Tissue distribution of phenylpropanoid metabolism in cotyledons of Raphanus sativus L.

The tissue distributions of sinapic acid esters (1-sinapoylglucose, sinapolyl-l-malate, 6,3-disinapoylsucrose), kaempferol glycosides, free malic acid and of the enzyme involved in the synthesis of sinapoyl-l-malate, 1-sinapoylglucose: l-malate sinapoyltransferase (SMT), have been investigated in cotyledons of Raphanus sativus L. seedlings. The kaempferol glycosides were mainly localized in the upper epidermis. The sinapoyl esters were found in all tissues, but differed markedly in their concentrations. While disinapoylsucrose was localized predominantly in the mesophyll, most sinapoylmalate was found in the epidermal layers, as was most SMT activity. Ultraviolet microscopy and microfluorospectrophotometry of isolated epidermal peels indicated that the epidermal sinapoyl esters were restricted to guard cells, guard mother cells and adjacent epidermal cells. Upon excitation by UV light (365 nm) these exhibited strong blue fluorescence with an emission maximum at about 480 nm. Our results indicate a highly tissue-and cell-specific secondary metabolism in Raphanus cotyledons and indicate that the biosynthesis of sinapoylmalate is intimately related to the malic-acid metabolism of the guard cells.Abbreviations HPLC high-performance liquid chromatography - SMT 1-sinapoylglucose: l-malate sinapoyltransferase     

Selecting for floral character associations in wild radish,Raphanus sativus L.

ESS models of reproductive allocation have been used extensively to explain patterns of floral diversity in angiosperms. These theoretical explorations assume that proportional allocation to pollen, ovules, and seeds, as well as to secondary features such as showy petals and nectar rewards, can evolve independently within the limits set by total resource availability. In populations of California wild radish, we have shown previously that petal size, a strong determinant of visitation by honey bee pollinators, is positively correlated with both pollen and nectar production, but not with ovule or seed number per flower. These phenotypic associations may reflect selection, environmental correlation, and/or genetic constraint. By exerting selection on the petal size : pollen number ratio over two generations, we eliminated the positive correlation between petal size and pollen production, with both characters showing significant change after a single selection episode. Once these two floral traits became uncoupled, nectar sugar production was significantly correlated only with petal size. Our results suggest that natural selection could readily alter reproductive allocation in these flowers, and that the phenotypic correlations observed in nature may be maintained by selection for effective reproductive phenotypes.     

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.     

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.     

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).     

Effects of Sucrose on Rooting and Senescence in Detached Raphanus sativus L. Cotyledons

Application of 0.5–4.0 per cent sucrose to excised radishcotyledons incubated in the light suppressed the formation ofroots, enhanced the rate of yellowing, and altered the locationof chlorophyll loss from the margin of the lamina to the petiolarregion in most of the cotyledons. These effects were not producedwhen osmotica such as mannitol or polyethylene glycol were used. A few cotyledons were able to root in the presence of sucroseand these showed enhanced yellowing at the margin. The numberof such cotyledons increased when they were incubated in waterfor the period during which primordium initiation begins, beforetransfer to sucrose. However, the pattern of yellowing in thepresence of sucrose was not affected by the repeated removalof the region of primordium development, suggesting that thepattern of senescence is not controlled by the formation ofroot primordia. The results are discussed in relation to the effects of lightand possible accumulation of carbohydrate on senescence androot formation in excised tissues.     

Changes in thiocyanate content in some radish Raphanus sativus L. cultivars during hypocotyl-root growth

Changes in the thiocyanate content in hypocotyl-roots and leaves of radish were observed in a two-year field experiment. Six cultivars were tested: early radish (Rex and Ostergruss Różowa), Japanese radish (Tokinashi and Minowase Summer Cross F₁), and winter radish (Monachijska Biała and Murzynka). A significant diversification in thiocyanate content among cultivars, plant parts, harvest dates and observation years was found. Early cultivars contained the least amount of these compounds, Murzynka — the greatest. The content of thiocyanates in leaves was 3 – 5 times higher than that in hypocotyl-roots. The changes in the thiocyanate content during root growth showed a constant rising tendency in the case of the leaves of all cultivars and the storage organs of Murzynka.     

Genetic control of the wavy shoots character in radish Raphanus sativus L

The results of genetic analysis of the wavy shoots character supported the hypothesis on the existence of two genes, Wsh1 and Wsh2, whose mutations caused wavy shoots in radish lines 16 and 26, respectively. As shown by analysis of joint inheritance with marker characters, wavy shoots were inherited independently of the biochemical markers (genes Lap1, Ep, Dia1, and Aat). Gene Wsh1 was shown to be linked to genes A, ac, and ar; the mode of inheritance depended on the cross combination. In hybrids with mutant line 16, plants with novel morphological defects, which were absent in the parental forms, were detected. Possible mechanisms of new morphological abnormalities and change of linkage in the combinations analyzed are discussed.     

Interrelationship between chromocentres and chiasmata in radish (Raphanus sativus L.)

A comparative study of the number and distribution of chromocentres in interphase nuclei and mean chiasma frequency at diakinesis has been made in three varietal populations of radish (Raphanus sativus L.), Scarlet Globe, Japanese White and Chinese White. The study showed a significant difference between the varietal populations in mean chiasma frequency and number of chromocentres (P<0.001), indicating that these nuclear characters are genotypically controlled. The correlation analysis revealed a significant negative correlation between chromocentres and chiasma frequency (r= -0.87). It was concluded that an increase in the amount of constitutive heterochromatin, as inferred by chromocentre counts, adversely affects the chiasma frequency and, consequently, genetic recombination in radish.     

QTL mapping of clubroot resistance in radish (Raphanus sativus L.)

A QTL analysis for clubroot resistance (CR) of radish was performed using an F₂ population derived from a crossing of a CR Japanese radish and a clubroot-susceptible (CS) Chinese radish. F₃ plants obtained by selfing of F₂ plants were used for the CR tests. The potted seedlings were inoculated and the symptom was evaluated 6 weeks thereafter. The mean disease indexes of the F₃ plants were used for the phenotype of the F₂. The results of two CR tests were analyzed for the presence of QTL. A linkage map was constructed using AFLP and SSR markers; it spanned 554 cM and contained 18 linkage groups. A CR locus was observed in the top region of linkage group 1 in two tests. Therefore, the present results suggest that a large part of radish CR is controlled by a single gene or closely linked genes in this radish population, although minor effects of other genomic areas cannot be ruled out. The CR locus was named Crs1. Markers linked to Crs1 showed sequence homology to the genomic region of the top of chromosome 3 of Arabidopsis, as in the case of Crr3, a CR locus in Brassica rapa. These markers should be useful for breeding CR cultivars of radish. As Japanese radishes are known to be highly resistant or immune to clubroot, these markers may also be useful in the introgression of this CR gene to Brassica crops.     

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。     

Assorbimento di saccarosio in ipocotili di ravanello (Raphanus sativus L.)

Abstract

In questo lavoro abbiamo determinato le caratteristiche di entrata del saccarosio in funzione della sua concentrazione esterna, incubando i pezzi in saccarosio triziato, in condizioni normali e di stress osmotico. Nelle parti apicali l'assorbimento di saccarosio esogeno è lineare in presenza di basse concentrazioni mentre non lo è ad alte concentrazioni. Per quanto riguarda le parti basali la linearità si mantiene per tutte le concentrazioni; inoltre nelle parti apicali l'assorbimento è maggiore che nelle basali e la differenza aumenta all' aumentare delle concentrazioni di saccarosio esterno. Abbiamo poi studiato l'assunzione di saccarosio con osmotici diversi in cui fosse presente anche saccarosio marcato. I risultati ottenuti dimostrano che le parti apicali stressate prendono meno saccarosio dei controlli, mentre nelle basi stressate l'assorbimento, pur essendo inferiore a quello degli apici, è superiore rispetto ai controlli. Le differenze nel trasporto del saccarosio negli apici e nelle basi potrebbero essere spiegate dal fatto che a parità di peso nelle zone apicali ci sono più cellule di dimensioni inferiori che non hanno completato la distensione cellulare e che essendo in attiva crescita sono metabolicamente più attive. Nelle zone basali è invece presente un maggiore numero di cellule morte (xilema completamente differenziato). Questi due fattori potrebbero determinare nelle parti apicali un maggiore assorbimento con prevalenza del trasporto attivo e nelle parti basali un assorbimento inferiore con prevalenza del trasporto per diffusione.     

萝卜单倍体培养

1 植物名称　萝卜(Raphanus Sativus),又名莱菔、芦菔. 2 材料类别　花药. 3 培养条件 (1)胚状体诱导:以改良的Keller为基本培养基,10%蔗糖、0.8%琼脂粉,pH 5.8,不附加任何激素.(2)诱导分化培养基:MS+ 0.2 mg·L-1 6-BA,2%蔗糖、0.8%琼脂条,pH 5.5.     

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.     

Phytochrome-regulated transfer of fructosidase from cytoplasm to cell wall in Raphanus sativus L. hypocotyls

The far-red absorbing form of phytochrome, P_fr, rapidly increases the rate of transfer of -fructosidase (E.C.3.2.1.26) from the cytoplasm to the cell wall in radish hypocotyls. Far-red light increases the level of enzyme in a particulate fraction: after two hours of light treatment, the particulate enzyme is associated almost exclusively with the endoplasmic reticulum. Transfer from the endoplasmic reticulum to the cell wall involves an incorporation into Golgi bodies and the plasmalemma: these membrane fractions were separated by centrifugation on a discontinuous sucrose density gradient and their degree of purity was determined by the use of known biochemical markers. With respect to -fructosidase, light controls, via P_fr: (1) the total amount, (2) the incorporation into the endoplasmic reticulum and (3) the transfer to the cell-wall. These three processes have different sensitivities to cycloheximide.Abbreviations -FFase -fructosidase - IDPase inosine diphosphatase - SGTase UDPG-sterol glucosyltransferase - NCRase NADPH-cytochrome c-oxydoreductase - NPA N-naphtylphtalamic acid - BSA bovine serum albumine     

Micropropagation of Raphanus sativus L. var. longipinnatus (Japanese radish) cv. Gungjung

Shoot cultures were established from seedling shoot tips of Raphanus sativus var. longipinnatus Bailey cv. Gungjung, (Japanese radish) cultured on a Murashige-Skoog medium supplemented with ca. 4.5–135 M kinetin or N⁶-benzyladenine. The latter cytokinin supported overall better growth, and 22.2 M was adopted for maintenance of established cultures. The nitrate: ammonium levels in the medium proved optimal for growth and shoot proliferation and both these parameters were significantly increased by addition of adenine sulfate or sodium phosphate. Rooting of excised shoots was achieved on auxin containing medium. Indole-3-butyric acid (ca. 5 or 10 M) also enhanced shoot growth. Plants were easily established in soil, appeared morphologically normal, and flowered.     

The Control of Root Initiation and Development in Detached Cotyledons of Sinapis alba L. and Raphanus sativus L.

The initiation and development of root primordia in detachedcotyledons of Sinapis alba (white mustard) and Raphanus sativus(radish) are studied, together with the inhibitory effects ofsucrose and mineral nutrients on these processes. Root primordium initiation on petioles of excised mustard cotyledonscultured in petridishes in water commenced after 3 days andwas completed after 5 days in culture, by which time a numberof the primordia had extended and emerged from the petiole.Both sucrose and mineral nutrient solution had an inhibitoryeffect which was most marked when the cotyledonswere culturedin the solution from the time of excision. The total numberof primordia initiated, their rate of development, and the finaltotal number of emerged roots were all reduced. The later thetime of transfer from water either to sucrose or to nutrient,the less marked the inhibition. Indeed, nutrient solution enhancedroot growth in mustard when cotyledons were transferred after5 days in water when root emergence had just commenced. The effects of sucrose and nutrients in relation to applicationbefore and after initial meristem formation has taken placeare discussed, together with the ways in which these two solutionsmay exert their effect on root initiation and development.     

Triadimefon enhances growth and net photosynthetic rate in NaCl stressed plants of Raphanus sativus L.

The effect of sodium chloride and triadimefon (TDM) on the chlorophyll (Chl) content, net photosynthetic rates (P_N), rate of transpiration (E), and intercellular CO₂ concentration (C_i) in Raphanus sativus was studied. The effect of NaCl salinity was partially ameliorated by TDM which caused increase in Chl content, P_N, and C_i. TDM also increased root dry matter production, decreased E, and increased the water use efficiency. This revised version was published online in June 2006 with corrections to the Cover Date.     

Thiocyanate content in relation to the quality features of radish Raphanus sativus L

Relationships between radish thiocyanate content and its dry weight, the content of sugar, protein, fibre, ascorbic acid, some minerals, the incidence of plant shooting, the firmness and pithiness of storage-roots, and the ratio of leaves to storageroot (wt/wt) were investigated. The analysis of linear correlation was based on numerous data from the 4-year field experiment with six radish cultivars and different sowing and harvest dates. The content of thiocyanate in radish roots was found to be positively correlated with their dry weight, and the content of total protein, crude fibre, and soluble sugar. A strong relationship was found between the content of thiocyanate and dry weight of radish leaves. The negative correlation between the thiocyanate content in the leaves and the firmness of storageroots and the positive correlation with their pithiness might indicate the translocation of this compounds into green plant parts during the ageing of root tissue. The root thiocyanate content and the percentage of shooting correlated significantly only in the case of Tokinashi. The closeness of relations between the ratio of leaves/storage-root and thiocyanate content, though in general small, was affected also by a cultivar. A similar effect was observed for the correlations between the thiocyanate contents in leaves and storage-roots.     

Triadimefon enhances growth and net photosynthetic rate in NaCl stressed plants of Raphanus sativus L.

The effect of sodium chloride and triadimefon (TDM) on the chlorophyll (Chl) content, net photosynthetic rates (P_N), rate of transpiration (E), and intercellular CO₂ concentration (C_i) in Raphanus sativus was studied. The effect of NaCl salinity was partially ameliorated by TDM which caused increase in Chl content, P_N, and C_i. TDM also increased root dry matter production, decreased E, and increased the water use efficiency.