Effects of Salt Stress on Seed Germination and Seedling Growth in Raphanus sativus L.

[Objective] This study aimed to investigate the effects of salt stress on seed germination and seedling growth in radish（Raphanus sativus L.）.[Method] First,effects of soaking seeds with clear water and different concentrations of saline solution were compared; then,different concentrations of salt solution （0.2％-1.0％） was used to treat radish during the whole germination period.[Result] soaking seeds with clear water can improve the germination indicators of radish seeds exposed to high salt stress; low salt concentration（0.2％,0.4％） had little effect on the growth indicators of radish seedlings,and 0.2％ salt concentration can even promote the growth of radish seedlings,however,the high salinity had a bad inhibition on seedling growth,the higher the salt concentration,the stronger the inhibition.[Conclusion] According to the comprehensive comparison,the order of the six varieties in salt-tolerance was as follows：Xinxiayuanbai＞W1029,Xiayuanbai ＞ Nanyangbailizhi,Chunyidianhong＞ Sijihong 3.

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。

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%.

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.

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.

Black radish root extract alleviates sodium valproate-induced liver damage via inhibiting mitochondrial membrane potential collapse and oxidative stress in mice

Objective:To explore the effect of black radish(Raphanus sativus L.var niger)root extract on liver enzymes,oxidative stress,and histopathological alterations in mice with sodium valproate-induced hepatotoxicity.Methods:Thirty-two mice were divided into four groups:the control group received drinking water by gavage,the second group was administered with 100 mg/kg of sodium valproate,the third group received 300 mg/kg of black radish root extract,and the fourth group was given both sodium valproate(100 mg/kg)and black radish root extract(300 mg/kg).After 28 days,the mice were euthanized,and serum levels of aspartate aminotransferase(AST),alanine aminotransferase(ALT),and alkaline phosphatase(ALP),along with liver malondialdehyde(MDA),reactive oxygen species(ROS),mitochondrial parameters,tumor necrosis factor-alpha(TNF-α)gene expression,and histopathological changes were assessed.Results:Sodium valproate caused hepatic damage in mice,characterized by elevated serum levels of liver enzymes,increased MDA and ROS levels and TNF-αgene expression,as well as histopathological alterations.The black radish root extract significantly alleviated sodium valproate-caused hepatic injury by decreasing the serum levels of ALT and AST,MDA,ROS,TNF-αgene expression,as well as mitochondrial impairment,but did not have a significant effect on sodium valproate-induced histopathological changes.Conclusions:The black radish root extract demonstrates protective effects against sodium valproate-induced liver injury,possibly through mitigating oxidative stress,mitochondrial impairment,and inflammatory mediator expression.

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。

Physiological and Phytosanitary Potentials of Coriander and Radish Seeds

Information on seed vigor of the vegetable crops is always important and necessary due to the increasing demand of high quality seeds for sowing and high-value commercial. The cultivation of these species, conducted intensively, should be established with seed high potential physiological and health for the development of a more productive and sustainable agriculture. The present study was conducted to evaluate the potential physiological of seed lots of radish and coriander. The experiment was conducted at the laboratory of seed analysis and greenhouse of the plant science department of the Federal University of Pelotas in South Brazil. Four radish seed lots, cultivar ＂Saxa＂, and four coriander seed lots, cultivar ＂Verdito＂ were used. Germination seed test, first count of germination, accelerated aging test, electrical conductivity and seedling emergence were used to evaluate the physiological quality of the lots. The experimental design was completely randomized design with five replications. Means were compared by Tukey test. The accelerated aging test was the most efficient test in assessing the physiological quality for both lots of radish seeds and coriander and this test provide coherent results with seedlings emergence.

萝卜SCPL3基因的克隆与生物信息学分析

采用RT-PCR技术克隆了萝卜(Raphanus sativus L.)SCPL3基因CDS序列,并对其进行生物信息学分析。结果表明,萝卜SCPL3基因CDS序列全长1 434 bp,编码477个氨基酸,分子式为C_(2493)H_(3762)N_(632)O_(709)S_(21),相对分子质量为54.6 kD,理论等电点为6.33,脂肪系数为81.30,不稳定系数为39.26,平均亲水值为-0.227,是稳定亲水蛋白;亚细胞定位预测该蛋白最有可能定位于液泡,存在1个信号肽和3个跨膜结构,含有SC族羧肽酶中的S10蛋白家族结构域;系统进化分析表明其与十字花科作物SCPL蛋白同源性较高。

萝卜抽薹基因连锁的AFLP和SCAR分子标记鉴定

本文运用AFLP分子标记技术,结合BSA法,对萝卜抽薹基因相连锁的AFLP分子标记进行研究,获得两个与萝卜耐抽薹基因相连锁的AFLP标记,遗传距离分别为14.6cM和9.1cM。序列测定和Blast分析表明,标记ACG-CAG(123bp)与拟南芥中锌指蛋白3(zinc finger protein3,ZFP3)的cDNA同源性为89%,期望值为6e-23;标记ACT-CTG(175bp)与拟南芥中的Copia类反转录转座子AtRE1基因的同源性为83%,期望值为4e-42。并将其中一个标记转化为简单易行的SCAR标记,遗传距离为7.5cM。

栽培萝卜种质亲缘关系的AFLP分析

利用筛选出的8对引物对56份来源于不同国家和地区的栽培萝卜种质的亲缘关系进行了AFLP分析,共扩增出327条带,其中多态性带128条,多态性位点百分率39.1%,显示出栽培萝卜种质之间存在着较丰富的遗传多样性。系统聚类分析将供试材料分为5类9组,主坐标分析将其分为4类7组,2种分类方法所获结果基本一致。基于分子标记的分类与种质的表型基本吻合。

不同钠盐浸种对NaCl胁迫下萝卜种子萌发的影响

[目的]为农作物的耐盐育种和栽培提供试验依据。[方法]通过不同钠盐浸种和NaCl溶液胁迫萌发试验,找出萝卜(Raphanus sativus L)种子抗盐的有效方法和耐盐程度。分别用浓度为200mmol/L的NaCl、NaHCO3、混合盐(NaHCO3,NaCl)3种溶液浸种,然后配置4种浓度的NaCl胁迫溶液:40、80、120、160mmol/L,分别对萝卜种子的萌发进行盐胁迫。[结果]混合盐浸种明显提高萝卜种子发芽指数和活力指数,促进幼苗生长。NaHCO3浸种明显抑制种子萌发,但明显促进侧根数目增多并提高了叶绿素的含量。促进种子萌发的影响因子顺序是:混合盐浸种>NaCl浸种>对照组>NaHCO3浸种。不同浸种方法处理,均表现出随NaCl胁迫溶液浓度升高,抑制种子萌发,而对幼苗生长的影响在NaCl胁迫溶液浓度较低时,随NaCl胁迫溶液浓度升高促进幼苗生长。相反,在NaCl胁迫溶液浓度较高时,随NaCl胁迫溶液浓度升高,抑制幼苗生长。[结论]混合盐浸种促进种子萌发和幼苗的生长。

NaCl胁迫对萝卜幼苗叶片生理特征及根尖细胞核形态的影响

对0.00(对照)～0.50 mol.L-1NaCl处理48 h后萝卜(Raphanus sativus L.)幼苗叶片的叶绿素含量、净光合速率、丙二醛(MDA)含量和相对电导率进行了测定,并观察了根尖细胞核形态变化和胁迫后幼苗生长的恢复状况。测定结果表明:经0.05 mol.L-1NaCl处理后,叶片的叶绿素含量和净光合速率均高于对照(0.00 mol.L-1NaCl);而用0.10～0.50 mol.L-1 NaCl处理后,叶绿素含量和净光合速率均随NaCl浓度的提高逐渐降低且与对照差异显著,其中,经0.35～0.50 mol.L-1NaCl胁迫处理后叶片净光合速率降至0.00μmol.m-2.s-1。随NaCl浓度的提高,MDA含量和相对电导率呈先快速上升而后逐渐下降的趋势,且均与对照差异显著;其中,经0.30 mol.L-1NaCl处理后MDA含量最高,经0.40 mol.L-1NaCl处理后相对电导率最高。观察结果显示:经0.05～0.25 mol.L-1NaCl处理后根尖细胞核由规则的圆形向各种不规则形状转变,染色质出现不同程度的浓缩,表现出不同程度的程序性细胞死亡特征;经0.30～0.50 mol.L-1NaCl处理后则出现中空细胞与部分正常细胞并存的现象。经0.05和0.10 mol.L-1 NaCl处理后,萝卜幼苗可恢复正常生长,且株高与对照无明显差异;而经0.25～0.50 mol.L-1NaCl处理后,幼苗均较难恢复正常生长,且随NaCl浓度的提高,幼苗出现枯萎、糜烂现象。研究结果显示:低浓度NaCl处理不仅对萝卜幼苗的生长、生理代谢及细胞核形态结构没有明显伤害反而有一定的促进作用,而高浓度NaCl胁迫可导致萝卜幼苗不可逆伤害。

NaCl胁迫下萝卜幼苗根系生理和细胞核形态的变化

以萝卜幼苗为材料,研究了0.15 mol/L NaCl胁迫处理不同时间其根系的生长速度、活力、过氧化物酶(POD)活性和相对电导率的变化,并观察了根尖细胞核形态的变化。结果表明,随着NaCl胁迫时间的延长,萝卜幼苗根系生长速度减慢,根系活力降低,胁迫时间超过48 h时,二者均迅速下降,表现得更为明显;萝卜幼苗根系POD活性和相对电导率则不断增大,胁迫48 h时达到最大,之后POD活性迅速降低,相对电导率随之降低;根尖细胞核由原来规则的圆形转变为各种不规则形状,出现不同程度的凋亡特征。

利用cDNA-AFLP分析大白菜、萝卜及其杂交种差异表达基因

为更好地理解十字花科蔬菜杂种优势的分子机理,对大白菜(Brassica rapa L. ssp. pekinensis)、萝卜(Raphanus sativus L.)及其杂交种的差异表达基因进行了初步研究。利用cDNA-AFLP技术对杂种F1及其双亲进行基因差异表达分析,共得到44个在杂种F1特异表达的差异片段,分属40个Unigene;通过Blast搜索,有43个差异片段可以在GenBank数据库中找到同源序列(包括EST序列),其中仅32个序列功能已知。这些已知功能基因参与了碳代谢、激素合成、转录、细胞周期、物质运输、信号转导、逆境响应和花发育等相关过程。

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。

NaCl胁迫对不同生育时期萝卜的影响

研究了NaCl胁迫对不同生育时期萝卜(Raphanus sativus L.)的影响。结果表明,萝卜种子萌发期和幼苗期对NaCl胁迫的反应较为敏感,100 mmol/L的NaCl胁迫下萝卜种子发芽势、发芽率、发芽指数、幼苗单株地上部鲜重和单株根鲜重显著降低,丙二醛(MDA)含量显著升高;150 mmol/L的NaCl胁迫下萝卜幼苗根系活力显著下降;成株期萝卜对150 mmol/L以内的NaCl胁迫反应不敏感。