洋葱AcLOX1基因的克隆及其表达分析

脂氧合酶(lipoxygenase,LOX)是植物十八碳酸代谢途径的关键酶,与种子老化密切相关.洋葱(Allium cepa)种子为短寿命种子.研究LOX基因与洋葱种子发育及老化的关系具有重要的意义.本研究以洋葱授粉后的子房和种子为材料,利用RT-PCR结合快速克隆cDNA末端(rapid-amplification of cDNA ends,RACE)技术,获得AcLOX1 (GenBank登录号:KU363822)基因全长.用生物信息学方法对其基因序列特征进行分析;利用RT-PCR和qRT-PCR分析AcLOX1在洋葱不同部位、种子发育过程以及种子储存过程中的表达模式.结果表明,该基因全长为2 847 bp,包含一个2 619 bp的开放阅读框,编码873个氨基酸.氨基酸序列分析表明,AcLOX1具有PLAT_LH2 (polycystin-1,lipoxygenase,alpha-toxin_lipoxygenase homology)和LOX两种结构域;与桉树(Eucalyptus grandis)、杏(Prunus dulcis)、葡萄(Vitis vinifera)等物种的LOX基因氨基酸序列同源性高达66％～73％.系统进化树分析表明,AcLOX1与其他物种的9-LOX聚在一起.时空表达模式表明,AcLOX1基因在洋葱各组织中均有表达,在授粉后5～25 d随着时间增长其表达量增加,但在授粉后25 d到成熟及储存一年的种子中少量持续表达.本实验初步证实了AcLOX1对洋葱种子发育具有促进作用且与种子老化相关联,为阐明洋葱种子短寿命的作用机制提供了理论依据.     

Effect of storage on fructan and fructooligosaccharide of onion (Allium cepa L.)

The purpose of this study was a comparative examination of the fructan and fructooligosaccharide (FOS) content of different varieties of onions (Allium cepa L. cv. Sturon, Hysam, Durco, Grano de Oro, and Caribo) and the changes produced during their commercial storage. In fresh onions, the Grano de Oro variety presented a remarkably different behavior, showing low contents of total fructans and FOS and high levels of reducing sugars. In the other varieties, Sturon, Hysam, Durco, and Caribo, fructans were the main carbohydrates, the lowest polymerized FOS being the major oligomer. Storage period caused in these varieties important increased levels of free fructose attributed to fructan hydrolysis. Maleic hydrazide treatment had no significant effect in avoiding the hydrolysis of fructans during storage conditions for the Sturon variety. Varieties with >16% dry matter or 15% soluble solids contents could be stored for 6 months at 0 degrees C and 60-65% relative humidity.     

Domestic processing of onion bulbs (Allium cepa) and asparagus spears (Asparagus officinalis): effect on flavonol content and antioxidant status.

Two commonly consumed plant foods, onion bulbs and asparagus spears, were subjected to typical domestic processing, including chopping, maceration, and boiling. The impact of these processes on flavonol content was assessed. Further, the consequences of these processes on the antioxidant capacity of the tissues were evaluated with the beta-carotene bleaching method. Chopping significantly affected rutin content in asparagus, yielding an 18.5% decrease in 60 min; but in onions, quercetin 3,4'-diglucoside (Q(DG)) and quercetin 4'-glucoside (Q(MG)) were virtually unaffected by chopping. Boiling for 60 min had more severe effects, as it caused overall flavonol losses of 20.6 and 43.9% in onions and asparagus, respectively. Chopping of tissues did not considerably influence the antioxidant capacity, but boiling did provoke notable changes.     

Effects of combined application of vermicompost and mineral fertilizer on the growth of Allium cepa L. and soil fertility

The promotion of organic farming involves curtailing extensive use of mineral fertilizers. The present study was aimed to compare the effects of vermicompost (10 Mg ha^–1), commercial mineral fertilizer (NPK—100:80:80), and their combination on (1) the growth of a major cash crop “onion” (Allium cepa L.) and (2) the changes that may have occurred in the amended soil. The experiment was a randomized complete block design with four replications during the crop‐growing season of 2008/09. Results showed significantly higher plant growth in the combined/mix treatment of vermicompost and NPK, as measured by the vegetative growth of bulbs, number and length of tillers per bulb, and fresh weight of bulbs and by the biochemical characteristics of the onion tillers/leaves (total chlorophyll, caretenoids, protein, and total sugar contents). Comparison of the mixed treatment as compared to the control showed increases in bulb size (54%), total number of bulbs per bed (52%), and fresh weight of all bulbs (198%). The chemical properties and enzyme activity of the amended soil also improved significantly in the combined treatment as compared to the application of vermicompost or the mineral fertilizer alone. Total organic C, microbial biomass C, and sulfate content were significantly higher in the mix treatment, with increases of 60%, 127%, and 126%, respectively, as compared to those of the chemical‐fertilizer‐alone treatment. Similarly, β‐glucosidase, alkaline phosphatase, and dehydrogenase were significantly higher by 145%, 91%, 71%, respectively, in the mix treatment as compared to those of fertilizer‐alone application. This study indicates that application of a combination of mineral fertilizer and vermicompost in the field can positively influence the biological properties and fertility of soils, and support better plant growth, when compared to the application of mineral fertilizer or vermicompost alone. The study suggests that this combined application can reduce the quantity and cost of mineral‐fertilizers application for bulbous‐crop cultivation by 50%, while also sustaining soil biological activity of tropical and subtropical soils.     

Modified method for rapid quantitation of S-alk(en)yl-L-cysteine sulfoxide in yellow onions (Allium cepa L.)

In this study, a rapid and sensitive method to determine the flavor precursors was established on the basis of modified Kubec's gas chromatography (GC) method. For the new method, the interference of halogen was removed by adding borohydride that replaced halide as a reducing reagent, and the analysis was completed within 25 min for the determination of S-alk(en)yl-L-cysteine sulfoxides (ACSOs). In comparison with the results of high-performance liquid chromatography (HPLC), the modified GC method is much better in the separation and detection of ACSOs in yellow onion bulbs than the original method. A small amount of 2-propenyl-L-cysteine sulfoxides (2-PeCSO) was detected with the modified GC method, and propyl-L-cysteine sulfoxides (PrCSO) were not detected. The results of GC determination indicated that 2-PeCSO probably was the hydrolysate of a byproduct or an isomer of gamma-glumtyl-1-propenyl-L-cysteine sulfoxides in biosynthesis process.     

Effect of Processing and Cooking Conditions on Onion ( Allium cepa L.) Induced Antiplatelet Activity and Thiosulfinate Content

Allium vegetables serve as sources of antiplatelet agents that may contribute to the prevention of cardiovascular disease. However, onion and garlic, the major Allium species, are usually cooked before consumption. Here, we examined the effect of cooking on onion in vitro antiplatelet activity (IVAA). Two different cooking systems (convection oven and microwaves) and several time-temperature variables were tested on whole bulbs, quarters of bulbs, and completely crushed bulbs, monitoring the degradation of sulfur antiplatelet compounds (e.g., thiosulfinates) by analysis of pyruvate levels. Although heating was, in general, detrimental for onion IVAA, the extent of this effect varied greatly, from unaffected antiplatelet activity (AA) (i.e., similar to raw onion) to a complete lost of activity, depending upon the manner in which onions were prepared prior to heating, the cooking method, and the intensity of the heat treatment. "Whole", "quarters", and "crushed" onions lost their IVAA after 30, 20, and 10 min of oven heating, respectively. The longer retainment of AA in intact bulbs was attributed to a later alliinase inactivation. Proaggregatory effects observed in samples subjected to the most intense oven and microwave heat treatments suggest that extensively cooked onions may stimulate rather than inhibit platelet aggregation. The efficacy of Allium species as antiplatelet agents, as affected by preparation and cooking conditions, is discussed.     

Quantification of Dry-Matter Accumulation and Nutrient Uptake Pattern of Short Day Onion (Allium cepa L.)

Understanding the physiology of dry-matter accumulation and nutrient uptake pattern of an onion crop is an essential criteria for optimizing fertilizer practices. An experiment was conducted to estimate the dry-matter accumulation and nutrient uptake pattern of short day onion. The dry-matter accumulation and nutrient uptake followed the sigmoid growth curve pattern. Uptake of nitrogen and potassium was slow for the first 15 days followed by rapid uptake from 15 to 60 days after transplanting and accounted for 76.2–78.2% of total uptake. Takeup of phosphorus, sulfur, manganese, zinc, and copper uptake was greater during 30 to 75 days after transplanting and accounted for 64.9–70.6% of total uptake. The deficiency of nutrients during these period reduces the crop yield significantly. The dry-matter accumulation and nutrient-uptake pattern reported in this study provide an opportunity for optimizing quantity and timing of nutrient application through fertilizers for short day onions.     

草莓叶主要挥发性物质的测定及其对草莓球腔菌的抑菌效果

为探究草莓叶片中挥发性物质对草莓球腔菌的抑菌作用,本研究测定了草莓叶片中主要挥发性物质,并以草莓球腔菌为处理对象,研究草莓叶片主要挥发性物质对其孢子萌发、菌丝生长以及线粒体膜电位的影响.结果表明,草莓叶片挥发性物质中的萜类化合物主要组分为芳樟醇、桃金娘烯醇,C6醛类化合物主要组分为己醛、2-己烯醛、反式-2-己烯醛、顺...     

盆栽条件下溶磷菌对花生生长的影响

利用本实验室在红壤茶园土中筛选出的高效溶磷菌B1,以红壤为基质进行花生盆栽试验,研究溶磷菌B1对花生生长的影响。试验设置接种溶磷菌(B1,苏云金芽孢杆菌)、直接添加草酸(OA)、磷酸二氢钾(KP)以及空白对照(CK)处理。结果表明溶磷菌在红壤中定殖能力强、存活性较好;与对照相比,接种溶磷菌能显著提高土壤有效磷含量,促进花生根瘤的形成以及地下部生长发育,增加根面积、根体积和根尖数,花生果实的双荚果比例提高5%,百仁重高出3.8 g,果实中粗蛋白的含量增加2.5%。各项指标都显示,接种溶磷菌的效果要显著好于直接添加草酸的效果,而与直接添加可溶性磷源的效果相当。因此开发有效的溶磷菌剂应用于大田生产是可行的。     

Chemical compositions and antioxidant/anti-inflammatory activities of steam distillate from freeze-dried onion ( Allium cepa L.) sprout

Freeze-dried onion sprout was steam-distilled, and the distillate was extracted with dichloromethane (volatile sample). Water sample I was obtained from the residual aqueous solution in the extractor. The filtrate and the methanol extract of filtrand from the residual aqueous solution in the steam distillation flask were named water sample II and methanol sample, respectively. Among the total of 71 components identified in the volatile sample, 24 were sulfur-containing compounds, which comprised 36.87% of the total volatile chemicals identified. The volatile sample inhibited hexanal oxidation for 40 days by >99% at levels >100 microg/mL. The volatile sample and water sample II exhibited moderate antioxidant activity in a malonaldehyde/gas chromatography assay and thiobarbituric acid assay, whereas water sample I did not show appreciable activity. The volatile sample, water sample I, and water sample II exhibited anti-inflammatory activity with a dose-related response in the lipoxygenase inhibitor screening assay. However, the methanol sample did not show appreciable activity in either antioxidant or anti-inflammatory tests. The results suggest that onion sprouts can be an excellent food source.     

Organically fertilized onions (Allium cepa L.): effects of the fertilizer placement method on quercetin content and soil nitrogen dynamics

Field-cured onions cv. Hyskin ( Allium cepa L.) supplied with organic nitrogen fertilizer were studied. The fertilizer was applied by broadcasting and harrowing, broadcasting and rotary cultivation, or placement between rows. Nitrogen dynamics were monitored throughout the growing season by soil sampling. Variation in quercetin content in the onion scales was analyzed by HPLC. The organically fertilized onions were compared with inorganically fertilized onions grown in the same field. Inoculation with arbuscular mycorrhizal fungi (AMF) in the row at sowing or during commercial transplant production was tested but did not significantly affect mycorrhizal root colonization levels in the field. Onions that received no fertilizer at all or that had fertilizer placed between rows had better establishment, probably due to more favorable soil nitrogen concentrations for seedling emergence. Broadcast application led to higher nitrogen concentration in the root zone, resulting in fewer but larger individual onions. Quercetin levels were not significantly altered as a result of nitrogen fertilizer source (inorganic or organic), application method, or mycorrhizal inoculation. However, variation between years was significant, with quercetin levels in 2004 almost twice as high as those in 2005.     

Quantitation of the intense aroma compound 3-mercapto-2-methylpentan-1-ol in raw and processed onions (Allium cepa) of different origins and in other Allium varieties using a stable isotope dilution assay

A stable isotope dilution assay was developed for the quantitation of the potent onion odorant 3-mercapto-2-methylpentan-1-ol (1) using mass chromatography and synthesized [(2)H(2)]-3-mercapto-2-methylpentan-1-ol as the internal standard. Application of the newly developed method on onions from different origins revealed amounts between 8 and 32 microg/kg in raw onions, whereas 34-246 microg was found in sliced, stored (50 min), and then cooked onions. In extracts prepared by simultaneous steam distillation-extraction the highest concentrations of 1 were formed, amounting to >1200 microg/kg. The much higher content of 3-mercapto-2-methylpentan-1-ol in cooked onions suggested its formation from specific, yet unkown, precursors enzymatically formed during cutting of raw onions. 1 was for the first time identified and also quantified in other Allium species such as chives, scallions, and leek, whereas surprisingly garlic and bear's garlic did not contain the aroma compound.     

Comparative investigation of concentrations of major and trace elements in organic and conventional Danish agricultural crops. 1. Onions (Allium cepa Hysam) and peas (Pisum sativum ping pong)

210 samples of onions (Allium cepa Hysam) from 11 conventionally and 10 organically cultivated sites and 190 samples of peas (Pisum sativum Ping Pong) from 10 conventionally and 9 organically cultivated sites in Denmark were collected and analyzed for 63 and 55 major and trace elements, respectively, by high-resolution inductively coupled plasma mass spectrometry. Sampling, sample preparation, and analysis of the samples were performed under carefully controlled contamination-free conditions. Comparative statistical tests of the element concentration mean values for each site show significantly (p < 0.05) different levels of Ca, Mg, B, Bi, Dy, Eu, Gd, Lu, Rb, Sb, Se, Sr, Ti, U, and Y between the organically and conventionally grown onions and significantly (p < 0.05) different levels of P, Gd, and Ti between the organically and conventionally grown peas. Principal component analysis (PCA) applied to the 63 elements measured in the individual onion samples from the 21 sites split up the sites into two groups according to the cultivation method when the scores of the first and third principal components were plotted against each other. Correspondingly, for peas, a PCA applied to the 55 elements measured as mean values for each site split up the 19 sites into two groups according to the cultivation method when the scores of the third and fourth principal component were plotted against each other. The methodology may be used as authenticity control for organic cultivation after further method development.     

牛羊胎盘免疫调节因子的提取及对PANC-Ⅰ增殖和迁移影响的比较

胎盘免疫调节因子(placenta immunoregulating factor,PF)是从胎盘提取的小分子多肽,对病毒性疾病、免疫缺陷疾病及恶性肿瘤等具有潜在的临床应用价值.为检测牛(Bos taurus)和绵羊(Ovis aries)胎盘免疫调节因对人(Homo sapiens)胰腺癌细胞(pancreatic cancer cells-Ⅰ,PANC-Ⅰ)增殖与迁移的影响,本研究对从屠宰场获得的牛和绵羊胎盘进行分离,利用双酶水解法制备PF,将质量浓度为100～2 000 ng/mL的牛、羊PF作用于体外培养的PANC-Ⅰ细胞,以3-(4,5-二甲基噻唑-2-基)-5-(3-羧甲酯基)-2-(4-磺苯基)-2H-四唑(3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium,MTS)法观察细胞增殖抑制率,应用流式细胞技术检测其对PANC-Ⅰ细胞周期的影响.并利用Transwell迁移实验检测添加PF对PANC-Ⅰ细胞迁移能力的影响,同时通过实时荧光定量PCR法检测肿瘤抑制基因(tumor suppressor gene,PS3)和细胞周期依赖性蛋白激酶抑制因子1A基因(cyclin-dependent kinaseinhibitor 1A,P21)的表达情况.结果显示,17.11 g绵羊胎盘制得0.588 mg PF,6.61 g牛胎盘制得0.312 mgPF,胎盘质量与所得PF的质量比约为20 000∶1.在体外培养的PANC-Ⅰ细胞培养液中添加PF,从形态上观察,牛羊PF对PANC-Ⅰ细胞没有明显的影响.绵羊PF处理后PANC-Ⅰ细胞增殖抑制率为9.75％～22.89％,牛PF处理后PANC-Ⅰ细胞增殖抑制率为-3.81％～23.56％,增殖抑制率最显著的绵羊PF浓度为500 ng/mL,牛PF浓度为1 000 ng/mL.绵羊PF在浓度为500 ng/mL时,PANC-Ⅰ细胞的PI和SPF最低,而牛PF各浓度下的胰腺癌细胞增殖指数(proliferation index,PI)和增殖活性(S-phase fraction,SPF)没有明显的变化.同时,牛羊PF对PANC-Ⅰ细胞的迁移能力无显著影响.牛羊PF使PANC-Ⅰ细胞P53表达上调而P21表达下调.上述结果表明,在一定浓度下,PF对体外培养的PANC-Ⅰ细胞增殖有负调控作用,且牛羊胎盘免疫调节因子对胰腺癌细胞的影响不同,可能因为其存在的活性成分有区别.本研究为胰腺癌的治疗提供了新的思路.     

氮、磷、钾肥对大葱商品性状及其产量的影响

应用311-A拟饱和最优回归设计,通过氮、磷、钾肥配比田间试验,建立大葱施用氮、磷、钾肥对大葱株高、葱白直径、产量及施肥利润的效应函数。结果表明,株高极大值氮、磷、钾优化施肥组合为:N 371.30、P2O5 157.50、K2O 309.58 kg/hm2,此时的株高为92.88 cm;葱白直径极大值氮、磷、钾优化施肥组合为:N 350.63、P2O5 157.50、K2O 225.00 kg/hm2,此时葱白的直径为1.769 cm;最高产量的氮、磷、钾优化施肥组合为:N 394.08、P2O5 193.62、K2O 225.00 kg/hm2,此时产量为55805.06 kg/hm2;最佳施肥利润的氮、磷、钾优化施肥组合为:N 391.35、P2O5 192.97、K2O 225.00 kg/hm2,其最佳施肥利润达136865.60元/hm2,经济产量达55802.74 kg/hm2。     

Growth,yield and bulb quality of onion (Allium cepa L.) in response to foliar application of boron and zinc

A field experiment was conducted to investigate the role of boron and zinc on growth, yield and quality of onion. There were eight treatment combinations consisted of four levels of boron (0, 0.1, 0.2 and 0.5%) and 4 levels of zinc (0, 0.1, 0.2 and 0.5%) were applied as foliar spraying. application of 0.5% boron significantly increased the growth (plant height, 63.93cm and number of leaves per plant, 7.25), yield (30.74 t ha^?1) and quality (total soluble solids, 13.45 ⁰B and pyruvic acid 5.94 µmol g^?1) of onion. Among various levels of zinc 0.5% exhibited the best growth (plant height, 67.25cm and number of leaf per plant, 7.75), yield (33.34 t ha^?1) and quality (total soluble solids, 14.57 ⁰B and pyruvic acid, 5.86 µmol g^?1) attributes of onion. These results suggest that the foliar application of boron and zinc significantly influenced the growth, yield and quality of onion.     

菌糠多糖-纳米硒复合物的制备及其对水稻促生效果的研究

为探究菌糠多糖-纳米硒复合物对水稻的促生效果,以茶树菇菌糠多糖为原料,采用抗坏血酸为还原剂制备菌糠多糖-纳米硒复合物,并初步探索其对水稻种子萌发及水稻幼苗生长的影响。结果表明,当茶树菇菌糠多糖占比为10%、Vc与亚硒酸钠的配比为4∶1、反应时间为4 h、反应温度为50℃时,可以获得粒径最小且稳定的菌糠多糖-纳米硒复合物。该条件制备的茶树菇菌糠多糖-纳米硒复合物有一定的体外抗氧化能力,对水稻生长具有明显促进作用,且其促生效果高于单一的菌糠多糖,其中对水稻根部发育的影响尤为显著。进一步研究表明,当水稻幼苗叶面喷施菌糠多糖-纳米硒复合物浓度为20 mg·L^-1时,根和叶中可溶性蛋白含量比对照分别提高了72.62%和27.80%,叶中可溶性糖含量增加了180.87%,叶绿素a、叶绿素b、类胡萝卜素含量分别增加了67%、58%、21%。叶面喷施对水稻的促生效果显著高于根部添加处理。本研究为菌糠多糖在农业领域的推广应用提供了理论与实践参考,同时也为利用外源性物质提高农作物生长提供了依据。     

长期施肥对旱地土壤有机磷及其组分的影响

通过15年长期定位试验,采用Bowman和Cole提出的土壤有机磷分级方法,研究了不同耕作施肥措施对旱地耕层土壤有机磷及其组分的影响。结果显示旱地土壤各个处理的活性有机磷含量低,占有机磷总量的比例低于3.6%,中活性有机磷为有机磷的主体,约占有机磷总量的73.37%-91.54%。休闲、撂荒及磷肥与其它肥料配合使用,均增加了耕层土壤的中活性有机磷含量,增加了中稳性有机磷含量及其比重;休闲、撂荒及磷钾提高了高稳性有机磷含量及比重,而氮磷钾及其与有机肥配合处理则降低了高稳性有机磷的含量。尽管不同耕作施肥管理均影响旱地土壤有机磷含量,但是施用有机肥对有机磷的增加贡献最大;撂荒与休闲相比,前者更有利于土壤有机磷的积累。