UV-B irradiation enhances the accumulation of beneficial glucosinolates induced by melatonin in Chinese kale sprout

Cruciferous sprout is a new form of vegetable product rich in bioactive compounds,especially glucosinolates.Previous studies have focused on increasing the accumulation of glucosinolates in cruciferous sprouts by applying different chemical regulators,with a particular focus on their contribution to nutritional quality and health benefits.Nevertheless,the effects of melatonin and UV-B irradiation on glucosinolate biosynthesis remain unclear.In this study,it was found that changes in melatonin concentrations significantly affected the contents of individual as well as total aliphatic and indolic glucosinolates.The 5μmol·L^(-1)melatonin was decided as the optimum concentration that could increase the content of beneficial glucosinolates including glucoraphanin and 4-methoxy glucobrassicin in Chinese kale sprouts.Notably,the enhancement of glucosinolate accumulation by melatonin treatment could be further amplified by UV-B irradiation.Furthermore,our results showed that R2R3-MYB transcription factor BoaMYB28 and BoaMYB51,which are central regulators of aliphatic and indolic glucosinolate biosynthesis respectively,were both involved in the regulation of glucosinolate biosynthesis by melatonin and UV-B irradiation.Additionally,the expression of glucosinolate biosynthetic genes,including BoaCYP79F1,BoaCYP83A1,BoaSUR1,BoaUGT74B1,BoaCYP79B2,BoaCYP79B3,and BoaCYP83B1 participated in the formation of core structures and BoaFMOGS-OX5,BoaAOP2,BoaCYP81F2,and BoaIGMT1 involved in the sidechain modification of aliphatic and indolic glucosinolate,was regulated by melatonin or UV-B irradiation.Taken together,these findings provide a potential strategy for improving the nutritional quality and resistance of Chinese kale sprouts.

Effects of fresh-cut and storage on glucosinolates profile using broccoli as a case study

Glucosinolates(GLS) contribute to the unique flavour, nutrition, and plant defence of the Cruciferous vegetables. Understanding the GLS changes through postharvest processing is essential for defined preservation. In this study, four different fresh-cut types, whole flower(W),floret(F), quarterly cut floret(QF) and shredded floret(FS) of broccoli, were stored for 0, 1, 2 and 3 day(s) to explore GLS responses to postharvest treatments. As a result, seven GLS were identified, mainly including glucoraphanin(RAA), neoglucobrassicin(NEO), and glucobrassicin(GBC)and accounting for 52.69%, 20.12% and 14.99% of the total GLS(21.92 ± 0.48) μmol · g ^(-1 )DW, respectively. FS had the sharpest decrease in GLS after three days of storage(6.55 ± 0.37) μmol · g-1DW, while QF had the least(10.16 ± 0.33) μmol · g ^(-1 )DW. All GLS components decreased over storage, except for 4-methoxyglucobrassicin(4 ME) in FS and QF, suggesting its key role in serious wound defence. The results suggested certain postharvest approaches influenced the flavour and nutrition of broccoli.

Diversity of glucosinolates among common Brassicaceae vegetables in China

Brassicaceae vegetables are an important traditional daily food in China and around the world, which provide nutrients and phytochemicals that are beneficial for human health. Among them, Brassica and Raphanus are widely cultivated and eaten, have been evolved and/or bred for special characteristics during the long history of cultivation. Epidemiological studies suggest that the health benefits of Brassicaceae vegetables are mainly associated with glucosinolates(GSLs) and their hydrolytic products. In this review, we discuss the diversity of common consumed Brassicaceae vegetables and their GSL composition in edible parts. We also discuss the diversity factors affecting GSL content, and the diversity roles and functions of GSL. The information in this review provides guidance for consumers to select vegetables with a high GSL content,optimum edible stages, suitable edible methods, and provides a theoretical basis for crop molecular breeding and market development of GSL products.

Glucosinolates and Their Hydrolysis Products in Arabidopsis thaliana Influence Performance and Feeding Choice of Pieris rapae and Spodoptera exigua

Glucosinolates and their hydrolysis products, found in plants of the order Brassicales, are well-known for their defensive properties against insect herbivores. Arabidopsis thaliana (Col-0) genetic lines with mutations that modify the type of glucosinolates (i.e. myb28myb29 and cyp79B2cyp79B3 are deficient in the production of aliphatic and indolyl glucosinolates, respectively) make it possible to test for the specific effects of these secondary chemicals on insect herbivores. The Pad3 mutant (deficient in camalexin), which has a role in resistance to pathogens, was also tested. Likewise, the effects of different glucosinolate hydrolysis products can be evaluated using genetically modified (GM) lines of the wild type Col-0 ecotype, which naturally produces isothiocyanates. These GM lines include the nitrile-producing 35S: ESP and the double knockout tgg1tgg2, which virtually lacks hydrolysis products. In both no-choice and choice experiments, the crucifer specialist Pieris rapae was virtually unaffected by differences in the type of glucosinolates or hydrolysis products. In contrast, the generalist insect Spodoptera exigua had statistically significant increases in pupae/adult weight and faster developmental times when reared on mutants deficient in the production of aliphatic and indolyl glucosinolates and their hydrolysis products. There were no differences in the performance of either insect species when reared on wild type Col-0 or Pad3. Results from feeding choice trials showed that Pieris rapae had no statistically significant preference for any of the genetic lines. In contrast, Spodoptera exigua had a significant feeding preference for the double mutant tgg1tgg2. This study provides evidence that variation in the type of glucosinolates and their hydrolysis products can influence insect performance and feeding choices, and that responses are species-specific.

The effect of glutathione on glucosinolate biosynthesis through the sulfur assimilation pathway in pakchoi associated with the growth conditions

Glucosinolates(GSLs) are a group of nitrogen-and sulfur-containing secondary metabolites, synthesized primarily in members of the Brassicaceae family, that play an important role in food flavor, plant antimicrobial activity, resistance to insect attack, stress tolerance, and human anti-cancer effects. As a sulfur-containing compound, glutathione has a strong connection with GSLs biosynthesis as a sulfur donor or redox system, and exists in reduced(glutathione;GSH) and oxidized(glutathione disulfide;GSSG) forms. However, the mechanism of GSH regulating GSLs biosynthesis remainds unclear. Hence, the exogenous therapy to pakchoi under normal growth condition and sulfur deficiency condition were conducted in this work to explore the relevant mechanism. The results showed that exogenous application of buthionine sulfoximine, an inhibitor of GSH synthesis, decreased the transcript levels of GSLs synthesis-related genes and transcription factors, as well as sulfur assimilation-related genes under the normal growth condition. Application of exogenous GSH inhibited the expression of GSLs synthesis-and sulfur assimilation-related genes under the normal condition, while the GSLs biosynthesis and the sulfur assimilation pathway were activated by exogenous application of GSH when the content of GSH in vivo of plants decreased owing to sulfur deficiency. Moreover,exogenous application of GSSG increased the transcript levels of GSLs synthesis-and sulfur assimilation-related genes under the normal growth condition and under sulfur deficiency. The present work provides new insights into the molecular mechanisms of GSLs biosynthesis underlying glutathione regulation.

Transcriptomics integrated with metabolomics reveals the mechanism of CaCl_(2)-HCl electrolyzed water-induced glucosinolate biosynthesis in broccoli sprouts

Glucosinolates are important phytochemicals in Brassicaceae.We investigated the effect of CaCl_(2)-HCl electrolyzed water(CHEW)on glucosinolates biosynthesis in broccoli sprouts.The results showed that CHEW treatment significantly decreased reactive oxygen species(ROS)and malondialdeh yde(MDA)contents in broccoli sprouts.On the the 8^(th)day,compared to tap water treatment,the the total glucosinolate content of broccoli sprouts with CHEW treatment increased by 10.6%and calcium content was dramatically enhanced from 14.4 mg/g DW to 22.7 mg/g DW.Comparative transcriptome and metabolome analyses revealed that CHEW treatment activated ROS and calcium signaling transduction pathways in broccoli sprouts and they interacted through MAPK cascades.Besides,CHEW treatment not only promoted the biosynthesis of amino acids,but also enhanced the expression of structural genes in glucosinolate synthesis through transcription factors(MYBs,bHLHs,WRKYs,etc.).The results of this study provided new insights into the regulatory network of glucosinolates biosynthesis in broccoli sprouts under CHEW treatment.

“GSS教学模式”在小学高段英语科普类绘本教学中的应用

英语绘本阅读有利于提升学生的绘本阅读能力和阅读效率,以及培养他们的阅读习惯。聚焦小学高段英语科普类绘本教学,从教师导读、自主阅读、分享阅读三个部分,介绍和展示如何运用“GSS教学模式”激发学生的阅读兴趣,提高他们的阅读能力,从而达到学科育人目标。

Solid state fermentation of rapeseed cake with Aspergillus niger for degrading glucosinolates and upgrading nutritional value

Background： Rapeseed cake is a good source of protein for animal feed but its utilization is limited due to the presence of anti-nutritional substances, such as glucosinolates （GIs）, phytic acid, tannins etc. In the present study, a solid state fermentation （SSF） using Aspergillus niger was carried out with the purpose of degrading glucosinolates and improving the nutritional quality of rapeseed cake （RSC）. The effects of medium composition and incubation conditions on the GIs content in fermented rapeseed cake （FRSC） were investigated, and chemical composition and amino acid in vitro digestibility of RSC substrate fermented under optimal conditions were determined. Results： After 72 h of incubation at 34℃, a 76.89% decrease in GIs of RSC was obtained in solid medium containing 70% RSC, 30% wheat bran at optimal moisture content 60% （w/w）. Compared to unfermented RSC, trichloroacetic acid soluble protein （TCA-SP）, crude protein and ether extract contents of the FRSC were increased （P〈 0.05） 103.71, 23.02 and 23.54%, respectively. As expected, the contents of NDF and phytic acid declined （P〈 0.05） by 9.12 and 44.60%, respectively. Total amino acids （TAA） and essential amino acids （EAA） contents as well as AA in vitro digestibility of FRSC were improved significantly （P 〈 0.05）. Moreover, the enzyme activity of endoglucanase, xylanase, acid protease and phytase were increased （P 〈 0.05） during SSF. Conclusions： Our results indicate that the solid state fermentation offers an effective approach to improving the quality of proteins sources such as rapeseed cake.

Effect of Nitrogen and Sulfur Supply on Glucosinolates in Brassica campestris ssp.chinensis

Glucosinolates （GSs） are a group of plant secondary metabolites containing abundant nitrogen （N） and sulfur （S） mainly in Brassica and have the beneficial effects on human health including anti-carcinogenic, cholesterol-reducing and other pharmacological effects. The objective of this study was to investigate the effect of different concentrations of N （5, 10, and 20 mmol L-a, denoted by N5, N10 and N20） and S （0,5, 1, and 2 mmol L^-1, denoted by S0.5, S1 and S2） on the yield and GSs in pakchoi （Brassica campestris L. ssp. chinensis var. communis） in hydroponics. Results showed that N10 and N20 significantly enhanced the yield compared with N5, however, N20 had a negative effect relative to N10. Only with N10 and N20 low S supply （S0.5） reduced the yield. The concentrations of aliphatic GSs, aromatic GS and total GSs were enhanced by N5 and indolyl GSs were enhanced by N20. S2 enhanced the concentration of individual GS and total GSs. The concentrations of indolyl GSs were maximized in N20S2 treatment, whereas the highest concentrations of aliphatic GSs, aromatic GS and total GSs were found in N5S2 treatment. Effects of N and S on aliphatic GSs were higher than on indolyl GSs. The results suggest that the accumulation of aliphatic GSs and aromatic GS could be enhanced by low N and high S and restricted by high N while that of indolyl GSs could be enhanced by high N and high S.

Identification of MAM1s in Regulation of 3C Glucosinolates Accumulation in Allopolyploid Brassica juncea

Allopolyploid Brassica juncea is particularly enriched in sinigrin,a kind of 3C aliphatic glucosinolates(GSLs),giving rise to characteristic taste after picking.However,the molecular mechanism underlying 3C aliphatic GSLs biosynthesis in this species remains unknown.In this study,we genome-widely identified GSLs metabolic genes,indicating different evolutionary rate of GSLs metabolic genes between subgenomes of B.juncea.Eight methythioalkylmalate synthase(MAMs)homologs were identified from B.juncea,in which six MAM1s were located in chloroplast and the other two were not detected with any expression.Furthermore,BjMAM1-4,BjMAM1-5,and BjMAM1-6 displayed higher expression levels in leaves than other tissues.Silenced expression analysis revealed that BjMAM1-4 and BjMAM1-6 function in 3C and 4C aliphatic GSLs accumulation.The specificity of the substrate selection for the second cycle reaction is much lower than that of the first cycle,suggesting these genes may preferentially catalyze 3C aliphatic GSLs biosynthesis.Our study provides insights into the molecular mechanism underlying the accumulation of 3C aliphatic GSLs,thereby facilitating the manipulation of aliphatic GSLs content in B.juncea.

Degradation of glucosinolates in rapeseed meal by Lactobacillus delbrueckii and Bacillus subtilis

Lactobacillus delbrueckii and Bacillus subtilis were employed as a new combination of strains to treat rapeseed meal by solid-state fermentation,aiming to efficiently degrade the glucosinolates,which are the main toxin in the meal.Single-factor tests and Response surface methodology(RSM)were used to optimize the fermentation parameters.Under the optimum fermentation parameters of 15%total injection volume of the mixture of Lactobacillus delbrueckii and Bacillus subtilis with a ratio of 2:1,bran content of16%,feed to water ratio of 1:1.5,fermentation temperature of 36°C and fermentation time of 72 h,the content of glucosinolates in rapeseed meal was decreased from 64.558μmol/g to 3.473μmol/g,reaching a high degradation rate(94.62%).The high detoxification rate by a consortium of Lactobacillus delbrueckii and Bacillus subtilis provides a bright application prospect in feed utilization of rapeseed meal.

Dissecting the genetic architecture of glucosinolate compounds for quality improvement in flowering stalk tissues of Brassica napus

Glucosinolates(GSLs) and their hydrolytic products contribute to the quality traits of rapeseed flowering stalk tissues, such as taste, flavor and anticarcinogenic properties(Glucoraphanin). However, little is known about the genetic mechanisms of GSL accumulation in rapeseed flowering stalks. In this study, the variation and genetic architecture of GSL metabolites in flowering stalk tissues were investigated for the first time among a panel of 107 accessions. All GSL compounds exhibited continuous and wide variations in the present population. Progoitrin,glucobrassicanapin and gluconapin were the most abundant GSL compounds. Five quantitative trait loci(QTL) significantly associated with three GSL compounds were identified by genome-wide association study. GRA_C04 was under selected during modern breeding, in which the ratio of lower GSL haplotype(HAP2) in the accessions bred before 1990(52.56%) was significantly lower than that after 1990(78.95%). Four candidate genes, BnaA01. SOT16, BnaA06. SOT17, Bna A06. MYB51a, and Bna A06. MYB51b, were identified in the GTL_A01 and 4OH_A06 regions.These findings provide new insights into GSL biosynthesis in flowering stalk tissues and facilitate quality improvement in rapeseed flowering stalks.

Analysis on Composition and Content of Glucosinolates in Broccoli Flowers and Leaves

Glucosinolate composition and content were evaluated in flowers and leaves of 12 different broccoli varieties. The results indicated that there were 9 glucosinolates in broccoli,namely Glucoiberin(IBE); Progoitrin(PRO);Sinigrin(SIN);Glucoraphanin(RAA);Gluconapin(NAP);4-Hydroxyglucobrassicin(4OH);Glucobrassicin(GBC);4-Methoxyglucobrassicin(4ME);Neoglucobrassicin(NEO).Total glucosinolate content in flowers was 1-5 times higher than in leaves. The predominant glucosinolate in broccoli was glucoraphanin.

Glucosinolates and Sinapine in Camelina Meal

Forty seven accessions of camelina (Camelina sativa L. Crantz) were analyzed for glucosinolates (GSLs) and sinapine in defatted meal. These antinutritional compounds are undesirable in camelina meal for use in animal feeding and therefore we show their variability to identify the best varieties for future breeding programs. Total GSLs ranged from 19.6 to 40.3 mmol Kg-1 dry weight (DW) with an average of 30.3 mmol kg-1 DW. Great variability has also been observed in the levels of individual GSLs (GSL1, GSL2 and GSL3), so that the content of GSL1 and GSL3 were not correlated to each other in the accessions of camelina. Five out of six winter forms of camelina showed low content of GSLs. Sinapine ranged from 1.09 to 4.75 g Kg-1 DW with an average of 2.57 g kg-1 DW. The sinapine content was not correlated with that of GSLs. The use of camelina meal is only limited by the presence of GSLs while sinapine content can be ignored in camelina varieties.

Identification and Analysis of Components of Glucosinolates in Succulent Roots and Leaves of Fruity Radish (Raphanus sativas L.)

The components of glucosinolates( GSL) in the succulent roots and leaves of three kinds of fruity radish( Xinlimei,Super Zhengyan,and Shawo) were identified and analyzed by high performance liquid chromatography( HPLC). Results showed that there were nine kinds of glucosinolates in the succulent roots and leaves of three kinds of fruity radish,including six kinds of aliphatic glucosinolates( 4-methyl sulfinyl-3-butenyl glucosinolates,2-allyl glucosinolates,4-methyl sulfinyl butyl glucosinolates,5-methyl sulfinyl amyl glucosinolates,4-methyl n-butyl glucosinolates,4-methyl thio-3-butenyl glucosinolates) and three kinds of indole glucosinolates( 4-methoxy methyl indole glucosinolates,3-methyl indole glucosinolates,1-methoxy methyl indole glucosinolates),but there were significant differences in the content of glucosinolates between different varieties and organs. The total content of glucosinolates in succulent roots of three kinds of fruity radish was22 472. 84,13 585. 86,and 28 200. 70 μg/g DM respectively,and more than 95% were aliphatic glucosinolates. The content of glucosinolates in succulent roots was 4. 56,2. 71,and 4. 55 times of that in leaves of three kinds of fruity radish. Main aliphatic glucosinolates in succulent roots and leaves of three kinds of fruity radish were 4-methyl sulfinyl-3-butenyl glucosinolates,accounting for 90. 11%-93. 92% and63. 03%-73. 72% of total content of glucosinolates.

Chemical Ionization Mass Spectrometry of Pertrimethylsilylated Desulphoglucosinolates

The chemical ionization mass spectra of ten different glucosinolates isolated from rape seed have been studied. More intense diagnostic ions were identified than in the electron impact mass spectra. The method is suitable for the structure analysis of mixture of glucosinolates.

基于硫苷种类和含量分析的高品质水果萝卜种质筛选

通过对根辣味、根甜味、根质地、空心、根光滑度、根疤痕等品质进行感官评价,从78份萝卜种质中筛选出12份品质优良的萝卜种质。随后利用LC-MS对这12份萝卜种质肉质根中硫苷的组分和含量进行鉴定和分析,最后通过主成分分析研究总硫苷与单个硫苷含量之间的相关性。结果表明,通过感官评价筛选的12份萝卜种质包括2份长白萝卜种质,3份圆白萝卜种质,3份长红萝卜种质,4份绿皮萝卜种质。在所试萝卜种质中,共鉴定出12种硫代葡萄糖苷,分别有脂肪族硫苷8种(GRA、GIB、GRH、GRE、GER、GBN、PRO、GAL)和吲哚族硫苷4种(GBR、NGBS、4HGBS、4MGBS)。其中,总硫苷含量最高的种质为F1005(22.947μmol/g),含量最低的是W1063(3.518μmol/g)。在成分上,主要的脂肪族硫苷为GRH,主要的吲哚族硫苷是NGBS。此外,在供试的12种萝卜种质中,脂肪族硫苷占总硫苷含量的95.78%~99.87%,吲哚族硫苷占总硫苷含量的0.16%~4.22%。各种质硫苷含量最高的都是GRH,占总硫苷含量的61.91%~93.54%,总硫苷含量与GRH、GRE含量呈极显著正相关,与NGBS含量呈显著负相关,且单一硫苷之间存在不同程度的相关性。共筛选出萝卜硫素前体硫苷GRA含量高的种质3份,分别为F1060、F1078和W1044,辣味物质主要硫苷GRH含量低的种质3份,分别为W1063、F1030和W1044。该研究结果为培育高品质优良萝卜新品种提供理论依据。

GSS系统治疗胸腰椎骨折并发症及防治

[目的]分析总结GSS（general spine system，GSS）系统治疗胸腰椎骨折时出现的并发症及其原因，以期找到更好的预防措施，减少治疗过程中并发症的发生。[方法]回顾性分析自2003年1月-2005年12月在本科接受胸腰椎骨折GSS系统内固定手术的83例患者治疗情况，术后所有患者均获得随访，平均随访时间22个月。[结果]并发症发生15例，发生率18．1％，其中定位错误3例，GSS螺钉进入椎间隙2例，椎体高度撑开不足4例，脑脊液漏1例，钉道松动2例，断钉2例，术后感染1例。分析并发症发生原因，主要为：（1）术前准备不充足；（2）患椎解剖结构变化；（3）GSS系统的设计缺陷；（4）技术因素。[结论]术前根据患椎的具体情况，制定完善的手术方案，术前准备工作充分，以及精确的手术操作，合理的术后康复过程，是降低GSS系统内固定失败发生率的有效手段。

通用型脊柱内固定系统(GSS)在腰椎不稳症治疗中的近期疗效观察

目的 :观察GSS对腰椎节段性不稳的固定及治疗效果。方法 :2 0 0 2年 10月～ 2 0 0 3年 9月 ,对 2 5例腰椎不稳症患者。其中 ,男 9例 ,女 16例 ,运用椎板减压、椎管后外侧植骨融合、GSS椎弓根螺钉内固定系统治疗 ,随访 6～ 18个月 (平均 15 7个月 )。结果 :术前JOA评分平均 4 48分的 2 5例患者 ,随访时 14 5分 (P <0 0 1) ,术后 14个月内动态X线检查显示GSS椎弓根螺钉固定患者植骨融合率 10 0 % ,融合节段稳定 ,临床症状改善优良率 96 0 %。结论 :后路运用GSS行椎管后外侧植骨融合内固定对腰椎不稳症患者是一种安全、有效的术式 。

基于EST和GSS序列的玉米未知微RNA的数据挖掘

miRNAs通过与靶基因互补位点配对结合,在转录后水平负性调控靶基因的表达。根据miRNA进化上的保守性,以拟南芥、水稻等已知的植物miRNAs为探针,与相关数据库中玉米表达序列标签(EST)和基因组序列(GSS)中的非编码序列比对,采用一系列的标准进行筛选,最后预测得到24个玉米miRNA前体,通过靶基因的预测共得到61个靶基因。通过生物信息学方法大大提高了人们发现miRNAs及其靶基因的效率,补充了玉米miRNA数据库的不足。