本地早蜜橘胚性愈伤组织的离体诱导

柑橘胚性愈伤组织诱导技术在柑橘种质资源离体保存、理化诱变、原生质体融合、外源基因转化等柑橘种质保存及新种质创制过程中有着重要的基础研究地位。目前随着柑橘分子生物育种技术的快速发展,胚性愈伤组织作为柑橘理化诱变、原生质体融合、外源基因转化的理想起始材料,其在柑橘种质创新研究中的基础应用作用更为明显。柑橘胚性愈伤组织目前主要可通过败育的成熟胚珠、未成熟胚珠、实生幼苗胚轴茎段、子叶以及花药培养等途径来诱导获得。     

蛋白酶抑制剂基因的杀虫机理及其应用

蛋白酶抑制剂（PI）是自然界含量最为丰富的蛋白种类之一，存在干所有生命体中（Laskowski，1980）。研究表明，这些蛋白酶抑制剂在植物对危害昆虫以及病原体侵染的天然防御系统中担当着重要角色（Ryan，1989）。昆虫饲喂实验发现，某些纯化的蛋白酶抑制剂具有明显的抗虫作用（Gatechouse等，1988；Chen等，1992）。利用蛋白酶抑制剂基因来提高植物的抗虫能力，已成为植物基因工程研究的一个热门领域。目前为止，在植物中发现有三类蛋白酶抑制剂：丝氨酸蛋白酶抑制剂，巯基蛋白酶抑制剂和金属蛋白酶抑制剂。其中对丝氨酸类蛋白酶抑制剂的…     

植物系统素研究进展

概述植物系统素的结构特点,综述系统素原、系统素的膜受体、系统素在伤信号转导中的作用等研究进展,展望系统素今后的研究方向。     

香菇发育生理研究进展

就温度、湿度、光照和通气等主要环境因子对香菇子实体发生和发育的影响,类脂物、蛋白酶抑制剂和子实体形成诱导物等外源添加物与子实体形成的关系以及与子实体形成相关的酶学研究进展进行了综述,并讨论了香菇发育生理研究的方向。     

天草杂柑胚乳培养研究初报

柑橘胚乳的离体培养是获得三倍体无籽柑橘的重要途径,关于柑橘胚乳再生植株诱导的培养时有报道,研究表明柑橘胚乳培养存在愈伤组织诱导率较低,愈伤组织分化不定芽困难等现状。本文针对这一现状以天草杂柑为对象进行研究,以期找到提高柑橘胚乳再生植株诱导率的新途径,提高柑橘胚乳培养成功率。     

不同生长调节因子对耐寒柑橘愈伤组织诱导效应研究

以耐寒柑橘子叶为试验材料,研究了不同浓度的2,4-D、NAA、6-BA与KT对柑橘子叶愈伤组织诱导的影响作用。结果表明:2,4-D、NAA、6-BA与KT对柑橘子叶愈伤组织诱导具有重要效应,其综合贡献率分别为86.4%、93.9%、93.9%与94.5%。4种生长调节因子对柑橘子叶愈伤组织诱导的膨大率、膨大速度、出愈率、出愈速度的效应大小分别为2,4-DNAA6-BAKT、KT2,4-DNAA6-BA、2,4-DNAA6-BAKT与2,4-DKTNAA6-BA。综合考虑测量指标效应,对于柑橘子叶愈伤组织诱导的最适生长调节因子组合配比为2,4-D 1.0mg/L+NAA 2.0mg/L+6-BA 0.5mg/L+KT 0.5mg/L。     

铁线莲品种Clematis ‘Bill MacKenzie’愈伤组织的诱导

以铁线莲Clematis‘Bill MacKenzie’优良单株的嫩茎为外植体,研究了不同灭菌时间、基本培养基类型对C.‘Bill MacKenzie’愈伤组织诱导的影响,并在初代愈伤诱导的基础上,研究了不同外源激素组合对C.‘Bill MacKenzie’愈伤组织诱导及状态的影响。结果表明:最佳的灭菌时间为12min;基本培养基类型为MS,诱导率为68.4%;不同的外源激素组合均能诱导出愈伤,但是愈伤状态不同,其中MS+1.0mg/L 6-BA+0.4mg/L IBA时愈伤状态最好。     

乙烯参与ALA-ABA/黑暗调控的苹果叶片气孔运动

以‘富士’苹果组培苗离体叶片下表皮为材料,运用不同试剂结合气相色谱、激光共聚焦扫描显微镜、光学显微镜和qRT-PCR等技术,从气孔开度、乙烯释放、保卫细胞H_2O_2及黄酮醇水平和基因表达等方面,研究了乙烯在5–氨基乙酰丙酸(ALA)抑制脱落酸(ABA)或黑暗诱导的气孔关闭过程中的作用。结果表明,ALA促进苹果叶片乙烯释放,而乙烯生物合成抑制剂氨氧乙基乙烯基甘氨酸(AVG)及乙烯受体抑制剂1–甲基环丙烯(1-MCP)削弱ALA对ABA或黑暗诱导气孔关闭的抑制效应。ALA上调乙烯生物合成及信号转导相关基因的表达。AVG削弱ALA抑制ABA或黑暗诱导的H_2O_2上升效应,也削弱ALA对保卫细胞黄酮醇积累的促进效应。据此认为,外源ALA通过上调基因表达,促进叶片保卫细胞中乙烯生物合成和信号转导,诱导黄酮醇积累,清除ABA或黑暗诱导增加的H_2O_2,抑制气孔关闭,从而为光合作用所需的气体交换打开通道。     

柑橘体细胞胚发生基因CsHB1特异肽段多克隆抗体的制备及其蛋白动态检测

【目的】构建柑橘HD-ZIP II转录因子CsHB1基因的原核表达系统,制备多克隆抗体,并检测抗体在‘伏令夏橙’胚性愈伤体胚诱导阶段中的特异性,为研究CsHB1基因在柑橘体细胞胚发生过程中的功能奠定基础。【方法】构建柑橘HD-ZIP II转录因子CsHB1基因的原核表达载体p GEX4T-CsHB1-N,转化大肠杆菌诱导目的融合蛋白的表达,并制备获得多克隆抗体anti-CsHB1-N。通过Western blot检测抗体在原核表达系统和柑橘体细胞胚诱导阶段的特异性,并分析体细胞胚诱导阶段CsHB1蛋白水平表达的动态变化。【结果】重组原核表达载体p GEX4T-CsHB1-N在Escherichia coli中高效表达出了分子质量约为49 ku的GST-CsHB1-N融合蛋白,并纯化获得多克隆抗体anti-CsHB1-N;经过Western blot分析表明,多克隆抗体可与‘伏令夏橙’愈伤体胚诱导阶段表达的目的蛋白特异结合;蛋白表达结果分析表明,在胚性愈伤组织中CsHB1蛋白呈现高的表达量,随着诱导培养的进行,呈现了先下降后上升的波动变化。【结论】多克隆抗体特异性好,可与‘伏令夏橙’胚性愈伤组织中目的蛋白特异性结合,可用于CsHB1基因功能分析。     

柑橘体细胞胚发生基因CsHB1特异肽段多克隆抗体的制备及其蛋白动态检测北大核心CSCD

【目的】构建柑橘HD-ZIP II转录因子CsHB1基因的原核表达系统,制备多克隆抗体,并检测抗体在‘伏令夏橙’胚性愈伤体胚诱导阶段中的特异性,为研究CsHB1基因在柑橘体细胞胚发生过程中的功能奠定基础。【方法】构建柑橘HD-ZIP II转录因子CsHB1基因的原核表达载体p GEX4T-CsHB1-N,转化大肠杆菌诱导目的融合蛋白的表达,并制备获得多克隆抗体anti-CsHB1-N。通过Western blot检测抗体在原核表达系统和柑橘体细胞胚诱导阶段的特异性,并分析体细胞胚诱导阶段CsHB1蛋白水平表达的动态变化。【结果】重组原核表达载体p GEX4T-CsHB1-N在Escherichia coli中高效表达出了分子质量约为49 ku的GST-CsHB1-N融合蛋白,并纯化获得多克隆抗体anti-CsHB1-N;经过Western blot分析表明,多克隆抗体可与‘伏令夏橙’愈伤体胚诱导阶段表达的目的蛋白特异结合;蛋白表达结果分析表明,在胚性愈伤组织中CsHB1蛋白呈现高的表达量,随着诱导培养的进行,呈现了先下降后上升的波动变化。【结论】多克隆抗体特异性好,可与‘伏令夏橙’胚性愈伤组织中目的蛋白特异性结合,可用于CsHB1基因功能分析。     

减氮配施脲酶/硝化抑制剂对冬瓜品质、产量和土壤氮磷淋失的影响

采用田间小区试验,研究了在氮肥减施基础上配施脲酶/硝化抑制剂对我国南方热带地区露地冬瓜产量、品质,以及土壤氮磷养分淋溶的影响.结果表明,与正常施肥处理相比,氮肥减施30％会导致冬瓜单果质量降低,减产约8.30％,而配施脲酶/硝化抑制剂后冬瓜增产17.03％,并且维持较低的亚硝酸盐含量和较高的品质;氮肥减施能够有效降低土...     

酸性蛋白酶抑制剂─—福菇肽在食用菌生产上的应用

酸性蛋白酶抑制剂─福菇肽能有好地抑制食用菌体内的酸性蛋白酶的活力，激活中性蛋白酶的活力，调节食用菌体内蛋白酶的代谢，具有促进子实体形成、增加子实体的数目、加速子实体生长的作用。将酸性蛋白酶抑制剂作为菇类增产剂应用于食用菌生产上表现出优良的生物学效应：促进生长效应、后劲效应与优质效应。有效地增加菇的产量、提高菇的品质，在大面积推广应用中展示出显著的经济效益与社会效益和广阔的应用前景。     

荞麦芽菜营养成分分析评价

 对甜荞(FagoFyrum esculentum)‘榆荞1号’和苦荞(F．tanaricum)‘榆6-21’籽粒萌发过程中胰蛋白酶抑制剂活性及荞麦芽的营养成分进行分析评价。结果表明：荞麦籽粒氨基酸均衡，但籽粒中含有大量胰蛋白酶抑制剂。萌发10 d后，荞麦芽中胰蛋白酶抑制剂活性消失或仅存痕量。利用氨基酸比值系数法评价显示荞麦芽菜的氨基酸较籽粒更为均衡。芽苗中芦丁含量较籽粒增加4～6倍，芽苗中含有丰富的维生素及有机酸。萌发可提高荞麦的营养保健价值。     

几种天然防腐剂对腌制小萝卜贮藏特性的效应

利用天然防腐剂植酸、乳酸链球菌素、洋葱熏蒸处理腌制小萝卜,研究天然防腐剂对腌制小萝卜在贮藏过程中总酸度、脆度、亚硝酸盐含量、颜色、感官及风味的影响。结果表明,植酸和乳酸链球菌素在贮藏1个月内有一定的防腐效果,但贮藏后期防腐作用不明显;洋葱熏蒸处理有一定的杀菌效果,熏蒸后真空包装的小萝卜贮藏性很好,没有出现生膜、胀袋现象,颜色和脆度也保持的较好,是一种很好的天然防腐方法。     

Advances in prevention of nonsteroidal anti-inflammatory drug-induced gastropathy by proton pump inhibitor

Nonsteroidal anti-inflammatory drugs (NSAIDs) are powerful in anti-inflammatory, analgesic and antirheumatic effects, and widely used in treating the corresponding diseases. Over the years, many new dosage forms and structures of NSAIDs appear since aspirin was developed 112 years ago. However, the universal use of NSAIDs produces unavoidable mucosal lesions in gastrointestinal tract. As yet, proton pump inhibitor (PPI) has been used in the treatment of gastropathy induced by NSAIDs. This article will focus on the advances in prevention of NSAIDs-induced gastropathy by proton pump inhibitor.     

珍稀食药用菌水提取物对糖苷酶的抑制

以5种食药用菌水提取物为试材,采用测定糖苷酶抑制率的方法,研究了5种珍稀食药用菌水提取物对糖苷酶的抑制作用,以期为进一步开发降血糖药物提供参考依据。结果表明:羊肚菌、桑黄、灵芝、蝉花、虫草水提取物对4种糖苷酶均有不同程度的抑制,且差异显著(P<0.05)。5种水提物中,桑黄水提物对蔗糖酶的抑制率最高(74.8%),略低于常用的降糖药阿卡波糖以及米格列醇,虫草水提物对蔗糖酶的抑制率最低(35.9%)。虫草水提物对α-淀粉酶的抑制能力最高(9.7%)且显著高于米格列醇(P<0.05),羊肚菌水提物对α-淀粉酶的抑制最低(1.1%)。羊肚菌水提物对α-葡萄糖苷酶显示出较好的抑制能力(15.4%),显著高于阿卡波糖和米格列醇(P<0.05),而灵芝水提物对α-葡萄糖苷酶的抑制水平最低(11.2%)。灵芝水提物对麦芽糖酶显示出较高的抑制能力(10.2%),显著高于米格列醇(P<0.05),而与阿卡波糖相当,桑黄水提物对麦芽糖酶抑制最差(4.6%)。5种水提物在20~60℃以及pH 6~8时对4种糖苷酶仍然保持较好的抑制能力。     

红毛五加种子特性及发芽抑制物研究

以红毛五加为试材,观察红毛五加种子形态,测定千粒重、种皮透水性,研究果实各部分水浸物对油菜种子发芽和胚根生长抑制活性的影响。结果表明:红毛五加种子较小,种皮较薄,种皮透水性良好,对发芽率没有影响;红毛五加种子成熟度极不均匀,种胚细小,不具备萌发能力,需经过形态后熟和生理后熟才能萌发;红毛五加果实各部浸提液,对油菜种子发芽、胚根生长有显著的抑制作用,随着浸泡量的增加,对油菜种子发芽抑制活性越强,胚根长度越短;红毛五加果实各部分水浸物对油菜种子发芽抑制活性,果肉最强,种皮次之,种仁最弱。     

Correlation between TAFI and blood fat and coagulation in the process of atherosclerosis

AIM: To investigate the relationship between the thrombin-activatable fibrinolysis inhibitor (TAFI) level in plasma and atherosclerosis, the TAFI level in plasma and blood fat and blood clotting index.METHODS: Totally 40 healthy male Wistar rats were randomly divided into 4 groups (n=10), control group, high lipid group, high lipid +vitamin D3 overload group, and high lipid +vitamin D3+endothelium injure group by treating the animals with normal diet, high lipid, high lipid+ vitamin D overload, and high lipid+vitamin D overload+endothelium injury, respectively, for inducing three stages of AS in rats. Then, the total cholesterol (TC), total triglyeride (TG), high density lipoprotein (HDL), low density lipoprotein (LDL), and prothrombin time (PT), activated partial thromboplasin time (APTT), fibrinogen (Fib), the activity of TAFI were measured.RESULTS: TC, TG, LDL-C, Fib and the activity level of TAFI in plasma in three model groups increased gradually compared with the control (P＜0.01). HDL-C, PT and APTT in plasma of three model groups decreased gradually compared with the control (P＜0.01 or P＜0.05). The activity of TAFI in plasma had positive correlation with TG, TC and Fib.CONCLUSION: The activity of TAFI increases in the progress of atherosclerosis. The activity of TAFI in plasma has a correlation with blood fat and coagulation, which indicates that TAFI might participate in the process of atherosclerosis, and TAFI is one of etiological factors of atherosclerosis.     

Effects of SAHA on hippocampal TLR4/MYD88 signaling and neuronal apoptosis after seizure in developing rats

AIM: To investigate the effect of vorinostat(suberoylanilide hydroxamic acid, SAHA), a histone deacetylase inhibitor, on seizure-induced brain damage in developing rats and its mechanism. METHODS: Male Sprague-Dawley rats(n=32) were randomly divided into control group, pentylenetetrazole(PTZ) group, PTZ+10 mg/kg SAHA group and PTZ+50 mg/kg SAHA group. Intraperitoneal injection of PTZ was used to induce rat seizure. SAHA was injected intraperitoneally 2 h before PTZ injection. The rats in different seizure stages were counted and mean seizure score was analyzed at 30~60 min after PTZ injection. Hippocampal tissues were sampled at 24 h after seizures. The expression of TLR4, MYD88, NF-κB P65 and IL-1β at mRNA and protein levels was detected by RT-qPCR and Western blot, respectively. The pathological changes of the brain tissues were observed by HE staining. The apoptotic neurons were observed by TUNEL staining. RESULTS: The mRNA and protein levels of TLR4, MYD88, NF-κB P65 and IL-1β, the apoptosis of neurons, the inflammation reaction and mean seizure score significantly increased after PTZ treatment(P<0.05), and these effects were attenuated by treatment with SAHA. Compared with PTZ+10 mg/kg SAHA group, PTZ+50 mg/kg SAHA group showed more significant protective effect against seizure-induced brain damage. CONCLUSION: Histone deacetylase inhibitor SAHA suppresses seizure-induced TLR4/MYD88 signaling and reduces apoptosis of neurons, suggesting a protective effect against brain damage associated with seizure in developing rats.     

Effect of RelB on proteasome inhibitor-induced maspin expression

AIM: To investigate the effects of RelB on proteasome inhibitor-induced maspin expression in prostate cancer cells. METHODS: Western blotting analysis was performed to examine endogenous and proteasome inhibitor(MG-132)-induced expression of RelA, RelB and maspin in prostate cancer cells. The expression profiles of RelB and maspin in human prostate cancer tissues were obtained by immunohistochemistry assay. RNA interference targeting RelB was performed in DU145 cells. The effects of RelB-silencing on maspin expression induced by MG-132 were detected by Western blotting. The cell viability was determined by PI staining and FACS analysis. RESULTS: RelB expression was increased in androgen-independent prostate cancer cell line DU145, while maspin expression was minimally detected. Among 10 tissue samples tested, a strong nuclear RelB staining and an absence of maspin expression were found in high-grade specimens (Gleason scores 4-5). RelB expression was reduced upon treatment with MG-132 for 24 h, which was coincided with the induction of maspin expression. RelB-silencing in DU145 cells by siRNA didn't influence the proteasome inhibitor-induced maspin expression. CONCLUSION: The expression of RelB is inversely correlated to maspin expression in androgen-independent prostate cancer cells and prostate cancer tissues. RelB expression is critical to the proteasome inhibitor-induced maspin expression.