Selenium distribution and nitrate metabolism in hydroponic lettuce(Lactuca sativa L.): Effects of selenium forms and light spectra

A deficiency in selenium(Se) in the human diet is a worldwide problem. The intake of Se-rich vegetables can be a safe way to combat Se deficiency for humans. However, most leafy vegetables can accumulate a high content of nitrates, which poses a potential threat to human health. Light is an important environmental factor that regulates the uptake and distribution of mineral elements and nitrogen metabolism in plants. However, the effects of Se forms and light conditions, especially light spectra, on the uptake and translocation of Se and on nitrate reduction are poorly understood. In this study, lettuce(Lactuca sativa L.) was treated with exogenous Se applied as selenate(10 mmol L^-1) and selenite(0.5 mmol L^-1) and grown under five different light spectra: fluorescent light(FL), monochromatic red LED light(R), monochromatic blue LED light(B), and mixed red and blue LED light with a red to blue light ratio at 4(R/B=4), 8(R/B=8), and 12(R/B=12), respectively. The effects of light spectra and Se forms on plant growth, photosynthetic performance, Se accumulation and nitrate reduction were investigated. The results showed that the light spectra and Se forms had significant interactions for plant growth, foliar Se accumulation and nitrate reduction. The Se concentration and nitrate content in the leaves were negatively correlated with the percentage of red light from the light sources. Compared to Se applied as selenite, exogenous Se applied as selenate was more effective in reducing nitrate via promoting nitrate reductase and glutamate synthase activities. The lowest nitrate content and highest plant biomass were observed under R/B=8 for both the selenate and selenite treatments. The significant effect of the light spectra on the root concentration factor and translocation factor of Se resulted in marked variations in the Se concentrations in the roots and leaves. Compared with FL, red and blue LED light led to significant decreases in the foliar Se concentration. The results from this study suggest that the light spectra can contribute to Se distribution and accumulation to produce vegetables with better food quality.

Effects of Priming in Combination with Fungicides on Germination and Infestation of Lettuce (Lactuca sativa L.)

Two seed lots of lettuce (Lactuca sativa L.) cv. Krlowa Majowych were primed in polyethylene glycol (PEG) 8000 at -1.25MPa for 7 days in dark at 20C. Two fungicide treatments combined with PEG were used to control fungi: 1) Funaben T(0.1%) and Apron 35 SD (0.02%) were added to -1.25 MPa PEG solution (PEG+F); 2) After seed priming and drying,Funaben T was mixed at 5 g kg-1 and Apron 35 SD at 1 g kg-1 of seeds (PEG then F dust). PEG, PEG+F and PEG then F dusttreatments reduced significantly thermodormancy and increased the seed germination at 35C in both lots. PEG then Fdust treatment had the best effect on reducing seed thermodormancy at 35C, and increased seed germination to 64.5% inlot 1 and 72.0% in lot 2 compared to 0 in untreated. PEG and PEG then F dust treatments have accelerated germinationspeed at 20 and 15C in both lots. There was a significant reduction in seed infestation in which the seeds were treatedwith PEG+F and PEG then F dust compared to untreated seeds and PEG primed seeds. PEG priming enhanced seedinfestation considerably by Alternaria alternata, Penicillium spp., Rhizopus spp. and Ulocladium spp. Mycologicalanalysis showed higher frequency of Alternaria alternata and Ulocladium spp. in untreated seeds at 20C. Priming incombination with fungicides significantly alleviated the infestation of fungi, especially Alternaria alternata infestation.

Bolting and Flowering Response of <i>Lactuca georgica</i>, a Wild Lettuce Relative, to Low Temperatures

To learn about the phenological adaptation of Lactuca georgica Grossh., a wild relative of domesticated lettuce, we studied seed sampled accessions obtained from individual plants at 19 locations throughout six regions in Armenia, and from two natural populations in Dagestan (Russian Federation) collected as bulk samples. The effects of various vernalization treatments on time to bolting, flowering and seed production time were investigated during four successive years at different growth stages of L. georgica plants. We demonstrate that low temperatures play a major role in stimulating the reproduction process of L. georgica plants. Our results would suggest that for L. georgica: 1) There is an obligatory (or nearly so) vernalization requirement;2) Plant age, vernalization duration, and genotype of original sample have a role in bolting and flowering regulation;3) Some plants behaved as typical annuals, responding to vernalization treatment at the seedling stage, but, most did not;4) Four months of vernalization could be adequate to reach bolting in plants with a developed vegetative rosette, for most—but not all—samples;5) In order to find the best solution for stimulating the reproductive process of multiple genotypes, it seems that further study should focus on about 4 - 6 months of vernalization at 4°C applied to plants of about 10 - 22 months old vegetative rosettes, with controlled post-vernalization condition;6) L. georgica germplasm could be used as a source for delayed bolting in breeding of domesticated lettuce varieties.

TDZ对莴苣(Lactuca sativa L.)器官发生及乙烯形成的影响

为了研究TDZ在植物器官发生中的生理作用，用不同浓度的TDZ替代MS中的BA，发现仅用1／50BA浓度（0．1mg／L）的TDZ处理，已相当于用BA处理所得芽数，可见它具有很强的细胞分裂素活性；但是TDZ又能显著促进乙烯形成，较低浓度时抑制莴苣的器官发生，可能此时乙烯起着主导作用。

生菜(Lactuca sativa var ro mana Hort.)下胚轴愈伤组织的诱导与植株再生

通过对生菜下胚轴的愈伤诱导和植株再生的研究 ,发现诱导愈伤组织时 ,MS培养基中加 0 .5mg/L 6 BA + 1mg/L 2 ,4 D + 0 .1mg/LNAA效果最好 ,其出愈率达 94 %;愈伤组织分化为芽时 ,在MS + 1.0mg/L 6 BA + 0 .0 5mg/L NAA中分化频率最高 ,达 10 0 %.再生苗可在 1/ 2MS+ 0 .5mg/L NAA中诱导生根成完整植株 .

Two new eudesmane sesquiterpenes from Lactuca sativa var.anagustata L

Two new eudesmane sesquiterpene lactones were isolated from the stalk of Lactuca sativa vat anagustata L and their structures were elucidated by means of spectroscopic methods, including 2D NMR （1H-1H COSY, HMBC and NOESY） as 1β-O-β-D- glucopyranosyl-4α-hydroxyl-5α, 6β, 11βH-eudesma-12, 6α-olide （1） and 1β-hydroxyl-15-O-（p-methoxyphenylacetyl）-5α, 6β, 11 βH-eudesma-3-en- 12, 6a-olide （2）.

Reduced nitrogen availability during growth improves quality in red oak lettuce leaves by minimizing nitrate content, and increasing antioxidant capacity and leaf mineral content

Overuse of N in lettuce production can lead to environmental problems caused by leaching and the accumulation of harmful nitrates in edible tissues. This study investigated the effect of applied nitrogen (N) concentrations between 40 and 2400 mg·L–1 on growth, nitrate accumulation, mineral leaf content, and antioxidant capacity in Oak Leaf lettuce cv. “Shiraz” grown under hydroponic conditions in Australia. Yield (g FW) increased with nitrogen (N) application rate up to 1200 mg·L–1, as did leaf N content, while C:N declined. Nitrogen Utilization Efficiency (NUtE) increased rapidly from 40 to 75 mg·L–1 applied N, leveling at 150 mg·L–1 with no subsequent effect of N concentrations between 400 and 2400 mg·L–1. Nitrate content rose significantly with increased N, particularly at 1200 and 2400 mg·L–1. Leaf total plant phenolic content (TPP) and antioxidant capacity (measured by ferric reducing antioxidant power—FRAP) were both maximal at 75 and 400 mg·L–1 applied N, while highest oxygen radical absorption capacity (ORAC) values were found in leaves supplied with low N (40 to 400 mg·L–1). Applied N as calcium nitrate also significantly affected leaf mineral content as B, Mg, Mn, and Zn significantly decreased with increasing N. These results indicate that N applications of 1200 mg·L–1 or higher can result in reduced antioxidant capacity and mineral content in lettuce leaves.

Lettuce production according to different sources of organic matter and soil cover

Lettuce (Lactuca sativa L.) is one of the leading vegetables produced by farmers in the Itabaiana region, and thus it is important to study means of sustainable management for the crop. The objective of this work was to evaluate the behavior of lettuce with different sources of organic matter and soil cover. Therefore, it was chosen to cultivate Veronica. The experiment was carried out in the experimental field plot of Project Small Farmer—Great Entrepreneur in the municipality of Itabaiana-SE, in 2010. The design was a randomized block design with four replications, with treatments consisting of five sources of organic matter (organic compost, chicken manure, cattle manure, sheep manure, and castor bean) and a control (no fertilizer), all with and without mulch, totaling 12 treatments. We evaluated the characteristics of plant diameter, plant height, fresh and dry root and shoot matter. We also evaluated the influence on the temperature and soil water tension. All productivity characteristics valuated showed the significant interaction between the type of fertilizer and the mulch, except for root dry weight, where only organic manure was a significant factor. When soil mulch was used, the best source of organic matter for growing lettuce was chicken manure, and in the impossibility of using this cover, we recommend the use of organic compost as a source of organic matter, as it showed the best result in the absence of coverage. The cover age also showed good results in relation to a temperature that promoted lower soil water tensions.

An Improved Method for Fractionation of Small Quantities of Lettuce Latex

While large quantities of latex can be handled either by standard extraction techniques such as Soxhlet extraction or accelerated solvent extraction (ASE), smaller samples on the order of 0.3 - 0.5 g require handling on a microscale. We collected latex from lettuce plants in microcentrifuge tubes and, after drying under vacuum, resuspended the dried sample in acetone by holding in an ultrasonic cleaner. The resulting fine suspension was readily extracted with acetone and toluene to provide fractions representing the resin and rubber content of the latex. Using this approach, we compared latex from stems of bolting lettuce and from the floral stem of lettuce plants. While both types of stems contained a similar percentage of resin, the rubber content of the bolting stems exceeded that of the floral stems.

Construction of Lactuca sativa Plastid Transformation Vector and High-level Expression of gfp Gene in Escherichia coli

Using genomic DNA of bolting-tolerant lettuce as a template,flanking fragments of lettuce plastid rpo A gene were amplified and cloned by PCR. Targeting the sites of these two fragments,homologous recombinant fragments of exogenous gene were integrated to construct lettuce plastid expression vector p Brpo AGFP,which harbored the expression cassette Prrn-gfp-aad A-Tpsb A. The results showed that the amplified flanking fragments were 1.2 and 1.1 kb in size. After sequencing,restriction digestion,ligation and transformation,lettuce plastid expression vector containing expression cassette Prrn-gfp-aad A-Tpsb A was constructed and confirmed by SDS-PAGE electrophoresis. The results of SDS-PAGE electrophoresis indicated that gfp gene was efficiently expressed under the regulation of plasmid specific promoter Prrn and terminator Tpsb A. GFP accounted for 45. 6% of total soluble proteins; inclusion bodies accounted for 47.5 % of bacterial proteins,which reached relatively high expression levels. The construction of lettuce plastid expression vector p Brpo A-GFP laid a solid foundation for establishment of subsequent lettuce plastid transformation system and genetic improvement of lettuce using various functional genes.

Production of Lettuce Seedlings with Association of Diazotrophic Bacteria and Humic Acids

The objective of this work was to verify the responses in agronomic and physiological characters of lettuce seedlings when submitted to different doses of humic acids and the presence or absence of inoculation of the bacterium Azospirillum brasilense. The experiment was carried out at the Protected Cultivation Station and Biological Control Teacher Mário César Lopes, belonging to the Experimental Stations Nucleus of the State University of the West of Paraná. The experimental design was a randomized block design, in a 5 × 2 factorial scheme, with four repetitions, totaling 40 plots. The first factor consisted of five solutions containing humic acid (100%, 75%, 50%, 25% and 0%) and the second factor was the presence or absence of the diazotrophic bacterium Azospirillum brasilense obtained by commercial product. Germination tests were carried out in addition to the determination of agronomic characters such as the number of fully expanded leaves, plant height, stem diameter, main root length, shoot dry mass, root dry mass, total dry mass, and Dickson quality index. The association of diazotrophic bacteria and humic acids did not influence the emergency rate, mean emergence time, mean speed of emergency, emergence speed index, plant height, and root length of lettuce seedlings. The dilutions of humic acids, regardless of inoculation with bacteria diazotrophic favored the development of the number of leaves, stem diameter and Dickson quality index. Inoculation of lettuce seeds with diazotrophic bacteria associated with the addition of humic acids, favored the development of the dry mass of root and total dry matter.

不同LED组合光质对莴苣灰霉病防御酶活性的影响

本试验在水培条件下,以LED红蓝光(红蓝光比例4∶1)为基础和对照,研究添加紫光、黄光、绿光组成的不同LED组合光质对叶用莴苣接种灰霉病菌后的病情指数以及接菌前后POD、PPO、PAL等相关防御酶活性的影响。结果表明,不同LED组合光质处理中,以添加紫光的LED红蓝紫处理最能降低莴苣灰霉病病情指数;防御酶活性上,红蓝光基础上添加紫光和黄光处理的POD、PAL、PPO、GLU活性较高,添加绿光、黄光处理的CHT活性较高。整体上看,在LED红蓝光基础上添加一定比例的紫光或黄光,不仅可以有效降低莴苣灰霉病的病情指数,还可以提高莴苣体内的防御酶活性。该结果可为莴苣生产中通过光环境控制来抑制灰霉病发生提供理论参考。

叶用莴苣热激蛋白基因LsHsp70-2711的克隆及高温胁迫下的功能分析

【目的】通过克隆Hsp70相关基因,并利用VIGS分析叶用莴苣热胁迫下Hsp70表达量和形态变化,为解析热激蛋白基因Hsp70在高温胁迫下的响应机制及分子机理奠定基础。【方法】通过同源克隆及RACE技术,获得叶用莴苣热激蛋白LsHsp70-2711的cDNA全长序列,利用实时荧光定量PCR(qRT-PCR)分析该基因在不同温度和不同高温时间下的叶用莴苣热敏品种‘P-S11’和耐热品种‘G-S59’的表达差异,确定基因和高温的相关性。根据VIGS技术,构建p TRV-LsHsp702711瞬时沉默载体,转化农杆菌GV1301,注射法侵染叶用莴苣叶片,三周后经PCR鉴定得到阳性植株。对照组和阳性组在基因表达和形态上进行对比,对照植株和阳性植株热胁迫和干旱处理后再次分析LsHsp70-2711的表达特性,观测形态变化。【结果】LsHsp70-2711 cDNA全长为2 226 bp,开放阅读框为2 154 bp,编码718个氨基酸,与拟南芥(NP_187864.1)、番茄(NP_001266213.1)、水母雪莲花(AAB99745.1)等物种的Hsp70同源性达到80%以上,证明此基因属于Hsp70家族。根据qRT-PCR结果,高温胁迫下该基因在两个品种中的表达均上调,在耐热品种中总体表达水平均高于热敏品种,耐热品种LsHsp70-2711的表达量最大值出现在42℃、60 min,37℃、60 min时热敏品种基因表达量达最大值,且在42℃高温下热敏品种‘P-S11’中基因表达随着胁迫时间的增加受到抑制,而耐热品种‘G-S59’则能长时间保持较高的表达水平,此结果和田间两品种间耐热差异表现相符合。亚细胞定位显示,LsHsp70-2711主要在细胞质中。将构建好的载体侵入叶用莴苣,鉴定后获得阳性植株。由定量PCR结果可知,未进行胁迫处理的阳性植株与对照株相比,LsHsp70-2711表达量下降,茎长明显增长。热胁迫和干旱处理后的阳性植株LsHsp70-2711表达量显著低于对照植株,高温处理对LsHsp70-2711的影响大于干旱胁迫。【结论】LsHsp70-2711属于Hsp70基因家族,其与叶用莴苣耐热性相关。研究结果为解析叶用莴苣热激蛋白LsHsp70-2711在叶用莴苣高温抽薹方面的功能提供了理论支持。

不同红蓝LED光照强度对叶用莴苣生长和品质的影响

以叶用莴苣品种永荣为试验材料,探讨了光照强度为100μmol·m-2·s-1(RB100)、200μmol·m-2·s-1(RB200)和300μmol·m-2·s-1(RB300)的红蓝LED光源对叶用莴苣生长与品质的影响,结果显示:与其他处理相比,RB100处理叶用莴苣植株高度、叶面积、根长、比叶面积等形态指标及可溶性蛋白含量显著增加;RB300处理叶用莴苣中VC含量最高,硝酸盐和叶绿素含量最低。结果表明:300μmol·m-2·s-1光照强度有利于提高叶用莴苣的品质,而100μmol·m-2·s-1光照强度有利于叶用莴苣的生长。

叶用莴苣热激蛋白90(LsHsp90)基因的克隆及其在热激下的表达

【目的】克隆叶用莴苣热激蛋白Hsp90基因并探讨其响应热胁迫的相关机制,为揭示叶用莴苣抗热分子机制奠定理论基础。【方法】采用同源克隆和RACE技术相结合,从叶用莴苣叶片总RNA中克隆LsHsp90 cDNA序列。通过实时荧光定量PCR(qRT-PCR)分析LsHsp90在叶用莴苣耐热品种Z36和热敏品种S106受37℃和42℃热激后叶片的表达差异。【结果】该基因cDNA序列全长2 330 bp,开放阅读框2 097 bp,编码698个氨基酸,推导的蛋白质分子量约79.8 kD。蛋白质结构预测及同源比对分析表明,LsHsp90基因编码蛋白含ATPase位点和Hsp90保守结构域,并与拟南芥(AY081302.1)、紫茎泽兰(EU269070.1)等多种物种的热激蛋白90高度同源;进化树分析表明,LsHsp90与紫茎泽兰Hsp90基因(EU269070.1)聚为一类。qRT-PCR分析表明,37℃热胁迫下,该基因在耐热品种和热敏品种的叶片中均上调表达;42℃高温胁迫下,热敏品种S106中下调表达,而耐热品种Z36上调表达。【结论】成功从叶用莴苣叶片中分离克隆到LsHsp90,该基因具有已知物种Hsp90基因的特征,该基因在热胁迫下有不同的表达特征,预示该基因其可能在抗高温胁迫中起重要作用。

生菜低温胁迫转录因子LsICE1的克隆、特征分析及其转化水稻

【目的】克隆生菜(Lactuca sativa L.)低温胁迫转录因子LsICE1,对其进行序列分析和水稻遗传转化,研究超表达LsICE1基因对水稻耐低温能力的影响。【方法】设计简并引物,利用RT-PCR技术获得生菜LsICE1基因保守区域,再通过SON-PCR技术获得LsICE1基因的5′端和3′端,拼接得到全长的cDNA。对该序列进行生物信息学分析,采用半定量RT-PCR研究LsICE1的低温表达模式。最后构建植物表达载体,利用农杆菌介导法对水稻进行遗传转化。通过比较低温处理后对照和转基因株系的存活率和生理指标,鉴定超表达LsICE1基因对水稻耐低温能力的调控作用。【结果】测序结果显示,拼接后的cDNA片段长1 622 bp,包含一个1 497 bp完整的开放阅读框,编码498个氨基酸残基,命名为LsICE1,GenBank登录号为HQ848932。半定量RT-PCR研究表明,LsICE1基因是冷诱导条件下差异表达的基因。进化树分析表明,LsICE1蛋白与葡萄的ICE1蛋白亲缘关系最近,处于同一进化分枝。PCR和RT-PCR分子检测证明,LsICE1基因已经整合到水稻基因组中。与对照相比,低温处理后超表达LsICE1基因的转基因株系存活率和脯氨酸含量明显增加,相对电导率和丙二醛含量积累速率明显下降。【结论】首次从生菜中克隆了低温胁迫转录因子LsICE1,超表达LsICE1基因水稻株系提高了抗低温胁迫能力。

叶用莴苣热激蛋白LsHsp70-3701基因的克隆及高温胁迫下的表达分析

为探讨热激蛋白Hsp70基因在高温胁迫下的响应机制及其分子机理,分析Hsp70基因与叶用莴苣耐热性的关系,本研究通过同源克隆及RACE技术,克隆了叶用莴苣热激蛋白Ls Hsp70-3701基因的c DNA全长序列,利用实时荧光定量PCR(qRT-PCR)分析该基因在叶用莴苣热敏品种P-S11和耐热品种G-S59中高温胁迫下的表达差异。该基因c DNA全长为2 191 bp,开放阅读框为1 950 bp,编码649个氨基酸,具有Hsp70家族特有的标签序列,与拟南芥、番茄、小麦等物种的Hsp70同源性达到90%以上。qRT-PCR分析显示,高温胁迫下该基因在2个品种中的表达均上调,在耐热品种中总体表达水平显著高于热敏品种,且在42℃高温下热敏品种P-S11中基因表达随着胁迫时间的增加受到抑制,而耐热品种G-S59中则能长时间保持较高的表达水平。结果表明,Ls Hsp70-3701基因可能与叶用莴苣耐热性相关。本研究为有效解决叶用莴苣高温抽薹问题提供了数据支持。

叶用莴苣热激蛋白LsHsp70-3701和LsHsp70-1707基因与抽薹特性的表达分析

[目的]探讨叶用莴苣热激蛋白Ls Hsp70-3701和Ls Hsp70-1707基因在抽薹前后的表达水平,分析Hsp70基因与叶用莴苣抽薹的相关性。[方法]采用实时荧光定量方法检测Ls Hsp70-3701和Ls Hsp70-1707基因在不同品种中的相对表达量,分析其差异性。[结果]Ls Hsp70-3701基因在热敏品种P-S11抽薹期的表达量显著高于抽薹前;在耐热品种G-S59抽薹初期的表达量与抽薹前无显著差异,但后期表达量显著降低。Ls Hsp70-1707基因在热敏品种P-S11抽薹初期表达量与抽薹前差异不显著,但其后表达水平显著高于抽薹前;在GS59中抽薹期的最大表达量低于抽薹前的最大表达量,但两者差异不显著,而其他时间的相对表达量却显著低于抽薹前最高水平。[结论]Ls Hsp70-3701和Ls Hsp70-1707基因与叶用莴苣抽薹具有一定的相关性,叶用莴苣抽薹是差异基因表达的结果。

共转化Atpsy和folE基因提高生菜类胡萝卜素和叶酸含量

通过农杆菌介导的遗传转化法将拟南芥八氢番茄红素合成酶基因(phytoene synthase,Atpsy)和按照拟南芥偏爱密码子人工合成的编码三磷酸腺苷环化水解酶(GTP cyclohydrolase,GCHI)的folE基因共同转入罗曼生菜中,通过Realtime-PCR、HPLC、干酪乳杆菌发酵法分别测定了转基因生菜中目标基因的表达量、叶黄素和β-胡萝卜素含量以及总叶酸含量。实验结果表明:转基因生菜中Atpsy基因表达量为野生型的43.54倍,folE基因表达量为野生型的38.05倍;转基因生菜植株叶黄素和β-胡萝卜素含量与野生型相比均有提高,含量最好的转基因植株L13中叶黄素含量为7.1mg/kg(鲜重,下同),是野生型对照6.8mg/kg的1.1倍,β-胡萝卜素含量为592.6mg/kg,是野生型对照196.9 mg/kg的3倍;叶酸含量最高是2 083.4 mg/kg,为野生型对照1 129.1mg/kg的1.85倍。该研究为通过基因工程手段改良类胡萝卜素和叶酸在植物体内的代谢途径,获得同时富含类胡萝卜素和叶酸的新型生菜品种打下了基础。