeRXam2 was involved in the resistance response to cassava disease. In addition, a MeRXam2-pET32a protein expression vector was constructed by homologous recombination, and the protein was induced and determined. After exogenous spraying of MeRXam2 protein on cassava leaf tissues, the number of bacteria was found significantly lower than that of the control strain, indicating that this protein could slow down bacterial infection. This founding might provide a theoretical basis and candidate protein for cassava molecular breeding.
Cassava (Manihot esculenta Crantz) has important economic value. Exploring the key genes of cassava is of great significance for improving cassava quality. To mine disease-resistant genes of cassava, a specific immune receptor-related gene resistant to Xam (MeRXam2) was cloned, and its protein structure was analyzed. The analysis was found that the coding sequence (CDS) region of this gene is
, Available online , doi: 10.15886/j.cnki.rdswxb.20240170
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The endoplasmic reticulum (ER) serves as a crucial site for protein processing in eukaryotic cells. Sec62, an essential component of the ER translocation complex, plays a significant role in growth, development, and stress regulation. An attempt was made to examine the transcription levels of the PcSec62 gene at various growth stages and during pathogenic processes. The PcSec62 gene was knocked out by using CRISPR/Cas9 gene editing to produce knockout mutants (ΔPcsec62) and a complementary strain (ΔPcsec62-C). The results showed that the transcriptional expression of PcSec62 significantly increased during the sporangia and infection stages. The ΔPcsec62 mutants exhibited notably reduced growth and sporulation abilities, alongside stunted hyphal growth. Additionally, the ΔPcsec62 strain showed significantly low tolerance to abiotic stress and reduced pathogenicity. These findings indicate that PcSec62 is involved in regulating the growth, development, abiotic stress responses, and pathogenicity of Phytophthora capsici.
The endoplasmic reticulum (ER) serves as a crucial site for protein processing in eukaryotic cells. Sec62, an essential component of the ER translocation complex, plays a significant role in growth, development, and stress regulation. An attempt was made to examine the transcription levels of the PcSec62 gene at various growth stages and during pathogenic processes. The PcSec62 gene was knocked out by using CRISPR/Cas9 gene editing to produce knockout mutants (ΔPcsec62) and a complementary strain (ΔPcsec62-C). The results showed that the transcriptional expression of PcSec62 significantly increased during the sporangia and infection stages. The ΔPcsec62 mutants exhibited notably reduced growth and sporulation abilities, alongside stunted hyphal growth. Additionally, the ΔPcsec62 strain showed significantly low tolerance to abiotic stress and reduced pathogenicity. These findings indicate that PcSec62 is involved in regulating the growth, development, abiotic stress responses, and pathogenicity of Phytophthora capsici.
, Available online , doi: 10.15886/j.cnki.rdswxb.20240149
Abstract:
The bactericidal effects of different antibiotics on Escherichia coli were analyzed by using fluorescence photoelectric microbial detection devices. The results prove that the antimicrobial analysis using fluorescence photoelectric microbial detection devices is equivalent to the K-B disk diffusion method. Compared to the traditional methods, the fluorescence photoelectric microbial detection method can be used to quickly identify microbial resistance in clinical specimens, especially of fastidious bacteria and other slow-growing strains, providing timely reference for the clinical application of antibiotics and drug resistance analysis of pathogens. The detection of the drug resistance of E. coli to 9 antibiotics showed that E. coli had the highest resistance to nalidixic acid, with a minimal inhibitory concentration (MIC) of 100 μg/mL. E. coli had the lowest resistance to cefotaxime, with a minimal inhibitory concentration of 0.2 μg/mL. Combination of nalidixic acid with ciprofloxacin, cefotaxime, or chloramphenicol indicated that the combination of 0.5 MIC concentration of nalidixic acid with 0.5 MIC concentration of ciprofloxacin had the best bactericidal effect.
The bactericidal effects of different antibiotics on Escherichia coli were analyzed by using fluorescence photoelectric microbial detection devices. The results prove that the antimicrobial analysis using fluorescence photoelectric microbial detection devices is equivalent to the K-B disk diffusion method. Compared to the traditional methods, the fluorescence photoelectric microbial detection method can be used to quickly identify microbial resistance in clinical specimens, especially of fastidious bacteria and other slow-growing strains, providing timely reference for the clinical application of antibiotics and drug resistance analysis of pathogens. The detection of the drug resistance of E. coli to 9 antibiotics showed that E. coli had the highest resistance to nalidixic acid, with a minimal inhibitory concentration (MIC) of 100 μg/mL. E. coli had the lowest resistance to cefotaxime, with a minimal inhibitory concentration of 0.2 μg/mL. Combination of nalidixic acid with ciprofloxacin, cefotaxime, or chloramphenicol indicated that the combination of 0.5 MIC concentration of nalidixic acid with 0.5 MIC concentration of ciprofloxacin had the best bactericidal effect.
, Available online , doi: 10.15886/j.cnki.rdswxb.20240140
Abstract:
Solanum lycopersicum is a model plant for research in genetics and molecular biology. Recently selection of tomato internal reference genes has been reported but with less types of exogenous hormones used for treatment. An attempt was hence made to select internal reference genes with stable expression in different tissues of tomato and under treatment with plant hormones. DANJ, EF-1α, ACT, UBI, APT, CAC, TIP41 and RPL8 were used as candidate internal reference genes, and selected under the 8 experimental conditions: exogenous hormone treatment with auxin, gibberellin, abscisic acid, cytokinin, salicylic acid, brassinolide and ethylene and various plant parts. The stability of the candidate reference genes was comprehensively evaluated using algorithms such as geNorm, NormFinder, BestKeeper, Delta CT and RefFinder. The stability of the reference genes was validated using the auxin-responsive gene SlGH3.4. The results showed that APT is the most stable reference gene expressed in auxin, gibberellin, abscisic acid, brassinosteroid treatment and various plant parts, that UBI is the most stable reference gene expressed in all samples under cytokinin and salicylic acid treatment, and that TIP41 is the most stable reference gene expressed under ethylene treatment. Finally, when APT, which has a relatively stable comprehensive ranking, was used as an internal reference gene, it was found that the expression level of SlGH3.4 gene showed a similar trend under IAA treatment conditions, while the less stable RPL8 gene failed to accurately correct the expression level of the target gene. All these results may provide theoretical support for the analysis of gene expression networks and molecular regulatory mechanisms in the response of tomato to exogenous hormone treatment.
Solanum lycopersicum is a model plant for research in genetics and molecular biology. Recently selection of tomato internal reference genes has been reported but with less types of exogenous hormones used for treatment. An attempt was hence made to select internal reference genes with stable expression in different tissues of tomato and under treatment with plant hormones. DANJ, EF-1α, ACT, UBI, APT, CAC, TIP41 and RPL8 were used as candidate internal reference genes, and selected under the 8 experimental conditions: exogenous hormone treatment with auxin, gibberellin, abscisic acid, cytokinin, salicylic acid, brassinolide and ethylene and various plant parts. The stability of the candidate reference genes was comprehensively evaluated using algorithms such as geNorm, NormFinder, BestKeeper, Delta CT and RefFinder. The stability of the reference genes was validated using the auxin-responsive gene SlGH3.4. The results showed that APT is the most stable reference gene expressed in auxin, gibberellin, abscisic acid, brassinosteroid treatment and various plant parts, that UBI is the most stable reference gene expressed in all samples under cytokinin and salicylic acid treatment, and that TIP41 is the most stable reference gene expressed under ethylene treatment. Finally, when APT, which has a relatively stable comprehensive ranking, was used as an internal reference gene, it was found that the expression level of SlGH3.4 gene showed a similar trend under IAA treatment conditions, while the less stable RPL8 gene failed to accurately correct the expression level of the target gene. All these results may provide theoretical support for the analysis of gene expression networks and molecular regulatory mechanisms in the response of tomato to exogenous hormone treatment.
, Available online , doi: 10.15886/j.cnki.rdswxb.20240158
Abstract:
Research on the central nervous system function of pests forms the foundation for developing precise behavior control technologies. Despite the significant role of neuron-labeling techniques based on genetic manipulation in this field, such techniques remain relatively scarce for non-model insects, such as the oriental fruit fly, Bactrocera dorsalis. In this context an attempt was made to identify four pan-neuronal expression genes in B. dorsalis with a view to laying the groundwork for constructing a neuron-labeling system for this species. The genomic structures of the pan-neuronal expression genes in B. dorsalis were identified and analyzed by employing bioinformatics and molecular biology to verify their full-length sequences and peripheral expression patterns. The results indicate that, by referring to four pan-neuronal expression genes from Drosophila, four homologous genes were identified in the B. dorsalis, namely BdornSyb, BdorSyt1, Bdorelav, and BdorBrp. The full genomic lengths of these four genes are 19,337 bp (5 exons, 4 introns), 26,884 bp (8 exons, 7 introns), 1,341 bp (1 exon), and 49,692 bp (14 exons, 13 introns), respectively. The domains of BdornSyb, BdorSyt1, and Bdorelav are highly conserved among closely related species. PCR cloning results indicated that the CDS sequence lengths of these four genes are all over 500 bp, consistent with the bioinformatics analysis results. Evolutionary and genomic structure analyses demonstrated that the four genes are highly conserved among Diptera insects. Expression pattern analysis revealed that all the four genes are expressed in the peripheral sensory organs of B. dorsalis, with three genes, BdornSyb, BdorSyt1 and BdorBrp, showing higher expression levels in the primary olfactory organs, the antennae, and the maxillary palp. The four genes identified are candidate pan-neuronal expression genes in B. dorsalis, providing a foundation for constructing a pan-neuronal labeling system for this species in the future.
Research on the central nervous system function of pests forms the foundation for developing precise behavior control technologies. Despite the significant role of neuron-labeling techniques based on genetic manipulation in this field, such techniques remain relatively scarce for non-model insects, such as the oriental fruit fly, Bactrocera dorsalis. In this context an attempt was made to identify four pan-neuronal expression genes in B. dorsalis with a view to laying the groundwork for constructing a neuron-labeling system for this species. The genomic structures of the pan-neuronal expression genes in B. dorsalis were identified and analyzed by employing bioinformatics and molecular biology to verify their full-length sequences and peripheral expression patterns. The results indicate that, by referring to four pan-neuronal expression genes from Drosophila, four homologous genes were identified in the B. dorsalis, namely BdornSyb, BdorSyt1, Bdorelav, and BdorBrp. The full genomic lengths of these four genes are 19,337 bp (5 exons, 4 introns), 26,884 bp (8 exons, 7 introns), 1,341 bp (1 exon), and 49,692 bp (14 exons, 13 introns), respectively. The domains of BdornSyb, BdorSyt1, and Bdorelav are highly conserved among closely related species. PCR cloning results indicated that the CDS sequence lengths of these four genes are all over 500 bp, consistent with the bioinformatics analysis results. Evolutionary and genomic structure analyses demonstrated that the four genes are highly conserved among Diptera insects. Expression pattern analysis revealed that all the four genes are expressed in the peripheral sensory organs of B. dorsalis, with three genes, BdornSyb, BdorSyt1 and BdorBrp, showing higher expression levels in the primary olfactory organs, the antennae, and the maxillary palp. The four genes identified are candidate pan-neuronal expression genes in B. dorsalis, providing a foundation for constructing a pan-neuronal labeling system for this species in the future.
, Available online , doi: 10.15886/j.cnki.rdswxb.20240073
Abstract:
Exploring the spatial ecological niche delineation between co-distributed proximate species is crucial to understand their coexistence in the community. A survey of two species of pheasants, Red Junglefowl (Gallus gallus jabouillei) and Chinese Francolin (Francolinus pintadeanus), were made at 105 and 79 distribution sites in Datian National Nature Reserve, Hainan, from March 2023 to January 2024, respectively, using the line transect method and satellite tracking technology. A maximum entropy model was used to predict the suitable habitats for these two species of pheasants by combining 12 environmental variables, and analyze their overlap. The results showed that there are differences in the distribution patterns of suitable habitats for the two species of pheasants. The suitable habitat for the Red Junglefowl was located in the north-western and central parts of the reserve, which are dominated by deciduous broad-leaved and scrub forests, while the suitable habitat of the Chinese Francolin was located in the northern part of the reserve, and was mainly in the grass area. The potential suitable habitats of the two species of pheasants were 633.18 hm2 and 337.31 hm2, accounting for 48.61% and 25.89% of the area of the reserve, respectively. The ecological overlap indices of the Red Junglefowl and the Chinese Francolin were low, with D and I values being 0.421 and 0.711, respectively, and the overlapping area of suitable habitat was 41.31 hm2, which accounted for 17.64% and 17.22% of the area of the suitable habitats for the Red Junglefowl and the Chinese Francolin, respectively. There were similarities and differences in the selection of environmental factors by the two species. Differences lie in the fact that the variables with the greatest influence on the habitat distribution of the two species of pheasants were the distance from the ranger path and the normalized difference vegetation index (NDVI), which are mainly manifested in the fact that the Red Junglefowl preferred deciduous broad-leaved forests and scrubs near the ranger paths, whereas the Chinese Francolin was more influenced by the NDVI; similarities lie in the fact that the variable with the second highest influence on the habitat distribution of the two species of pheasants was the distance from the water source, which was manifested in the fact that the Red Junglefowl and the Chinese Francolin preferred to distribute within 400 m and 500～1000 m from water sources, respectively. This survey reveals a spatial sympatric niche relationship between the Red Junglefowl and the Chinese Francolin, and helps to further understand the interspecific competition and coexistence mechanisms of tropical pheasants.
Exploring the spatial ecological niche delineation between co-distributed proximate species is crucial to understand their coexistence in the community. A survey of two species of pheasants, Red Junglefowl (Gallus gallus jabouillei) and Chinese Francolin (Francolinus pintadeanus), were made at 105 and 79 distribution sites in Datian National Nature Reserve, Hainan, from March 2023 to January 2024, respectively, using the line transect method and satellite tracking technology. A maximum entropy model was used to predict the suitable habitats for these two species of pheasants by combining 12 environmental variables, and analyze their overlap. The results showed that there are differences in the distribution patterns of suitable habitats for the two species of pheasants. The suitable habitat for the Red Junglefowl was located in the north-western and central parts of the reserve, which are dominated by deciduous broad-leaved and scrub forests, while the suitable habitat of the Chinese Francolin was located in the northern part of the reserve, and was mainly in the grass area. The potential suitable habitats of the two species of pheasants were 633.18 hm2 and 337.31 hm2, accounting for 48.61% and 25.89% of the area of the reserve, respectively. The ecological overlap indices of the Red Junglefowl and the Chinese Francolin were low, with D and I values being 0.421 and 0.711, respectively, and the overlapping area of suitable habitat was 41.31 hm2, which accounted for 17.64% and 17.22% of the area of the suitable habitats for the Red Junglefowl and the Chinese Francolin, respectively. There were similarities and differences in the selection of environmental factors by the two species. Differences lie in the fact that the variables with the greatest influence on the habitat distribution of the two species of pheasants were the distance from the ranger path and the normalized difference vegetation index (NDVI), which are mainly manifested in the fact that the Red Junglefowl preferred deciduous broad-leaved forests and scrubs near the ranger paths, whereas the Chinese Francolin was more influenced by the NDVI; similarities lie in the fact that the variable with the second highest influence on the habitat distribution of the two species of pheasants was the distance from the water source, which was manifested in the fact that the Red Junglefowl and the Chinese Francolin preferred to distribute within 400 m and 500～
, Available online , doi: 10.15886/j.cnki.rdswxb.20230143
Abstract:
The rubber tree (Hevea brasiliensis) is an important economic crop in tropical areas of China, and its related industries play an important role in the local agricultural economy. The powdery mildew fungus Erysiphe quercicola is the most severe disease infecting H. brasiliensis. The effector proteins secreted by phytopahogenic microbes play important roles in the pathogenesis, but how effector proteins promote the infection by E. quercicola is less documented, which has limited disease resistance breeding programme and disease control strategies. The E. quercicola has 24 highly conserved candidate effector proteins, which has homologous in many other powdery mildews. It is possible that these proteins are required for the pathogenicity of powdery mildew. In order to find the important virulence factor among the 24 CSEPs, two conserved effector proteins were selected for functional analysis and found that only the effector CSEP00565 suppressed hypersensitive responses induced by Phytophthora infestans INF1 when these effectors were expressed in Nicotiana benthamiana. The effector CSEP00565 suppressed callose deposition induced by fungal chitin and bacteria flg22 when this effector was expressed in Arabidopsis thaliana, further supporting that CSEP00565 has a function to inhibit plant immunity. When CSEP00565 was further silenced by using spray-induced gene, the pathogenicity of the fungus significantly decreased. And the expression level of CSEP00565 was up-regulated at the haustorium formation stage (24 h) after inoculation with E. quercicola. Further research has discovered that the signal peptide of CSEP00565 possesses the activity of direct protein secretion. All these results indicate that CSEP00565 is a key virulence factor in promoting the infection by E. quercicola, which is conducive to understanding of the molecular mechanisms underlying H. brasiliensis-E. quercicola interaction.
The rubber tree (Hevea brasiliensis) is an important economic crop in tropical areas of China, and its related industries play an important role in the local agricultural economy. The powdery mildew fungus Erysiphe quercicola is the most severe disease infecting H. brasiliensis. The effector proteins secreted by phytopahogenic microbes play important roles in the pathogenesis, but how effector proteins promote the infection by E. quercicola is less documented, which has limited disease resistance breeding programme and disease control strategies. The E. quercicola has 24 highly conserved candidate effector proteins, which has homologous in many other powdery mildews. It is possible that these proteins are required for the pathogenicity of powdery mildew. In order to find the important virulence factor among the 24 CSEPs, two conserved effector proteins were selected for functional analysis and found that only the effector CSEP00565 suppressed hypersensitive responses induced by Phytophthora infestans INF1 when these effectors were expressed in Nicotiana benthamiana. The effector CSEP00565 suppressed callose deposition induced by fungal chitin and bacteria flg22 when this effector was expressed in Arabidopsis thaliana, further supporting that CSEP00565 has a function to inhibit plant immunity. When CSEP00565 was further silenced by using spray-induced gene, the pathogenicity of the fungus significantly decreased. And the expression level of CSEP00565 was up-regulated at the haustorium formation stage (24 h) after inoculation with E. quercicola. Further research has discovered that the signal peptide of CSEP00565 possesses the activity of direct protein secretion. All these results indicate that CSEP00565 is a key virulence factor in promoting the infection by E. quercicola, which is conducive to understanding of the molecular mechanisms underlying H. brasiliensis-E. quercicola interaction.
, Available online , doi: 10.15886/j.cnki.rdswxb.20240111
Abstract:
As an important part of Hainan's agriculture, rice industry in Hainan has unique development advantages, high-quality tropical natural conditions and abundant light and heat resources. The current situation of rice industry in Hainan was analyzed in terms of planting area, production, rice varieties and branding. The analysis showed that there are major problems in rice production in Hainan, such as high cost, low yield and no premium. In order to solve these problems and promote the green and efficient development of Hainan's rice industry, a series of countermeasures are proposed, including promoting intelligent mechanised planting to reduce costs, relying on scientific and technological innovations to improve rice yield, implementing green production modes to improve the quality and efficiency of rice, as well as strengthening brand building and market promotion. The comprehensive application of these countermeasures aims to improve the market competitiveness of Hainan rice, achieve a win-win situation in terms of ecological and economic benefits, and promote the sustainable development of rice industry in Hainan.
As an important part of Hainan's agriculture, rice industry in Hainan has unique development advantages, high-quality tropical natural conditions and abundant light and heat resources. The current situation of rice industry in Hainan was analyzed in terms of planting area, production, rice varieties and branding. The analysis showed that there are major problems in rice production in Hainan, such as high cost, low yield and no premium. In order to solve these problems and promote the green and efficient development of Hainan's rice industry, a series of countermeasures are proposed, including promoting intelligent mechanised planting to reduce costs, relying on scientific and technological innovations to improve rice yield, implementing green production modes to improve the quality and efficiency of rice, as well as strengthening brand building and market promotion. The comprehensive application of these countermeasures aims to improve the market competitiveness of Hainan rice, achieve a win-win situation in terms of ecological and economic benefits, and promote the sustainable development of rice industry in Hainan.
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