蔗糖水解酶注释基因XC0805与野油菜黄单胞菌8004菌株的蔗糖利用及致病相关

 野油菜黄单胞菌(Xanthomonas campestris pv.campestris,Xcc)是造成十字花科植物减产的重要病原菌,其通过水孔或伤口进入植物体内并在维管束中快速繁殖。光合产物(蔗糖)的运输主要依靠植物维管组织,同时蔗糖也是维管组织重要的能量来源。本研究利用生物信息学和遗传学等方法对Xcc中参与蔗糖利用的基因进行了分析,并研究了这些基因与其致病力的关系。Xcc 8004菌株中注释的参与蔗糖分解蛋白的编码基因有XC1002、XC1642、 XC1645和XC0805。 XC1002和XC1642的编码产物属于糖苷水解酶GH97蛋白超家族,而XC1645和XC0805的编码产物则属于GH13家族。突变分析发现除XC0805外,其他3个都不影响Xcc的蔗糖利用能力。接种实验表明,XC0805参与Xcc的侵染或在植物体内的生长,其他3个不参与致病过程。另外,Xcc 8004中预测参与蔗糖转运的基因(XC0806和XC0807)突变并不影响细菌的蔗糖利用和致病力,表明其可能存在其他的蔗糖转运通路。上述结果说明XC0805可能是Xcc 8004主要的蔗糖水解酶基因,且在致病中起重要作用。     

滞绿突变对大豆叶片蔗糖代谢相关基因表达的影响

为研究大豆滞绿突变对叶片衰老过程中蔗糖代谢相关基因表达产生的影响，以晋大滞绿1号及其亲本晋大74号为试验材料，测定了大豆开花后至成熟期间叶片可溶性糖含量的变化趋势，并对比了不同基因型大豆品种叶片衰老过程中蔗糖代谢相关基因的表达差异。结果表明:1)花后至成熟期间，2个供试大豆品种可溶性总糖含量均呈波动升高然后降低的变化趋势。花后14至42天，晋大滞绿1号可溶性总糖含量高于晋大74号；2)花后29至42天，晋大滞绿1号蔗糖磷酸合成酶基因SPS4，转化酶基因CInv、CWInv2以及蔗糖合酶基因SS1、SS2-2表达水平显著高于非滞绿品种晋大74号；己糖激酶和果糖激酶基因Hxk和Frk表达量也高于非滞绿品种晋大74号；3)除 SUT4-1外，其余6个蔗糖转运体基因SUTs的表达模式具有明显的品种特异性。花后29至42天，晋大滞绿1号叶片SUTs表达水平较高。综上，滞绿突变对于大豆蔗糖代谢相关基因的表达产生了明显的影响，特别是在鼓粒初期和中期等大豆籽粒形成的关键时期，滞绿品种晋大滞绿1号蔗糖代谢相关基因的表达更加活跃。     

乙酰羟酸合酶抑制剂类除草剂的植物抗性机制

乙酰羟酸合酶(AHAS)抑制剂类除草剂已被广泛用于农业生产,然而使用过程中可能对部分敏感农作物产生药害,因此创制对不同类别除草剂具有抗性的一系列作物新品种至关重要。本文将从AHAS抑制剂类除草剂的类别与特点、AHAS靶酶的特性及其在支链氨基酸合成中的作用、除草剂的靶标抗性与非靶标抗性机制等方面分析国内外研究现状与未来发展动态,以期为农作物除草剂抗性性状的遗传改良和开发应用提供参考。     

茶树GGPS基因家族的克隆、生物信息学和表达分析

牻牛儿基牻牛儿基焦磷酸合酶(Geranylgeranyl diphosphate synthase,GGPS)是萜类合成途径的结构酶,对植物生长发育具有重要意义。本研究通过RACE和RT-PCR方法克隆得到5条潜在的茶树GGPS序列,分别命名为CsGGPS1-4和CsGGPS9,其中CsGGPS9存在3条等位基因,分别是CsGGPS9-1、CsGGPS9-2和CsGGPS9-3,在系统进化树上与其他基因分成两支。蛋白质序列分析表明,茶树GGPS家族成员都具有polyprenyl_synt结构域,不存在信号肽序列。亚细胞定位预测结果显示,CsGGPS1、CsGGPS2和CsGGPS4定位在叶绿体上,CsGGPS3和CsGGPS9定位在线粒体上。通过Swiss Model进行三维建模,结合"three-floor"模型对茶树GGPS家族成员的功能进行预测,预测结果显示,CsGGPS1、CsGGPS2和CsGGPS4是GGPS;CsGGPS3是异源二聚体形式的牻牛儿基焦磷酸合酶的小亚基;CsGGPS9的催化主产物是碳链数大于30的异戊烯基焦磷酸。q RT-PCR分析表明,CsGGPS1整体表达丰度较低,仅在一芽二叶中表达量稍高;CsGGPS2在茶树各个组织中均有表达,在花中表达量最高,且花发育过程中表达量先上升后下降;CsGGPS3在叶和幼根中的表达量高于花,花发育过程中表达平稳;CsGGPS4在茶树各个组织中表达量数值相近,在花发育过程中表达量变化趋势与CsGGPS2相同;CsGGPS9的表达量在成熟叶中显著低于幼嫩叶片。     

Effects of taurine-zinc on learning and memory abilities of vascular dementia mice

AIM:To observe the effects of taurine-zinc (TZC) on the learning and memory abilities of vascular dementia (VD) mice and to investigate the related mechanism. METHODS:The mice were randomly divided into model group, sham group, and TZC at 50 mg/kg, 100 mg/kg and 200 mg/kg groups. The mice in drug groups were given TZC by gavage at 10 mL/kg once daily. The mice in sham group and model group were given equal volume of distilled water. VD mice were established by intercepting both common carotid arteries and bleeding at caudal vein after 14 d of gavage. The levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were detected by ELISA. The levels of inducible nitric oxide synthase (iNOS) and nitric oxide (NO) were measured via spectrophotometer. Step-down test and Morris water maze test were used to examine the abilities of learning and memory in the mice. RESULTS:TZC at 50 mg/kg, 100 mg/kg and 200 mg/kg reduced the levels of TNF-α, IL-1β, iNOS and NO in the brain tissues. In the water maze test, TZC at 100 mg/kg and 200 mg/kg significantly decreased the error times and latency compared with model group. In the step-down test, the escape latency was prolonged and error times were lowered significantly by treatment with TZC at 50 mg/kg, 100 mg/kg and 200 mg/kg as compared with model group. CONCLUSION:TZC improves the abilities of learning and memory, which might be related to the reduction of TNF-α, IL-1β, iNOS and NO levels in VD mice.     

Changes in photosynthesis,growth, and sugar content of commercial sugarcane cultivars and Erianthus under flood conditions

Sugarcane (Saccharum spp.) is an economical crop in the tropical and subtropical countries. However, because of global climate change, flooding has become problematic, particularly during the rainy season, in Thailand. We investigated the effects of floods on three commercial sugarcane cultivars, namely NiF8, U-thong 6 (UT6), and U-thong 9 (UT9), as well as Erianthus spp. Growth was assessed using a pot experiment in a glasshouse with two treatments: (1) control and (2) 60 d of flooding followed by 30 d of normal conditions. In comparison with control, during prolonged flooding, Erianthus showed greatly decreased CO₂ assimilation, whereas NiF8, UT6, and UT9 showed slightly declined CO₂ assimilation. Growth in plants subjected to 60 d of flooding was less influenced by floods while sucrose content was not affected except in UT6. During flooding, some roots died, resulting in plants compensating adventitious roots to offset the negative effects of root death and to assist them in maintaining their growth, which appeared from the submerged nodes, with different characteristics for each cultivar. However, 30 d after draining, roots remained damaged, while adventitious roots died, resulting in lesser growth as compared with the control, but it did not significantly affect sucrose content and sugar yield. This study suggests that sugarcane plants need to produce the adventitious roots to compensate their roots’ death during flooding and require time to recover their root system after flooding for obtaining the optimum yield and quality at harvest.     

亚洲玉米螟幼虫膜结合海藻糖酶基因RNAi效应

海藻糖是昆虫的血糖，为昆虫提供能量；海藻糖酶催化海藻糖分解为葡萄糖，是几丁质生物合成的原料。围食膜（peritrophic membranes，PMs）是昆虫消化道特有结构，对昆虫消化食物、保护肠道表皮细胞具有重要作用；几丁质是PMs的重要组分。海藻糖酶（trehalase，Tre）和几丁质合成酶（chitin synthase，CHS）是几丁质合成途径的第1个和最后1个酶。本研究通过饲喂亚洲玉米螟（Orstrinia furnacalis，Asian corn borer，ACB）膜结合海藻糖酶（OfMT）基因特异的干扰dsRNA，研究RNAi对ACB幼虫中肠CHSB（OfCHSB）基因表达及幼虫发育的影响。发现处理48 h后，OfMT基因和OfCHSB基因表达量分别下降了52%和53%，幼虫发育迟缓。通过对处理及对照ACB幼虫中肠石蜡切片进行苏木精-伊红染色（hematoxylin-ethanol，HE染色）和几丁质标记，发现血腔内脂肪体组织减小、中肠围食膜组织中几丁质含量减少。推测OfMT基因有可能成为ACB的生物防治的靶标基因。     

Roles of protein phosphatase 4 in down-regulation of eNOS Ser633 phosphorylation induced by palmitic acid in human umbilical vein endotheli?al cells

AIMTo investigate the roles of protein phosphatase 4 (PP4) in down-regulation of endothelial nitric oxide synthase (eNOS) Ser633 phosphorylation induced by palmitic acid (PA). METHODSHuman umbilical vein endothelial cells (HUVECs) were treated with PA at 25 μmol/L, 50 μmol/L, 100 μmol/L and 200μmol/L for 36 h, or treated with PA at 100 μmol/L for 12 h, 24 h, 36 h and 48 h. Protein phosphatase 2A (PP2A) family inhibitor fostriecin (FST, 20 nmol/L) or okadaic acid (OA, 5 nmol/L) was selected to pretreat the HUVECs for 30 min. Protein phosphatase 4 catalytic subunit (PP4c) siRNA or protein phosphatase 2A catalytic subunit (PP2Ac) siRNA was transfected into the HUVECs. The protein expression levels of of eNOS, PP4c and PP2Ac, as well as the level of eNOS Ser633 phosphorylation, were detected by Western blot. The intracellular nitric oxide (NO) content was measured by DAF-FM DA. RESULTS(1) Compared with control group, the levels of eNOS Ser633 phosphorylation were decreased in PA groups in which the HUVECs were treated with 25 μmol/L, 50 μmol/L, 100 μmol/L and 200 μmol/L PA for 36 h (P<0.05) and 100 μmol/L PA for 24 h, 36 h and 48 h (P<0.05). No significant difference in the level of total eNOS protein expression among all the groups was observed. (2) Compared with control group, both FST and OA pretreatment reversed the reduction of eNOS Ser633 phosphorylation (P<0.05) and the decrease in intracellular NO content (P<0.05) induced by PA. No significant difference in the level of total eNOS protein expression among all the groups was observed. (3) Compared with si-Control group, the PP4c protein expression was significantly reduced (P<0.05), while the level of eNOS Ser633 phosphorylation was significantly increased in si-PP4c group (P<0.05). Although the levels of PP2Ac protein expression declined significantly (P<0.05), the level of eNOS Ser633 phosphorylation remained unchanged in si-PP2Ac group. No significant differencein the level of total eNOS protein expression among all the groups was found. CONCLUSION PA significantly reduces the level of eNOS Ser633 phosphorylation and the content of NO in the HUVECs, which may be due to PA inducing the activation of the PP2A family member PP4 rather than PP2A.     

中国野生毛葡萄芪合酶基因VqSTS11和VqSTS23调控抗白粉病的研究

利用同源克隆法得到抗白粉病的中国野生毛葡萄‘丹凤–2’芪合酶基因VqSTS11和VqSTS23的开放阅读框，并构建过表达载体；通过器官发生途径诱导不抗白粉病的欧洲葡萄‘无核白’分生愈伤组织并采用农杆菌介导法对其进行转化；经过PCR检测和Western blot鉴定，获得了5株VqSTS11超量表达植株和3株VqSTS23超量表达植株；人工接种白粉菌发现，转基因植株中白粉菌菌丝生长速度慢，孢子萌发受到一定的抑制，并且转基因植株中芪合酶基因相对表达量升高，抗病相关基因表达上调，芪类物质含量积累增多。本研究结果进一步证明中国野生毛葡萄‘丹凤–2’芪合酶基因VqSTS11和VqSTS23对白粉菌具有抗性，‘丹凤–2’可以为改良欧洲葡萄品种抗病性提供抗病基因，作为抗病育种的资源。