花生溶血磷脂酸酰基转移酶基因的克隆与表达分析

溶血磷脂酰基转移酶(LPAT)是植物油脂合成途径的一个关键酶,在植物油脂品质改良和提高种子含油量方面具有重要的应用价值。本研究通过构建花生种子全长cDNA文库,结合大规模EST测序和功能注释,从花生中克隆了溶血磷脂酸酰基转移酶基因,命名为AhLPAT。该基因cDNA全长1 629 bp,对应的基因组序列5 531 bp,由11个外显子和10个内含子组成,内含子剪接方式符合GT-AG剪接规则。根据编码区预测AhLPAT编码一条387个氨基酸组成的多肽,预测分子量为43.2 kD,等电点为9.42。AhLPAT蛋白含有一个典型的酰基转移酶保守功能结构域以及溶血磷脂酰基转移酶相似的保守区域。该蛋白的氨基酸序列与已报道的其他物种LPAT蛋白序列有较高的一致性。AhLPAT与旱金莲、油菜、海甘蓝、蓖麻和拟南芥的LPAT蛋白氨基酸相似性依次为90%、89%、89%、88%和87%。系统进化分析表明,AhLPAT与拟南芥AtLPAT2亲缘关系较近,且同属于内质网型LPAT蛋白。RT-PCR分析表明,AhLPAT基因在花生根、茎、叶、花、果针和种子中均有表达,在花生开花后50~60 d,果针和种子中的表达量最高,且AhLPAT的表达量与花生种子含油量积累速率变化一致,二者显著相关(r=0.63,P<0.05)。推测AhLPAT基因在花生种子油脂合成中起重要作用。     

New mutants of winter rapeseed (Brassica napus L.) with changed fatty acid composition

Seeds of the winter oilseed rape (Brassica napus L.) line PN 3756/93 were treated with ethyl methanesulphonate to induce mutations in the fatty acid biosynthetic pathway. The seed mutagenic treatment was repeated in the M₂ generation. After treatments, individual seed and plant selections were made for changes in fatty acid composition during several generations of inbreeding. Self‐pollinated plants with changed fatty acid compositions were inbred to obtain genetically homozygous and stable mutant lines. Two mutants, M‐10453 and M‐10464, with increased levels of oleic acid (approximately 76%) and reduced linoleic and linolenic acid contents (8.5% and 7.5%, respectively) were selected. Gene or genes controlling desaturation of oleic acid were probably mutated in these plants. The third mutant, M‐681 had a very low linolenic acid content (approximately 2.6%) and increased linoleic acid content (approximately 26%). This would suggest the occurrence of mutations in genes controlling linoleic acid desaturation. The results of selection work during several generations showed that the environment had substantial influence on the composition of seed oil. This made the search for mutants with modify fatty acid compositions difficult. The induced mutants are not directly usable as new varieties, but can be used as parents in crosses for the development of high quality rapeseed varieties.     

类猪圆环病毒因子P1核酸链型和极性的研究

研究报道类猪圆环病毒因子P1的核酸链型和极性的研究结果。采用氯化铯平衡密度梯度离心获得较为纯净的病毒粒子后,提取病毒DNA,经S1核酸酶消化、特异性单引物第一轮PCR扩增结合常规第二轮PCR扩增等研究,结果表明,类猪圆环病毒因子P1为单股、负链基因组。     

Vertical advection of extracellular DNA by water capillarity in soil columns

The fate of extracellular DNA in the environment concerns both the fate of transgenes from genetically modified organisms and the evolution of active bacteria capable of incorporating this DNA into their genomes. This study addressed the possibility that DNA, like other organic molecules, could move vertically in the capillary fringe of groundwater aquifers. The targeted gene fragment used here was the 35S-nptII sequence, which was below detection levels in controls. Initial microcosm studies detected the DNA target molecule by PCR during the entire experiment. The vertical advection of water and DNA were monitored for a period of 3 days in soil columns. DNA was added as a water solution at the bottom of the unsaturated soil column, and then DNA-free water was added at the bottom after 12 and 24 h. After the addition of the DNA solution, capillary water rose 4 cm within the soil column and the target DNA was detected up to that height. After 60 min, the entire soil column (10 cm) was wetted and the target sequence was detected up to a height of 7.5 cm. After the second wetting (12 h later), the target sequence was detected up to the top of the soil column (10 cm). However, after the third wetting (24 h later), the marker sequence was only found at heights from 0.5 to 4 cm. Results clearly show the vertical movement of DNA due the capillary rise and suggest the possibility of DNA degradation within the soil column.     

水氮耦合对滁菊产量和品质的影响

采用二因素二次回归旋转组合设计,以盆栽试验方式研究了水、氮因子对滁菊产量和品质的影响,探讨了滁菊人工栽培适宜的水氮管理模式。研究结果表明,水氮耦合对滁菊花产量、叶片水提总黄酮和氯原酸含量、滁菊花水提总黄酮和氯原酸含量均有极显著影响,水氮间存在显著交互作用。氮肥因子对滁菊花产量、总黄酮含量、氯原酸含量的影响大于水分因子。当土壤水分水平为0.284 1,氮肥水平为0.403 7时,即田间持水量80%,氮肥用量0.257g/kg时,滁菊花产量最高,达18.09g/株;当土壤水分和氮肥用量均为-1.414水平时,即田间持水量50%,氮肥用量为0g/kg时,滁菊花总黄酮含量和氯原酸含量最大。从滁菊花产量、品质角度综合考虑,滁菊生育前期水氮管理以中水中氮较为适宜,滁菊现蕾开花期以低水低氮较为适宜。     

植酸对霍山石斛类原球茎悬浮培养细胞生长和多糖合成的影响

研究了植酸对霍山石斛类原球茎悬浮培养细胞生长、多糖合成和培养基中主要营养成分消耗的影响,并分析了细胞中可溶性还原糖、可溶性蛋白质、丙二醛含量以及过氧化物酶和多酚氧化酶的活性。结果表明,植酸能抑制过氧化物酶和多酚氧化酶的活性,提高细胞活力,从而促进细胞生长和多糖合成,以2.5g/L浓度的植酸效果最好,培养36d时,类原球茎干重为29.4g/L,多糖产量为2.06g/L。动力学分析表明,添加2.5g/L的植酸还有利于碳、氮、磷等营养物质的吸收。     

基于长江三角洲地表水典型土壤铜、锌环境容量探究

我国水污染日益严重,Ⅰ类地表水地区较少,目前可供饮用的Ⅲ类水成为研究土壤地表水体系污染状况的热点。选取具有代表性的长江三角洲地区4种母质（河相、湖相、海相沉积物和沟沟堆积物）发育而成的土壤,通过加入不同量的外源铜、锌金属培育污染土样,使其分别达到国家土壤环境质量三级标准的0.25、0.5、1.0、1.5、2.0、2.5倍。在此基础上研究雨水和模拟酸雨条件下,对模拟污染的土样进行解吸,并对解吸曲线进行拟合,在基于地表水Ⅲ类水质标准的铜、锌环境容量的基础上,计算了4种土壤在不同情况下的土壤环境容量,并通过探讨国家土壤标准的吻合性,提出针对不同母质的土壤,对其重金属的土壤污染临界值进行适当的调整,以达到Ⅲ类水标准,保障人类的安全与健康。结果表明,在模拟雨水情况下,基于地表水Ⅲ类标准的4种土壤的环境容量分别为：SEBC-06：铜10002mg·kg^-1,锌848.1mg·kg^-1;SEBC-19：铜1162mg·kg^-1,锌612.9mg·kg^-1;SEBC-20：铜9796mg·kg^-1,锌1047mg·kg^-1;SEBC-25：铜720mg·kg^-1,锌180.2mg·kg^-1。     

Effect of salvianolic acid B on vasodilatory function,NF-κB activation and inflammatory cytokine expression in diabetic rats

AIM: To investigate the ameliorative effect of salvianolic acid B on vasodilatory function in diabetic rats and the possible mechanisms. METHODS: SD rats (n=40) were fed on high-sugar and high-fat diet for 4 weeks, followed by a single intraperitoneal injection of streptozotocin (40 mg/kg). The rats with random blood glucose level over 16.7 mmol/L were considered diabetic and randomly allocated to 3 groups, namely model group, low dose (80 mg·kg^-1·d^-1) of salvianolic acid B group and high dose (160 mg·kg^-1·d^-1) of salvianolic acid B group. The rats in salvianolic acid B groups were intragastrically administered with corresponding doses of salvianolic acid B for 6 weeks. Vasodilatory function was measured as endothelium-dependent and-independent vasodilation of the aortic rings. The primary histopathological changes of aorta were observed by HE staining. Serum levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and C-reactive protein (CRP) were measured by ELISA. The levels of total antioxidant capacity, malondialdehyde (MDA) and nitric oxide (NO) in aortic tissues were evaluated by colorimetric assays. The protein levels of intercellular adhesion molecule-1 (ICAM-1) and monocyte chemotactic protein-1 (MCP-1), and the activation of nuclear factor-κB (NF-κB) were determined by Western blot. RESULTS: Treatment with salvianolic acid B evidently ameliorated endothelium-dependent diastolic function and pathological changes of aorta in diabetic rats (P<0.05 or P<0.01). Supplementation with salvianolic acid B resulted in significant increases in NO content and total antioxidant capacity in aortic tissues, accompanied by marked decreases in the level of MDA in aorta tissues and the serum levels of IL-6, TNF-α and CRP (P<0.05 or P<0.01). Salvianolic acid B markedly down-regulated NF-κB p65 nuclear translocation and protein expression of ICAM-1 and MCP-1 in aorta tissues (P<0.05 or P<0.01). CONCLUSION: Salvianolic acid B effectively ameliorates endothelium-dependent diastolic function of aorta in diabetic rats, which might be attributed to suppression of NF-κB activation and subsequent expression of inflammatory cytokines. The beneficial effect of salvianolic acid B on vascular endothelium might be derived from its antioxidant capacity.     

Mechanisms of Aluminum Tolerance in Phosphoenolpyruvate Carboxylase Transgenic Rice

Transgenic rice, in which C4-phosphoenolpyruvate carboxylase (PEPC) is overexpressed, was grown in hydroponic culture to evaluate its tolerance for high aluminum (Al) content in relation to organic acid synthesis and exudation into the rhizosphere. With regard to root growth in short-term experiments, PEPC-transgenic rice was more tolerant of Al than the wild type. At increased Al levels in the root medium, root tips of the transgenic rice accumulated less Al than those of the wild type. Aluminum-induced oxalate exudation from roots occurred at increased rates in the transgenic line. Overexpression of C4-PEPC drastically increased PEPC activity in leaves of the transgenic rice, causing higher organic acid concentrations in leaves and roots. The increased organic acid concentrations seemed to cause higher exudation rates of oxalate from roots, leading to a detoxification of Al in the root apoplast and rhizosphere.     

套袋对鸭梨果实香气及糖酸组分的影响

采用静态顶空和气相色谱-质谱联用技术、高效液相色谱技术,研究了生长期套袋对鸭梨果实挥发性成分、糖酸组分等风味物质的组成和含量的影响。结果表明:套袋导致鸭梨香气物质种类减少和相对百分含量降低。套袋鸭梨采收时测定的香气成分有3类18种,其中酯类15种、烯类2种、醇类1种;不套袋鸭梨采收时测定的香气成分有3类26种,其中酯类23种、烯类2种、醇类1种。鸭梨的主要香气成分是乙酸己酯、己酸乙酯、丁酸乙酯。套袋和不套袋鸭梨均检测到3种糖、4种有机酸组分,其中糖组分主要为果糖,其次为蔗糖、葡萄糖,有机酸组分主要为苹果酸,其次为乙酸、柠檬酸、草酸,但不同处理间含量存在差异。