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Effects of Honeysuckle Chlorogenic Acid on Secretory Enzymes,Lipoxygenase A4,and Biochemical Indicators inModel Mice with Aluminum Induced Alzheimer's Disease
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作者 Jialing HUANG Cuijuan HUANG +8 位作者 Xiaomian TANG Weiming ZHOU Hanchao FENG Cuiping WEI Oufei HUANG Haipeng ZHANG Song NONG Shuqiu ZHANG Fuyu PAN 《Medicinal Plant》 2024年第5期35-38,43,共5页
[Objectives]To explore the effects of chlorogenic acid from honeysuckle on the secretion enzymes,lipoxygenase A4(LXA4),and blood biochemical indicators in mice with aluminum induced Alzheimer's disease(AD).[Method... [Objectives]To explore the effects of chlorogenic acid from honeysuckle on the secretion enzymes,lipoxygenase A4(LXA4),and blood biochemical indicators in mice with aluminum induced Alzheimer's disease(AD).[Methods]Chlorogenic acid was extracted from hon-eysuckle by ultrasound assisted alcohol extraction method.Seventy mice were randomly divided into normal group,model group,and low,me-dium and high dose groups of honeysuckle chlorogenic acid.All the mice in each group except for the normal group were given maltol aluminum by intraperitoneal injection to establish models of aluminum induced AD,continuously injected for 5 d and stopped for 2 d,totally poisoned for 8 weeks.Starting from the 5^(th) week of poisoning,the low,medium and high dose groups of honeysuckle chlorogenic acid were given honeysuck-le chlorogenc acid solution 40,80 and 160 mg/kg by gavage,respectively,while the normal group and the model group were fed with an equal volume of distilled water,all once daily,continuously gavaged until the end of the 8^(th) week.At the end of the experiment,the learning memory ability of the mice was tested by Y-type waler maze,and the number of tests required to reach the learning standard,the number of memory er-rors in 20 tests and the error rate of the mice were recorded.The brains of mice were taken to determine the contents of β-secretase,α-secre-tase,γ-secretase,LXA4 and acetylcholinesterase(AchE)in the homogenates of brain tissues by ELISA,and their blood was taken to deter-mine the biochemical indexes.[Results]Compared with the normal group,the number of learning tests,number of memory errors,error rate and the contents of β-secretase,γ-secretase and AchE in brain tissue of the mice in the model group were all significantly increased(all P<0.05),the contents of LXA4 in brain tissue were significantly decreased(all P<0.05),and the contents of α-secretase did not change significantly(all P>0.05);compared with the model group,the number of learning tests,the number of memory errors,the error rate and the content of β-secretase,γ-secretase and AchE in brain tissue were all significantly reduced(all P<0.05),the content of LXA4 in brain tissue of the high dose group of honeysuckle chlorogenic acid was significantly increased(P<0.05),and there was no significant change in the content of α-secretase in brain tissue of all groups of honeysuckle chlorogenic acid(all P>0.05).Compared with the normal group,the levels of blood glucose,TC,TG,ALT,BUN,Cr and UA in the model group and the levels of TC,TG and BUN in the low-and medium-dose groups of honeysuckle chlorogenic acid were significantly increased(all P<0.05),and the level of HDL-C in the model group and the levels of UA in the medium-and high-dose groups of honeysuckle chlorogenic acid were significantly decreased(all P<0.05);compared with the model group,the levels of blood glucose,ALT,BUN,UA in each group of honeysuckle chlorogenic acid,the levels of TC and Cr in medium and high dose groups of honeysuckle chlorogenic acid,and the level of TG in the high dose group of honeysuckle chlorogenic acid were all signifi-cantly lower(all P<0.05),while the level of HDL-C in the medium and high dose groups of honeysuckle chlorogenic acid and the level of to-tal protein in the high dose group of honeysuckle chlorogenic acid were all significantly higher(all P<0.05).[Conclusions]Chlorogenic acid from honeysuckle may improve AD induced by aluminum exposure via regulating related secretory enzymes,LXA4,and various biochemi-cal indicators. 展开更多
关键词 Alzheimer's disease Chlorogenic acid SECRETASE lipoxygenase A4
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萝卜硫素通过调节ALOX5/NF-κB信号通路调控巨噬细胞糖酵解抑制糖尿病肾病进展 被引量:1
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作者 乌日娜 丁海东 +2 位作者 常宏 孙娜娜 张磊 《安徽医科大学学报》 CAS 北大核心 2024年第3期390-397,共8页
目的探讨萝卜硫素(SFN)调节花生四烯酸5-脂氧合酶基因(arachidonic acid 5-lipoxygenase,ALOX5)/核因子kappa B(NF-κB)信号通路调节巨噬细胞糖酵解对糖尿病肾病(DN)进展的影响。方法生物信息学分析SFN治疗DN的靶基因。使用30 mmol/L高... 目的探讨萝卜硫素(SFN)调节花生四烯酸5-脂氧合酶基因(arachidonic acid 5-lipoxygenase,ALOX5)/核因子kappa B(NF-κB)信号通路调节巨噬细胞糖酵解对糖尿病肾病(DN)进展的影响。方法生物信息学分析SFN治疗DN的靶基因。使用30 mmol/L高葡萄糖(HG)处理人近端肾小管上皮细胞系(HK-2细胞)诱导体外DN模型。将HK-2细胞分为如下组:正常糖(NG)组、HG组、HG+SFN(3 mmol/L)组、HG+ALOX5组、HG+SFN(3 mmol/L)+ALOX5组、HG处理的巨噬细胞+HK-2细胞组、HG+SFN(3 mmol/L)处理的巨噬细胞+HK-2细胞组、HG+ALOX5转染处理的巨噬细胞+HK-2细胞组、HG+SFN(3 mmol/L)+ALOX5转染处理的巨噬细胞+HK-2细胞组。CCK-8检测细胞活力,原位末端脱氧核苷酸转移酶标记(TUNEL)法检测细胞凋亡;葡萄糖和乳酸试剂盒检测各组细胞中葡萄糖和乳酸水平;Western blot检测各组细胞中ALOX5、NF-κB以及糖酵解相关蛋白己糖激酶-2(HK2)、丙酮酸激酶M2(PKM2)、葡萄糖转运蛋白1(GLUT1)的表达;使用链脲佐菌素(STZ)构建DN小鼠模型,DN小鼠给与SFN(0.5 mg/kg)治疗;检测小鼠各项生化指标,HE染色检测肾组织病理变化;Western blot检测小鼠肾脏巨噬细胞中糖酵解相关蛋白己糖激酶-2(HK2)、丙酮酸激酶M2(PKM2)、葡萄糖转运蛋白1(GLUT1)的表达。结果生物信息学分析结果显示ALOX5是SFN治疗DN的靶基因。与HG组相比,SFN处理增强HK-2细胞活力并抑制细胞凋亡(P<0.05);同时,SFN处理抑制HG诱导的巨噬细胞糖酵解相关蛋白的表达,减弱巨噬细胞介导的HK-2细胞损伤(P<0.05);Western blot结果表明SFN抑制ALOX5和NF-κB的表达(P<0.05);小鼠实验结果显示,SFN治疗改善DN小鼠肾功能和肾组织病理学改变,抑制肾组织中巨噬细胞糖酵解相关蛋白的表达(P<0.05)。结论SFN通过抑制ALOX5/NF-κB信号通路抑制巨噬细胞糖酵解从而改善DN进展。 展开更多
关键词 萝卜硫素 糖尿病肾病 巨噬细胞 糖酵解 花生四烯酸5-脂氧合酶 NF-ΚB信号通路
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基于5-LOX/LTB4信号通路探讨黄芩苷/栀子苷对PM_(2.5)暴露致血管内皮功能障碍的干预作用及其机制 被引量:1
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作者 孙欠欠 张行行 +5 位作者 赵麓 赵安东 史永恒 王川 刘继平 王斌 《中南药学》 CAS 2024年第2期307-314,共8页
目的基于5-LOX/LTB4信号通路探究黄芩苷/栀子苷(BC/GD)对PM_(2.5)诱导的血管内皮功能障碍的改善作用及其机制。方法利用离体肌张力描记技术测定BC/GD对不同状态下血管环张力的影响。将32只大鼠随机分为对照组、PM_(2.5)组、30 mg·k... 目的基于5-LOX/LTB4信号通路探究黄芩苷/栀子苷(BC/GD)对PM_(2.5)诱导的血管内皮功能障碍的改善作用及其机制。方法利用离体肌张力描记技术测定BC/GD对不同状态下血管环张力的影响。将32只大鼠随机分为对照组、PM_(2.5)组、30 mg·kg^(-1)BC/GD组和60 mg·kg^(-1)BC/GD组,利用气管滴注法构建PM_(2.5)暴露模型,持续2个月,造模1个月后灌胃给予对应药物,持续1个月,末次给药后,HE染色观察肠系膜动脉血管内皮状态;化学发光法检测肠系膜动脉活性氧(ROS)水平;硝酸还原酶法检测一氧化氮(NO)水平;ELISA法检测血清炎症因子肿瘤坏死因子α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、转化生长因子β1(TGF-β1)、白三烯B4(LTB4)水平;Western blot法检测5-脂氧酶(5-LOX)、内皮细胞一氧化氮合酶(eNOS)、诱导型一氧化氮合酶(iNOS)蛋白表达。结果BC/GD对基础状态的肠系膜动脉血管环张力无明显影响,但能明显舒张由5-HT预收缩的血管环;一氧化氮合酶抑制剂L-NAME、环氧合酶抑制剂INDO或L-NAME+INDO均能明显抑制BC/GD的血管舒张作用,但L-NAME的抑制作用更明显;50、100 mg·mL^(-1)PM_(2.5)能降低血管环对乙酰胆碱(ACh)的舒张血管效应,而BC/GD能明显改善PM_(2.5)诱导的舒张效应减弱。与对照组相比,PM_(2.5)组大鼠肠系膜动脉内皮完整性严重受损、内皮皱缩,大鼠血清中TNF-α、IL-1β、IL-6、LTB4水平显著升高、TGF-β1水平显著降低,肠系膜动脉组织中5-LOX、iNOS蛋白表达明显增加,eNOS蛋白表达降低,ROS水平升高,NO水平降低。与PM_(2.5)组相比,BC/GD能改善内皮损伤程度和完整性,可显著降低血清中TNF-α、IL-1β、IL-6、LTB4水平,升高TGF-β1水平;明显降低肠系膜动脉5-LOX、iNOS表达,升高eNOS蛋白表达;降低ROS水平,升高NO水平。结论BC/GD可通过抑制5-LOX/LTB4信号通路表达,从而降低ROS水平,抑制炎性损伤,上调eNOS表达,下调iNOS表达,升高血管中NO水平,改善PM_(2.5)诱导的血管内皮功能障碍。 展开更多
关键词 PM_(2.5) 黄芩苷 栀子苷 内皮功能障碍 5-lox/LTB4信号通路
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Prokaryotic Expression, Purification and Characterization of a Novel Rice Seed Lipoxygenase Gene OsLOX1 被引量:8
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作者 WANG Ren SHEN Wen-biao +3 位作者 LIU Ling-long JIANG Ling ZHAI Hu-qu WAN Jian-min 《Rice science》 SCIE 2008年第2期88-94,共7页
Lipoxygenase (LOX, EC1.13.11.12) is a key enzyme during the degradation of lipids in animals and even plants, and also the first key enzyme responsible for the biosynthesis of jasmonate. To purify and characterize t... Lipoxygenase (LOX, EC1.13.11.12) is a key enzyme during the degradation of lipids in animals and even plants, and also the first key enzyme responsible for the biosynthesis of jasmonate. To purify and characterize the OsLOX1 gene from rice seeds, the entire coding region of the OsLOX1 gene was inserted into an expression vector pET30a(+) and transformed into Escherichia coil BL21 (DE3). Expression of the fusion protein was successfully induced by isopropyl-β-D- thiogalactopyranoside (IPTG) and the purified recombinant protein was obtained by His.Bind Kits. Further assay showed that the purified recombinant protein exhibited the LOX activity. The optimum pH was 4.8 (acetate buffer) and the optimum temperature was 30℃ for the above enzyme. Thus, the recombinant might confer an available usage for the synthesis of jasmonate in vitro, and also provides a possibility for elucidating the inter-relationship between the primary structure of the plant seed lipoxygenase protein and its physiological functions. 展开更多
关键词 rice seed lipoxygenase gene prokaryotic expression PURIFICATION CHARACTERIZATION
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外源茉莉酸对脂氧合酶基因LOX3敲除的粳稻防御响应的影响
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作者 杨婧 苏顺雨 +5 位作者 赵添琦 李永杰 唐萍 左如斌 刘崇兰 杨静 《南方农业学报》 CAS CSCD 北大核心 2024年第2期397-410,共14页
【目的】揭示外源茉莉酸(Jasmonic acid,JA)对脂氧合酶基因LOX3敲除水稻株防御响应的影响,为利用外源JA等激发子提高农作物因抗性相关基因丧失导致的水稻防御响应降低问题提供基础数据。【方法】以感病粳稻丽江新团黑谷(LTH)及其LOX3敲... 【目的】揭示外源茉莉酸(Jasmonic acid,JA)对脂氧合酶基因LOX3敲除水稻株防御响应的影响,为利用外源JA等激发子提高农作物因抗性相关基因丧失导致的水稻防御响应降低问题提供基础数据。【方法】以感病粳稻丽江新团黑谷(LTH)及其LOX3敲除水稻株系(Δlox3#1、Δlox3#10和Δlox3#20)为材料,开展水稻株高、分蘖数和百粒重等表型分析、过氧化物酶(POD)、活性氧(ROS)和木质素等抗菌化合物活性(含量)测定,以及防御相关基因表达分析;进一步对供试水稻进行稻瘟菌接种,调查供试水稻稻瘟病症状,检测稻瘟菌接种水稻中抗菌化合物活性(含量)及防御相关基因表达等,并观察稻瘟菌在水稻中的侵染进程;在此基础上,使用外源JA处理稻瘟菌接种36和48 hpi时的Δlox3敲除水稻株和LTH,分析JA对水稻稻瘟病症状及稻瘟菌侵染的水稻中抗菌化合物活性(含量)和防御相关基因表达等的影响,同时观察JA对稻瘟菌侵染进程的影响。【结果】Δlox3敲除水稻株种子发芽速度快于野生型,但株高、分蘖数和百粒重等与野生型相近;稻瘟菌接种的Δlox3敲除水稻株稻瘟病症状较稻瘟菌接种的野生型水稻严重,且胼胝质数、POD活性及木质素含量降低,ROS含量和死细胞数增加,OsWRKY45、OsPR1a和OsPOX1基因下调幅度和稻瘟菌菌丝定殖量明显高于稻瘟菌接种野生型水稻;外源JA可有效减缓稻瘟菌接种36和48 hpi的Δlox3敲除水稻株稻瘟病症状,且JA提高了稻瘟菌接种36和48 hpi的Δlox3敲除水稻株中胼胝质数、POD活性和木质素含量以及防御相关基因OsWRKY45、OsPR1a和OsPOX1表达水平,而降低了OsRbohB表达量及ROS含量和细胞死亡数,同时限制了稻瘟菌菌丝扩展;其中JA对稻瘟菌接种48 hpi的Δlox3敲除水稻株稻瘟病症状减轻效果优于JA对稻瘟菌接种36 hpi的Δlox3敲除水稻株,主要体现在JA使稻瘟菌接种48 hpi的Δlox3敲除水稻株中抗菌化合物增加幅度、防御相关基因表达水平、细胞死亡数和稻瘟菌菌丝扩展受限等优于JA处理稻瘟菌接种36 hpi的Δlox3敲除水稻株。【结论】LOX3基因敲除提升了水稻种子发芽速度而降低了水稻抗瘟性,外源JA则减缓LOX3敲除导致的水稻防御响应降低,提高了水稻抗瘟性。JA在减缓农作物因抗性基因持久性短而造成的抗性降低或丧失方面具有潜在的应用前景。 展开更多
关键词 水稻 稻瘟菌 脂氧合酶基因 茉莉酸 防御响应
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结肠癌中ALOX12表达与临床病理特征及预后的关系分析
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作者 付志强 高启行 +1 位作者 郭文文 何震宇 《南京医科大学学报(自然科学版)》 CAS 北大核心 2024年第5期643-648,共6页
目的:探讨花生四烯酸12-脂氧合酶(arachidonate 12-lipoxygenase,ALOX12)在结肠癌患者癌组织中的表达及意义。方法:回顾性收集257例结肠癌患者肿瘤及瘤旁组织,免疫组化法检测ALOX12的表达,比较肿瘤及瘤旁组织内表达差异并分析其表达与... 目的:探讨花生四烯酸12-脂氧合酶(arachidonate 12-lipoxygenase,ALOX12)在结肠癌患者癌组织中的表达及意义。方法:回顾性收集257例结肠癌患者肿瘤及瘤旁组织,免疫组化法检测ALOX12的表达,比较肿瘤及瘤旁组织内表达差异并分析其表达与患者临床病理特征的相关性。随访并探讨ALOX12表达与患者5年总生存率的关系,并采用Kaplan-Meier法绘制生存曲线。运用单因素及多因素Cox回归综合分析影响结肠癌患者5年总生存率的因素。结果:与瘤旁组织相比,ALOX12在肿瘤中阳性表达率更高(63.8%vs.33.1%),差异具有统计学意义(P<0.001)。ALOX12表达与患者年龄、性别、肿瘤部位均无明显相关性(P>0.05),而与TNM分期、淋巴结转移有显著相关性(P<0.05)。ALOX12高表达患者的5年总生存率明显低于低表达患者(P<0.001)。此外,单因素和多因素Cox回归分析结果显示年龄、TNM分期、淋巴结转移、ALOX12表达是影响患者5年总生存率的独立影响因素(P<0.05)。结论:ALOX12高表达与结肠癌TNM分期、淋巴结转移相关,可作为预测患者病情及预后的潜在指标。 展开更多
关键词 结肠癌 花生四烯酸12-脂氧合酶 铁死亡 临床病理特征 预后
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Variants of the arachidonate 5-lipoxygenase-activating protein (ALOX5AP) gene and risk of ischemic stroke in Han Chinese of eastern China 被引量:12
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作者 Gannan Wang Yao Wang +4 位作者 Hao Sun Weijuan Cao Jing Zhang Hang Xiao Jinsong Zhang 《The Journal of Biomedical Research》 CAS 2011年第5期319-327,共9页
Variants of the arachidonate 5-1ipoxygenase-activating protein (ALOX5AP) gene have been suggested to play an important role in the pathogenesis of atherosclerosis and ischemic stroke. This study was aimed to explore... Variants of the arachidonate 5-1ipoxygenase-activating protein (ALOX5AP) gene have been suggested to play an important role in the pathogenesis of atherosclerosis and ischemic stroke. This study was aimed to explore the association of ALOX5AP variants with ischemic stroke risk in Han Chinese of eastern China. A total of 690 ischemic stroke cases and 767 controls were recruited. The subjects were further subtyped according to the Trial of Org 10172 in Acute Stroke Treatment (TOAST) criteria. On the basis of that, two polymorphisms of the ALOX5AP gene (rs10507391 and rs12429692) were determined by TaqMan genotyping assay. In addition, plasma leukotriene B4 (LTB4) levels were analyzed in these subjects. There was no evidence of association between the two variants of ALOX5AP and the risk of ischemic stroke or its TOAST-subtypes. Haplotype analysis and stratification analysis according to sex, age, body mass index, hypertension, and diabetes also showed negative association. Analysis of LTB4 levels in a subset of cases and controls revealed that LTB4 levels were significantly higher in ischemic stroke cases than in the controls (70.06± 14.75 ng/L vs 57.34±10.93 ng/L; P = 0.000) and carriers of the T allele of the rs10507391 variant were associated with higher plasma LTB4 levels (P = 0.000). The present study suggests there is no association of the two polymorphisms in the ALOX5AP gene with ischemic stroke risk in Han Chinese of eastern China. 展开更多
关键词 arachidonate 5-lipoxygenase-activating protein ischemic stroke VARIANTS leukotriene B4 risk factors
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JA-mediated MYC2/LOX/AOS feedback loop regulates osmotic stress response in tea plant
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作者 Junyan Zhu Hongrong Chen +5 位作者 Lu Liu Xiaobo Xia Xiaomei Yan Xiaozeng Mi Shengrui Liu Chaoling Wei 《Horticultural Plant Journal》 SCIE CAS CSCD 2024年第3期931-946,共16页
Osmotic stress caused by low-temperature,drought and salinity was a prevalent abiotic stress in plant that severely inhibited plant development and agricultural yield,particularly in tea plant.Jasmonic acid(JA)is an i... Osmotic stress caused by low-temperature,drought and salinity was a prevalent abiotic stress in plant that severely inhibited plant development and agricultural yield,particularly in tea plant.Jasmonic acid(JA)is an important phytohormone involving in plant stress.However,underlying molecular mechanisms of JA modulated osmotic stress response remains unclear.In this study,high concentration of mannitol induced JA accumulation and increase of peroxidase activity in tea plant.Integrated transcriptome mined a JA signaling master,MYC2 transcription factor is shown as a hub regulator that induced by mannitol,expression of which positively correlated with JA biosynthetic genes(LOX and AOS)and peroxidase genes(PER).CsMYC2 was determined as a nuclei-localized transcription activator,furthermore,ProteinDNA interaction analysis indicated that CsMYC2 was positive regulator that activated the transcription of CsLOX7,CsAOS2,CsPER1 and CsPER3via bound with their promoters,respectively.Suppression of CsMYC2 expression resulted in a reduced JA content and peroxidase activity and osmotic stress tolerance of tea plant.Overexpression of CsMYC2 in Arabidopsis improved JA content,peroxidase activity and plants tolerance against mannitol stress.Together,we proposed a positive feedback loop mediated by CsMYC2,CsLOX7 and CsAOS2 which constituted to increase the tolerance of osmotic stress through fine-tuning the accumulation of JA levels and increase of POD activity in tea plant. 展开更多
关键词 Camellia sinensis Jasmonic acid(JA) MYC2 transcription factor lipoxygenase(lox) Osmotic stress Peroxidase(POD) Reactive oxygen species(ROS)
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CgLOX4基因在长牡蛎幼虫发育中的表达特征
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作者 胡悦 冷金源 +2 位作者 孙洁洁 王玲玲 宋林生 《大连海洋大学学报》 CAS CSCD 北大核心 2024年第1期38-47,共10页
为研究长牡蛎(Crassostrea gigas)LOX4基因(CgLOX4)在幼虫发育中的表达特征,采用PCR技术扩增CgLOX4的全长cDNA序列,利用荧光定量PCR和整体免疫荧光技术检测CgLOX4在幼虫不同发育时期的表达特征。结果表明:CgLOX4的cDNA全长为1 862 bp,... 为研究长牡蛎(Crassostrea gigas)LOX4基因(CgLOX4)在幼虫发育中的表达特征,采用PCR技术扩增CgLOX4的全长cDNA序列,利用荧光定量PCR和整体免疫荧光技术检测CgLOX4在幼虫不同发育时期的表达特征。结果表明:CgLOX4的cDNA全长为1 862 bp,开放阅读框为834 bp,可编码277个氨基酸,等电点为8.36,预测其氨基酸序列只含有一个保守的HOX结构域;CgLOX4在长牡蛎成体各组织中均有表达,其中在闭壳肌中的表达量显著高于肝胰腺、鳃、血淋巴、性腺和唇瓣组织(P<0.05);CgLOX4在所检测的早期胚胎中均有表达,其中在受精卵和4细胞期的表达量相对较高;CgLOX4阳性信号主要分布在担轮幼虫壳形成区、D型幼虫内脏团和壳顶幼虫消化腺中。研究表明,CgLOX4是长牡蛎早期发育过程中较早激活的转录因子之一,可能在调控幼虫壳形成和消化器官形成等方面发挥重要作用。 展开更多
关键词 长牡蛎 lox4 幼虫发育 分布特征
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基于LOX-1/NOX2信号通路探讨槲皮素抑制中性粒细胞胞外诱捕网形成机制
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作者 刘思 罗纲 +3 位作者 王君畅 应琳瑶 王严 肖琳 《中国药理学通报》 CAS CSCD 北大核心 2024年第11期2047-2054,共8页
目的探讨溶血磷脂酰胆碱(lysophosphatidylcholine,LPC)诱导中性粒细胞胞外诱捕网(neutrophil extracellular traps,NETs)形成的分子机制及槲皮素的干预作用。方法使用LPC诱导大鼠原代中性粒细胞,构建NETs形成的体外模型并进行槲皮素干... 目的探讨溶血磷脂酰胆碱(lysophosphatidylcholine,LPC)诱导中性粒细胞胞外诱捕网(neutrophil extracellular traps,NETs)形成的分子机制及槲皮素的干预作用。方法使用LPC诱导大鼠原代中性粒细胞,构建NETs形成的体外模型并进行槲皮素干预,检测NETs形成情况及相关通路蛋白表达水平。结果100 mg·L^(-1) LPC可在体外诱导NETs形成,其与LOX-1/NOX2通路相关蛋白表达上调有关,槲皮素干预可有效降低LOX-1和NOX2蛋白表达水平并减少NETs形成。结论槲皮素通过调控LOX-1/NOX2信号通路抑制NETs形成,发挥对相关炎症性疾病的保护作用。 展开更多
关键词 炎症 溶血磷脂酰胆碱 中性粒细胞胞外诱捕网 槲皮素 lox-1 NOX2
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血必净通过调节miR-145-5p/LOX信号通路对H_(2)O_(2)诱导的心肌细胞氧化应激及凋亡的影响
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作者 李英 冯凯 +4 位作者 王文霞 严丽娟 张国旗 刘志艳 周来来 《陕西中医药大学学报》 2024年第5期97-102,共6页
目的探讨血必净对H_(2)O_(2)诱导的心肌细胞氧化损伤及凋亡的影响及其分子机制。方法培养H9C2细胞后将其随机分为对照组(Control)、模型组(Model)和血必净组(XBJ)3组,除Control组外,其余组的H9C2细胞用H_(2)O_(2)诱导构建氧化应激损伤... 目的探讨血必净对H_(2)O_(2)诱导的心肌细胞氧化损伤及凋亡的影响及其分子机制。方法培养H9C2细胞后将其随机分为对照组(Control)、模型组(Model)和血必净组(XBJ)3组,除Control组外,其余组的H9C2细胞用H_(2)O_(2)诱导构建氧化应激损伤细胞模型,然后XBJ组用12.5 g·L^(-1)、25 g·L^(-1)和50 g·L^(-1)浓度的XBJ处理24 h。CCK-8法检测不同浓度的XBJ对细胞活力的影响;生化试剂盒检测肌酸激酶同工酶(CK-MB)、丙二醛(MDA)、超氧化物歧化酶(SOD)及还原型谷胱甘肽(GSH)的活性;流式检测H9C2细胞凋亡率;qRT-PCR检测miR-145-5p、LOX mRNA的表达;Western blot检测赖氨酰氧化酶蛋白(lysyloxidase,LOX)的表达;双荧光素酶实验检测miR-145-5p与LOX的靶向结合关系。结果不同浓度XBJ处理后,细胞活性呈梯度性显著增高;与对照组相比,模型组细胞CK-MB、MDA的活性显著增加(P<0.01),SOD、GSH活性显著降低,细胞凋亡率显著提高,miR-145-5p表达水平显著降低,LOX水平显著增加(P<0.01);经血必净处理后上述指标的水平得到有效改善(P<0.05)。结论XBJ能通过提高抗氧化应激能力,抑制细胞凋亡并调控miR-145-5p/LOX信号通路保护H_(2)O_(2)诱导的心肌细胞损伤。 展开更多
关键词 血必净 心肌细胞 氧化应激 细胞凋亡 miR-145-5p/lox通路
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Activities of lipoxygenase and phenylalanine ammonia lyase in poplar leaves induced by insect herbivory and volatiles 被引量:3
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作者 胡增辉 张雯 +2 位作者 沈应柏 付怀军 苏晓华 《Journal of Forestry Research》 SCIE CAS CSCD 2009年第4期372-376,I0007,I0008,共7页
A study was conducted to explore the defense response in woody plants after insect herbivory. The activities of two enzymes, lipoxygenase (LOX), a key enzyme ofjasmonate (JA) pathway, and phenylalanine ammonia lya... A study was conducted to explore the defense response in woody plants after insect herbivory. The activities of two enzymes, lipoxygenase (LOX), a key enzyme ofjasmonate (JA) pathway, and phenylalanine ammonia lyase (PAL), a rate-limiting enzyme of phenyl- propanoid pathway, were measured in the leaves of one-year-old poplar (Populus simonii × P. pyramidalis 'Opera 8277') cuttings after Clostera anachoreta larvae attack. The results show that the increased activities of LOX and PAL were found not only in the leaves wounded by C. anachoreta larvae but also in their tipper systemic leaves, indicating that JA and phenylpropanoid pathways were activated, and the defense response was stimulated systemically. The increase in LOX and PAL activities in neighboring intact poplar cuttings sug- gested that there exists the interplant communication between poplar plants mediated by the herbivore-induced volatiles. Methyl jasmonate (MeJA) was also proved to be an airborne signal to induce defense response in P simonii × P pyramidalis 'Opera 8277' cuttings. 展开更多
关键词 Clostera anachoreta larvae lipoxygenase methyl jasmonate phenylalanine ammonia lyase Populus simonii × p pyramidalis 'Opera 8277'cuttings
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钩吻脂氧合酶基因GeLOX1的鉴定及低温胁迫表达分析 被引量:1
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作者 尤垂淮 谢津津 +7 位作者 张婷 崔天真 孙欣路 臧守建 武奕凝 孙梦瑶 阙友雄 苏亚春 《生物技术通报》 CAS CSCD 北大核心 2023年第11期318-327,共10页
近年来,钩吻(Gelsemium elegans)的药用和饲用价值日益凸显,但钩吻在生长过程中不耐低温,挖掘其低温响应基因,为钩吻的抗寒育种研究奠定基础。植物中,脂氧合酶(lipoxygenase, LOX)在种子老化、抗逆境胁迫等方面的生理生化过程中有重要... 近年来,钩吻(Gelsemium elegans)的药用和饲用价值日益凸显,但钩吻在生长过程中不耐低温,挖掘其低温响应基因,为钩吻的抗寒育种研究奠定基础。植物中,脂氧合酶(lipoxygenase, LOX)在种子老化、抗逆境胁迫等方面的生理生化过程中有重要影响。基于课题组构建的钩吻转录组数据库,挖掘响应低温胁迫的钩吻LOX基因,运用RT-PCR技术,从中克隆到一条GeLOX1的cNDA全长序列,对其进行生物信息学、亚细胞定位、基因表达、原核表达及平板胁迫等分析。结果显示,GeLOX1所编码蛋白的氨基酸长度为761 aa,蛋白相对分子质量为87.00 kD,预测为不稳定的亲水性蛋白,含有28个丝氨酸磷酸化位点,22个苏氨酸磷酸化位点和9个酪氨酸磷酸化位点。进化树分析结果表明,GeLOX1属于9-LOX家族的成员。亚细胞定位检测结果显示,GeLOX1蛋白定位于细胞质中。实时荧光定量PCR分析发现,GeLOX1在钩吻的根中高表达,且其在4℃低温胁迫下的表达量呈现下调的趋势。经原核表达诱导后,GeLOX1的重组蛋白在约111 kD处出现目标条带,且重组蛋白的积累量在诱导8 h时达到峰值。此外,平板胁迫试验表明,GeLOX1的原核表达菌株相较于对照组对低温胁迫更敏感。钩吻GeLOX1能够应答低温胁迫。 展开更多
关键词 钩吻 脂氧合酶(lox) 序列分析 低温胁迫 表达分析
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Lipoxygenase Activity and Its Relationship with Traits of Whole-mill Color in Wheat 被引量:1
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作者 包晓婷 颛孙湘溪 +3 位作者 郑文寅 张文明 郭文善 姚大年 《Agricultural Science & Technology》 CAS 2016年第10期2242-2246,2255,共6页
130 wheat varieties are selected as materials to test the LOX activity, whiteness, yellowness, carotenoids content and RVA parameters, which are to be analyzed by variance analysis, correlation analysis, cluster analy... 130 wheat varieties are selected as materials to test the LOX activity, whiteness, yellowness, carotenoids content and RVA parameters, which are to be analyzed by variance analysis, correlation analysis, cluster analysis and classified according to the origins. The result shows there are highly significant variations in LOX activity from varieties; LOX activity is positively correlated with whiteness, and significantly and negatively correlated with yellowness. Cluster analysis applying longest distance method based on LOX activity clusters all the varieties or lines into three major groups. There are great differences between the averages of LOX activity from varieties. LOX activities are discussed in the application of nutritional quality improvement in wheat. 展开更多
关键词 WHEAT lox WHITENESS Yellowness Carotenoids Correlation analysis Cluster analysis
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12/15-Lipoxygenase inhibition counteracts MAPK phosphorylation in mouse and cell culture models of diabetic peripheral neuropathy 被引量:11
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作者 Roman Stavniichuk Alexander A. Obrosov +3 位作者 Viktor R. Drel Jerry L. Nadler Irina G. Obrosova Mark A. Yorek 《Journal of Diabetes Mellitus》 2013年第3期101-110,共10页
Background: Increased mitogen-activated protein kinase (MAPK) phosphorylation has been detected in peripheral nerve of human subjects and animal models with diabetes as well as high-glucose exposed human Schwann cells... Background: Increased mitogen-activated protein kinase (MAPK) phosphorylation has been detected in peripheral nerve of human subjects and animal models with diabetes as well as high-glucose exposed human Schwann cells, and have been implicated in diabetic peripheral neuropathy. In our recent studies, leukocytetype 12/15-lipoxygenase inhibition or gene deficiency alleviated large and small nerve fiber dysfunction, but not intraepidermal nerve fiber loss in streptozotocin-diabetic mice. Methods: To address a mechanism we evaluated the potential for pharmacological 12/15-lipoxygenase inhibition to counteract excessive MAPK phosphorylation in mouse and cell culture models of diabetic neuropathy. C57Bl6/J mice were made diabetic with streptozotocin and maintained with or without the 12/15-lipoxygenase inhibitor cinnamyl-3,4-dihydroxy-α-cyanocinnamate (CDC). Human Schwann cells were cultured in5.5 mMor30 mMglucose with or without CDC. Results: 12(S) HETE concentrations (ELISA), as well as 12/15-lipoxygenase expression and p38 MAPK, ERK, and SAPK/JNK phosphorylation (all by Western blot analysis) were increased in the peripheral nerve and spinal cord of diabetic mice as well as in high glucose-exposed human Schwann cells. CDC counteracted diabetes-induced increase in 12(S)HETE concentrations (a measure of 12/15-lipoxygenase activity), but not 12/15-lipoxygenase overexpression, in sciatic nerve and spinal cord. The inhibitor blunted excessive p38 MAPK and ERK, but not SAPK/ JNK, phosphorylation in sciatic nerve and high glucose exposed human Schwann cells, but did not affect MAPK, ERK, and SAPK/JNK phosphorylation in spinal cord. Conclusion: 12/15-lipoxygenase inhibition counteracts diabetes related MAPK phosphorylation in mouse and cell culture models of diabetic neuropathy and implies that 12/15-lipoxygenase inhibitors may be an effective treatment for diabetic peripheral neuropathy. 展开更多
关键词 Diabetes lipoxygenase NEUROPATHY Schwann Cells MITOGEN-ACTIVATED Protein KINASE
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Effects of Lipoxygenase Null Genes of Soybean in Controlling Beany-flavor of Soymilk and Soyflour 被引量:5
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作者 MA Hao, GUAN Chun-yun,HE Xiao-ling, ZHANG Guo-zheng and DIN An-lin(College of Plant Sciences and Technology , Hunan Agricultural University, Changsha 410128 ,P. R. China Soybean ResearchInstitute , Nanjing Agricultural University, Nanjing 210095 ,P.R. China Institute of Crop Breeding and Cultivation ,Chinese Academy of Agricultural Sciences , Beijing 100081 ,P.R. China ) 《Agricultural Sciences in China》 CAS CSCD 2002年第9期965-971,共7页
The flavor of the soymilk and soyflour obtained from the lipoxygenase mutant isolines was concentrated by simultaneous distillation and extraction (SDE), and its constituents were identified by gas chro-matography (GC... The flavor of the soymilk and soyflour obtained from the lipoxygenase mutant isolines was concentrated by simultaneous distillation and extraction (SDE), and its constituents were identified by gas chro-matography (GC) and gas chromatography-mass spectrometry. Results showed that the same 24 flavor constituents were isolated in both soymilk and soyflour, and most of them were aldehydes and alcohols. Lox2 was most responsible for the production of the volatile and beany-flavor components, and Lox1 less responsible. Lox3 was least responsible and can reduce the yield of hexanal. Either Lx1 or Lx2 could significantly reduce the volatile and beany-flavor, and Lx3 could significantly increase the yield of hexanal. Primary and secondary interactions existed among the null mutant genes, and the major effects and interactions could be affected by processing conditions. The isoline with triple lipoxygenase null genes yielded the least volatile and beany-flavor components, and the isoline without the lipoxygenase gene Lx3 produced the greatest amount of the volatile and beany-flavor components. The amounts of volatile and beany-flavor components produced by the other isolines were between that of the isoline with triple lipoxygenase null genes and the isoline without lipoxygenase gene Lx3. According to the correlation analysis, the hexanal amount could be used as an index in evaluating the importance of lipoxygenase isozymes in the yield of beany-flavor compounds, and the effects of the different types of lipoxygenase null mutants in controlling beany-flavor compounds. The cultivars with triple lipoxygenase null genes will be a quality raw material for soy food processing. 展开更多
关键词 Glycine max (L.) Merrill lipoxygenase null gene Gas chromatography and gas chromato-graphy-mass chromatography Beany-flavor
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脂氧合酶基因LOX6克隆及其表达与鲜切芋头褐变的相关性 被引量:1
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作者 袁晓 杨盼迪 +3 位作者 林冲 朱云娜 王光 王斌 《山东农业科学》 北大核心 2023年第12期17-25,共9页
细胞膜脂质过氧化是引起鲜切果蔬褐变的重要原因,脂氧合酶(LOX)在膜脂过氧化反应中具有重要作用。为探究脂氧合酶基因LOX6表达与鲜切芋头褐变的相关性,本研究以芋头球茎cDNA为模板,克隆了LOX6基因的cDNA全长序列,并分析了其在鲜切芋头... 细胞膜脂质过氧化是引起鲜切果蔬褐变的重要原因,脂氧合酶(LOX)在膜脂过氧化反应中具有重要作用。为探究脂氧合酶基因LOX6表达与鲜切芋头褐变的相关性,本研究以芋头球茎cDNA为模板,克隆了LOX6基因的cDNA全长序列,并分析了其在鲜切芋头褐变过程中的表达模式。结果表明,芋头LOX6编码序列全长2 751 bp,编码916个氨基酸残基。不同植物LOX6氨基酸序列之间的相似性不是很高,芋头LOX6与同属天南星科的马蹄莲ZaLOX6的亲缘关系最近。随着鲜切芋头褐变加重,LOX6表达量逐渐增加,LOX6表达水平与褐变指标△E呈显著正相关(R2=0.72);外源香草酸处理显著抑制鲜切芋头褐变,且显著下调LOX6基因的表达。表明LOX6表达与鲜切芋头褐变具有明显相关性,抑制其表达可抑制鲜切芋头褐变。LOX6基因启动子中含有丰富的逆境响应相关元件,表明鲜切处理诱导LOX6表达可能是促进鲜切芋头褐变的重要原因。本研究结果可为不易褐变的芋头品种培育提供重要的基因资源,并为优化鲜切芋头褐变防控技术提供帮助。 展开更多
关键词 鲜切芋头 褐变 脂氧合酶基因lox6 低温贮藏 膜脂过氧化
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Generation of seed lipoxygenase-free soybean using CRISPR-Cas9 被引量:15
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作者 Jie Wang Huaqin Kuang +5 位作者 Zhihui Zhang Yongqing Yang Long Yan Mengchen Zhang Shikui Song Yuefeng Guan 《The Crop Journal》 SCIE CAS CSCD 2020年第3期432-439,共8页
Beany flavor induced by three lipoxygenases(LOXs, including LOX1, LOX2, and LOX3)restricts human consumption of soybean. It is desirable to generate lipoxygenase-free new mutant lines to improve the eating quality of ... Beany flavor induced by three lipoxygenases(LOXs, including LOX1, LOX2, and LOX3)restricts human consumption of soybean. It is desirable to generate lipoxygenase-free new mutant lines to improve the eating quality of soybean oil and protein products. In this study, a pooled clustered regularly interspaced short palindromic repeats(CRISPR)-CRISPRassociated protein 9(Cas9) strategy targeting three GmLox genes(GmLox1, GmLox2, and GmLox3) was applied and 60 T_0 positive transgenic plants were generated, carrying combinations of sg RNAs and mutations. Among them, GmLox-28 and GmLox-60 were gmlox1 gmlox2 gmlox3 triple mutants and GmLox-40 was a gmlox1 gmlox2 double mutant.Sequencing of T_1 mutant plants derived from GmLox-28, GmLox-60, and GmLox-40 showed that mutation in the GmLox gene was inherited by the next generation. Colorimetric assay revealed that plants carrying different combinations of mutations lost the corresponding lipoxygenase activities. Transgene-free mutants were obtained by screening the T_2 generation of lipoxygenase-free mutant lines(GmLox-28 and GmLox-60). These transgeneand lipoxygenase-free mutants could be used for soybean beany flavor reduction without restriction by regulatory frameworks governing transgenic organisms. 展开更多
关键词 lox Generation of seed lipoxygenase-free soybean using CRISPR-Cas9 CRISPR
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Effect of Ethylene on Polygalacturonase,Lipoxygenase and Expansin in Ripening of Tomato Fruits 被引量:2
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作者 薛照辉 寇晓虹 +2 位作者 罗云波 朱本忠 许文涛 《Transactions of Tianjin University》 EI CAS 2009年第3期173-177,共5页
Fruit ripening is a complex process and is regulated by many factors. Ethylene and polygalacturonase (PG), lipoxygenase (LOX), expansin (EXP) are all critical regulating factors in fruit ripening and softening p... Fruit ripening is a complex process and is regulated by many factors. Ethylene and polygalacturonase (PG), lipoxygenase (LOX), expansin (EXP) are all critical regulating factors in fruit ripening and softening process. With antisense ACS tomato, Nr mutant tomato and cultivated tomato as materials, Northern blot hybridization showed that PG, LeEXP1 and LOXexpressed differently in different parts of cultivated tomato fruit during ripening, which was related to fruit ripening. The ripening process of columella and radial pericarp was faster than pericarp. In both Nr mutant and antisense ACS transgenic tomato fruit, expression levels ofPG, LeEXPI and LOXwere generally lower than those in cultivated fruit but still related to fruit ripening. The expression levels ofPG, LeEXP1 and LOX increased in the mature green tomato fruits after 0.5 h treatment with ethylene (100 μL/L). These results indicate that gene expression ofPG, LeEXP1 and LOXwere positively regulated by ethylene. The time and cumulative effect of the concentration exists in the expression of PG regulated by ethylene. The regulation of LOX expression mainly depended on the fruit development after great amount of ethylene was produced. PG played a major role in ripening and softening of tomato fruit, and cooperated with the regulation of EXP and LOX. 展开更多
关键词 TOMATO ripening and softening ETHYLENE EXPANSIN lipoxygenase POLYGALACTURONASE gene expression
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高油酸低脂氧合酶(FAD-LOX)多基因编辑大豆优异种质创制 被引量:1
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作者 何梓莹 于莉莉 +4 位作者 邱红梅 谷勇哲 洪慧龙 高华伟 邱丽娟 《大豆科学》 CAS CSCD 北大核心 2023年第5期513-523,共11页
大豆是植物油和蛋白质的主要来源。不饱和脂肪酸的含量是影响植物油品质的主要因素,油酸是不饱和脂肪酸中含量最丰富的成分之一,具有较强的氧化稳定性和热稳定性。增加油酸的含量可以提高大豆油的耐储存性,也有利于人体健康。GmFAD2-1A... 大豆是植物油和蛋白质的主要来源。不饱和脂肪酸的含量是影响植物油品质的主要因素,油酸是不饱和脂肪酸中含量最丰富的成分之一,具有较强的氧化稳定性和热稳定性。增加油酸的含量可以提高大豆油的耐储存性,也有利于人体健康。GmFAD2-1A和GmFAD2-1B是大豆编码脂肪酸去饱和酶,调控油酸向亚油酸转化的关键基因。脂氧合酶是大豆种子中的抗营养因子之一。它在酶促氧化的条件下产生豆腥味,限制了大豆产品的应用。为满足不同人群的需要和降低加工成本,有必要培育无脂氧合酶的突变品系。大豆中GmLOX基因(包括GmLOX1、GmLOX2、GmLOX3)编码脂氧合酶。脂氧酶也能催化亚油酸氧化生成脂质氢过氧化物,然后形成豆腥味。因此,降低亚油酸的含量有助于减少豆腥味的形成。同时编码脂氧酶也能延缓豆油的酸败。本研究利用CRISPR/Cas9对中吉602的GmFAD2-1A、GmFAD2-1B、GmLOX1、GmLOX2和GmLOX3基因进行编辑。通过农杆菌介导的大豆遗传转化,在T2代分离到7个具有不同等位变异的优异新种质。与中吉602相比,突变体种子中脂氧合酶活性显著降低,油酸含量提高至81.7%~83.5%。本研究为有效开发大豆种质资源以满足不断变化的市场需求提供了新材料。 展开更多
关键词 大豆品质 油酸 脂氧合酶 CRISPR/Cas9
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