The artemisinin accumulation in the hairy root cultures of Artemisia annua L. was enhanced via a treatment of three fungal elicitors separately ( Verticillium dahliae Kleb., Rhizopus stolonifer (Ehrenb. ex ...The artemisinin accumulation in the hairy root cultures of Artemisia annua L. was enhanced via a treatment of three fungal elicitors separately ( Verticillium dahliae Kleb., Rhizopus stolonifer (Ehrenb. ex Fr.) Vuill and Colletotrichum dematium (Pers.) Grove). Among these three elicitors, V. dahliae had the highest inducing efficiency, but none of them manifests any noticeable effects on the cell growth of the hairy root cultures. The artemisinin content of the hairy root cultures treated with V. dahliae elicitor was 1.12 mg/g DW, which was 45% higher than the control (0.77 mg/g DW). The results showed that elicitation was dependent on the elicitor concentration, the incubation period and the physiological stage at which the hairy root cultures were treated. In addition, the authors found that for V. dahliae , the optimum concentration was 0.4 mg carbohydrate per millilitre medium, the strongest response of A. annua hairy root cultures to the elicitation was at the late exponential growth stage, and the highest artemisinin content of the hairy root cultures was on the 4th day post treatment.展开更多
To optimize the technique of rapid propagation of Dendrobium hybrida seedlings and to explore a hormone-free tissue culture method for D.hybrida,six kinds of mycorrhizal fungi which were isolated from the wild orchids...To optimize the technique of rapid propagation of Dendrobium hybrida seedlings and to explore a hormone-free tissue culture method for D.hybrida,six kinds of mycorrhizal fungi which were isolated from the wild orchids were made into fungal elicitors.These fungal elicitors were added into the DE medium with concentrations of 40,60 and 80 ml/L,respectively.After a 90-d culturing,the effects of fungal elicitors on the growth of D.hybrida cultivar ‘088＇ tissue culture seedlings were studied.The results showed that treatment 13（T13） extremely significantly increased the fresh weight,but other treatment groups had no significant effects.In addition,T1,T5,T9,T11 and T13 extremely significantly influenced the contents of chlorophyll a and chlorophyll a ＋ b.However,T1 and T11 had extremely significantly effect on the content of chlorophyll b.Combining the effects on fresh weight and chlorophyll content,it could be concluded T13（40 ml/L of Y05） has promoting effects on the growth of D.hybrida tissue culture seedlings.展开更多
【目的】明确山田胶锈菌(Gymnosporangium yamadae)吸器中效应蛋白GyHGSRE1的基础生物学功能,为进一步揭示山田胶锈菌效应蛋白作用的分子机制,制定持久的锈病防控策略奠定基础。【方法】基于前期获得的山田胶锈菌吸器转录组数据筛选具...【目的】明确山田胶锈菌(Gymnosporangium yamadae)吸器中效应蛋白GyHGSRE1的基础生物学功能,为进一步揭示山田胶锈菌效应蛋白作用的分子机制,制定持久的锈病防控策略奠定基础。【方法】基于前期获得的山田胶锈菌吸器转录组数据筛选具有高表达丰度的效应蛋白;通过MEME(http://meme-suite.org/)、AI(https://drug.ai.tencent.com)等在线网站预测GyHGSRE1的蛋白质结构;利用荧光定量PCR检测GyHGSRE1基因在山田胶锈菌侵染过程中的表达水平,以酵母菌分泌系统验证GyHGSRE1信号肽的分泌功能,并通过农杆菌介导的瞬时表达技术分析GyHGSRE1在本氏烟(Nicotiana benthamiana)和苹果(Malus domestica)叶片中的功能。【结果】在山田胶锈菌吸器转录组数据中获得了FPKM(fragments per Kb per million mapped reads)值为117.92、富含甘氨酸和丝氨酸的小分子分泌蛋白GyHGSRE1。其N端具有1个富含丝氨酸motif的信号肽。qRT-PCR显示GyHGSRE1基因在山田胶锈菌的吸器形成阶段以及性、锈孢子形成及发育阶段均上调表达,并定位于本氏烟叶片细胞的细胞质和细胞核,能够诱导细胞坏死并激发基础免疫防御反应,但含有信号肽的全长效应蛋白GyHGSRE1能够诱导苹果叶片细胞坏死,去除信号肽后诱导细胞坏死的能力减弱。【结论】富含甘氨酸和丝氨酸的非典型效应蛋白GyHGSRE1定位于植物细胞的细胞质和细胞核,能够诱导本氏烟及苹果叶片细胞坏死,因此可能具有广谱性的激发子活性。GyHGSRE1在山田胶锈菌侵染苹果叶片的定殖和孢子发育阶段显著表达从而发挥功能,其信号肽可能决定着对寄主特异性识别的相关功能。展开更多
The Northeast region is the main producer of guava in Brazil,generating employment and income.However,water availability means that producer’s resort to using water with high salinity,which harms plant development,es...The Northeast region is the main producer of guava in Brazil,generating employment and income.However,water availability means that producer’s resort to using water with high salinity,which harms plant development,especially during the seedling formation phase.The adoption of techniques that mitigate the deleterious effect of salinity is increasingly necessary,such as the use of elicitors such as ascorbic acid.The purpose of this study was to analyze the morphophysiology of guava seedlings under saline and ascorbic acid levels.The study was carried out by applying treatments composed of five saline levels(SL=0.3;1.3;2.3;3.3 and 4.3 dS m^(-1))and four levels of ascorbic acid—AA(0,200,400,and 600 mg L^(-1)),in a 5×4 factorial arrangement,adopting a randomized block design.Gas exchange and growth of guava seedlings are limited from 0.3 dS m^(-1).Using 400 mg L^(-1)of AA reduces damage from salinity on stomatal conductance,transpiration,and net assimilation rate up to the estimated SL of 1.80 dS m^(-1).In contrast,AA level 412 mg L^(-1)increased instantaneous water use efficiency up to the salinity of 2.3 dS m^(-1).AA level of 600 mg L^(-1)attenuated salt stress effects on leaf area and height/stem diameter ratio up to SL of 2.05 dS m^(-1).The number of leaves and the absolute and relative growth rates were stimulated by AA under the lowest saline level.展开更多
Callus cultures of Origanum vulgare L. were established from leaf discus on Murashige and Skoog （MS） medium containing different levels of growth regulators, i.e., 2,4-Dichlorophenoxyacetic acid （2,4-D）, Naphthale...Callus cultures of Origanum vulgare L. were established from leaf discus on Murashige and Skoog （MS） medium containing different levels of growth regulators, i.e., 2,4-Dichlorophenoxyacetic acid （2,4-D）, Naphthalene acetic acid （NAA）, Benzyl Adenine （BA） and Kinetin （Kn） and incubated under dark condition. Callus tissues were employed to study the influence of abiotic elicitors on the production of thymol. Constant weights of callus （300 mg） were cultured on accumulation medium treated separately with each one of elicitors used （50 g/L sucrose, 200 mg/L NaC1 and 50 or 100 mg/L proline）. The fresh and dry weights of callus were recorded after six weeks. The result indicated that maximum production of fresh and dry callus weight were 1,014 mg and 46.20 mg respectively achieved at 0.5 mg/L 2,4-D and 3 mg/L BA adding to the medium. Dry callus tissues were extracted with 70% methanol and analyzed by HPLC to determine the concentrations of thymol. The addition of abiotic elicitors to MS medium caused significant reduction in fresh weight of callus compared with control treatment. The concentration of thymol in the callus cultured on control treatment was 146.6 ppm. The data showed that 50 or 100 mg/L proline produced the highest yield of thymol 181.48 ppm and 174.58 ppm respectively, followed by sucrose 162.9 ppm, whereas the treatment with NaCI caused reduction in thymol concentration to percentage of 50.56% compared with the control.展开更多
In this study, using Taxus cuspidata as a raw material, we obtained stable high-yielding cell lines by subculturing and quantified paclitaxel content using ultra- sonic extraction combined with TLC-UV spectrophotom- e...In this study, using Taxus cuspidata as a raw material, we obtained stable high-yielding cell lines by subculturing and quantified paclitaxel content using ultra- sonic extraction combined with TLC-UV spectrophotom- etry. In single factor and multiple factors tests to optimize design and study the effects of elicitors, precursors, and metabolic inhibitors on paclitaxel production by Taxus cuspidata cells, paclitaxel production reached 4.32 mg/L when 100 μmol/L methyl jasmonate, 20 mg/L salicylic acid, 400 mg/L phenylalanine and 2 mg/L gibberellin （GA3） were added to the culture medium of suspension cells. When adding metabolic adjustment factors on the 7th day of culture, extra- and intracellular paclitaxel production was the highest at 4.855 mg/L, paclitaxel release rate was 10.48 %, fresh mass and paclitaxel production of cell increased, respectively, by 6.08 and 11.57 %. By controlling the anabolism of paclitaxel, paclitaxel yield was significantly improved.展开更多
为探讨真菌诱导子调控紫草素合成的分子机制,以新疆紫草无菌苗为试材,经尖孢镰刀菌、立枯丝核菌制备成的诱导子生物诱导后,对其根部进行RNA测序和分析。结果表明,与对照组比较,尖孢镰刀菌试验组差异表达基因1735个;立枯丝核菌试验组差...为探讨真菌诱导子调控紫草素合成的分子机制,以新疆紫草无菌苗为试材,经尖孢镰刀菌、立枯丝核菌制备成的诱导子生物诱导后,对其根部进行RNA测序和分析。结果表明,与对照组比较,尖孢镰刀菌试验组差异表达基因1735个;立枯丝核菌试验组差异表达基因1043个。GO(gene ontology)分析发现,2个试验组差异表达基因主要富集在细胞过程、细胞组分中膜和分子功能中催化活性等生物学过程。KEGG(kyoto encyclopedia of genes and genomes)分析发现,2个试验组在植物与病原菌互作、植物激素信号转导途径和苯丙素合成等通路均有大量差异表达基因富集。2个试验组差异表达情况相同的转录因子主要包括bHLH、AP2/ERF-ERF和LOB等,并发现参与紫草素合成及其正向调控的AeGHQH、AeDSH1、AeAP、AePAL、AeDI2、AePGT、AeHMGR、AeG10H基因在2个试验组均上调表达,其中尖孢镰刀菌试验组上调表达较显著。以上结果从分子水平探究了新疆紫草对真菌诱导子生物诱导的响应机制,为未来真菌诱导子应用于新疆紫草种植生产奠定了理论基础。展开更多
Salt stress is one of the most harmful environmental stresses in recent times and represents a significant threat to food security. Soil salinization is caused by spontaneous natural processes of mineral dissolution a...Salt stress is one of the most harmful environmental stresses in recent times and represents a significant threat to food security. Soil salinization is caused by spontaneous natural processes of mineral dissolution and human activities such as inappropriate irrigation practices. Natural geological progressions like weathering of rocks, arid climate, and higher evaporation, as well as anthropogenic activities, including the use of brackish water for irrigation, and poor tillage operations, are the foremost causes of soil salinization. Typical characteristics of saline soils are salt stress, high pH, and lack of organic carbon, as well as low availability of nutrients. Disruption of precipitation patterns as well as high average annual temperatures due to climate change additionally negatively affects the process of soil salinization. Productivity and ability to support crop growth are reduced on saline soil. Salinity-induced stress reduces plant growth by modulating the antioxidative system and nutrient orchestration. The aim of this work is to show that the mentioned problems can be alleviated in several ways such as the addition of biochar, exogenous application of several elicitors, seed priming, etc. Research has shown that the addition of biochar can significantly improve the recovery of saline soil. The addition of biochar has no significant effect on soil pH, while the cation exchange capacity of the soil increased by 17%, and the electrical conductivity of the saturated paste extract decreased by 13.2% (depends on the initial salinity and the type of biochar raw material). Moreover, biochar enriched with silicon increases the resistance of bananas to salt stress. In addition, exogenous application of several elicitors helps plants to alleviate stress by inducing stress-related physicochemical and molecular changes (selenium, sulfur, silicon, salicylic acid). Finally, seed priming showed positive effects on metabolomics, proteomics and growth of plants subjected to abiotic stress. Priming usually involves immersing the seed in a solution for a period of time to induce physiological and metabolic progression prior to germination.展开更多
文摘The artemisinin accumulation in the hairy root cultures of Artemisia annua L. was enhanced via a treatment of three fungal elicitors separately ( Verticillium dahliae Kleb., Rhizopus stolonifer (Ehrenb. ex Fr.) Vuill and Colletotrichum dematium (Pers.) Grove). Among these three elicitors, V. dahliae had the highest inducing efficiency, but none of them manifests any noticeable effects on the cell growth of the hairy root cultures. The artemisinin content of the hairy root cultures treated with V. dahliae elicitor was 1.12 mg/g DW, which was 45% higher than the control (0.77 mg/g DW). The results showed that elicitation was dependent on the elicitor concentration, the incubation period and the physiological stage at which the hairy root cultures were treated. In addition, the authors found that for V. dahliae , the optimum concentration was 0.4 mg carbohydrate per millilitre medium, the strongest response of A. annua hairy root cultures to the elicitation was at the late exponential growth stage, and the highest artemisinin content of the hairy root cultures was on the 4th day post treatment.
基金Supported by National Nonprofit Institute Research Grant of CATAS-TCGRI(1630032014027)Special Fund for Agro-scientific Research in the Public Interest(201203071)Special Project for Scientific and Technological Achievements Demonstration and Promotion in Hainan Province(SQ2013CGSF0006)~~
文摘To optimize the technique of rapid propagation of Dendrobium hybrida seedlings and to explore a hormone-free tissue culture method for D.hybrida,six kinds of mycorrhizal fungi which were isolated from the wild orchids were made into fungal elicitors.These fungal elicitors were added into the DE medium with concentrations of 40,60 and 80 ml/L,respectively.After a 90-d culturing,the effects of fungal elicitors on the growth of D.hybrida cultivar ‘088＇ tissue culture seedlings were studied.The results showed that treatment 13（T13） extremely significantly increased the fresh weight,but other treatment groups had no significant effects.In addition,T1,T5,T9,T11 and T13 extremely significantly influenced the contents of chlorophyll a and chlorophyll a ＋ b.However,T1 and T11 had extremely significantly effect on the content of chlorophyll b.Combining the effects on fresh weight and chlorophyll content,it could be concluded T13（40 ml/L of Y05） has promoting effects on the growth of D.hybrida tissue culture seedlings.
文摘【目的】明确山田胶锈菌(Gymnosporangium yamadae)吸器中效应蛋白GyHGSRE1的基础生物学功能,为进一步揭示山田胶锈菌效应蛋白作用的分子机制,制定持久的锈病防控策略奠定基础。【方法】基于前期获得的山田胶锈菌吸器转录组数据筛选具有高表达丰度的效应蛋白;通过MEME(http://meme-suite.org/)、AI(https://drug.ai.tencent.com)等在线网站预测GyHGSRE1的蛋白质结构;利用荧光定量PCR检测GyHGSRE1基因在山田胶锈菌侵染过程中的表达水平,以酵母菌分泌系统验证GyHGSRE1信号肽的分泌功能,并通过农杆菌介导的瞬时表达技术分析GyHGSRE1在本氏烟(Nicotiana benthamiana)和苹果(Malus domestica)叶片中的功能。【结果】在山田胶锈菌吸器转录组数据中获得了FPKM(fragments per Kb per million mapped reads)值为117.92、富含甘氨酸和丝氨酸的小分子分泌蛋白GyHGSRE1。其N端具有1个富含丝氨酸motif的信号肽。qRT-PCR显示GyHGSRE1基因在山田胶锈菌的吸器形成阶段以及性、锈孢子形成及发育阶段均上调表达,并定位于本氏烟叶片细胞的细胞质和细胞核,能够诱导细胞坏死并激发基础免疫防御反应,但含有信号肽的全长效应蛋白GyHGSRE1能够诱导苹果叶片细胞坏死,去除信号肽后诱导细胞坏死的能力减弱。【结论】富含甘氨酸和丝氨酸的非典型效应蛋白GyHGSRE1定位于植物细胞的细胞质和细胞核,能够诱导本氏烟及苹果叶片细胞坏死,因此可能具有广谱性的激发子活性。GyHGSRE1在山田胶锈菌侵染苹果叶片的定殖和孢子发育阶段显著表达从而发挥功能,其信号肽可能决定着对寄主特异性识别的相关功能。
基金supported by CNPq(National Council for Scientific and Technological Development—Processo:151057/2024-9),CAPES(Coordination for the Improvement of Higher Education Personnel)financial code—001,and UFCG(Universidade Federal de Campina Grande).
文摘The Northeast region is the main producer of guava in Brazil,generating employment and income.However,water availability means that producer’s resort to using water with high salinity,which harms plant development,especially during the seedling formation phase.The adoption of techniques that mitigate the deleterious effect of salinity is increasingly necessary,such as the use of elicitors such as ascorbic acid.The purpose of this study was to analyze the morphophysiology of guava seedlings under saline and ascorbic acid levels.The study was carried out by applying treatments composed of five saline levels(SL=0.3;1.3;2.3;3.3 and 4.3 dS m^(-1))and four levels of ascorbic acid—AA(0,200,400,and 600 mg L^(-1)),in a 5×4 factorial arrangement,adopting a randomized block design.Gas exchange and growth of guava seedlings are limited from 0.3 dS m^(-1).Using 400 mg L^(-1)of AA reduces damage from salinity on stomatal conductance,transpiration,and net assimilation rate up to the estimated SL of 1.80 dS m^(-1).In contrast,AA level 412 mg L^(-1)increased instantaneous water use efficiency up to the salinity of 2.3 dS m^(-1).AA level of 600 mg L^(-1)attenuated salt stress effects on leaf area and height/stem diameter ratio up to SL of 2.05 dS m^(-1).The number of leaves and the absolute and relative growth rates were stimulated by AA under the lowest saline level.
文摘Callus cultures of Origanum vulgare L. were established from leaf discus on Murashige and Skoog （MS） medium containing different levels of growth regulators, i.e., 2,4-Dichlorophenoxyacetic acid （2,4-D）, Naphthalene acetic acid （NAA）, Benzyl Adenine （BA） and Kinetin （Kn） and incubated under dark condition. Callus tissues were employed to study the influence of abiotic elicitors on the production of thymol. Constant weights of callus （300 mg） were cultured on accumulation medium treated separately with each one of elicitors used （50 g/L sucrose, 200 mg/L NaC1 and 50 or 100 mg/L proline）. The fresh and dry weights of callus were recorded after six weeks. The result indicated that maximum production of fresh and dry callus weight were 1,014 mg and 46.20 mg respectively achieved at 0.5 mg/L 2,4-D and 3 mg/L BA adding to the medium. Dry callus tissues were extracted with 70% methanol and analyzed by HPLC to determine the concentrations of thymol. The addition of abiotic elicitors to MS medium caused significant reduction in fresh weight of callus compared with control treatment. The concentration of thymol in the callus cultured on control treatment was 146.6 ppm. The data showed that 50 or 100 mg/L proline produced the highest yield of thymol 181.48 ppm and 174.58 ppm respectively, followed by sucrose 162.9 ppm, whereas the treatment with NaCI caused reduction in thymol concentration to percentage of 50.56% compared with the control.
基金supported by development plan project during ‘‘the 12th Five Year Plan’’ Nation Science and Technology in rural area(No.2012AA10A506-04 and No.2013AA103005-04)Changchun City science and technology development program(No.2014174)Changchun City science and technology support program(No.2014NK002)
文摘In this study, using Taxus cuspidata as a raw material, we obtained stable high-yielding cell lines by subculturing and quantified paclitaxel content using ultra- sonic extraction combined with TLC-UV spectrophotom- etry. In single factor and multiple factors tests to optimize design and study the effects of elicitors, precursors, and metabolic inhibitors on paclitaxel production by Taxus cuspidata cells, paclitaxel production reached 4.32 mg/L when 100 μmol/L methyl jasmonate, 20 mg/L salicylic acid, 400 mg/L phenylalanine and 2 mg/L gibberellin （GA3） were added to the culture medium of suspension cells. When adding metabolic adjustment factors on the 7th day of culture, extra- and intracellular paclitaxel production was the highest at 4.855 mg/L, paclitaxel release rate was 10.48 %, fresh mass and paclitaxel production of cell increased, respectively, by 6.08 and 11.57 %. By controlling the anabolism of paclitaxel, paclitaxel yield was significantly improved.
文摘为探讨真菌诱导子调控紫草素合成的分子机制,以新疆紫草无菌苗为试材,经尖孢镰刀菌、立枯丝核菌制备成的诱导子生物诱导后,对其根部进行RNA测序和分析。结果表明,与对照组比较,尖孢镰刀菌试验组差异表达基因1735个;立枯丝核菌试验组差异表达基因1043个。GO(gene ontology)分析发现,2个试验组差异表达基因主要富集在细胞过程、细胞组分中膜和分子功能中催化活性等生物学过程。KEGG(kyoto encyclopedia of genes and genomes)分析发现,2个试验组在植物与病原菌互作、植物激素信号转导途径和苯丙素合成等通路均有大量差异表达基因富集。2个试验组差异表达情况相同的转录因子主要包括bHLH、AP2/ERF-ERF和LOB等,并发现参与紫草素合成及其正向调控的AeGHQH、AeDSH1、AeAP、AePAL、AeDI2、AePGT、AeHMGR、AeG10H基因在2个试验组均上调表达,其中尖孢镰刀菌试验组上调表达较显著。以上结果从分子水平探究了新疆紫草对真菌诱导子生物诱导的响应机制,为未来真菌诱导子应用于新疆紫草种植生产奠定了理论基础。
文摘Salt stress is one of the most harmful environmental stresses in recent times and represents a significant threat to food security. Soil salinization is caused by spontaneous natural processes of mineral dissolution and human activities such as inappropriate irrigation practices. Natural geological progressions like weathering of rocks, arid climate, and higher evaporation, as well as anthropogenic activities, including the use of brackish water for irrigation, and poor tillage operations, are the foremost causes of soil salinization. Typical characteristics of saline soils are salt stress, high pH, and lack of organic carbon, as well as low availability of nutrients. Disruption of precipitation patterns as well as high average annual temperatures due to climate change additionally negatively affects the process of soil salinization. Productivity and ability to support crop growth are reduced on saline soil. Salinity-induced stress reduces plant growth by modulating the antioxidative system and nutrient orchestration. The aim of this work is to show that the mentioned problems can be alleviated in several ways such as the addition of biochar, exogenous application of several elicitors, seed priming, etc. Research has shown that the addition of biochar can significantly improve the recovery of saline soil. The addition of biochar has no significant effect on soil pH, while the cation exchange capacity of the soil increased by 17%, and the electrical conductivity of the saturated paste extract decreased by 13.2% (depends on the initial salinity and the type of biochar raw material). Moreover, biochar enriched with silicon increases the resistance of bananas to salt stress. In addition, exogenous application of several elicitors helps plants to alleviate stress by inducing stress-related physicochemical and molecular changes (selenium, sulfur, silicon, salicylic acid). Finally, seed priming showed positive effects on metabolomics, proteomics and growth of plants subjected to abiotic stress. Priming usually involves immersing the seed in a solution for a period of time to induce physiological and metabolic progression prior to germination.
