Anaerobic Accumulation of gamma-Aminobutyric Acid and Alanine in Radish Leaves (Raphanus sativus, L.)

In leaves, the anaerobic accumulation of alanine was accompanied by a loss of aspartate, and these changes preceded γ-aminobutyrate accumulation and glutamate loss. Changes in keto acid content did not appear to be the cause of amino acid changes. Accumulation of γ-aminobutyrate was due to acceleration of glutamate decarboxylation and arrest of γ-aminobutyrate transamination. Changes in enzyme content did not explain the changes in reaction rates in vivo. Most of the aspartate may be converted anaerobically to alanine via oxalacetate and pyruvate.     

alpha-l-Arabinofuranosidase from Radish (Raphanus sativus L.) Seeds

An α-l-arabinofuranosidase has been purified 1043-fold from radish (Raphanus sativus L.) seeds. The purified enzyme was a homogeneous glycoprotein consisting of a single polypeptide with an apparent molecular weight of 64,000 and an isoelectric point value of 4.7, as evidenced by denaturing gel electrophoresis and reversed-phase or size-exclusion high-performance liquid chromatography and isoelectric focusing. The enzyme characteristically catalyzes the hydrolysis of p-nitrophenyl α-l-arabinofuranoside and p-nitrophenyl β-d-xylopyranoside in a constant ratio (3:1) of the initial velocities at pH 4.5, whereas the corresponding α-l-arabinopyranoside and β-d-xylofuranoside are unsusceptible. The following evidence was provided to support that a single enzyme with one catalytic site was responsible for the specificity: (a) high purity of the enzyme preparation, (b) an invariable ratio of the activities toward the two substrates throughout the purification steps, (c) a parallelism of the activities in activation with bovine serum albumin and in heat inactivation of the enzyme as well as in the inhibition with heavy metal ions and sugars such as Hg²⁺, Ag⁺, l-arabino-(1→4)-lactone, and d-xylose, and (d) results of the mixed substrate kinetic analysis using the two substrates. The enzyme was shown to split off α-l-arabinofuranosyl residues in sugar beet arabinan, soybean arabinan-4-galactan, and radish seed and leaf arabinogalactan proteins. Arabinose and xylose were released by the action of the enzyme on oat-spelt xylan. Synergistic action of α-l-arabinofuranosidase and β-d-galactosidase on radish seed arabinogalactan protein resulted in the extensive degradation of the carbohydrate moiety.     

A beta-Galactosidase from Radish (Raphanus sativus L.) Seeds

A basic β-galactosidase (β-Galase) has been purified 281-fold from imbibed radish (Raphanus sativus L.) seeds by conventional purification procedures. The purified enzyme is an electrophoretically homogeneous protein consisting of a single polypeptide with an apparent molecular mass of 45 kilodaltons and pl values of 8.6 to 8.8. The enzyme was maximally active at pH 4.0 on p-nitrophenyl β-d-galactoside and β-1,3-linked galactobiose. The enzyme activity was inhibited strongly by Hg²⁺ and 4-chloromercuribenzoate. d-Galactono-(1→4)-lactone and d-galactal acted as potent competitive inhibitors. Using galactooligosaccharides differing in the types of linkage as the substrates, it was demonstrated that radish seed β-Galase specifically split off β-1,3- and β-1,6-linked d-galactosyl residues from the nonreducing ends, and their rates of hydrolysis increased with increasing chain lengths. Radish seed and leaf arabino-3,6-galactan-proteins were resistant to the β-galase alone but could be partially degraded by the enzyme after the treatment with a fungal α-l-arabinofuranosidase leaving some oligosaccharides consisting of d-galactose, uronic acid, l-arabinose, and other minor sugar components besides d-galactose as the main product.     

Draft Sequences of the Radish (Raphanus sativus L.) Genome

Radish (Raphanus sativus L., n = 9) is one of the major vegetables in Asia. Since the genomes of Brassica and related species including radish underwent genome rearrangement, it is quite difficult to perform functional analysis based on the reported genomic sequence of Brassica rapa. Therefore, we performed genome sequencing of radish. Short reads of genomic sequences of 191.1 Gb were obtained by next-generation sequencing (NGS) for a radish inbred line, and 76,592 scaffolds of ≥300 bp were constructed along with the bacterial artificial chromosome-end sequences. Finally, the whole draft genomic sequence of 402 Mb spanning 75.9% of the estimated genomic size and containing 61,572 predicted genes was obtained. Subsequently, 221 single nucleotide polymorphism markers and 768 PCR-RFLP markers were used together with the 746 markers produced in our previous study for the construction of a linkage map. The map was combined further with another radish linkage map constructed mainly with expressed sequence tag-simple sequence repeat markers into a high-density integrated map of 1,166 cM with 2,553 DNA markers. A total of 1,345 scaffolds were assigned to the linkage map, spanning 116.0 Mb. Bulked PCR products amplified by 2,880 primer pairs were sequenced by NGS, and SNPs in eight inbred lines were identified.     

Adenosine Phosphates in Germinating Radish (Raphanus sativus L.) Seeds

Changes in concentrations of adenosine phosphates (AMP, ADP, and ATP), oxygen utilization, and fresh weights were measured during the first 48 hours after imbibition of water by quiescent radish seeds (Raphanus sativus L.) at 22.5 C. The changes in ATP concentrations, oxygen utilization, and fresh weights followed a triphasic time course, characterized by a rapid initial increase, which extended from 0 to approximately 1.5 hours, a lag phase from 1.5 to 16 hours, and a sharp linear increase from 16 to 48 hours. In unimbibed seeds, the concentrations of ATP, ADP, and AMP were <0.1, 0.9, and 2.2 nmoles/seed, respectively. After imbibition of water by the quiescent seeds, for 1 hour, the ATP concentration had increased to 2.5, and ADP and AMP concentrations had decreased to 0.3 and 0.1 nmole/seed, respectively. These early changes occurred also in seeds maintained under anaerobic conditions (argon), or when treated with either 5 mm fluoroacetate, or 5 mm iodoacetate. The concentrations of ADP and AMP did not change significantly from 1 to 48 hours. The termination of the lag phase at 16 hours correlated with radicle emergence. Cell division in the radicles was initiated at approximately 28 hours. ATP concentrations in seeds maintained under argon or treated with fluoroacetate remained relatively constant from approximately 2 to 48 hours. In contrast, the ATP concentration of iodoacetate-treated seeds decreased curvilinearly from 4 to 48 hours. Oxidative phosphorylation was estimated to have contributed 15, 20, and 65% of the pool ATP at 1.5, 16, and 48 hours, respectively.     

In Vitro Antifungal Activity of a Radish (Raphanus sativus L.) Seed Protein Homologous to Nonspecific Lipid Transfer Proteins

A basic 9-kD protein was purified from seeds of radish (Raphanus sativus L.). The 43 amino-terminal amino acids show extensive sequence identity with nonspecific lipid transfer proteins from other plant species. The radish seed nonspecific lipid transfer protein-like protein inhibits the growth of several fungi in vitro.     

利用氟乐灵进行同源四倍体萝卜种质创制研究

本研究应用除草剂氟乐灵处理两叶一心幼苗生长点,进行同源四倍体萝卜种质诱导,对变异植株进行形态、细胞学鉴定和花粉母细胞染色体数目鉴定。结果表明,应用 0.2 mmol/L 和 1.0 mmol/L 氟乐灵处理,6个萝卜品种都获得同源四倍体植株,10 mmol/L 处理仅在 Nau-zhqh 得到同源四倍体;其中 0.2 mmol/L处理 Nau-dy 和 1.0 mmol/L 处理 Nau-xbch 获得四倍体最高诱导率(40%);四倍体种质与二倍体种质相比,形态性状、气孔大小、保卫细胞内叶绿体数目、花器官大小、花粉粒大小及花粉萌发率都存在显著差异,将形态、气孔鉴定和染色体计数结合可以准确确定变异株的倍性。研究表明利用氟乐灵诱导是进行萝卜同源四倍体种质创新的有效途径之一。本研究应用除草剂氟乐灵处理两叶一心幼苗生长点,进行同源四倍体萝卜种质诱导,对变异植株进行形态、细胞学鉴定和花粉母细胞染色体数目鉴定。结果表明,应用 0.2 mmol/L 和 1.0 mmol/L 氟乐灵处理,6个萝卜品种都获得同源四倍体植株,10 mmol/L 处理仅在 Nau-zhqh 得到同源四倍体;其中 0.2 mmol/L处理 Nau-dy 和 1.0 mmol/L 处理 Nau-xbch 获得四倍体最高诱导率(40%);四倍体种质与二倍体种质相比,形态性状、气孔大小、保卫细胞内叶绿体数目、花器官大小、花粉粒大小及花粉萌发率都存在显著差异,将形态、气孔鉴定和染色体计数结合可以准确确定变异株的倍性。研究表明利用氟乐灵诱导是进行萝卜同源四倍体种质创新的有效途径之一。     

Arabinogalactan-Proteins from Primary and Mature Roots of Radish (Raphanus sativus L.)

Organ-specific variations in blood group H-like activity were observed in developing radish plants. A temporary increase in serological activity was found to occur in the roots at the earlier stages of development. Arabinogalactan-proteins (AGPs) were isolated from primary and mature roots, and investigated for changes in their physicochemical properties, structure, and serological activities. These root AGPs were composed mainly of l-arabinose and d-galactose but were distinguishable from each other in their contents of l-fucose as well as of protein and hydroxyproline. The structures of the carbohydrate moieties of the root AGPs were essentially similar to those of AGPs isolated from seeds and mature leaves in that they consisted of consecutive (1→3)-linked β-d-galactosyl backbone chains having side chains of (1→6)-linked β-d-galactosyl residues, to which α-l-arabinofuranosyl residues were attached in the outer regions. One prominent feature of the primary root AGPs was that they contained appreciable amounts of l-fucose, which was presumably responsible for expression of the serological activity. In their immunological reactions with rabbit anti-radish leaf AGP antibody, the root AGPs were shown to share common antigenic determinant(s) with those of seed and leaf AGPs.     

Growth and Development of Radish (Raphanus sativus, L.) Under Selected Light Environments

Plants of radish (Raphanus sativus L.) were grown under selectedlight conditions in controlled environmental chambers in orderto monitor the role of photoperiod, irradiance level and inputlight energy in plant development. Results indicated that thedaily input of light energy was the most important light factoraffecting leaf development while photoperiod and irradiancelevel had the major influences on storage organ development.Distribution of assimilates to leaves and storage organs variedunder different light regimes with long photoperiods and highirradiances producing the largest storage organs. Once initiated,the rate of storage organ growth was similar under all testedlight environments. Raphanus sativus L., radish, growth, development, light, photoperiod, assimilate distribution, storage organ     

春萝卜抽薹过程中内源激素含量变化分析

采用酶联免疫吸附法(ELISA)测定了春萝卜抽薹过程中顶端生长点处嫩叶的赤霉素(GA₄)、生长素(IAA)、细胞分裂素(iPA)和脱落酸(ABA)等含量的变化情况。研究结果表明,在萝卜抽薹过程中,GA₄含量在现蕾点存在明显的峰值,表明GA₄与现蕾抽薹密切相关,起着主导作用;内源ABA含量变化与GA₄变化趋势基本一致;与GA₄和ABA含量相比,抽薹过程中叶片IAA含量相对较低;iPA含量在现蕾期逐步增加,至抽薹中后期出现一高峰值。研究发现iPA/ABA值增大,GA₄/iPA和GA₄/IAA值变小有利于花芽分化后萝卜植株抽薹开花,表明内源激素之间的平衡对萝卜抽薹开花也起着重要作用。     

Inheritance and Molecular Marker for Flowering Time in Radish (Raphanus sativus L.)

Plant Molecular Biology Reporter - The mixed inheritance model involving major genes and polygenes was used to analyze the inheritance of radish flowering time trait in the B1, B2, F1, and F2...     

肉质色不同萝卜遗传多样性的SSR分子标记分析

利用微卫星(SSR)标记技术,从600对SSR引物中筛选86对扩增条带清晰的引物,检测了来自我国不同地区37个肉质不同颜色萝卜品种的遗传多样性。86对引物共扩增到976个条带,每对引物扩增出2～17个条带,平均为10.7个,其中多态性条带892个,多态性条带比例为91.39%;共检测出753个基因型,每对引物检测2～20个基因型,平均8.7个,其中有效基因型443.99,有效基因型比例为58.96%;Shannon多态性指数变幅为0.44～2.77,平均1.76。当相似系数为0.81时,可将供试萝卜分成3类,第I类包括6份白色肉质萝卜,第II类包括3份红色肉质萝卜和6份白色肉质萝卜,第III类包括22份红色肉质萝卜。各红色肉质萝卜品种间的遗传相似系数有97%大于0.80,而各白色肉质萝卜品种间的遗传相似系数有91%大于0.80。红色肉质萝卜遗传多样性略低于白色肉质萝卜,红色肉质萝卜与白色肉质萝卜间平均相似系数为0.83,说明不同肉色的萝卜间亲缘关系较密切,在分类上红色肉质萝卜可能是白色萝卜的一个变种。     

Heavy Metal Phytotoxicity to Radish (Raphanus sativus L.) in a Digested Sludge-amended Gangetic Alluvium

A limiting factor in land application of sewage sludge is the resultant heavy metal accumulation in soils followed by biomagnification in the food chain, posing a potential hazard to animal and human health. In view of this fact, pot experiments were conducted to evaluate the effect of digested sludge application to soil on phytotoxicity of heavy metals such as Cd, Cr, Ni, and Pb to radish (Raphanus sativus L.) plants. Increasing sludge levels resulted in increased levels of DTPA-extractable heavy metals in the soil. Cadmium was the dominant metal extracted by DTPA followed by Ni, Pb, and Cr. The extractability of metals by DTPA tended to decrease from the first to the second crop. Dry matter yield of radish increased significantly as a function of increasing sludge treatments. Soil application of sludge raised the concentration of one or more heavy metals in plants. Shoots contained higher concentrations of Cd, Cr, and Ni than the roots of radish plants. Shoot concentrations of Cd, Cr, Ni, and Pb were within the tolerance levels of this crop at all rates of sludge application. Shoot as well as root concentration of Cd was above 0.5 mg kg^?1, considered toxic for human and animal consumption. The levels of DTPA-extractable Cd and Ni were less correlated while those of Cr and Pb were more correlated with their respective shoot and root contents. The results emphasize that accumulation of potentially toxic heavy metals in soil and their build-up in vegetable crops should not be ignored when sludge is applied as an amendment to land.     

Genetic Diversity of Radish (Raphanus sativus L.) Germplasm Resources Revealed by AFLP and RAPD Markers

Genetic diversity of 56 radish accessions, representing nearly all the typical types and origins of cultivated radish germplasms conserved in the National Mid-term Genebank for Vegetables of China, was assessed with amplified fragment length polymorphism (AFLP) and random amplified polymorphic DNA (RAPD) markers. A total of 72 and 128 polymorphic bands were generated by the 12 selected RAPD primers and eight AFLP primer combinations respectively. A moderate correlation with the value of r = 0.66 was observed between AFLP and RAPD markers. The total 200 polymorphic bands were integrated to assess the genetic diversity of 56 radish accessions. The Jaccard similarity coefficients between the accessions varied from 0.30 to 0.83 with the mean of 0.54. Cluster analysis classified the germplasms into three groups of var. hortensis Becker, var. sativus, and var. niger Kerner. The three-dimensions scatter plot of principle coordinate analysis (PCA) further divided var. hortensis Becker germplasms into two separate groups. The results indicated that the genetic diversity harbored among var. hortensis Becker germplasms was very abundant, which could be further exploited for radish genetic improvement.     

Diversity,abundance and foraging behaviour of insect pollinators in Radish (Raphanus raphanistrum subsp. sativus L.)

Insect pollinators and flowering plants have co-evolved together for millions of years, developing a wide array of mutualistic interactions. Several cultivated plant species depend exclusively on insect-mediated pollination services for their subsistence, reproduction and survival. Radish, Raphanus raphanistrum subsp. sativus, a popular edible root vegetable (Family: Brassicaceae) completely depends on pollinators as self-pollination is totally absent due to sporophytic self-incompatibility. The present study was conducted at the experimental fields of the ICAR-Indian Institute of Horticultural Research, Bengaluru, to understand the diversity and foraging behaviour of insect pollinators in R. sativus. Continuous survey during the flowering period revealed that a total of fifteen insect species that belong to three insect orders (Hymenoptera, Diptera and Lepidoptera) were found to visit R. sativus flowers. The most abundant species was Apis florea L., followed by Apis dorsata F., Tetragonula iridipennis (Smith), Apis cerana F., Lassioglossum sp. and Ischiodon scutellaris F. Floral visitors differed significantly in terms of their visitation frequency, with A. florea (0.76 ± 0.22 visits/flower/ min) being the most frequent visitor followed by A. dorsata (0.66 ± 0.21 visits/flower/ min) and T. iridipennis (0.59 ± 0.14 visits/flower/30 min). The maximum visitation rate was recorded for A. dorsata (7.8 ± 1.40 flowers visited/min) followed by A. cerana (6 ± 1.15). The maximum stay time on individual flower was recorded for Lassioglossum sp. (48.2 ± 3.67 sec) followed by T. iridipennis (44.2 ± 4.24 sec). The maximum numbers of pollen grains were deposited by A. dorsata (206.70 ± 56.45) followed by A. cerana (151.20 ± 31.84). All the floral visitors except A. florea made contact with stigma on every single visit (100 %). While, only 53 per cent of total visitation by the Apis florea led to stigma contact. Further, nectar robbing behaviour was majorly exhibited by A. florea and Delta conoideum compared to all other insect pollinators observed. Based on the overall pollinator effectiveness per day A. dorsata was found to be the most effective single pollinator species.     

Purification and Characterization of L-Alanine: 4,5-Dioxovalerate (Glyoxylate) Aminotransferase from Radish (Raphanus sativus L.) Seedlings

L-Alanine:4,5-dioxovalerate (DOVA) aminotransferase was purified131-fold and characterized from greening seedlings of radish(Raphanus sativus L.). The enzyme was shown to be identicalwith alanine:glyoxylate aminotransferase. The rate of activityof DOVA aminotransferase was 15 times less than that of glyoxylateaminotransferase. Its molecular weight was estimated to be approximately123,000 with two identical subunits, and exhibited a single,broad pH optimum at 8.0. DOVA aminotransferase activity wascompetitively inhibited by glyoxylate. A kinetic study of theenzyme at different alanine concentrations suggested a pingpong reaction mechanism. The K_m values for DOVA and L-alaninewere 0.71 and 1.7 mM, respectively. The activity ratio of transamination under various conditions,the cellular localization of the enzyme and the lack of correlationbetween the activity of this enzyme and chlorophyll synthesis,indicate that DOVA aminotransferase in radish is not involvedin 5-aminolevulinate synthesis. (Received July 7, 1984; Accepted September 11, 1984)     

萝卜种质资源对芜菁花叶病毒的抗性鉴定与评价

病毒病是危害萝卜作物的主要病害之一,芜菁花叶病毒(TuMV)作为萝卜病毒病的主要毒源,给萝卜生产造成严重经济损失。由于缺乏精准鉴定,萝卜育种中缺乏稳定可靠的抗TuMV材料。本研究就国家蔬菜种质资源中期库提供的来自中国25个省份125个县市和其他3个国家的150份代表性萝卜种质资源对TuMV的抗性进行了苗期鉴定和ELISA检测,筛选出23份抗病材料,其中1份表现免疫。这些材料对萝卜抗病毒病基因定位和新品种培育具有重要意义。     

Acylated anthocyanins from red radish (Raphanus sativus L.)

Twelve acylated anthocyanins were isolated from the red radish (Raphanus sativus L.) and their structures were determined by spectroscopic analyses. Six of these were identified as pelargonidin 3-O-[6-O-(E)-feruloyl-2-O-beta-D-glucopyranosyl]-(1-->2)-beta-D-glucopyranoside]-5-O-(beta-D-glucopyranoside), pelargonidin 3-O-[6-O-(E)-caffeoyl-2-O-(6-(E)-feruloyl-beta-D-glucopyranosyl)-(1-->2)-beta-D-glucopyranoside]-5-O-(beta-D-glucopyranoside), pelargonidin 3-O-[6-O-(E)-p-coumaroyl-2-O-(6-(E)-caffeoyl-beta-D-glucopyranosyl)-(1-->2)-beta-D-glucopyranoside]-5-O-(beta-D-glucopyranoside), pelargonidin 3-O-[6-O-(E)-feruloyl-2-O-(6-(E)-caffeoyl-beta-D-glucopyranosyl)-(1-->2)-beta-D-glucopyranoside]-5-O-(beta-D-glucopyranoside), pelargonidin 3-O-[6-O-(E)-p-coumaroyl-2-O-(6-(E)-feruloyl-beta-D-glucopyranosyl)-(1-->2)-beta-D-glucopyranoside]-5-O-(beta-D-glucopyranoside), and pelargonidin 3-O-[6-O-(E)-feruloyl-2-O-(2-(E)-feruloyl-beta-D-glucopyranosyl)-(1-->2)-beta-D-glucopyranoside]-5-O-(beta-D-glucopyranoside).