Firmness is one of the most important fruit quality traits in strawberries.The postharvest shelf life of this soft fruit is highly limited by the loss of firmness,where cell wall disassembly plays an important role.Pr...Firmness is one of the most important fruit quality traits in strawberries.The postharvest shelf life of this soft fruit is highly limited by the loss of firmness,where cell wall disassembly plays an important role.Previous studies demonstrated that the polygalacturonase FaPG1 has a key role in remodelling pectins during strawberry softening.In this study,FaPG1 knockout strawberry plants have been generated using the CRISPR/Cas9 system delivered via Agrobacterium tumefaciens.Ten independent lines,cv.“Chandler”,were obtained,and all of them were successfully edited as determined by PCR amplification and T7 endonuclease assay.The targeted mutagenesis insertion and deletion rates were analyzed using targeted deep sequencing.The percentage of edited sequences varied from 47%up to almost 100%,being higher than 95%for seven of the selected lines.Phenotypic analyses showed that 7 out of the eight lines analyzed produced fruits significantly firmer than the control,ranging from 33 to 70%increase in firmness.There was a positive relationship between the degree of FaPG1 editing and the rise in fruit firmness.Minor changes were observed in other fruit quality traits,such as colour,soluble solids,titratable acidity or anthocyanin content.Edited fruits showed a reduced softening rate during postharvest,displayed a reduced transpirational water loss,and were less damaged by Botrytis cinerea inoculation.The analysis of four potential off-target sites revealed no mutation events.In conclusion,editing the FaPG1 gene using the CRISPR/Cas9 system is an efficient method for improving strawberry fruit firmness and shelf life.展开更多
目的:探讨腹腔镜胃癌D2根治术患者经协同护理及患者自评-主观全面评定(patient-generated subjective global assessment,PG-SGA)个体化营养干预的效果。方法:选择蚌埠医科大学第一附属医院2020年8月—2021年11月收治的107例腹腔镜胃癌D...目的:探讨腹腔镜胃癌D2根治术患者经协同护理及患者自评-主观全面评定(patient-generated subjective global assessment,PG-SGA)个体化营养干预的效果。方法:选择蚌埠医科大学第一附属医院2020年8月—2021年11月收治的107例腹腔镜胃癌D2根治术患者,随机分为参照组(n=54,行常规护理)和试验组(n=53,行协同护理+基于PG-SGA个体化营养干预)。对比两组患者的心理状态及生活质量。结果:试验组干预后汉密尔顿焦虑量表(Hamilton anxiety scale,HAMA)、汉密尔顿抑郁量表(Hamilton depression scale,HAMD)评分均较参照组低,健康调查简表(the MOS item short form health survey,SF-36)各维度评分均较参照组高,差异均有统计学意义(P<0.05)。结论:协同护理联合基于PG-SGA个体化营养干预应用于腹腔镜胃癌D2根治术患者,能够有效改善患者的心理状态,提升其生活质量。展开更多
In this study, 1 500 bp PG gene promoter was amplified from leaves of tomato cultivar ＇ Zhongshu No. 4＇. The bioinformatic analysis of PG promoter sequences was conducted. PG promoter elements were predicted and ana...In this study, 1 500 bp PG gene promoter was amplified from leaves of tomato cultivar ＇ Zhongshu No. 4＇. The bioinformatic analysis of PG promoter sequences was conducted. PG promoter elements were predicted and analyzed by PLANTCARE and PLACE. The results showed that tomato PG promoter contained multiple c/s-acting regulatory elements such as typical basic elements TATA-Box and CAAT-Box, light responsive elements 3-AF1 binding site, ATl-motif, ATCT- motif, Box 4, Box I, GA-motif, GTl-motif, Spl and MRE, heat stress-responsive element HSE, ethylene-responsive element ERE, meristem-specifie regulatory element CCGTCC-box, endosperm expression-related regulatory elements GCN4_motif and Skn-l_motif, defense and stress responsiveness element TC-rich repeats, and circadian control-related element circadian, indicating that the expression of tomato PG gene is related to light, temperature, hormone, stress and other factors. This study laid the foundation for subsequent research about regulation of PG gene expression in plants.展开更多
基金supported by the Ministerio de Ciencia,Innovación y Universidades and FEDER EU funds(grant numbers AGL2017-86531-C2-1R and PID2020-118468RB-C21)the University of Malaga(grant number B1-2020_09)+1 种基金CS-R was awarded a PhD Fellowship from the Ministerio de Ciencia,Innovación y Universidades(PRE2018-085509)PhD Program Advanced Biotechnology,University of Málaga.
文摘Firmness is one of the most important fruit quality traits in strawberries.The postharvest shelf life of this soft fruit is highly limited by the loss of firmness,where cell wall disassembly plays an important role.Previous studies demonstrated that the polygalacturonase FaPG1 has a key role in remodelling pectins during strawberry softening.In this study,FaPG1 knockout strawberry plants have been generated using the CRISPR/Cas9 system delivered via Agrobacterium tumefaciens.Ten independent lines,cv.“Chandler”,were obtained,and all of them were successfully edited as determined by PCR amplification and T7 endonuclease assay.The targeted mutagenesis insertion and deletion rates were analyzed using targeted deep sequencing.The percentage of edited sequences varied from 47%up to almost 100%,being higher than 95%for seven of the selected lines.Phenotypic analyses showed that 7 out of the eight lines analyzed produced fruits significantly firmer than the control,ranging from 33 to 70%increase in firmness.There was a positive relationship between the degree of FaPG1 editing and the rise in fruit firmness.Minor changes were observed in other fruit quality traits,such as colour,soluble solids,titratable acidity or anthocyanin content.Edited fruits showed a reduced softening rate during postharvest,displayed a reduced transpirational water loss,and were less damaged by Botrytis cinerea inoculation.The analysis of four potential off-target sites revealed no mutation events.In conclusion,editing the FaPG1 gene using the CRISPR/Cas9 system is an efficient method for improving strawberry fruit firmness and shelf life.
文摘目的:探讨腹腔镜胃癌D2根治术患者经协同护理及患者自评-主观全面评定(patient-generated subjective global assessment,PG-SGA)个体化营养干预的效果。方法:选择蚌埠医科大学第一附属医院2020年8月—2021年11月收治的107例腹腔镜胃癌D2根治术患者,随机分为参照组(n=54,行常规护理)和试验组(n=53,行协同护理+基于PG-SGA个体化营养干预)。对比两组患者的心理状态及生活质量。结果:试验组干预后汉密尔顿焦虑量表(Hamilton anxiety scale,HAMA)、汉密尔顿抑郁量表(Hamilton depression scale,HAMD)评分均较参照组低,健康调查简表(the MOS item short form health survey,SF-36)各维度评分均较参照组高,差异均有统计学意义(P<0.05)。结论:协同护理联合基于PG-SGA个体化营养干预应用于腹腔镜胃癌D2根治术患者,能够有效改善患者的心理状态,提升其生活质量。
基金Supported by Natural Science Foundation of Yunnan Province(2011FB049)National Natural Science Foundation of China(31260481,31460516)+1 种基金Fund of Yunnan Education Department(2013Y251)Development Fund of the Department of Life Science and Technology,Kunming University(GXKM201505)
文摘In this study, 1 500 bp PG gene promoter was amplified from leaves of tomato cultivar ＇ Zhongshu No. 4＇. The bioinformatic analysis of PG promoter sequences was conducted. PG promoter elements were predicted and analyzed by PLANTCARE and PLACE. The results showed that tomato PG promoter contained multiple c/s-acting regulatory elements such as typical basic elements TATA-Box and CAAT-Box, light responsive elements 3-AF1 binding site, ATl-motif, ATCT- motif, Box 4, Box I, GA-motif, GTl-motif, Spl and MRE, heat stress-responsive element HSE, ethylene-responsive element ERE, meristem-specifie regulatory element CCGTCC-box, endosperm expression-related regulatory elements GCN4_motif and Skn-l_motif, defense and stress responsiveness element TC-rich repeats, and circadian control-related element circadian, indicating that the expression of tomato PG gene is related to light, temperature, hormone, stress and other factors. This study laid the foundation for subsequent research about regulation of PG gene expression in plants.
