Citrus canker, an epidemic quarantine disease caused by Xanthomonas axonopodis pv. citri, has brought a great damage in citrus production worldwide. Herein, a rice PRR (pattern recognition receptor) gene Xa21 togeth...Citrus canker, an epidemic quarantine disease caused by Xanthomonas axonopodis pv. citri, has brought a great damage in citrus production worldwide. Herein, a rice PRR (pattern recognition receptor) gene Xa21 together with GUS reporter gene and hygromycin phosphotransferase gene (HPT) was introduced into Anliucheng sweet orange (Citrus sinensis Osbeck) via Agrobacterium-mediated transformation of embryogenic callus. The transgenic calluses were screened on MT basal medium containing hygromycin (HYG) and detected by histochemical GUS staining. The transgenic plantlets were recovered through somatic embryogenesis pathway. The regenerated plantlets were accustomed to and maintained in the greenhouse. The transgene integration of recovered plantlets was identiifed by PCR and Southern blot hybridization. It showed that all the transgenic plantlets tested had undergone single copy integration, the expression of Xa21 in eight different transgenic lines detected by qRT-PCR can be divided into three grades, high for T5 and T6, middle for T4 and low for the rest. The tolerance to citrus canker disease of the three recovered transgenic lines T2, T4 and T6 was assessed by in vitro pin-puncture inoculation. The results showed that all the three transgenic lines conferred improved resistance to citrus canker bacterium infection and the T4 transgenic line displayed the highest resistance. The mechanism and feasibility of rice Xa21 in triggering innate immunity in citrus was brielfy discussed.展开更多
Citrus tristeza virus (CTV) causes economically important losses to the citrus industry worldwide. Mild strain cross protection (MSCP) against tristeza has hardly been practised due to mixed infection of different...Citrus tristeza virus (CTV) causes economically important losses to the citrus industry worldwide. Mild strain cross protection (MSCP) against tristeza has hardly been practised due to mixed infection of different CTV-strains and little background of its molecular biology in China. For better cognition on CTV, 192 sweet orange samples collected from eight provinces (Chongqing, Sichuan, Fujian, Hunan, Guangxi, Yunnan, Guangdong and Jiangxi) were tested by direct tissue blot immuno-assay (DTBIA), and 158 of them were tested positively, which therefore were subjected to coat protein gene (CPG)/Hinf Ⅰ restriction fragment length polymorphism (RFLP) analysis. Sample bulks were compared between Chongqing and Fujian by some statistical data, including ratios of single infection and mixed infection to local samples, proportions of CTV isolates with single RFLP groups, and rates of each RFLP group. The simplified analysis of samples from the other six provinces were then conducted. This study suggests that CTV isolates with CPG/Hinf Ⅰ RFLP groups Ⅲ and Ⅰ are the main epidemic ones in China, and mixed infection of CTV in fields are popular. Based on observation of severity of stem-pitting symptom in field trees, CTV isolates with CPG/Hinf Ⅰ RFLP groups Ⅲ and Ⅰ caused severe stem-pittings in sweet oranges in China.展开更多
Silver nanoparticles are considerecl as good antimicrobial agent. AgNPs were synthesized by mixing silver nitrate solution with citrus sinesb extract for 2 h at 37 and analyzed by UV-visible spectra, SEM, XRD, and FT...Silver nanoparticles are considerecl as good antimicrobial agent. AgNPs were synthesized by mixing silver nitrate solution with citrus sinesb extract for 2 h at 37 and analyzed by UV-visible spectra, SEM, XRD, and FTIR. AgNPs were tested against B. subtilis, Shigello, S. oureus, ond E. coli. Minimum inhibitory concentration of AgNPs was 20 I^g/mL for B. subtilis and Shigello and 30 I^g/mL for S. oureus and E. coll. Antibiofilm activity (80% to 90%) was observed at 25 IJg/mL. AgNPs were stable for five months with sustained an'timicrobial activity. Biosynthesized AgNPs can bE: used to inhibit food poisoning microbial growth.展开更多
基金financially supported by the National HighTech R&D Program of China (863, 2011AA100205)the National Natural Science Foundation of China (31125024)
文摘Citrus canker, an epidemic quarantine disease caused by Xanthomonas axonopodis pv. citri, has brought a great damage in citrus production worldwide. Herein, a rice PRR (pattern recognition receptor) gene Xa21 together with GUS reporter gene and hygromycin phosphotransferase gene (HPT) was introduced into Anliucheng sweet orange (Citrus sinensis Osbeck) via Agrobacterium-mediated transformation of embryogenic callus. The transgenic calluses were screened on MT basal medium containing hygromycin (HYG) and detected by histochemical GUS staining. The transgenic plantlets were recovered through somatic embryogenesis pathway. The regenerated plantlets were accustomed to and maintained in the greenhouse. The transgene integration of recovered plantlets was identiifed by PCR and Southern blot hybridization. It showed that all the transgenic plantlets tested had undergone single copy integration, the expression of Xa21 in eight different transgenic lines detected by qRT-PCR can be divided into three grades, high for T5 and T6, middle for T4 and low for the rest. The tolerance to citrus canker disease of the three recovered transgenic lines T2, T4 and T6 was assessed by in vitro pin-puncture inoculation. The results showed that all the three transgenic lines conferred improved resistance to citrus canker bacterium infection and the T4 transgenic line displayed the highest resistance. The mechanism and feasibility of rice Xa21 in triggering innate immunity in citrus was brielfy discussed.
基金supported by the National Natural Science Foundation of China(30471205)under the program"Comparison Research on Strains of Citrus tristeza virus."
文摘Citrus tristeza virus (CTV) causes economically important losses to the citrus industry worldwide. Mild strain cross protection (MSCP) against tristeza has hardly been practised due to mixed infection of different CTV-strains and little background of its molecular biology in China. For better cognition on CTV, 192 sweet orange samples collected from eight provinces (Chongqing, Sichuan, Fujian, Hunan, Guangxi, Yunnan, Guangdong and Jiangxi) were tested by direct tissue blot immuno-assay (DTBIA), and 158 of them were tested positively, which therefore were subjected to coat protein gene (CPG)/Hinf Ⅰ restriction fragment length polymorphism (RFLP) analysis. Sample bulks were compared between Chongqing and Fujian by some statistical data, including ratios of single infection and mixed infection to local samples, proportions of CTV isolates with single RFLP groups, and rates of each RFLP group. The simplified analysis of samples from the other six provinces were then conducted. This study suggests that CTV isolates with CPG/Hinf Ⅰ RFLP groups Ⅲ and Ⅰ are the main epidemic ones in China, and mixed infection of CTV in fields are popular. Based on observation of severity of stem-pitting symptom in field trees, CTV isolates with CPG/Hinf Ⅰ RFLP groups Ⅲ and Ⅰ caused severe stem-pittings in sweet oranges in China.
文摘Silver nanoparticles are considerecl as good antimicrobial agent. AgNPs were synthesized by mixing silver nitrate solution with citrus sinesb extract for 2 h at 37 and analyzed by UV-visible spectra, SEM, XRD, and FTIR. AgNPs were tested against B. subtilis, Shigello, S. oureus, ond E. coli. Minimum inhibitory concentration of AgNPs was 20 I^g/mL for B. subtilis and Shigello and 30 I^g/mL for S. oureus and E. coll. Antibiofilm activity (80% to 90%) was observed at 25 IJg/mL. AgNPs were stable for five months with sustained an'timicrobial activity. Biosynthesized AgNPs can bE: used to inhibit food poisoning microbial growth.