Hesperidin is a dihydroflavonoids, accounting for more than 50% of the total flavonoids in Citri Reticulatae Pericarpium(CRP) of traditional Chinese medicine. It is an effective antioxidant and free radical scavenger ...Hesperidin is a dihydroflavonoids, accounting for more than 50% of the total flavonoids in Citri Reticulatae Pericarpium(CRP) of traditional Chinese medicine. It is an effective antioxidant and free radical scavenger that has anti-inflammatory, antiviral and hypoglycemic properties.The latest studies reported that hesperidin has a potential for novel coronavirus resistance. However, little is known about the synthesis regulation and accumulation site of hesperidin in plants. In this study, hesperidin synthase gene Crc1,6RhaT was cloned, and the protein can be completely transformed flavanone-7-O-glucoside into hesperidin in vitro and in vivo. Studies on biological characteristics of ovary walls and exocarps showed that the relative expression levels of the Crc1,6RhaT gene and protein decreased gradually with the development of citrus fruits, and the relative content of hesperidin firstly increased, then sequentially decreased. In situ hybridization results further revealed that Crc1,6RhaT transcription was mainly concentrated in the secretory cavity cells, which are revealed to be the site of flavonoid synthesis.Immunocytochemistry localization results showed that the Crc1,6RhaT was mainly located in the endoplasmic reticulum, nucleus and vacuole of secretory cells. We inferred that the Crc1,6RhaT was synthesized in the endoplasmic reticulum, then was transported into the vacuoles through enlarged vesicles at the end of the endoplasmic reticulum. Our results not only revealed that Crc1,6RhaT may be involved in the synthesis of hesperidin of the main bioactive substance in the medicinal plant Citrus reticulata ‘Chachi' fruit, but also provided novel insights into the main subcellular sites of hesperidin biosynthesis in vacuoles.展开更多
[Objectives]To investigate the effects and mechanism of fruit immersion treatment with salicylic acid(SA),calcium chloride(CaCl_(2))solutions at different concentrations or their combinations on post-harvest preservat...[Objectives]To investigate the effects and mechanism of fruit immersion treatment with salicylic acid(SA),calcium chloride(CaCl_(2))solutions at different concentrations or their combinations on post-harvest preservation of Gonggan Mandarin(Citrus reticulata)fruit.[Methods]13 experiments(2 storage temperatures,3 control treatments,3 concentrations of SA solution,4 concentrations of CaCl_(2) solution and 3 combinations of SA+CaCl_(2) solution for fruit immersion treatment)were conducted for 90 d.Gonggan fruit weight loss rate,rotting rate,disease index,disease type,soluble solids and organic acids content,and the differences in the activity of six enzymes(APX,PPO,CHI,CAT,SOD,PAL)related to fruit disease resistance were detected regularly.[Results]With the increase of storage days,the weight loss rate,rotting rate and disease index increased,the content of soluble solids and organic acids decreased,the activity of APX,PPO,CHI and SOD increased,and the activity of CAT and PAL decreased.However,the increase or decrease of fruit loss at low temperature was the smallest,and the increase or decrease of fruit at natural room temperature was the largest.Gonggan fruit cleaning before storage is an effective preservation and fresh-keeping technology,but its preservation and fresh-keeping effect is far less than that of fruit immersion by SA,CaCl_(2) and SA+CaCl_(2) compound solution.The effect of fruit immersion by SA solution was greater than that of fruit immersion by CaCl_(2) solution;the best concentration of SA alone was 6.0 mmol/L,which was not significantly different from 3.0 mmol/L,but the effect of 9.0 mmol/L decreased;the best concentration of CaCl_(2) alone was 6.0 mmol/L,which was significantly better than 3.0 and 9.0 mmol/L;the optimal concentration combination of SA+CaCl_(2) compound solution for fruit immersion treatment was 6.0 mmol/L SA+3.0 mmol/L CaCl_(2).[Conclusions]The fruit immersion by SA,CaCl_(2) and their compound solutions induced the increase of activity of six enzymes related to disease resistance in Gonggan fruit,and PPO and CHI were judged to be two key enzymes based on response intensity.展开更多
Manual fruit thinning(MFT)in fruit trees has been previously shown to increase fruit size and enhance fruit quality,but the effect of MFT on Ponkan(Citrus reticulata Blanco)and the underlying mechanisms remain poorly ...Manual fruit thinning(MFT)in fruit trees has been previously shown to increase fruit size and enhance fruit quality,but the effect of MFT on Ponkan(Citrus reticulata Blanco)and the underlying mechanisms remain poorly understood.In this study,efforts were made to elucidate how MFT influences the fruit quality of Ponkan.The results showed that MFT substantially increased fruit size and elevated fruit total soluble solids in comparison with the fruit from the unthinned trees(used as control).Expression analyses demonstrated that m RNA abundance of three important sugar transporter genes,including CrSUT1,CrSTP1 and CrTMT1,was evidently elevated in the flesh of thinned fruit when compared with those of the control.In addition,MFT prominently up-regulated the transcript levels of various auxin and gibberellin(GA)biosynthesis and signaling genes,including CrYUC6,CrAUX/IAA,CrGA20ox1 and CrGA3ox1.Concurrently,the contents of endogenous IAA and GA3,measured at 90 d after fruit thinning,were notably elevated in the fruit from trees with the thinning treatment relative to the control,although no difference was detected in the two groups before the thinning manipulation.Taken together,these results indicate that manual fruit thinning could greatly improve fruit quality,which may be attributed to promoting fruit expansion due to the increased auxin levels and expediting sugar accumulation through the up-regulation of sugar transporter genes.展开更多
Citrus reticulata (Mandarin Orange), commonly known as “Sweet Orange”, is one of the most difficult plants to improve through traditional breeding approaches as it poses various biological limitations that greatly h...Citrus reticulata (Mandarin Orange), commonly known as “Sweet Orange”, is one of the most difficult plants to improve through traditional breeding approaches as it poses various biological limitations that greatly hinder the cultivar improvement. In the present study, using the fresh seed of native orange as explant, an efficient, reproducible, regeneration method was developed through in vitro organogenesis. Mature, healthy and dehusked seeds were treated with Murashige and Skoog, (MS) media containing 3% sucrose, 0.7% agar supplemented with different concentrations and combinations of phytohormones. The highest calli initiation (93.3% ± 0.5%) responses were observed on MS medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) at 3.0 mg/L followed by 2,4-D at 3.5 mg/L (86.7% ± 1.75%) in this experiment. Maximum shoot regeneration (86.7% ± 3.35%) responses were reported using MS medium supplemented with the combination of 6-benzylaminopurine (BAP) at 3.0 mg/L and 1-naphthaleneacetic acid (NAA) at 2.0 mg/L. MS medium supplemented with NAA at 1.0 mg/L showed the best rooting (80% ± 2.89%) response in comparison to (70% ± 5.20%) indole-3-butyric acid (IBA) at 1.0 mg/L. The regenerated plantlets were acclimatized in pots containing sterile garden soil mixture to examine their response in natural conditions.展开更多
Huanglongbing （HLB, or citrus greening） is the most destructive disease of citrus, which is associated with Candidatus Liberibacter asiaticus （Las）. Few management options are available, aside from preventive meas...Huanglongbing （HLB, or citrus greening） is the most destructive disease of citrus, which is associated with Candidatus Liberibacter asiaticus （Las）. Few management options are available, aside from preventive measures such as removing infected plants, planting disease-free seedlings, and managing the insect vector. In this study, we assessed the efficacy of thermotherapy against HLB under controlled greenhouse conditions. A total of 60 two-year-old, graft-infected Citrus reticulata Blanco plants were used. The plants were randomly divided into three groups （45℃, 48℃, and untreated control）, with five plants/replicate （rep） and four reps/treatment. The treated plants were placed in phytotrons for a 4-h treatment session, repeated once per week for three consecutive weeks. Disease remission was observed eight weeks post-treatment. Real-time PCR assays revealed that Las titers in HLB-affected seedlings were significantly reduced in both 45 and 48℃ treatments four weeks after treatment, with the exception of eight plants. In contrast, Las titers in the untreated control plants increased significantly during the same period, with a maximum increase of 28-fold. Except for seven plants, Las titers in the new flushes of treated plants decreased more than 90% eight weeks after treatment. Las titers in mature leaves of treated plants decreased 56 and 60% in average at 45 and 48℃, respectively, eight weeks after treatment. The HLB symptoms and Las titer of seedings were markedly alleviated eight weeks after treatment in both 45 and 48℃ treatments. Our results laid a good foundation for the further development of citrus free-disease seedling cultivation and Huanglongbing control in the field. The whole plants were replaced for scion or branch in previous as the research object in this study, and the expression of Huanglongbing symptoms combined with real-time polymerase chain reaction （PCR） were used to evaluate the effect of heat treatment in the greenhouse.展开更多
基金supported by the National Natural Science Foundation of China (Grant No.32270381)Natural Science Foundation of Guangdong (Grant No.2022A1515011086)+2 种基金Key Realm R&D Program of Guangdong Province (Grant No.2020B020221001)Provincial Special Fund for Modern Agriculture Industry Technology Innovation Teams (Grant No.2019KJ125)Research Fund of Maoming Branch,Guangdong Laboratory for Lingnan Modern Agriculture (Grant No.2022KF009)。
文摘Hesperidin is a dihydroflavonoids, accounting for more than 50% of the total flavonoids in Citri Reticulatae Pericarpium(CRP) of traditional Chinese medicine. It is an effective antioxidant and free radical scavenger that has anti-inflammatory, antiviral and hypoglycemic properties.The latest studies reported that hesperidin has a potential for novel coronavirus resistance. However, little is known about the synthesis regulation and accumulation site of hesperidin in plants. In this study, hesperidin synthase gene Crc1,6RhaT was cloned, and the protein can be completely transformed flavanone-7-O-glucoside into hesperidin in vitro and in vivo. Studies on biological characteristics of ovary walls and exocarps showed that the relative expression levels of the Crc1,6RhaT gene and protein decreased gradually with the development of citrus fruits, and the relative content of hesperidin firstly increased, then sequentially decreased. In situ hybridization results further revealed that Crc1,6RhaT transcription was mainly concentrated in the secretory cavity cells, which are revealed to be the site of flavonoid synthesis.Immunocytochemistry localization results showed that the Crc1,6RhaT was mainly located in the endoplasmic reticulum, nucleus and vacuole of secretory cells. We inferred that the Crc1,6RhaT was synthesized in the endoplasmic reticulum, then was transported into the vacuoles through enlarged vesicles at the end of the endoplasmic reticulum. Our results not only revealed that Crc1,6RhaT may be involved in the synthesis of hesperidin of the main bioactive substance in the medicinal plant Citrus reticulata ‘Chachi' fruit, but also provided novel insights into the main subcellular sites of hesperidin biosynthesis in vacuoles.
基金Supported by Guangdong Science and Technology Plan Project(2014A020208143)。
文摘[Objectives]To investigate the effects and mechanism of fruit immersion treatment with salicylic acid(SA),calcium chloride(CaCl_(2))solutions at different concentrations or their combinations on post-harvest preservation of Gonggan Mandarin(Citrus reticulata)fruit.[Methods]13 experiments(2 storage temperatures,3 control treatments,3 concentrations of SA solution,4 concentrations of CaCl_(2) solution and 3 combinations of SA+CaCl_(2) solution for fruit immersion treatment)were conducted for 90 d.Gonggan fruit weight loss rate,rotting rate,disease index,disease type,soluble solids and organic acids content,and the differences in the activity of six enzymes(APX,PPO,CHI,CAT,SOD,PAL)related to fruit disease resistance were detected regularly.[Results]With the increase of storage days,the weight loss rate,rotting rate and disease index increased,the content of soluble solids and organic acids decreased,the activity of APX,PPO,CHI and SOD increased,and the activity of CAT and PAL decreased.However,the increase or decrease of fruit loss at low temperature was the smallest,and the increase or decrease of fruit at natural room temperature was the largest.Gonggan fruit cleaning before storage is an effective preservation and fresh-keeping technology,but its preservation and fresh-keeping effect is far less than that of fruit immersion by SA,CaCl_(2) and SA+CaCl_(2) compound solution.The effect of fruit immersion by SA solution was greater than that of fruit immersion by CaCl_(2) solution;the best concentration of SA alone was 6.0 mmol/L,which was not significantly different from 3.0 mmol/L,but the effect of 9.0 mmol/L decreased;the best concentration of CaCl_(2) alone was 6.0 mmol/L,which was significantly better than 3.0 and 9.0 mmol/L;the optimal concentration combination of SA+CaCl_(2) compound solution for fruit immersion treatment was 6.0 mmol/L SA+3.0 mmol/L CaCl_(2).[Conclusions]The fruit immersion by SA,CaCl_(2) and their compound solutions induced the increase of activity of six enzymes related to disease resistance in Gonggan fruit,and PPO and CHI were judged to be two key enzymes based on response intensity.
基金supported by the Hubei Provincial Agriculture Research System,Chinathe Special Program for Technology Innovation of Hubei Province,China(2020BBA036)the Hubei Provincial Agricultural Science and Technology Innovation,China。
文摘Manual fruit thinning(MFT)in fruit trees has been previously shown to increase fruit size and enhance fruit quality,but the effect of MFT on Ponkan(Citrus reticulata Blanco)and the underlying mechanisms remain poorly understood.In this study,efforts were made to elucidate how MFT influences the fruit quality of Ponkan.The results showed that MFT substantially increased fruit size and elevated fruit total soluble solids in comparison with the fruit from the unthinned trees(used as control).Expression analyses demonstrated that m RNA abundance of three important sugar transporter genes,including CrSUT1,CrSTP1 and CrTMT1,was evidently elevated in the flesh of thinned fruit when compared with those of the control.In addition,MFT prominently up-regulated the transcript levels of various auxin and gibberellin(GA)biosynthesis and signaling genes,including CrYUC6,CrAUX/IAA,CrGA20ox1 and CrGA3ox1.Concurrently,the contents of endogenous IAA and GA3,measured at 90 d after fruit thinning,were notably elevated in the fruit from trees with the thinning treatment relative to the control,although no difference was detected in the two groups before the thinning manipulation.Taken together,these results indicate that manual fruit thinning could greatly improve fruit quality,which may be attributed to promoting fruit expansion due to the increased auxin levels and expediting sugar accumulation through the up-regulation of sugar transporter genes.
文摘Citrus reticulata (Mandarin Orange), commonly known as “Sweet Orange”, is one of the most difficult plants to improve through traditional breeding approaches as it poses various biological limitations that greatly hinder the cultivar improvement. In the present study, using the fresh seed of native orange as explant, an efficient, reproducible, regeneration method was developed through in vitro organogenesis. Mature, healthy and dehusked seeds were treated with Murashige and Skoog, (MS) media containing 3% sucrose, 0.7% agar supplemented with different concentrations and combinations of phytohormones. The highest calli initiation (93.3% ± 0.5%) responses were observed on MS medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) at 3.0 mg/L followed by 2,4-D at 3.5 mg/L (86.7% ± 1.75%) in this experiment. Maximum shoot regeneration (86.7% ± 3.35%) responses were reported using MS medium supplemented with the combination of 6-benzylaminopurine (BAP) at 3.0 mg/L and 1-naphthaleneacetic acid (NAA) at 2.0 mg/L. MS medium supplemented with NAA at 1.0 mg/L showed the best rooting (80% ± 2.89%) response in comparison to (70% ± 5.20%) indole-3-butyric acid (IBA) at 1.0 mg/L. The regenerated plantlets were acclimatized in pots containing sterile garden soil mixture to examine their response in natural conditions.
基金funded by the United States Department of Agriculture (USDA)-APHIS-PPQ-CPHST and North Carolina State University joint project (2012-0195-01)the Special Fund for Agro-Scientific Research in the Public Interest, China (201003067-05)
文摘Huanglongbing （HLB, or citrus greening） is the most destructive disease of citrus, which is associated with Candidatus Liberibacter asiaticus （Las）. Few management options are available, aside from preventive measures such as removing infected plants, planting disease-free seedlings, and managing the insect vector. In this study, we assessed the efficacy of thermotherapy against HLB under controlled greenhouse conditions. A total of 60 two-year-old, graft-infected Citrus reticulata Blanco plants were used. The plants were randomly divided into three groups （45℃, 48℃, and untreated control）, with five plants/replicate （rep） and four reps/treatment. The treated plants were placed in phytotrons for a 4-h treatment session, repeated once per week for three consecutive weeks. Disease remission was observed eight weeks post-treatment. Real-time PCR assays revealed that Las titers in HLB-affected seedlings were significantly reduced in both 45 and 48℃ treatments four weeks after treatment, with the exception of eight plants. In contrast, Las titers in the untreated control plants increased significantly during the same period, with a maximum increase of 28-fold. Except for seven plants, Las titers in the new flushes of treated plants decreased more than 90% eight weeks after treatment. Las titers in mature leaves of treated plants decreased 56 and 60% in average at 45 and 48℃, respectively, eight weeks after treatment. The HLB symptoms and Las titer of seedings were markedly alleviated eight weeks after treatment in both 45 and 48℃ treatments. Our results laid a good foundation for the further development of citrus free-disease seedling cultivation and Huanglongbing control in the field. The whole plants were replaced for scion or branch in previous as the research object in this study, and the expression of Huanglongbing symptoms combined with real-time polymerase chain reaction （PCR） were used to evaluate the effect of heat treatment in the greenhouse.
