Identification of key genes regulating the synthesis of quercetin derivatives in Rosa roxburghii through integrated transcriptomics and metabolomics

Rosa roxburghii fruit is rich in flavonoids, but little is known about their biosynthetic pathways. In this study, we employed transcriptomics and metabolomics to study changes related to the flavonoids at five different stages of R. roxburghii fruit development. Flavonoids and the genes related to their biosynthesis were found to undergo significant changes in abundance across different developmental stages, and numerous quercetin derivatives were identified. We found three gene expression modules that were significantly associated with the abundances of the different flavonoids in R. roxburghii and identified three structural UDP-glycosyltransferase genes directly involved in the synthesis of quercetin derivatives within these modules. In addition, we found that RrBEH4, RrLBD1 and RrPIF8could significantly increase the expression of downstream quercetin derivative biosynthesis genes. Taken together,these results provide new insights into the metabolism of flavonoids and the accumulation of quercetin derivatives in R. roxburghii.

Effect of fermented Rosa roxburghii Tratt fruit juice on gut microbiota in a dextran sulfate sodium-induced colitis mouse model

This study mainly investigated the regulatory effect of Rosa roxburghii Tratt fruit juice fermented by Lacticaseibacillus paracasei SR10-1(LAB-RRTJ)on modulating gut microbiota in dextran sulfate sodium(DSS)-induced ulcerative colitis in mice.Compared to control group,DSS induction decreased body weight of mice,indexes of Shannon,Simpson,Chao1 and Faith_pd,and increased disease activity index(DAI)and levels of interleukin 1β(IL-1β),IL-6,tumor necrosis factorα(TNF-α)and interferon-γ(IFN-γ);And this induction also led to an increase in Proteobacteria,Verrucomicrobia and Actinobacteria at phylum level,harmful bacterial species richness at genus level,and relative richness of S.sciuri,Desulfovibrio C21_c20,R.gnavus and Akkermansia muciniphila at species level,and a decrease in Firmicutes at phylum level and relative richness of B.acidifaciens in mice.LAB-RRTJ increased body weight of mice with DSS induced ulcerative colitis(UC)and indexes of Shannon,Simpson,Chao1 and Faith_pd,reduced DAI and the content of four infl ammatory factors and improved gut microbiota imbalance in DSS induced UC mice.Besides,the number of operational taxonomic units(OTUs)increased,α-diversity andβ-diversity were restored and similar to those in mice in the control group after LAB-RRTJ treatment.Compared with the positive drug treatment group,LAB-RRTJ has a better effect on regulating gut microbiota diversity in colitis mice.Correlation analysis showed that infl ammatory factors were positively correlated with harmful bacteria and negatively correlated with beneficial bacteria which commonly found in some colitis mice.Taken together,our study demonstrated that LAB-RRTJ could alleviate DSS-induced colitis in mice through the modulation of infl ammatory cytokines and gut microbiota composition.

Chloroplast DNA-based genetic variation of Rosa roxburghii in Southwest China:Phylogeography and conservation implications

Rosa roxburghii Tratt.is a well-known commercial horticultural crop in China with nutritional and medicinal value.Wild germplasms of this species are mainly distributed in Southwest China but the population is decreasing due to continuous exploitation,habitat destruction,and fragmentation.Therefore,assessing the genetic diversity and phylogeography is essential for efficient conservation.Herein,two chloroplast intergenic spacers(trnL-trnF and accD-psaI)were investigated in 255 individuals from 29 R.roxburghii populations and 18 haplotypes(H1–H18)were identified.High levels of haplotype diversity(Hd=0.829)and nucleotide diversity(π=1.3×10^(−3))were detected in these populations.Also,the genetic variation representing 86.4%of the total variation was detected by an analysis of molecular variance.A significant correlation was established between genetic divergence and geographic distance by the Mantel test(r=0.204,P=0.04,9999 permutations),suggesting the isolation-by-distance model.A significantly higher Nst than Gst(Nst=0.257,Gst=0.136,P<0.05)indicated the phylogeographic structure of R.roxburghii.Further phylogeographic analysis revealed rapid range expansion in the population,probably between 647073 and 217848 years ago.The primary processes shaping the genetic patterns of the R.roxburghii populations included restricted gene flow with isolation distance within clades 1-8,2-3,and overall,contiguous expansion within clades 1-3 and 3-2,past fragmentation,and/or long-distance colonization within clades 1-9 and 2-2.Conservation priority should be given to the core populations GZ,FQ,DF,DS,xy,AL,LC,PB,and XY in the Yunnan-Guizhou Plateau,NZ and MX in the Qingling-Bashan mountains,and MN in the Hengduan mountains,where an in situ preservation and management strategy should be applied.

Changes of Endogenous Hormone Contents in Fruit, Seeds and Their Effects on the Fruit Development of Rosa roxburghii

The fruit of cultivar Guinong5 (Rosa roxburghii Tratt.) was used to investigate the regularity of its growth and development, and the changes of endogenous hormone contents during the period of the growth and development. The result showed that contents of GA 1+3, IAA and ZRs in the flower receptacle of young fruit were high and then decreased, contents of GA 1+3and IAA in seeds increased with its development until maturity. The contents of ZRs and ABA in the flower receptacle of young fruits and seeds were high, and then had a tendency of descent with the development of fruits. The contents of GA 1+3, IAA, ZRs and ABA in flower receptacle of young fruit with normal fertilization were much higher than those of in non-fertilization fruits. In the flower receptacle of malformed fruits, the contents of GA 1+3, IAA and ZRs in normally developing flower receptacle were significantly higher than those in malformed fruits. The growth and development of fruits depended largely on the seeds, development because the seeds in the fruits with non-fertilization failed to be formed or even formed but irreproducible, which would lead to the decrease of GA 1+3, IAA contents and the abnormal development of fruits. Therefore the fruits deformed and dropped inevitably.

Determination of 13 Trace Elements in Anshun Thorn Pear (Rosa roxburghii) and Golden Thorn Pear (Rosa sterilis S.D.Shi) by ICP-MS

[Objectives] This study was conducted to establish an analytical method for the establishment of 13 trace elements in Anshun thorn pear(Rosa roxburghii) and golden thorn pear(Rosa sterilis S. D. Shi). [Methods] Nitric acid was used as the digestion system for microwave digestion. Inductively coupled plasma mass spectrometry(ICP-MS) was used to determine the trace elements in thorn pear and golden thorn pear. [Results] Be, Ti, V, Cr, Mn, Co, Ni, As, Sr, Mo, Cd, Sb and Hg were determined by ICP-MS method in thorn pear and golden thorn pear. The correlation coefficients of the regression equations of the elements were higher than 0.996; the precision RSD was lower than 2.87%; and the recovery ranged from 94.3% to 109.8%. [Conclusions] The method has the advantages of high sensitivity, good accuracy and simultaneous detection of multiple elements, and provides a reference method and theoretical basis for determining trace elements in thorn pear and golden thorn pear and other fruits.

Rosa roxburghii-edible fungi fermentation broth attenuates hyperglycemia, hyperlipidemia and affects gut microbiota in mice with type 2 diabetes

Type 2 diabetes(T2D)is a worldwide chronic disease,which is associated with metabolic disorders and gut microbiota imbalance.The anti-diabetic effects of two fermentation broth composed of Rosa roxburghii and edible fungi in different proportions(FBA and FBB)were studied using a high-fat diet combined with streptozotocin-induced type 2 diabetes mouse model.Our results revealed that intervention with FBA and FBB could ameliorate body weight loss,blood glucose,insulin,blood lipids,morphology of the kidney and liver,and oxidative stress levels in diabetic mice.Furthermore,FBA and FBB interventions significantly improved the community richness and diversity of gut microbiota.Supplementation with FBA and FBB increased the relative abundance of some beneficial and SCFAs-producing genus such as Bifidobacterium,Prevotella,Ruminococcus,Clostridium,and Allobaculum,and also increased the levels of SCFAs in the colonic contents.These results indicated that FBA and FBB can be used as promising functional foods for preventing or alleviating diabetes through modulating gut microbiota and production of SCFAs.

Improved physicochemical and functional properties of dietary fiber from Rosa roxburghii pomace fermented by Bacillus natto

Rosa roxburghii juice is of increasing importance in recent years and has become a favorite fruit juice in the market due to its extremely high Vc content.Meanwhile,a large amount of pomace from juice process is usually treated as waste.However,the pomace still contains a lot of nutrients and has great reuse value.In this study,the Rosa roxburghii pomace was fermented with Bacillus natto to develop a new healthy food ingredient rich in high quality dietary fiber(soluble dietary fiber,SDF).Under the optimal fermentation process conditions(inoculum concentration 4%,temperature 38.7°C,time 12 h,and pH 4.97),SDF content of R.roxburghii pomace increased from 7.68%to 10.68%.SEM and FTIR analysis revealed that the fermentation can lead to degradation of cellulose and hemicellulose,resulting in the formation of more porous lax structure,the related functions such as water holding,oil holding,substance adsorption and cation exchange capacity were obviously increased.In addition,19 phytochemicals from R.roxburghii pomace were quantified by UPLC-QqQ-MS/MS,which demonstrated the fruit pomace also could be used as a functional ingredient in food field.

Characterization of Amino Acid Composition in Fruits of Three Rosa roxburghii Genotypes

Rosa roxburghii Tratt., is an important commercial horticultural crop in China due to its nutritional and medicinal value. To date, the economic production of a single predominant cultivar of R. roxburghii(‘Guinong 5') consumes 30 000 hm^2 in China. Here, we characterize the amino acid composition of ‘Guinong 5' and two novel genotypes(AL11, GP6) that prove valuable and deserve wide application. Here, 17 amino acids(acid hydrolysis of proteins) were detected in ‘Guinong 5', AL11 and GP6 fruits, with Glutamine and Asparagine being abundant, and L-cystine being absent. The essential amino acids and total amino acids were the highest in AL11, while Asparagine was most abundant in GP6. For the free amino acid, 18 amino acids and 9 metabolites were detected in ‘Guinong 5', AL11 and GP6 fruits. γ-aminobutyric acid, L-arginine and ornithine were the most abundant in ‘Guinong 5', while phosphoserine and taurine were the highest in AL11. In comparison, GP6 had the highest levels of L-asparagine, an essential amino acid, and the highest total N containing compounds.

贵州野生刺梨叶黄酮的纯化工艺研究

研究旨在通过超声波提取法利用60%乙醇提取贵州野生刺梨叶片中的总黄酮,通过静态吸附试验研究了5种大孔吸附树脂对总黄酮的动态吸附特性,筛选出了对刺梨叶黄酮具有良好吸附和解吸性能的大孔吸附树脂。以吸附率和解吸率作为考察指标,对大孔树脂纯化刺梨叶黄酮的提取工艺进行优化。结果显示,大孔吸附树脂AB-8具有较好的吸附以及解吸性能,当纯化工艺为径高比1∶2(2BV)、上样液浓度0.6 g/L、控制流速2 mL/min时,AB-8大孔吸附树脂可快速有效地吸附刺梨叶中的黄酮。进一步使用55%乙醇80 mL,保持洗脱流速为2 mL/min时可以快速充分地洗脱AB-8大孔吸附树脂吸附的黄酮。在该纯化条件下,刺梨叶黄酮的纯度从3.368 mg/g提高至6.679 mg/g。研究表明,该工艺条件适于刺梨叶黄酮的纯化。

刺梨籽对肉兔消化代谢、血浆生化指标、屠宰性能及肉品质的影响

本试验旨在研究刺梨籽对肉兔消化代谢、血浆生化指标、屠宰性能及肉品质的影响。采用完全随机区组试验设计,将96只体重相近、体况良好的5周龄断奶新西兰白兔分为4组,每组12个重复,每个重复2只。对照组饲喂不添加刺梨籽的基础饲粮,试验组分别饲喂添加120、240和360 mg/kg刺梨籽的试验饲粮。饲养试验持续77 d,其中预试期14 d,正试期63 d。结果显示:与对照组相比,1)饲粮添加不同水平刺梨籽均可显著升高酸性洗涤纤维和粗脂肪表观消化率(P<0.05);添加240 mg/kg刺梨籽可显著提高干物质与氮表观消化率(P <0.05);添加120 mg/kg刺梨籽可显著提高代谢能和总能表观消化率(P<0.05)。2)饲粮中添加不同水平刺梨籽均可显著增强肉兔血浆超氧化物歧化酶活性(P<0.05),显著降低血浆葡萄糖含量(P <0.05);添加240或360 mg/kg刺梨籽可显著提升血浆总抗氧化能力和尿素氮、白蛋白含量(P<0.05);同时,添加360 mg/kg刺梨籽可显著降低血浆丙二醛含量(P<0.05)。3)饲粮中添加不同水平刺梨籽对肉兔屠宰性能和器官指数均无显著影响(P>0.05)。4)饲粮中添加不同水平刺梨籽均可显著上调肌肉pH_(45 min)(P<0.05),添加240 mg/kg刺梨籽可显著降低肌肉pH_(24 h)(P<0.05)。由此可见,饲粮中添加240 mg/kg刺梨籽可改善肉兔对饲粮中养分的消化利用,增强血浆抗氧化能力,对屠宰性能与肉品质无不良影响。

不同提取工艺对刺梨中维生素C提取量的影响研究

本文以柠檬酸为提取剂,分别采用浸提法、超声辅助法、微波辅助法提取刺梨果肉中维生素C,并利用紫外分光光度计测定维生素C提取量。结果表明,超声辅助提取法的提取率最高,在超声功率为80 W、超声时间为8 min条件下,维生素C的提取量为35.9 mg·g^(-1)。

高维生素C刺梨饼干配方工艺优化及其体外抗氧化作用

以刺梨为基本原料,制作一款高维生素C(VC)含量的刺梨饼干,并对其配方工艺进行优化,对其体外抗氧化活性进行评价.采用4因素4水平正交试验设计,对影响刺梨饼干制作中VC含量和稳定性的4个关键因素烘焙温度、烘焙时间、苹果酸用量和红花籽油用量进行优化;采用不同种类的普通食用油等量替代优化配方中的红花籽油,观察并对比普通食用油和红花籽油对产品中VC含量的影响;通过测定产品对羟基自由基和超氧阴离子自由基的清除率及对脂质过氧化的抑制率来评价产品的体外抗氧化活性.结果得到最优的产品配方工艺条件为:烘焙温度为140℃,烘焙时间为20 min,苹果酸用量为0.75%,红花籽油用量为8%.产品中的红花籽油采用普通食用油替代后,起酥油的VC损失率最大,比红花籽油高4.8倍以上,普通植物食用油的VC损失率也比红花籽油高89%以上.按优化后的配方工艺制得的刺梨饼干产品中VC含量达210 mg/100 g,质量稳定.优化后的产品和等剂量VC对照组对羟基自由基和超氧阴离子自由基都有良好的清除作用,对脂质过氧化有良好的抑制作用,并且优化后的产品效果都显著优于等剂量的VC对照组.

刺梨果醋发酵过程中理化特性和风味特征的变化

为探究刺梨果醋发酵过程中成分变化,本文以刺梨为原料,采用全液态发酵技术,经酒精发酵和醋酸发酵制备果醋,动态监测发酵过程中的理化特性,并采用固相微萃取-气相色谱-质谱联用技术(Headspace solid-phase microextraction-gaschromatography-mass spectrometry,HS-SPME-GC-MS)和气味活性值(Odoractivityvalue,OAV)分析挥发性风味成分。结果表明,随着发酵时间的进行,可溶性固形物、pH、总糖及还原糖均呈下降趋势,而总酸和V_(C)含量持续增加,由原汁到醋酸发酵结束分别从1.86 g/100 mL和956.82 mg/100 mL增加至6.79 g/100 mL和1275.88 mg/100 mL。有机酸的含量不断变化,草酸、奎宁酸、丙酮酸、抗坏血酸、乳酸、乙酸及富马酸呈现不同程度的显著增加(P<0.05),而甲酸、柠檬酸、马来酸和琥珀酸含量变化小。共检测到92种挥发性物质,结合OAV分析,10种挥发性物质被确定为刺梨果醋中OAV≥1的关键香气物质,即壬醛、乙醛、异丁酸乙酯、丁酸乙酯、2-甲基丁酸乙酯、乙醇、异戊醇、叶醇、芳樟醇及苯乙醇。其中,丁酸乙酯和叶醇对刺梨果醋香气贡献最大,是刺梨果醋具有青草香、青苹果香的特征风味物质。研究结果为有效控制发酵条件、揭示发酵刺梨果醋的理化特性和风味特征,形成刺梨果醋品质评价方法、制定质量标准提供理论基础和实践指导。

四种喷雾器械对刺梨白粉病施药效果比较研究

为筛选出适合刺梨病虫害防治采用的高效低耗新型植保喷雾器械,本研究比较了手动喷雾器、电动喷雾器、汽油喷雾器以及植保无人机等4种植保喷雾器械喷施30%吡唑醚菌酯悬浮剂对刺梨白粉病的田间防控效果,及4种植保喷雾器械的作业效率和作业成本。研究结果表明,4种喷雾器械在施药后7和14 d的防控效果均>80%。在喷雾器械工作效率方面,无人机作业效率最高,手动喷雾器作业效率最低。在喷雾器械作业成本方面,电动喷雾器成本最低为345元/hm2,手动喷雾器作业成本最高,为495元/hm2。本研究结果为刺梨病虫害防控喷雾器械的选择提供了科学指导。

黔产刺梨VC的提取及纯化工艺研究

以黔产刺梨汁为原料,采用95%乙醇提取制备刺梨维生素C(VC)粗提液,再经HPD100大孔树脂分离纯化,并以刺梨VC的损失率为指标进行了刺梨VC提取物的浓缩方法、干燥方法筛选,发现减压浓缩法、真空干燥法更加适用于刺梨VC提取物制备;本文所建立的刺梨VC提取物的制备方法简便、有效,可为培育“贵州刺梨”优质产品奠定基础,助力贵州刺梨产业发展。

姜蛋白脱除刺梨果汁中单宁的工艺研究

为有效脱除刺梨果汁(Rosa roxburghii juice)中的单宁,降低其涩味改善口感,该研究以刺梨果汁为对象,采用化学沉淀法,以姜蛋白为单宁脱除剂,并以单宁脱除率和维生素C(VC)保留率为考察指标,采用单因素实验和正交试验优化姜蛋白脱除单宁工艺确定最优工艺。结果表明:(1)姜蛋白脱除刺梨果汁单宁的最优工艺条件为液固比30∶1.2(mL∶g),刺梨果汁pH 3.0,搅拌温度5℃,搅拌时间30 min。(2)由正交试验分析可知,各因素对刺梨果汁脱除单宁的影响程度依次为液固比>搅拌温度>刺梨果汁pH>搅拌时间。(3)在最优工艺条件下,单宁脱除率为(47.451±0.608)%,VC保留率为(75.904±1.244)%。(4)在最优工艺条件下,果汁透光率从(8.44±0.662)%提高到(92.47±0.397)%,涩味明显改善,同时丰富了刺梨果汁风味。该研究结果为解决刺梨果汁深加工行业面临的共性关键技术问题提供了一个新思路和新工艺技术路线基础,也为拓展生姜资源的综合利用奠定了一定技术基础。

体外产气法评价刺梨果渣对肉羊瘤胃体外发酵参数的影响

研究旨在利用体外产气法评价不同水平刺梨果渣的肉羊瘤胃体外发酵参数,为开发利用刺梨果渣饲料资源奠定基础。以4只成年山羊作为瘤胃液供体羊,体外发酵底物为动物饲养试验中肉羊采食的3组饲粮(对照组:不含刺梨果渣;处理1组:含12%刺梨果渣;处理2组:含24%刺梨果渣),测定产气量、发酵参数、纤维素酶活性。结果显示:处理1组6 h累计产气量(GP_(6 h))显著高于对照组(P<0.05),3组间其余时间点产气量差异不显著(P>0.05)。处理1组总挥发性脂肪酸、乙酸含量具有高于对照组、处理2组的趋势(P=0.08);3组间其余发酵参数[氨态氮(NH_(3)-N)、干物质降解率(DMD)和有机物降解率(OMD)]无显著差异(P>0.05)。3组间瘤胃纤维素酶(纤维二糖酶、羧甲基纤维素酶、木聚糖酶)活性不受刺梨果渣水平的影响,差异不显著(P>0.05)。由此可见,刺梨果渣对肉羊瘤胃体外发酵没有负面影响,可作为肉羊饲料使用,适量水平(12%)的刺梨果渣可提高产气量,促进瘤胃发酵,但仍需结合生产实际情况考虑其对肉羊生产性能的影响。

刺梨SOD基因家族鉴定与表达模式分析

【目的】超氧化物歧化酶(SOD)是植物体内氧自由基的天然清除剂,对于保护植物免受环境胁迫起着关键作用,了解其在刺梨响应干旱胁迫中的作用,为深入研究刺梨SOD基因家族的功能提供基础。【方法】通过生物信息学对刺梨SOD基因家族进行系统鉴定和分析,对其理化性质、染色体位置、基因结构、亚家族进化、顺式作用元件和WGCNA进行全面分析,并利用RT-qPCR分析刺梨SOD基因家族在干旱胁迫下的表达模式。【结果】刺梨SOD基因家族有9个成员,包括4个Cu/ZnSOD基因、3个MnSOD基因和2个FeSOD基因,分布在5条染色体上。基因结构显示,同一亚家族成员的基序组成较为相似,但内含子/外显子排列和数量差异较大。亚家族进化分析发现,MnSOD亚家族较原始,其在所有蛋白质位置都表现出高度保守性,其次是FeSOD和Cu/ZnSOD,Cu/ZnSOD亚家族各序列之间差异较大,又可以分为3个亚类。顺式作用元件分析显示,该家族基因参与多种植物激素、生长发育以及胁迫响应,特别是RrCSD2启动子区含有类黄酮生物合成元件(MBSI)。进一步通过转录组和WGCNA分析发现,RrCSD2所在模块的基因显著富集到黄酮和黄酮醇生物合成、类黄酮生物合成和花生四烯酸代谢等途径,表明RrCSD2可能参与类黄酮代谢过程。RT-qPCR结果显示,在干旱胁迫下,除RrMSD1和RrMSD2的表达水平显著下降外,其余7个基因的表达水平总体呈现上升趋势,特别是RrCSD2和RrCSD3的表达水平显著上调。【结论】刺梨SOD基因在干旱胁迫中起到重要作用。

刺梨复合果茶的制备及品质研究

以刺梨、柠檬、金桔、赤藓糖醇为原料,以维生素C(VC)含量和感官评分为评价指标,进行刺梨复合果茶的制备工艺及品质研究。各原料经干燥工艺处理后,采用混料设计中的单纯形格子设计确定刺梨复合果茶的最佳配方,并对刺梨复合果茶的感官指标、理化指标以及抗氧化能力进行分析。结果表明,刺梨复合果茶的最佳配方是:刺梨70%、柠檬5%、金桔5%、赤藓糖醇20%,该果茶产品的维生素C含量为7.09 g/100 g、总酸含量为80.62 g/kg、p H为3.46,DPPH自由基清除率为98.43%、羟基自由基清除率为13.71%。刺梨复合果茶的维生素C含量较高、抗氧化能力较强,具有较好的品质。

刺梨叶茶的品质分析及功能研究

以刺梨树春季嫩叶为原料,采用3种传统工艺分别制备得到刺梨叶绿茶、红茶、白茶,并对其进行品质分析及功能研究。采用感官评价结合模糊数学评价,通过层次分析(AHP)法确定多酚、黄酮、感官评分权重,计算刺梨叶茶的综合评分,并对刺梨叶茶的抗氧化能力和体外降血糖能力进行分析。结果表明,3种刺梨叶茶的综合评价为刺梨叶绿茶(19.26)>刺梨叶白茶(16.96)>刺梨叶红茶(12.57)。以市售荷叶茶为对照,抗氧化能力的测试中,3种刺梨叶茶及荷叶茶的1,1-二苯基-2-三硝基苯肼(DPPH)、2,2’-联氮-双(3-乙基苯并噻唑啉-6-磺酸)(ABTS)、羟基自由基清除率及铁还原能力分别为刺梨叶绿茶89.84%、97.96%、76.24%、0.910;刺梨叶红茶87.35%、67.63%、62.32%、0.573;刺梨叶白茶83.60%、75.89%、60.05%、0.531;荷叶茶66.94%、56.31%、98.35%、0.180。降血糖功能研究中,3种刺梨叶茶及荷叶茶的α-葡萄糖苷酶、α-淀粉酶活性抑制率分别为刺梨叶绿茶96.57%、21.75%;刺梨叶红茶89.90%、23.11%;刺梨叶白茶92.52%、22.97%;荷叶茶92.05%、18.82%。综上,3种刺梨叶茶的相关指标基本都优于市售荷叶茶,表明其具有较好的抗氧化能力和降血糖能力,其中刺梨叶绿茶的效果最好。