Identification and Molecular Mapping of the Rs Dm R Locus Conferring Resistance to Downy Mildew at Seedling Stage in Radish(Raphanus sativus L.)

Downy mildew （DM）, caused by the fungus Peronospora parasitica, is a destructive disease of radish （Raphanus sativus L.） worldwide. Host resistance has been considered as an attractive and environmentally friendly approach to control the disease. However, the genetic mechanisms of resistance in radish to the pathogen remain unknown. To determine the inheritance of resistance to DM, F1, F2 and BC1F1 populations derived from reciprocal crosses between a resistant line NAU-dhp08 and a susceptible line NAU-qtbjq-06 were evaluated for their responses to DM at seedling stage. All F1 hybrid plants showed high resistance to DM and maternal effect was not detected. The segregation for resistant to susceptible individuals statistically iftted a 3：1 ratio in two F2 populations （F2（SR） and F2（RS））, and 1：1 ratio in two BC1F1 populations, indicating that resistance to DM at seedling stage in radish was controlled by a single dominant locus designated as RsDmR. A total of 1 972 primer pairs （1 036 SRAP, 628 RAPD, 126 RGA, 110 EST-SSR and 72 ISSR） were screened, and 36 were polymorphic between the resistant and susceptible bulks, and consequently used for genotyping individuals in the F2 population. Three markers （Em9/ga24370, NAUISSR826700 and Me7/em10400） linked to the RsDmR locus within a 10.0 cM distance were identiifed using bulked segregant analysis （BSA）. The SRAP marker Em9/ga24370 was the most tightly linked one with a distance of 2.3 cM to RsDmR. These markers tightly linked to the RsDmR locus would facilitate marker-assisted selection and resistance gene pyramiding in radish breeding programs.

不同灌溉方式下秋冬萝卜(Raphanus Sativus L.)的生长分析

本文对两种不同灌溉方式(滴灌和沟灌)下的秋冬萝卜(RaphanusSativusL.)生长情况进行了比较研究.结果表明:秋冬萝卜的生长、产量与灌溉方式有较密切关系.与沟灌方式相比,滴灌下萝卜的相对生长速率、净同化率、叶面积比率、比叶面积、叶重比、根冠比等指标以及产量都明显提高;采用滴灌方式,也有利于萝卜品质的提高.

萝卜(Raphanus sativus L.)种质莱菔子素含量分析与评价

莱菔子素是由萝卜中萝卜甙水解形成的一种异硫氰酸酯类物质，在植物抗性和人类抗癌方面具有重要作用。本试验利用已优化的HPLC检测方法测定了93份萝卜种质肉质根的莱菔子素含量，分析了不同来源地和不同类型萝卜种质莱菔子素含量分布。试验发现不同萝卜种质莱菔子素含量存在显著差异，其含量分布范围为34．445～1446．9mg／kg-DW，最高含量约是最低含量的42倍；红皮白肉和绿皮白肉类型的萝卜种质莱菔子素平均含量较高，华东地区的萝卜种质莱菔子素平均含量显著高于其他来源地的萝卜种质。试验初步获得莱菔子素含量较高的萝卜种质2份，为进一步试验研究提供了良好的材料。

Developing Stable Cultivar through Microspore Mutagenesis in ×<i>Brassicoraphanus koranhort</i>, Inter-Generic Allopolyploid between <i>Brassica rapa</i>and <i>Raphanus sativus</i>

A stable progeny was developed through induced mutation, using microspore culture, of the hybrid (F1F1) produced by crossing a newly synthesized, unstable allopolyploid (F1) and a stable cultivar, BB#1(F1) in xBrassicoraphanus. An F1F1 plant was subjected to the induced mutation system established during production of BB#1. Morphological characteristics of the progeny such as color, and leaf number and length, differed from those of BB#1. The bolting time of the progeny in spring cropping was very late compared to BB#1, allowing it to be grown to an adult plant in spring. Genomic in situ hybridization analysis of pollen mother cells at prophase I identified 19 bivalents, 10 from Brassica rapa and 9 from Raphanus sativus. The glucoraphenin content was almost identical to that of BB#1. Two cultivars are available in the baemoochae crop now. These results indicate that induced mutation using microspore culture is a viable method of stabilizing intergeneric allopolyploids between B. rapa and R. sativus.

The Cumulative Effects of Sewage Sludge Compost on Raphanus sativus L: Growth and Soil Properties

The main goal of this research study was to establish the physicochemical characteristics of sewage sludge, and the nutrients needed to enhance the properties of soil with a view to exploiting were potential for agriculture. The example used was the cultivation of radish (Raphanus sativus). The findings are intended as a contribution towards making agriculture sustainable. This study was carried out to assess the response of radish (Raphanus sativus L.) to doses of sewage sludge applied in medium texture soils, and the socioeconomic contributions that such cultivation of radish would make. Levels of 0.25 and 75 ton/ha of sewage sludge plus the equivalent dose of 25 ton/ha of sewage sludge vermin compost by California Red were used after making a randomized experimental design with three replicates. 30 days after sowing radish seeds, the following data were collected: the number of leaves per plant (NLP);plant height (PH in cm);root diameter (RD) and tubercle production (TP ton/ha). In addition, the pH, EC-electric conductivity (EC), and soil organic matter (SOM) were determined. Statistical analysis showed that 25 and 50 tons/ha gave the best results for the parameters assessed. The estimated increase in family mean income from selling radish crops fertilized with sewage sludge is around 17%.

Ameliorating effects of Raphanus sativus leaves on sodium arsenite-induced perturbation of blood indices in Swiss albino mice

Objective: To evaluate the ameliorating effects of Raphanus sativus leaves(RSL)against sodium arsenite(Sa)-induced adverse effects through mice experiments.Methods: Swiss albino mice were divided into four equal groups: control, Sa, RSL,RSL + Sa. Sa(10 mg/kg body weight/day), and powder form of RSL(50 mg/kg body weight/day) were provided as food supplement orallty. Blood indices were measured using commercially available kits through colorimetric methods.Results: It was observed that lactate dehydrogenase, alkaline phosphatase, alanine aminotransferase, and aspartate aminotransferase activities were significantly(P < 0.05)higher in Sa-treated mice than those in the control group. RSL significantly reduced Sainduced elevation of the activities of these enzymes in serum significantly(P < 0.05).Serum butyrylcholinesterase activity and high density lipoproteins cholesterol levels in Satreated mice were significantly(P < 0.05) lower than the control group, and the food supplementation of RSL could significantly(P < 0.05) prevent the reduction of Sa-mediated serum butyryl cholinesterase activity and high density lipoproteins cholesterol levels. RSL could also reduce the Sa-induced elevation of serum urea level significantly(P < 0.05).Conclusions: Results of this study suggest the protective or ameliorating effects of RSL on Sa-induced perturbation of blood indices are related to the hepatic, cardiovascular and kidney dysfunction. Therefore, RSL may be useful to reduce arsenic toxicity in human in the future.

Molecular Cloning and Characterization of Pollen Development Related Gene RsMF2 from Raphanus sativus L.

In the paper, the full length cDNA of RsMF2 gene, homologous with the BcMF2 gene encoding pollen-specificpolygalacturonase of Chinese cabbage-pak-choi (Brassica campestris L. ssp. chinensis Makino) was cloned from Raphanussativus L. cv. Yuanbai by PCR, with a pair of primer designed according to the coding sequence of BcMF2. The largestopening reading frame of RsMF2 gene is 1 266 bp in length and encodes a protein of 421 amino acids with a predictedmolecular mass of 43.9 kDa. Sequence analysis revealed that it has three potential N-glycosylation sites and onepolygalacturonase active position (RVTCGPGHGLSVGS). And the first 32 amino acids of the predicted RsMF2 proteinform a N-terminal hydrophobic domain which displays the properties of a signal peptide. The predicted secondarystructure composition for the protein has 6.9% helix, 42.0% sheet and 51.1% loop. Four domains which are highly conservedin the whole plant and fungal PGs is present in RsMF2. Phylogenetic analysis showed that RsMF2 falls into the categoryof clade-C, which includes PGs related to pollen. These results indicate that RsMF2 may act as polygalacturonase relatedto pollen development.

In Silicon Cloning and Bioinformatics Analysis of the Raphanus Sativus WUS Gene

The complete coding sequence ofRaphanus sativusWUS gene was obtained by usingArabidopsis thalianagene se-quence (NM_127349) as a probe in silico cloning. Then the hydrophilicity, secondary structure and advanced structure of WUS protein inRaphanus sativusare analyzed by using bioinformatics methods. The results show that the cDNA was 1298 bp, with no intron, contains an open reading frame of 936bp, encoding 312aa protein. The protein coded byRaphanus sativusgene showed 74% similarity toArabidopsis thaliana.

A Preliminary Study on the Quality Standard of Raphanus sativus L. Produced in Guizhou Province

[Objectives]This study was conducted to analyze the Raphanus sativus L.medicinal materials from different origins and batches in Guizhou Province to explore its quality standards.[Methods]The plant morphology,material shape and microscopic characteristics of R.sativus were identified;and qualitative identification was performed by the TLC method.[Results]The plant morphology,material shape and microscopic characteristics of the R.sativus medicinal materials were described;and the TLC identification method of the R.sativus medicinal materials was established,with clear spots,good separation,and good specificity.[Conclusions]The method is accurate,simple,highly sensitive,interference-free,has good reproducibility,and can be used as a data support for the formulation of quality standards for R.sativus medicinal materials.

Remineralization of a Dystric Ferralsol Using Basalt and Tephra Dusts,Effective Microorganisms Manure and NPK 20-10-10 for Rad­ish(Raphanus sativus)Production in Bamougoum(Cameroon Western Highlands)

This paper studies the effect of basalt and tephra dusts,as alternatives to chemical fertilizers,on soil fertility and Radish(Raphanus sativus)production.The experiment was conducted in the field and in the labo­ratory in the years 2017 and 2018 on two separate plots so as to annul residual effects of fertilizers).The experimental design in the field was a randomized complete block design(56 m^(2)),including five treatments and three replications:control(T_(0)),basalt dust(T_(1)),tephra dust(T_(2)),effective micro-organism(EM)fertilizer(T_(3))and NPK 20-10-10(T_(4)).The main results show the following decreasing trend based on yield:T_(1)>T_(3)>T_(0)>T_(4)>T_(2).The best yields appear in T_(1) and T_(3) probably because they supplied the highest levels of soil nutrients to match the needs of the crops.Although T_(2) plants performed poorly,soil properties like pH.H_(2)O(6.14 to 6.49),sum of exchangeable bases,base saturation,available phosphorus and cation balance were improved after tephra treatment.T_(2) plants might have performed poorly due to intrinsic properties of the tephra dust like low availability of trace elements compared to T_(1) and T_(3).T_(4) plants show the highest number of leaves,leaf area index and plant height.The Fe,Mn,Cu and Zn levels in bulbs and leaves will not pose danger of toxicity to human upon consumption and could serve as nutri­ent supplement for children and expectant mothers.The most profitable treatment is T_(1) permitting to recommend the popularization of basalt dust for radish cultivation as an alternative to chemical fertilizers.

Establishing VIGS and CRISPR/Cas9 techniques to verify RsPDS function in radish

Virus-induced gene silencing(VIGS)and clustered regularly interspaced short palindromic repeats/CRISPR-associated protein(CRISPR/Cas)systems are effective technologies for rapid and accurate gene function verification in modern plant biotechnology.However,the investigation of gene silencing and editing in radish remains limited.In this study,a bleaching phenotype was generated through the knockdown of RsPDS using tobacco rattle virus(TRV)-and turnip yellow mosaic virus(TYMV)-mediated gene silencing vectors.The TYMV-mediated gene silencing efficiency was higher than the TRV-based VIGS system in radish.The expression level of RsPDS was significantly inhibited using VIGS in'NAU-067'radish leaves.The rootless seedlings of‘NAU-067'were infected with Agrobacterium rhizogenes using the 2300GN-Ubi-RsPDS-Cas9 vector with two target sequences.Nine adventitious roots were blue with GUs staining,and four of these adventitious roots were edited at target sequence 1 of the RsPDS gene as indicated by Sanger sequencing.Furthermore,albino lines were generated with A.tumefaciens-mediated transformation of radish cotyledons.Five base substitutions and three base deletions occurred at target sequence 2 in Line 1,and three base insertions and three base substitutions occurred at target sequence 1 in Line 2.This study shows that VIGS and CRISPR/Cas9 techniques can be employed to precisely verify the biological functions of genes in radish,which will facilitate the genetic improvement of vital horticultural traits in radish breeding programs.

萝卜开花基因RsFLC3的克隆与表达分析

植物开花春化途径中关键抑制因子FLC基因属于MADS-box基因家族,为探索萝卜开花抑制基因FLC3的功能,以抽薹开花差异较大的萝卜品种YZH(易抽薹)和XHT(耐抽薹)为研究材料,对萝卜开花基因进行生物信息学分析,克隆了萝卜RsFLC3基因,并进行亚细胞定位和两个萝卜品种不同时期的qRT-PCR分析。结果表明,共鉴定出萝卜开花相关基因638个,亚细胞定位显示该基因编码的蛋白定位于细胞核,RsFLC3基因在萝卜抽薹开花过程中表达量整体呈下降趋势,但不同品种间大部分时期的表达量差异较大。

萝卜叶多酚的超声波辅助提取工艺优化及抗氧化活性研究

为提高萝卜叶资源的利用率,并发掘其潜在应用价值,探究了萝卜叶多酚的超声波辅助提取工艺,并对其抗氧化活性进行了评估。通过在单因素实验基础上引入Box-Behnken响应面法,对萝卜叶多酚的超声波辅助提取工艺进行了优化。通过体外实验的评估,进一步验证了其抗氧化能力。结果表明,在超声温度为68℃、液料比为32 mL/g、乙醇体积分数为63%以及超声时间为31 min的最佳工艺条件下,萝卜叶多酚可以被高效地提取,其提取率达到了60.84 mg/g,且相对误差低至0.73%,充分证实了响应面分析方法优化该提取工艺的有效性和可行性。萝卜叶多酚展现出一定的抗氧化特性,对DPPH自由基、O_(2)^(-)自由基的半抑制质量浓度(half maximal inhibitory concentration,IC_(50))分别为78.91、403.55 mg/L,为进一步研发其作为天然植物源抗氧化剂提供了参考。

萝卜(Raphanus sativus)Dof基因家族全基因组鉴定分析

Dof(DNA-binding with one finger)蛋白是一类在植物中起着关键作用的转录因子。本研究对萝卜Dof转录因子基因家族进行了系统进化关系、保守模体、染色体位置分布及不同组织表达模式分析。研究发现,萝卜基因组中鉴定出的63个Dof基因,分布于9条染色体上,可以分为9个亚族。十字花科(Brassicaceae)四种作物系统进化树的分析表明,该家族基因在植物中具有高度同源性。对其在不同组织和发育时期的表达模式研究发现,该基因家族具有一定的时空表达特异性。基因RsaDof03和RsaDof25在各个组织中表达量均相对较高,预测其参与萝卜不同的发育过程。本研究为进一步研究萝卜Dof基因家族的功能提供科学依据。

莱菔及莱菔子的药理作用及临床应用研究进展

莱菔及莱菔子是我国常见的中药,其中莱菔作为“药食同源”的传统中药,在我国应用甚广。二者具有消食除胀、降气化痰等功效,常在临床应用于咳嗽、哮喘等呼吸道疾病,并用于治疗食积、便秘、术后胃肠功能恢复等消化道疾病。近年来针对莱菔及莱菔子的药理作用及临床研究日趋增多,在药理作用方面具有抗肿瘤、降脂保肝、降糖、抗氧化、抗炎、抑菌、抗焦虑、心血管保护及对中枢神经的保护作用。在临床应用中,除了根据其中药药性所开发的具有对呼吸道及消化道的作用,还具有一定的抗炎和对泌尿系的功效。文章通过收集分析总结莱菔及莱菔子的最新文献研究,针对莱菔及莱菔子的药理作用及临床应用做一综述,旨在为今后合理地开发及应用提供更多的理论依据。

萝卜籽酶解制备萝卜硫素工艺优化及其体外消化研究

为探究酶解法制备萝卜籽萝卜硫素的最佳工艺及其消化特性。采用红萝卜种子为原料,以萝卜硫素得率为指标,通过单因素实验和响应面优化试验得出最佳酶解工艺条件。进一步采用体外模拟胃肠消化模型,探究萝卜籽乙酸乙酯提取物萝卜硫素含量及其抗氧化活性的变化规律。得到萝卜籽制备萝卜硫素的最佳酶解条件为:酶解时间22 min,酶解温度40℃,VC添加量0.8 mg/g,在此最佳工艺条件下,萝卜硫素得率为2.11±0.02 mg/g,与预测值(2.14 mg/g)接近,相对误差为1.4%,酶解工艺切实可行。体外消化实验结果表明,胃肠消化液中萝卜硫素保留率分别为79.64%,76.22%;与未消化提取物比较,胃消化液中ABTS+、DPPH、OH三种自由基清除率均显著降低(P<0.05),分别为54.23%、33.90%、15.07%,而肠消化液与胃消化液中自由基清除率均无显著性差异(P>0.05)。本研究丰富了酶法制备萝卜硫素方面的研究,将为萝卜籽开发及高附加值利用提供科学依据。

干旱胁迫下萝卜种子萌发期蛋白的双向电泳分析

种子萌发是植物生长、发育的最初阶段,也是容易遭到包括干旱在内的非生物胁迫的重要时期。蛋白组学是解析植物响应逆境胁迫的主要策略之一,双向电泳分离是蛋白组学的重要手段和环节。本研究以“春不老”萝卜为材料,利用聚乙二醇-6000(PEG-6000)模拟干旱胁迫,对干旱胁迫下萝卜种子萌发早期无子叶胚蛋白进行双向电泳分离和蛋白图谱差异分析。采取直接对萝卜无子叶胚体样品进行裂解和上样的方法,可得到分辨度和重复性较好的双向电泳蛋白图谱;对干旱胁迫下的萝卜种子萌发早期无子叶胚蛋白表达图谱进行差异分析,共检测和匹配265个萝卜蛋白点,响应干旱胁迫的蛋白质点有28个,其中13个显著上调表达,15个显著下调表达。研究结果表明,部分萝卜种子萌发期蛋白响应了干旱胁迫,为后续的蛋白质谱分析和响应蛋白的鉴定工作打下良好基础。本研究采用的对植物样品直接裂解和上样的方法,也为其他植物蛋白的双向电泳分析提供了借鉴。

萝卜ERF11基因克隆与序列分析

采用RT-PCR技术克隆了心里美萝卜ERF11基因CDS序列,并对其进行生物信息学分析。结果表明,RsERF11基因开放阅读框全长507 bp,无内含子,编码168个氨基酸,相对分子量约为41 513 u,理论等电点为5.18,属于亲水性蛋白质,亚细胞定位预测其主要定位于细胞核。RsERF11含有1个AP2保守结构域,不含跨膜结构域和信号肽。系统进化分析表明,该蛋白质与甘蓝型油菜(Brassica napus)、甘蓝(Brassica oleracea)和白菜(Brassica rapa)等十字花科物种的同源性较高。