Radish Accessions Research Articles

Induction of Glucoraphasatin Biosynthesis Genes by MYB29 in Radish (Raphanus sativus L.) Roots.

Glucoraphasatin (GRH) is a specific aliphatic glucosinolate (GSL) that is only abundant in radish (Raphanus sativus L.). The gene expression regulating GRH biosynthesis in radish is still poorly understood. We employed a total of 59 radish accessions to analyze GSL profiles and showed that GRH was specific and predominant among the aliphatic GSLs in radish roots. We selected five accessions roots with high, moderate and low GSL biosynthesis, respectively, to conduct a comparative transcriptome analysis and the qRT-PCR of the biosynthesis genes for aliphatic GSLs. In this study, among all the accessions tested, roots with the accession RA157-74 had a high GRH content and showed a significant expression of the aliphatic GSL biosynthesis genes. We defined the genes involved in the GRH biosynthesis process and found that they were regulated by a transcription factor (RSG00789) at the MYB29 locus in radish roots. We found 13 aliphatic GSL biosynthesis genes regulated by the RSG00789 gene in the GRH biosynthesis pathway.

Identification of genome-wide single-nucleotide polymorphisms among geographically diverse radish accessions.

Radish (Raphanus sativus L.) is cultivated around the world as a vegetable crop and exhibits diverse morphological and physiological features. DNA polymorphisms are responsible for differences in traits among cultivars. In this study, we determined genome-wide single-nucleotide polymorphisms (SNPs) among geographically diverse radish accessions using the double-digest restriction site-associated DNA sequencing (ddRAD-Seq) method. A total of 52,559 SNPs was identified in a collection of over 500 radish accessions (cultivated and wild) from East Asia, South and Southeast Asia, and the Occident and Near East. In addition, 2,624 SNP sites without missing data (referred to as common SNP sites) were identified among 510 accessions. Genetic diversity analyses, based on the common SNP sites, divided the cultivated radish accessions into four main groups, each derived from four geographical areas (Japan, East Asia, South and Southeast Asia, and the Occident and Near East). Furthermore, we discuss the origin of cultivated radish and its migration from the West to East Asia. SNP data generated in this work will facilitate further genetic studies on the radish breeding and production of DNA markers.

Genetic and histological characterization of downy mildew resistance at the cotyledon stage in Raphanus sativus L.

Downy mildew (DM; Hyaloperonospora brassicae) is one of the most serious diseases in radish and responsible for high production losses. The objectives of this study were (1) to identify new sources of DM resistance at the cotyledon stage; (2) to determine the inheritance of the resistance; and (3) to clarify the histological and cellular response involved in DM infection. We screened 200 radish accessions for DM resistance at the cotyledon stage. Radish accessions Rd001, Rd004, Rd048 and Rd150 showed a complete resistance response (hypersensitive reaction), and accession Rd193 exhibited a partial resistance with confined sporulation. The highly susceptible accession Rd197 was then crossed with all resistant genotypes for a genetic analysis. F2 segregation analysis suggested that cotyledon resistance in Rd001, Rd004, Rd048 and Rd150 is conferred by a single dominant gene while resistance in Rd193 is polygenic. Genetic data suggests Rd193 resistance might be conferred by two dominant genes with complementary action, althought further experiments are needed to confirm this hypothesis. To conduct a histological study using light and fluorescence microscopy, we collected cotyledons from three genotypes with either differing levels of resistance (Rd004 and Rd193) and high susceptibility (Rd197) at ten different time points, from 3 h to 7 days post inoculation. Tissue necrosis was responsible for the inhibition and reduction of H. brassicae growth in Rd004 and Rd193 within 24 h of inoculation. For susceptible accession Rd197, the first evidence of necrotic cells appeared 48 h post inoculation. Callose deposition associated with intercellular H. brassicae growth occurred 3 h post inoculation for all genotypes and was the lowest in resistant accession Rd004. Cotyledon resistance to DM identified in radish is of high commercial interest and has the potential to be used in breeding programs.

AHAS Trp574Leu substitution in Raphanus sativus L.: screening, enzyme activity and fitness cost.

Feral radish (Raphanus sativus L.) is a problematic weed that has become resistant to acetohydroxyacid synthase (AHAS) inhibitor herbicides due to the Trp574Leu mutation. An AHAS gene mutation that causes herbicide resistance may have negative pleiotropic effects on plant fitness. This study reports the effects of the Trp574Leu mutation on AHAS activity and reproductive traits of R. sativus. Eight of 17 feral radish accessions presented individuals resistant to metsulfuron-methyl at 0.5% to >90.0% and all the resistant individuals analyzed showed the Trp574Leu mutation. Without herbicide selection, the AHAS activity was 3.2-fold higher in the susceptible accession than in the resistant one. The resistant accession was >9000-fold more resistant to metsulfuron-methyl and imazethapyr than the susceptible accession. Under low intraspecific competition during two growing seasons, AHAS-resistant feral radish accessions showed 22-38% and 21-47% lower seed numbers and yield per plant than the susceptible accession. This is the first report of fitness cost associated with the AHAS Trp574Leu mutation in R. sativus populations. This fitness cost could reduce frequency of the resistant allele without herbicide selection. © 2018 Society of Chemical Industry.

AGRO-BIOLOGICAL ASSESSMENT OF RADISH ACCESSIONS AT NOVOSIBIRSK AREA

The result of collaboration work between Siberian Research Institute of Plant Cultivation and Breeding, the branch of The Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences (ICG SB RAS) and the company OOO Geterozisnaya Selectia on ecological trial and assessment of economically valuable traits were obtained in promising radish accessions. The data on influence of sowing terms on yielding ability, formation of flower bearing stalks was analyzed and morphological descriptions for radish accessions were given. It was shown that the development of marketable radish roots can be possible in condition of long lasting light period. When sowing in the second decade of June less number of flower stalks formed on the plants than on the control plants of ‘Raniy Krasniy’. The sowing in the first decade of August did not give any plants with flower stalks in accessions 17-12 and 17-13. The highest yield at first sowing term was observed in accession 17-12 at second sowing term in accession 17-13, 5.6 and 6.1 kg/m2 respectively. The accession 17-12 was distinguished by root weight, 19.2 g. The highest dry matter content (5.31%) was observed in accession 17-11; the highest total sugar content (2.18%) was in accession 17-12; the highest ascorbic acid content (25.1 mg %) was in accession 17-13. The comparative analysis of data on summer and autumn sowing terms showed that when sowing in summer time, at long lasting light period the radish accessions were characterized by the more number of plants with flower stalks; more developed leaf apparatus; larger roots and enriched biochemical composition as compared with the accessions sown in autumn time. All accessions tested belonged to early-maturing group, where the root formation takes for 22-25 days. For economically valuable traits the accessions 17-12 and 17-13 were selected out.

EVALUATION OF ESTERASE POLYMORPHISMS IN MATURE SEEDS OF RADISH (RAPHANUS SATIVUS L.) ACCESSIONS OF VIR COLLECTION

A biochemical evaluation of 25 radish accessions (Raphanus sativus L.) on esterase isozymes of mature seeds has been carried out. The results of the experiments showed a wide range of diversity among the genotypes based on electrophoretic zones of esterase isoenzymes. The revealed isoenzyme complex of esterases was represented by eight isoforms with molecular weights from 37.7 kD to 57.6 kD. All accessions were divided into 13 electrophoretic zymotypes, differing from each other by the presence or absence of definite zones. The most often observed electrophoretic zymo-type is Gr. 1, which includes 24% of the total number of accessions evaluated. There are 8 zymotypes (Gr. 6 Gr. 13) with a frequency of occurrence 4%. Three groups (Gr. 2 – Gr. 4) had the same frequency of occurrence – 12%. Zimotype of Gr. 5 containes the maximum number of zones – 8. 2 zimotypes – Gr. 3 and Gr. 12 had the smallest number of 4 zones. Two zones of esterases – zones 7 and 8 (Мr 39.7кD and Мr 37.7 kD, respectively) were monomorphic. The remaining six zones were polymorphic, i.e. could be absent in some zimotypes. The frequency of occurrence of each zone in different zymotypes has varied from 6.58% to 17.11%. As results of this research the accessions that were selected can become the most promising parent forms for future genetic and selection studies of this culture.

Cytological and morphological analysis of hybrids between Brassicoraphanus, and Brassica napus for introgression of clubroot resistant trait into Brassica napus L.

Interspecific hybridization is a powerful tool for improvement of crop species, it has the potential to broaden the genetic base and create new plant forms for breeding programs. Synthetic allopolyploid is a widely-used model for the study of genetic recombination and fixed heterosis in Brassica. In Brassica napus breeding, identification and introgression of new sources of clubroot resistance trait from wild or related species into it by hybridization is a long-term crop management strategy for clubroot disease. Radish (Raphanus sativus L.) is a close relative of the Brassica and most radish accessions are immune to the clubroot disease. A synthesized allotetraploid Brassicoraphanus (RRCC, 2n = 36) between R. sativus cv. HQ-04 (2n = 18, RR) and Brassica oleracea var. alboglabra (L.H Bailey) (2n = 18, CC) proved resistant of multiple clubroot disease pathogen P. brassicae. To predict the possibility to transfer the clubroot resistance trait from the RR subgenome of allotetraploid Brassicoraphanus (RRCC, 2n = 36) into Brassica napus (AACC, 2n = 38), we analyzed the frequency of chromosome pairings in the F1 hybrids produced from a cross between B. napus cv. HS5 and the allotetraploid, characterize the genomic composition of some backcrossed progeny (BC1) using GISH, BAC-FISH and AFLP techniques. The level of intergenomic pairing between A and R genomes in the F1 hybrid was high, allosyndetic bivalents formed in 73.53% PMCs indicative of significant level of homeologous recombination between two genomes and high probability of incorporating chromosomal segments/genes from R-genome into A/C-genomes. The BC1 plants inherited variant extra R chromosomes or fragments from allotetraploid as revealed by GISH and AFLP analysis. 13.51% BC2 individuals were resistant to clubroot disease, and several resistance lines had high pollen fertility, Overall, the genetic material presented in this work represents a potential new genetic resource for practical use in breeding B. napus clubroot resistant cultivars.

THE USE THE GENETICALLY DIFFICULTLY INHERITED TRAIT OF PURPLE ROOT COLOR IN BREEDING PROGRAM FOR THE COMPLICATED TRAIT IN RADISH

The understanding the nature of trait inheritance in any crops is that determines the quality of results in breeding program. According to reference on previous publication, it is known that phenotypic manifestation of purple root color in radish was caused by regulatory interrelationship mechanisms of genetic control that is difficult to be used directly in breeding program. From literature sources and on the basis of their own research work the authors have proven the practice to maintain the trait in generations, and implementations of development of purple radish breeding accessions have been presented. At first stage of breeding program the selection of initial breeding accessions was carried out, where 14 varieties (red x white) were regarded on the basis of top-crosses to obtain F1 and F2 progenies to be analyzed. Thus, four best combinations from crossing were chosen with 100% of hybridity. Through analysis of hybrids for individual progenies the hybrid population F1 of radish ‘Konfeti’ with different root colors was developed. As result of the individual inbreeding selection on seed plants with pigmented stems and the colored flower rim, the stable breeding accession with purple root was obtained. Thus, in breeding practice in radish it was succeeded to obtain the stably inheriting purple root color in radish accessions, variety ‘Siniiy Iniey’.

Reproductive traits and molecular evidence related to the global distribution of cultivated radish (Raphanus sativus L.)

Reproductive characteristics frequently vary geographically, and reproductive success is critical for survival and local adaption in cultivated species. To document such variation in radish (Raphanus sativus L.), we examined reproductive traits and neutral genetic markers in 64 wild and landrace accessions varying in geographic origin. Flowering time tended to increase with latitude of origin, with southeastern Asian accessions (the lowest latitude) flowering earliest. Accessions from eastern Asia, which encompasses the greatest latitudinal range (18° and 54°N) of the regions we examined, exhibited the greatest variation in flowering time, clustering into a single group according to neutral genetic markers. Cluster analysis of genetic data divided the radish accessions into European and Asian groups. These groups were further subdivided into six, and two subgroups of Europe could be distinguished by root skin color. Although the accessions tended to cluster by geographic origin, those from central, southern, and western Asia clustered with both European and Asian accessions. Our results suggest that artificial selection of flowering time as well as root skin color has played important roles in local adaptation and increases in the genetic diversity of radish landraces in different geographic regions.

Broad resistance to acetohydroxyacid-synthase-inhibiting herbicides in feral radish (Raphanus sativus L.) populations from Argentina.

Soon after the commercial release of sunflower cultivars resistant to imidazolinone herbicides, several uncontrolled feral radish (Raphanus sativus L.) populations were found in south-eastern Buenos Aires, Argentina. These populations were studied in field, glasshouse and laboratory experiments aiming to characterise their resistance profile and to develop management tools. Three feral radish accessions were highly resistant to ten active ingredients of five families of acetohydroxyacid synthase (AHAS)-inhibiting herbicides. Sequence analysis of the AHAS gene detected a Trp574Leu mutation in all resistant accessions. One accession with an intermediate level of resistance was heterozygous for this mutation, probably owing to gene exchange with a susceptible subpopulation located in the field margin. Herbicide-resistant and herbicide-susceptible radish could be controlled in sunflower by alternative herbicides. This is the first report of feral radish with resistance to herbicides belonging to all the AHAS-inhibiting herbicide families, conferred by Trp574Leu mutation in the AHAS gene. An appropriate herbicide rotation with alternative herbicides such as fluorochloridone or aclonifen and an increase in the diversity of cropping systems are important for minimising the prevalence of these biotypes.

Genetic diversity and evolutionary relationship analyses within and among Raphanus species using EST-SSR markers

We screened 626 pairs of EST-SSR primers from the radish marker database using 37 different cultivars. Of the primer pairs, 89.3 % (559) were polymorphic. The average polymorphism information content (PIC) value per primer was 0.39, ranging from 0.07 to 0.73. Fifty EST-SSR markers were designated core markers according to PCR band reproducibility and stability, high polymorphism levels, easily discriminated alleles, and an even distribution of molecular markers over each radish chromosome. Genetic diversity and the evolutionary relationships of 93 radish accessions, representing nearly all typical Raphanus germplasms, were assessed with 50 core EST-SSR markers. A total of 254 alleles at 52 loci (4.88 alleles per locus on average), with a mean PIC value of 0.55, were detected. The wild radish had the largest range of variation, followed by the East Asian big long radish and the European small radish. Unweighted pair group method with arithmetic mean cluster analysis classified the 93 germplasms into four groups. The first group included most accessions belonging to R. sativus, while seven wild Raphanus strains were separated into the remaining groups. In the R. sativus group, the subspecies var. hortensis Becker, var. raphanistroides Makino, var. caudatus Hooker and Anderson, var. sativus L., var. niger Kerner, and var. oleiferus Metzg were sorted into separate subgroups. Population genetics structure analyses show that the accessions were attributed to three populations: (1) Asia cultivated and East Asian wild, (2) Europe cultivated, and (3) wild Raphanus species. Herein, we discuss the origin and ancestry of cultivated radishes based on these results.

Physiology, anatomy, and cell membrane thermostability selection of leafy radish (Raphanus sativus var. oleiformis Pers.) with different tolerance under heat stress

Leafy radish (Raphanus sativus L. var. oleiformis Pers.), a temperate crop, is usually heat-sensitive. Line 9911-9 is a heat-tolerant line bred for summer vegetables in Taiwan. Understanding the heat tolerance mechanism of Line 9911-9 may facilitate future breeding programs. We conducted a comparative study on growing plants of heat-tolerant Line 9911-9, moderately tolerant Line 9611, and heat-sensitive ‘Taichung No. 1’ at 40/35°C and 25/20°C to elucidate the heat tolerance mechanism of leafy radish. After 28-day of acclimation, the maintenance of net photosynthetic rate (Pn) and chlorophyll fluorescence (Fv/Fm) accompanying a double increase of stomatal conductance (gs) and transpiration rate (E) in Line 9911-9 revealed that the heat-tolerant line had adjusted physiologically to be more efficient in heat dispersing. Morphologically, Line 9911-9 developed more xylem vessels, an enlarged stomatal aperture by 5-fold, and an increased stomatal opening to 93%, which contributed 6-fold increase in the occupancy of total stomata aperture on leaf without altering stomata density under high temperature stress. Furthermore, the occupancy was correlated with gs and E with a coefficient of 0.87 and 0.98, respectively, confirming that stomatal factors play a crucial role in heat tolerance in leafy radish. Regarding the heat shock response, Line 9911-9 could have sustained over 9 days at 40/35°C, whereas Line 9611 was maintained over 3 days, with ‘Taichung No. 1’ for only 1 day. Line 9911-9 was able to maintain its cellular membrane stability by inducing the activities of superoxide dismutase (SOD) and catalase (CAT) to scavenge reactive oxygen species (ROS) within the first 24h and detoxify ROS unbalance under high temperature stress. These results indicated that Line 9911-9 was able to lower the non-stomatal limitation effect on Pn. A high correlation (R2=0.89) between cotyledon relative injury at 50°C for 30min in 21 leafy radish accessions and their yield reduction ratio under high temperature stress suggests that the CMT of cotyledon may be used as a reliable index for selecting heat-tolerant leafy radish.

Resistance gene analogue isolation and RGA-based marker development for identifying downy mildew resistance in radish (Raphanus sativus L.)

Resistance Gene Analogs (RGAs)-based molecular markers can significantly enhance the breeding efficiency for disease resistance in various crops. However, RGAs isolation and RGA-based marker development have not been conducted in radish. In this study, two prevalent approaches, PCR amplification with degenerate primers and data-mining of radish EST databases, were used to isolate radish resistance gene candidate sequences. A total of 26 RsRGAs with high homology to known R-genes or RGAs were obtained by PCR amplification. Meanwhile, 257 R-gene-like unigenes/ESTs were identified using data-mining method. In total, 115 out of 124 designed RGA-specific primer pairs can successfully amplified the target bands. To explore the RGA genetic variability, 32 radish accessions were genotyped and 269 RGA loci were successfully identified. It was found that 44 RGA primers only generated monomorphic band and the remaining 71 RGA primers generated 225 RGA polymorphic bands among the 32 radish accessions. Based on RGA marker analysis, 32 radish accessions were clustered into four main groups at the similarity index of 0.68, which was in a high accordance with disease index from a downy mildew evaluation. This study might be the first report on candidate R-genes identification and RGA-based markers development in radish. These findings will facilitate the disease resistance identification and speed up the genetic improvement of DM resistances in radish breeding programs.

Novel and useful genic-SSR markers from de novo transcriptome sequencing of radish (Raphanus sativus L.)

Simple sequence repeats (SSRs) are highly polymorphic and co-dominant markers, providing an important genomic resource for genetic research. Recently, large-scale transcriptome sequencing has become a reliable and efficient approach for the identification and development of new genic-SSR markers and has been successfully conducted in a few important plant species. However, SSR development based on transcriptome sequencing remains limited in radish (Raphanus sativus L.). In the present study, from a total of 73,084 unigenes and 150,455 contigs which were assembled from 71.95 million Illumina sequence reads of a radish taproot library, a collection of 11,928 genic-SSR loci were successfully identified in 11,311 unigene sequences. Trinucleotide repeats were the most abundant repeat units, as in many other plants, with a frequency of 52 %. Furthermore, a total of 5,503 genic-SSR primers were developed, from which 1,052 SSR primers were synthesized, and a subset of 823 (78.23 %) primers could generate stable bands. Moreover, 67 selected informative genic-SSR markers were used to determine the genetic diversity of 32 radish genotypes, in which the polymorphism information content values ranged from 0.49 to 0.89. For effective cultivar identification, a novel strategy called manual cultivar identification diagram was employed. Thirty-two radish accessions were clearly separated by six genic-SSR markers. Additionally, the cross-species/genera transferability of these SSRs was further validated in nine relatives in Brassicaceae. These results suggested that the novel genic-SSR markers, as a basis for future genetic linkage and gene tagging analysis, could be very valuable in facilitating genetic mapping, marker-assisted selection and comparative genome analysis.

Comprehensive analysis of expressed sequence tags from cultivated and wild radish (Raphanus spp.)

BackgroundRadish (Raphanus sativus L., 2n = 2× = 18) is an economically important vegetable crop worldwide. A large collection of radish expressed sequence tags (ESTs) has been generated but remains largely uncharacterized.ResultsIn this study, approximately 315,000 ESTs derived from 22 Raphanus cDNA libraries from 18 different genotypes were analyzed, for the purpose of gene and marker discovery and to evaluate large-scale genome duplication and phylogenetic relationships among Raphanus spp. The ESTs were assembled into 85,083 unigenes, of which 90%, 65%, 89% and 89% had homologous sequences in the GenBank nr, SwissProt, TrEMBL and Arabidopsis protein databases, respectively. A total of 66,194 (78%) could be assigned at least one gene ontology (GO) term. Comparative analysis identified 5,595 gene families unique to radish that were significantly enriched with genes related to small molecule metabolism, as well as 12,899 specific to the Brassicaceae that were enriched with genes related to seed oil body biogenesis and responses to phytohormones. The analysis further indicated that the divergence of radish and Brassica rapa occurred approximately 8.9-14.9 million years ago (MYA), following a whole-genome duplication event (12.8-21.4 MYA) in their common ancestor. An additional whole-genome duplication event in radish occurred at 5.1-8.4 MYA, after its divergence from B. rapa. A total of 13,570 simple sequence repeats (SSRs) and 28,758 high-quality single nucleotide polymorphisms (SNPs) were also identified. Using a subset of SNPs, the phylogenetic relationships of eight different accessions of Raphanus was inferred.ConclusionComprehensive analysis of radish ESTs provided new insights into radish genome evolution and the phylogenetic relationships of different radish accessions. Moreover, the radish EST sequences and the associated SSR and SNP markers described in this study represent a valuable resource for radish functional genomics studies and breeding.

Variation of Glucosinolates in Wild Radish (Raphanus raphanistrum) Accessions

Glucosinolate composition was determined in wild radish accessions from eight states in the northeastern and southern United States to determine the variability of production among accessions. Glucosinolates were evaluated from roots, leaves, flowers, primary, and secondary branches. Seventeen glucosinolates were identified, with glucoerucin, glucoraphenin, glucobrassicin, and gluconasturtiin contributing 90% to 100% of the total glucosinolates. Flowers contained the highest glucosinolate concentrations, 12.07 to 55.36 μmol/g, but flowers contributed only 5.3 to 21.3% to the total glucosinolates. Of the eight accessions, the Mississippi accession produced significantly higher levels of total glucosinolates and glucosinolates which can be degraded to isothiocyanates per plant, totals of 618.97 and 563.53 μmol/plant, respectively. Total plant biomass did not differ between accessions indicating a difference in the ability of the Mississippi accession to produce glucosinolates. Further studies are needed to determine if this accession would consistently produce higher glucosinolate levels under different environmental conditions.

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.

Inter-accession variation in floral morphology in radish (Raphanus sativus L.)

We investigated variation in floral morphology among 36 accessions of radish (Raphanus sativus L.). We found wide, continuous inter-accession variation in 15 traits related to the size of the floral organs and the stigma position relative to the anthers of long stamens. A principal-component analysis of the 15 floral traits also showed wide variation among accessions. A large proportion of this variation was accounted for by lengths of stamens and the petal tube. In addition, the phenotypic stabilities with respect to the macro-environment (as affected by different sowing dates) and the micro-environment (among the flowers within a plant) differed between traits. Correlation analyses using all traits suggest the existence of genetic linkages or pleiotropies among floral organs. Our results indicate that floral morphologies varied greatly among radish accessions and they could be affected by the macro- and micro-environment. To explore the relationship between floral morphology and pollination efficiency, we compared the number of pollen grains deposited on the stigmas in the presence and absence of insect pollinators using three accessions with very different floral morphology. Because an accession with a higher stigma position than anthers might avoid self-pollination and an accession with a large stigma size received more pollen grains, floral morphology may affect pollination efficiency. Because the effect of pollination efficiency on seed production is well known, our results suggest that radish floral morphology affects seed production through its effect on pollination efficiency.

Genetic variation in radish (Raphanus sativus L.) germplasm from Pakistan using morphological traits and RAPDs

Genetic diversity of 30 radish (Raphanus sativus L.) accessions was investigated at the phenotypic level with morphological characters and at the DNA level using the random amplified polymorphic DNA (RAPD) technique. Thirty-six morpho-physiological traits were recorded from seedling stage to harvest. The 31 primers used generated 202 RAPD bands, of which 158 (78.2%) were polymorphic. Multivariate procedures were used to classify the germplasm on the basis of phenotypic traits and RAPD fragments. Dendrograms were generated for the Euclidean distance from the morphological data and the Nei's genetic distance from the RAPD markers. Phenotypically, all the accessions were classified into four major groups corresponding to the different forms of cultivated radish. The morphological diversity existing within each of these groups suggested that they should be discriminated into the three botanical convarieties, sativusT (large-rooted), caudatus (pod-type) and oleifer (oilseed-type). Clustering of the accessions did not show any pattern of association between the morphological characters and the collection sites. Instead, landrace groups were associated with their morphological similarities and horticultural uses. On the other hand, the intra-specific genetic relationships of several accessions based on RAPD analysis were related primarily to their collection sites rather than to their phenotypic affinities. The level of polymorphism exhibited by the various convarieties could be exploited in genetic mapping populations to tag economically important traits. These genotypes also could serve as a useful germplasm source for root, leaf, pod and seed. This preliminary study of traditional radish landraces from Pakistan provides useful information regarding their horticultural potential.