Oilseed Brassica Species Research Articles

Genome-wide analysis of Catalase gene family reveal insights into abiotic stress response mechanism in Brassica juncea and B. rapa.

Environmental stresses affect the yield and productivity of Brassica crops. Catalases are important antioxidant enzymes involved in reducing excess hydrogen peroxide produced by environmental stresses. In the present study, nine and seven CAT family members in two oilseed Brassica species (B. juncea and B. rapa) were identified with complete characterization based on gene and protein structure. Phylogenetic classification categorized CAT proteins into three classes and differentiated the monocot and dicot-specific CAT proteins. Further, the gene and protein characterizations revealed a high degree of conservation across the CAT family members. Differences were observed in the CAT-HEME binding affinity in CAT1, CAT2, and CAT3 isozymes, which could suggest their differential enzyme activities in different conditions. Furthermore, protein-protein interaction with other antioxidant proteins suggested their coordinated role in ROS scavenging mechanisms. Notably, the differential gene expression of BjuCATs and BraCATs and CAT enzyme activities suggested their crucial roles in major abiotic stresses faced by Brassica species. Promoter analysis in BjuCATs and BraCATs suggested the presence of abiotic-stress responsive cis-regulatory elements. Gene regulatory network analysis suggested miRNA and TF mediated stress response in BjuCATs and BraCATs. CAT family screening and characterization in Brassica sp. has established a basic ground for further functional validation in abiotic and heavy-metal stresses which can help in developing stress tolerant crops.

The Global Assessment of Oilseed Brassica Crop Species Yield, Yield Stability and the Underlying Genetics.

The global demand for oilseeds is increasing along with the human population. The family of Brassicaceae crops are no exception, typically harvested as a valuable source of oil, rich in beneficial molecules important for human health. The global capacity for improving Brassica yield has steadily risen over the last 50 years, with the major crop Brassica napus (rapeseed, canola) production increasing to ~72 Gt in 2020. In contrast, the production of Brassica mustard crops has fluctuated, rarely improving in farming efficiency. The drastic increase in global yield of B. napus is largely due to the demand for a stable source of cooking oil. Furthermore, with the adoption of highly efficient farming techniques, yield enhancement programs, breeding programs, the integration of high-throughput phenotyping technology and establishing the underlying genetics, B. napus yields have increased by >450 fold since 1978. Yield stability has been improved with new management strategies targeting diseases and pests, as well as by understanding the complex interaction of environment, phenotype and genotype. This review assesses the global yield and yield stability of agriculturally important oilseed Brassica species and discusses how contemporary farming and genetic techniques have driven improvements.

Floral volatiles may influence honey bee visitations in oilseed Brassica species

ABSTRACT Understanding foraging preferences of honey bees (Apis mellifera L.) is essential to maneuver pollinator movement to enhance outcrossing and hence the crop yields. In this communication, we report the role of floral volatile organic compounds (VOCs) in structuring foraging decisions of honey bees. Supercritical fluid extraction of VOCs, coupled with gas chromatography-mass spectrometry, revealed 75 VOCs. Of these, 19 were strong attractants and 8 were strong repellants. Moreover, we detected significant, but variable, species- or genome-level differences in floral scents. Honey bees avoided Brassica juncea cv. PBR357 (0.81 bee visits/sq m/min) and B. carinata cv. PC5 (0.77 bee visits/sq m/min), which possessed peak concentrations of antifeedants or repellents (dibutyl phthalate and 9-octadecenoic acid) in their floral volatiles. B. napus cvs. GSC6 and GSC7 attracted the highest number of bee visitations (1.07 and 0.99 bee visits/sq m/min, respectively), as these possessed VOCs with honey-fruity odor in high amounts and negligible levels of antifeedants. Our results highlight that variability in floral reward chemistry should be considered when breeding crops that are frequently visited upon by bee pollinators to support bee pollinators, in general, and honey industry, in particular.

Oilseed Brassica Species Diversification and Crop Geometry Influence the Productivity, Economics, and Environmental Footprints under Semi-Arid Regions

The article presents the findings of three-year field experiments conducted during 2017–2020 on the productivity, economics, and environmental footprints of the oilseed Brassica (OSB) with species diversification and crop geometry alterations in semi-arid regions of India. The objectives of the field experimentation was to assess the system of mustard intensification (SMI) in enhancing productivity and profitability with ensuring fewer environmental footprints. The results revealed that Brassica carinata gave a maximum seed productivity (3173.8 kg ha−1) and net returns (US$ 1141.72 ha−1) under a crop geometry of 60 cm × 60 cm. Further, an increase of 38% and 54% in seed yield and net returns from B. carinata was observed over the existing traditional Brassica juncea with conventional crop geometry. The maximum energy output was also recorded from B. carinata (246,445 MJ ha−1). The broader crop geometry (60 cm × 60 cm) also resulted in maximum energy output. The environmental footprint was lesser due to increased carbon gain (CG), carbon output (CO), and carbon production efficiency (CPE) and lower greenhouse gas intensity (GHGi) in B. carinata. However, the maximum water-use efficiency (WUE) was recorded in B. juncea (19.15 kg per ha-mm), with a minimum water footprint (WFP), whereas, greater crop geometry (60 cm × 60 cm) resulted in lower WFPs and better irrigation water use. Enhanced seed yield, economics, and fewer environmental footprints were observed at broader crop geometry in B. carinata over remaining OSBs.

A comprehensive Vis-NIRS equation for rapid quantification of seed glucosinolate content and composition across diverse Brassica oilseed chemotypes

The globally cultivated Brassica crops contain high deliverable concentrations of health-promoting glucosinolates. Development of a Visible-Near InfraRed Spectroscopy (Vis-NIRS) calibration to profile different glucosinolate components from 641 diverse Brassica juncea chemotypes was attempted in this study. Principal component analysis of HPLC-determined glucosinolates established the distinctiveness of four B. juncea populations used. Subsequently, modified partial least square regression based population-specific and combined Vis-NIRS models were developed, wherein the combined model exhibited higher coefficient of determination (R2; 0.81–0.97) for eight glucosinolates and higher ratio of prediction determination (RPD; 2.42–5.35) for seven glucosinolates in B. juncea populations. Furthermore, range error ratio (RER > 4) for twelve and RER > 10 for eight glucosinolates make the combined model acceptable for screening and quality control. The model also provided excellent prediction for aliphatic glucosinolates in four oilseed Brassica species. Overall, our work highlights the potential of Vis-NIR spectroscopy in estimating glucosinolate content in the economically important Brassica oilseeds.

Embryo rescue response and genetic analyses in interspecific crosses of oilseed Brassica species

Embryo rescue technique is widely utilized to save developing embryo from abortion in interspecific hybridization of plants. In Brassica species, success of an embryo rescue depends on the age of embryo, developmental stage of embryo, and media composition. In this study, ten crosses were conducted between diploid oilseed species B. rapa × tetraploid B. napus and B. rapa × tetraploid B. juncea along with their reciprocal crosses. The immature embryos from parents and crosses were cultured at 14 d after pollination in MS and ½ MS medium without any growth regulators for direct embryogenesis. Embryos were found at different stages of development during isolation, from globular to cotyledonary, indicating that rate of embryo development depends on genotype. Embryos isolated at the cotyledonary stage showed the highest regeneration percentage (av. 83.29%) and required the lowest time to regenerate (5.8 d) compared with another type of embryo, torpedo stage, cultured. Embryo rescued in ½ MS media accounted for 10–51%, 4.0–61%, 6.0–21%, and 5.0–43% greater shoot length, root length, number of leaves, and days to flowering, respectively, in different cross combinations compared with full-strength MS media. The results indicated that the rate of regeneration from embryo is higher in ½ MS media compared with full-strength MS media. Percent regeneration from the cultured embryos differed significantly in different cross combinations. Days required for regeneration from embryos, length of siliqua, and the number of ovules per siliqua accounted for high heterosis and broad sense heritability. The findings of this experiment would be useful in developing cost-effective embryo rescue protocol and desired interspecific hybrids through embryo rescue in Brassica species.

Diversity in Alternaria brassicae (Berk.) Sacc. isolates and characterization of host response in different oilseed Brassica species

Alternaria brassicae (Berk.) Sacc., a necrotrophic fungus devastating oilseed Brassica crops in India, causes up to 47% reduction in seed yield. So the study is conducting to develop the knowledge about the variability of A. brassicae in terms of cultural, morphological, pathogenic characters and their effects on different Brassica species of rapeseed mustard, for minimizing the losses caused by A. brassicae and help to stabilize the rapeseed mustard production. In variability study, all the isolates showed cultural, morphological variability. Average conidial length was maximum in isolate Ab2 (21.7μ) followed by Ab4 (20.7μ) and minimum in Ab8 (15.6μ). Septation in conidia, both horizontal and vertical also showed variation in different isolates. Finally it was revealed that smallest size of conidia in isolates Ab8 and longest size of conidia in isolates Ab4 were observed. On solid medium, maximum sporulation was observed in Ab7 followed while minimum sporulation recorded in Ab8. On liquid medium, maximum sporulation was observed in Ab7 while minimum sporulation recorded in Ab8. All the isolates showed the variability in qualitative characters i.e. spot colour, presence or absence of central point, concentric rings and yellow halo and as well as in quantitative characters i.e. incubation period and Percent Disease Index.

Nondestructive Phenomic Tools for the Prediction of Heat and Drought Tolerance at Anthesis in Brassica Species.

Oilseed Brassica species are vulnerable to heat and drought stress, especially in the early reproductive stage. We evaluated plant imaging of whole plant and flower tissue, leaf stomatal conductance, leaf and bud temperature, photochemical reflectance index, quantum yield of photosynthesis, and leaf gas exchange for their suitability to detect tolerance to heat (H) and/or drought (D) stress treatments in 12 Brassica genotypes (G). A replicated factorial experiment was set up with 7 d of stress treatment from the beginning of anthesis with various levels of three factors H, D, and G. Most phenomics tools detected plant stress as indicated by significant main effects of H, D, and H×D. Whole plant volume was highly correlated with fresh weight changes, suggesting that whole plant imaging may be a useful surrogate for fresh weight in future studies. Vcmax, the maximum carboxylation rate of photosynthesis, increased rapidly on day 1 in H and H+D treatments, and there were significant interactions of G×H and G×D. Vcmax of genotypes on day 1 in H and H+D treatments was positively correlated with their harvested seed yield. Vcmax on day 1 and day 3 were clustered with seed yield in H and H+D treatments as shown in the heatmaps of genotypic correlations. TPU, the rate of triose phosphate use, also showed significant positive genotypic correlations with seed yield in H+D treatments. Flower volume showed significant interactions of G×H and G×D on day 7, and flower volume of genotypes on day 7 in H was positively correlated with their harvested seed yield. There were few interactions of G×H or G×D for leaf stomatal conductance, leaf and bud temperature, photochemical reflectance index, and quantum yield of photosynthesis. Vcmax, TPU, and volume of flowers are potential nondestructive phenomic traits for heat or combined heat and drought stress tolerance screening in Brassica germplasm.

Analysis of present status, production constraints and future research strategies in Oilseed Brassica species

Rapeseed-mustard crops in India comprise traditionally grown indigenous species, namely toria (Brassica campestris L. var. toria), brown sarson (Brassica campestris L. var. brown sarson), yellow sarson (Brassica campestris L. var. yellow sarson), Indian mustard (Brassica juncea L. Czern and Coss.), black mustard (Brassica nigra) and taramira (Eruca sativa/ vesicaria Mill.), which have been grown since about 3,500 BC along with non-traditional species like gobhi sarson (Brassica napus L.) and Ethiopian mustard or karan rai (Brassica carinata A. Braun).

Non-destructive phenomic tools for the prediction of heat and drought tolerance at anthesis in Brassica species

Abiotic stresses lead to an array of physiological and biochemical changes in crops which affect their growth and development leading to grain yield reductions. Oilseed Brassica species are vulnerable to heat and drought stress, especially in the early reproductive stage. We evaluated the use of several non-destructive phenomic tools to predict heat and drought tolerance at anthesis in three Brassica species, including five B. rapa, five B. napus and two B. juncea genotypes. Heat, drought or combined heat and drought treatments were applied for seven days from the beginning of anthesis, following which plants were returned to normal temperatures and well-watered conditions. Leaf and floral bud temperatures increased relative to ambient, and leaf conductance decreased in response to seven days of drought or combined treatments, but not to heat or control treatments. The drought and/or heat treatment significantly affected three photosynthesis-related parameters: the maximum carboxylation rate allowed by Rubisco (Vcmax), the photosynthetic electron transport rate and the rate of triose phosphate use. One day after the beginning of the heat and combined heat and drought treatments, Vcmax increased to more than double the basal level but remained low in the drought and control treatments. Vcmax on the first day of heat and combined treatments was positively correlated to the harvested seed yield of genotypes (r = 0.670 and 0.878, respectively), suggesting that a rapid increase in Vcmax in response to heat stress during anthesis was a positive predictor of final grain yield. The tissue volume of the whole plant, detected by plant growth imaging using a digital imaging system, was positively correlated with the above-ground fresh weight of genotypes after seven days in all treatments and was, therefore, an excellent surrogate for above-ground biomass of the whole plant. The volume of the yellow flowers after seven days in all treatments was positively correlated with the harvested seed yield of genotypes, and is a potential non-destructive phenomic screen for heat and drought stress tolerance in Brassica germplasm.

SSR marker variations in Brassica species provide insight into the origin and evolution of Brassica amphidiploids

BackgroundOilseed Brassica represents an important group of oilseed crops with a long history of evolution and cultivation. To understand the origin and evolution of Brassica amphidiploids, simple sequence repeat (SSR) markers were used to unravel genetic variations in three diploids and three amphidiploid Brassica species of U’s triangle along with Eruca sativa as an outlier.ResultsOf 124 Brassica-derived SSR loci assayed, 100% cross-transferability was obtained for B. juncea and three subspecies of B. rapa, while lowest cross-transferability (91.93%) was obtained for Eruca sativa. The average % age of cross-transferability across all the seven species was 98.15%. The number of alleles detected at each locus ranged from one to six with an average of 3.41 alleles per primer pair. Neighbor-Joining-based dendrogram divided all the 40 accessions into two main groups composed of B. juncea/B. nigra/B. rapa and B. carinata/B. napus/B. oleracea. C-genome of oilseed Brassica species remained relatively more conserved than A- and B-genome. A- genome present in B. juncea and B. napus seems distinct from each other and hence provides great opportunity for generating diversity through synthesizing amphidiploids from different sources of A- genome. B. juncea had least intra-specific distance indicating narrow genetic base. B. rapa appears to be more primitive species from which other two diploid species might have evolved.ConclusionThe SSR marker set developed in this study will assist in DNA fingerprinting of various Brassica species cultivars, evaluating the genetic diversity in Brassica germplasm, genome mapping and construction of linkage maps, gene tagging and various other genomics-related studies in Brassica species. Further, the evolutionary relationship established among various Brassica species would assist in formulating suitable breeding strategies for widening the genetic base of Brassica amphidiploids by exploiting the genetic diversity present in diploid progenitor gene pools.

A simple spectrophotometric method for estimating total glucosinolates in mustard de-oiled cake

ABSTRACTGlucosinolates are anti-nutritional factors present abundantly in the seed meal fraction of oilseed Brassica species. They are found in varying levels among different genotypes. Those genotypes containing less than 30 µmol/g are considered low/zero glucosinolate type and are preferred for edible purposes due to low pungency. Twenty two different genotypes were taken for the analysis of glucosinolates by spectrophotometry. A regression model was obtained using Ordinary Least Square technique which predicted a formula. Total glucosinolates (µmol/g) = 1.40 + 118.86 × A425, where A425 is the absorbance at 425 nm. The total glucosinolate content obtained by the prediction formula when compared with HPLC data showed a correlation coefficient of 0.942. This high correlation between the two data sets validated the developed methodology. This method also simplifies the estimation of total glucosinolates by excluding the use of HPLC or other sophisticated instruments.

Identification and expression analysis of glucosinolate biosynthetic genes and estimation of glucosinolate contents in edible organs of Brassica oleracea subspecies.

Glucosinolates are anti-carcinogenic, anti-oxidative biochemical compounds that defend plants from insect and microbial attack. Glucosinolates are abundant in all cruciferous crops, including all vegetable and oilseed Brassica species. Here, we studied the expression of glucosinolate biosynthesis genes and determined glucosinolate contents in the edible organs of a total of 12 genotypes of Brassica oleracea: three genotypes each from cabbage, kale, kohlrabi and cauliflower subspecies. Among the 81 genes analyzed by RT-PCR, 19 are transcription factor-related, two different sets of 25 genes are involved in aliphatic and indolic biosynthesis pathways and the rest are breakdown-related. The expression of glucosinolate-related genes in the stems of kohlrabi was remarkably different compared to leaves of cabbage and kale and florets of cauliflower as only eight genes out of 81 were expressed in the stem tissues of kohlrabi. In the stem tissue of kohlrabi, only one aliphatic transcription factor-related gene, Bol036286 (MYB28) and one indolic transcription factor-related gene, Bol030761 (MYB51), were expressed. The results indicated the expression of all genes is not essential for glucosinolate biosynthesis. Using HPLC analysis, a total of 16 different types of glucosinolates were identified in four subspecies, nine of them were aliphatic, four of them were indolic and one was aromatic. Cauliflower florets measured the highest number of 14 glucosinolates. Among the aliphatic glucosinolates, only gluconapin was found in the florets of cauliflower. Glucoiberverin and glucobrassicanapin contents were the highest in the stems of kohlrabi. The indolic methoxyglucobrassicin and aromatic gluconasturtiin accounted for the highest content in the florets of cauliflower. A further detailed investigation and analyses is required to discern the precise roles of each of the genes for aliphatic and indolic glucosinolate biosynthesis in the edible organs.

Molecular mechanism of manipulating seed coat coloration in oilseed Brassica species

Yellow seed is a desirable characteristic for the breeding of oilseed Brassica crops, but the manifestation of seed coat color is very intricate due to the involvement of various pigments, the main components of which are flavonols, proanthocyanidin (condensed tannin), and maybe some other phenolic relatives, like lignin and melanin. The focus of this review is to examine the genetics mechanism regarding the biosynthesis and regulation of these pigments in the seed coat of oilseed Brassica. This knowledge came largely from recent researches on the molecular mechanism of TRANSPARENT TESTA (tt) and similar mutations in the ancestry model plant of Brassica, Arabidopsis. Some key enzymes in the flavonoid (flavonols and proanthocyanidin) biosynthetic pathway have been characterized in tt mutants. Some orthologs to these TRANSPARENT TESTA genes have also been cloned in Brassica species. However, it is suggested that some alterative metabolism pathways, including lignin and melanin, might also be involved in seed color manifestation. Polyphenol oxidases, such as laccase, tyrosinase, or even peroxidase, participate in the oxidation step in proanthocyanidin, lignin, and melanin biosynthesis. Moreover, some researches also suggested that melanic pigment in black-seeded Brassica was several fold higher than in yellow-seeded Brassica. Although more experiments are required to evaluate the importance of lignin and melanin in seed coat browning, the current results suggest that the flavonols and proanthocyanidin are not the only roles affecting seed color.

Construction of a genetic linkage map and QTL analysis of erucic acid content and glucosinolate components in yellow mustard (Sinapis alba L.).

BackgroundYellow mustard (Sinapis alba L.) is an important condiment crop for the spice trade in the world. It has lagged behind oilseed Brassica species in molecular marker development and application. Intron length polymorphism (ILP) markers are highly polymorphic, co-dominant and cost-effective. The cross-species applicability of ILP markers from Brassica species and Arabidopsis makes them possible to be used for genetic linkage mapping and further QTL analysis of agronomic traits in yellow mustard.ResultsA total of 250 ILP and 14 SSR markers were mapped on 12 linkage groups and designated as Sal01-12 in yellow mustard. The constructed map covered a total genetic length of 890.4 cM with an average marker interval of 3.3 cM. The QTL for erucic content co-localized with the fatty acid elongase 1 (FAE1) gene on Sal03. The self-(in)compatibility gene was assigned to Sal08. The 4-hydroxybenzyl, 3-indolylmethyl and 4-hydroxy-3-indolylmethyl glucosinolate contents were each controlled by one major QTL, all of which were located on Sal02. Two QTLs, accounting for the respective 20.4% and 19.2% of the total variation of 2-hydroxy-3-butenyl glucosinolate content, were identified and mapped to Sal02 and Sal11. Comparative synteny analysis revealed that yellow mustard was phylogenetically related to Arabidopsis thaliana and had undergone extensive chromosomal rearrangements during speciation.ConclusionThe linkage map based on ILP and SSR markers was constructed and used for QTL analysis of seed quality traits in yellow mustard. The markers tightly linked with the genes for different glucosinolate components will be used for marker-assisted selection and map-based cloning. The ILP markers and linkage map provide useful molecular tools for yellow mustard breeding.

A simple and rapid procedure for identification of seed coat colour at the early developmental stage of Brassica juncea and Brassica napus seeds

With 4 figures and 2 tablesAbstractThe yellow seed is a desirable trait in oilseed Brassica species. However, seed coat colour cannot be observed until the maturation of the seed. Identification of seed coat colour at early seed development will allow for the early detection of yellow‐seeded individual plants in the segregating populations at the same generation and thus speed up yellow‐seeded rapeseed breeding programme. Colour of seed coats from Brassica juncea and Brassica napus can precisely be determined as early as 15 days after pollination by staining with 0.5% vanillin–HCl solution for 5 min, which is shown to be simple, rapid and reliable. Flowering duration of an individual plant usually lasts for approximately 25–35 days in Brassica species and so the seed coat colour of an individual plant can be determined 10–20 days before the end of its flowering. Through this technique, breeding for the yellow‐seeded trait can now be advanced one generation earlier by vanillin staining than by conventional visual observation of testa colour of mature Brassica seeds.

Differential morphological and physiological responses of two oilseed Brassica species to a new herbicide ZJ0273 used in rapeseed fields

Weeds are considered as a major threat to the production of oilseed Brassica crops. The use of herbicides that are safe for crops and effective in controlling weeds is crucial for the agronomists and farmers. Propyl 4-(2-(4,6-dimethoxypyrimidin-2-yloxy)benzylamino)benzoate (ZJ0273), a derivative of 2-pyrimidinyloxy- N-aryl benzoate, is a new herbicide used in the rapeseed field. To evaluate the tolerance of Brassica species against this new herbicide, two cultivars of rapeseed Brassica napus cv. ZS 758 and Brassica rapa cv. Xiaoyoucai were tested by a foliar spray of ZJ0273 at the rate of 100, 500 and 1000 mg/L and a currently used ALS (acetolactate synthase)-inhibiting herbicide bispyribac-sodium (BS) at the rate of 100 mg/L. The results showed that both the cultivars of Brassica were less affected by ZJ0273 as compared to BS. Increasing level of ZJ0273 herbicide from 100 to 1000 mg/L increased the stress for the plants of both the cultivars as indicated by enhanced accumulation of malondialdehyde content. The activities of ALS and antioxidant enzymes (superoxide dismutase and peroxidase), soluble protein and sugar contents, photosynthetic system (SPAD value, photosynthetic rate and chlorophyll fluorescence) as well as the agronomic characters also declined consistently with each successive increase in ZJ0273 concentration. In general, the plants treated with 100 mg/L ZJ0273 recovered from the herbicide stress after 28 days. B. napus showed more tolerance than B. rapa to the new herbicide. Nevertheless, BS application at 100 mg/L did not allow the plants of both the cultivars to recover from the herbicidal stress.

Crop Ferality and Volunteerism

Crop Ferality and Volunteerism

Comparative Evaluation of RAPD, ISSR and Anchored-SSR Markers in the Assessment of Genetic Diversity and Fingerprinting of Oilseed Brassica Genotypes

Forty-two genotypes representing oilseed Brassica species were analyzed for the level of genetic diversity and molecular identity using Random Amplified Polymorphic DNA (RAPD), Inter-Simple Sequence Repeat (ISSR) and 5'-Anchored Simple Sequence Repeat (ASSR) markers. DNA profiles revealed high degree of interspecific polymorphism, while the level was considerably low within a species, particularly in B. juncea. The UPGMA clusters clearly delineated genotypes of the respective Brassica species. Comparison of cophenetic matrices indicated a high degree of correspondence between dendrograms generated by different marker systems. A minimum of 10 random primers (approximately 105 bands) were required for the RAPD profiles to generate the expected cluster. Comparatively less number of primers was required to do the same in case of ISSR (4 primers) and ASSR (3 primers). The principal component analysis revealed similar genetic relationship among the genotypes as in cluster analysis. Although none of the DNA profiles could individually identify all the B. juncea genotypes, a combined DNA profile consisting 125 markers from the informative primers of all the three DNA marker systems could do the same. A positive correlation was found among the marker utility parameters (calculated for individual primers of different marker systems) such as marker index (MI), resolving power (Rp) and discrimination coefficient (D) with the number of genotypes identified by each primer with a few exceptions. Single plant analysis for a set of five B. juncea varieties revealed absence of intra-varietal heterogeneity in case of ASSR profiles, thereby suggesting its utility in varietal identification and differentiation.

Medium, Explant and Genotype Factors Influencing Shoot Regeneration in Oilseed Brassica spp.

AbstractThe effects of culture media, explants and genotypes on shoot regeneration in oilseed Brassica species were examined in this study. The maximum shoot regeneration frequency was obtained in Murashige and Skoog medium supplemented with 3 mg l−1 6‐benzylaminopurine and 0.15 mg l−1 1‐naphthaleneacetic acid. The addition of 2.5 mg l−1 AgNO3 was very beneficial to shoot regeneration in B. napus and Ag2S2O3 (10 mg l−1) was even superior to AgNO3 (2.5 mg l−1). Explant age, explant type and carbon source also significantly affected shoot regeneration. Four‐day‐old seedlings of cotyledonary explants showed the maximum shoot regeneration frequency and number of shoots per explant. Of the four explants – peduncles, hypocotyls, cotyledons and leaf petioles – cotyledons produced the highest shoot regeneration frequency (56.67 %). Four carbon sources – glucose, maltose, starch and sucrose – were compared for their respective effects on shoot regeneration from cotyledonary explants. Sucrose appeared to be the best carbon source for shoot regeneration with the highest shoot regeneration frequency (76.00 %). Considerable variation in shoot regeneration from cotyledonary explants was observed both between and within Brassica species. The shoot regeneration frequency ranged from 10.00 % for cv. R5 (B. rapa) to 83.61 % for cv. N1 (B. napus). Two B. napus, one B. carinata and one B. juncea cultivars exhibited shoot regeneration frequency higher than 70 %. In terms of the number of shoots produced per explant, B. rapa showed the highest variation, ranging from 5.64 for cv. R3 to 1.33 for cv. R5. Normal plantlets were regenerated from all induced shoots and developed normally. The regenerated plants were fertile and identical with the source plants.