Effect of glucose on glucosinolates,antioxidants and metabolic enzymes in Brassica sprouts

The purpose of this study was to investigate the effect of glucose on glucosinolates, antioxidants and metabolic enzymes in Brassica sprouts. After glucose treatment, total glucosinolates, phenolics and anthocyanin contents and antioxidant activity were significantly enhanced in Chinese kale and pak choi sprouts, while only the anthocyanin and ascorbic acid contents were increased in radish sprouts. These results indicate that glucose treatment has selectively improved the nutritional compounds in different Brassica sprouts.     

Germination of Passiflora mollissima (Kunth) L.H.Bailey, Passiflora tricuspis Mast. and Passiflora nov sp. seeds

Passiflora mollissima, Passiflora tricuspis and Passiflora nov sp. are three passion fruit species occurring in Bolivia. Germination percentages and rates were determined for 11 different treatments. Per species, germination of 100 seeds was monitored every 3 days, during 90 days. Germination started after 9 days and 50% of final germination was reached within a month or less. Successful, recommended methods for P. mollissima are removing the basal point of seeds (27% germination) or removing the basal point in combination with pre-soaking seeds for 48 h in 50 ppm GA₃ (18%). Pre-soaking seeds for 24 h in 400 ppm GA₃ (42%) and removal of the basal point in combination with pre-soaking seeds for 48 h in 50 ppm GA₃ (36%) are suggested methods to improve germination of P. nov sp. Removing the apical point of P. tricuspis seeds resulted in maximal germination (57%). No unique treatment gave satisfactory results for the three species tested. Exogenous dormancy, probably a combination of mechanical and chemical dormancy is present in the three species studied. Presence of physical dormancy was found in P. mollissima.     

Regeneration from leaf explants and protoplasts of Brassica oleracea var. botrytis (cauliflower)

Adventitious shoot regeneration and protoplast isolation and culture were examined from leaf explants of in vitro shoot cultures of several cauliflower (Brassica oleracea var. botrytis) cultivars, sourced from Europe and Australia, was investigated with the aim to develop improved nuclear and plastid transformation protocols for this vegetable crop. Eight out of 10 cultivars regenerated shoots from at least 79% of leaf explants. Mesophyll protoplasts from leaves gave high yields and division frequencies. Growth of shoot cultures in large glass vessels with vented lids was the key factor in obtaining high protoplast division frequencies of up to 71% and at least 70% of protoplast calluses regenerating shoots.     

Heterosis for mineral elements in single cross-hybrids of cabbage (Brassica oleracea var. capitata L.)

Cabbage (Brassica oleracea var. capitata L.), a cross-pollinated crop, was observed to exhibit strong heterosis for high yield; better plant stand; early maturity; larger and more uniform heads; uniformity in head compactness; and disease tolerance in F₁ hybrids. However, information on the heterosis for mineral elements such as Fe, Zn, Cu, Mn, K and Ca although is not available to the best of our knowledge but it is important as it is likely to influence the plant and subsequently the human nutrition. Therefore, an attempt was made to estimate the heterosis for mineral elements in cabbage. Significant mean square for parents and hybrids was observed for all minerals under study which indicated the prevalence of sufficient variation. The parents 83-2, Pride of Asia, Red Cabbage, AC-204 and MR-1 were found to have the potential for use in cabbage quality breeding programme as they exhibited higher hybrid effects for Fe, Zn, Cu and Mn content. The single cross-hybrids, i.e. 83-2 × AC-204; Pride of Asia × C-2 and Pride of Asia × Red Cabbage; Pride of Asia × MR-1; 83-2 × Red Cabbage; and Pride of Asia × AC-204 and 83-2 × MR-1 were the best for Fe and Zn; Fe and Cu; Zn and Mn; Cu and Zn; and Cu, respectively. This study revealed clearly that none of the hybrids excelled for all the minerals suggesting the significance and need for multiple crossing breeding approaches, i.e. three way cross-hybrid, double cross-hybrid, population improvement, synthetics, composites, etc., for increasing the mineral concentration in cabbage head, i.e. “Breeding Cabbage for Higher Mineral” (Biofortification) without losing the vigour advantage for yield and other traits of economic importance to combat mineral deficiencies in human beings and plant systems.     

Changes in kale (Brassica oleracea L. var. acephala) carotenoid and chlorophyll pigment concentrations during leaf ontogeny

There has been recent market interest in “baby” salad greens. However, little information exists on the nutritional differences between immature “baby” greens and produce traditionally sold at the fully mature stage. Kale (Brassica oleracea L. var. acephala D.C.) contains high levels of lutein and β-carotene, which possess important human health properties. Kale was grown in a controlled environment and pigments were measured in young (<1 week), immature (1–2 weeks), mature (2–3 weeks), fully developed (3–4 weeks) and senescing (>4 weeks) leaves using high-performance liquid chromatography (HPLC). Significant differences were observed for all pigments during leaf development. Pigment accumulation followed a quadratic trend, with maximums occurring between the 1st and 3rd week of leaf age. The highest concentrations of lutein measured 15.1 mg/100 g fresh mass and occurred in 1–2 week old leaves. The remaining pigments reached maximum levels at 2–3 weeks (β-carotene at 11.6 mg/100 g; chlorophyll a at 251.4 mg/100 g; and chlorophyll b at 56.9 mg/100 g fresh mass). Mature fully expanded kale leaves accumulated higher carotenoid concentrations than immature or “baby” leaves, with senescent leaves having the lowest carotenoid concentrations. Harvesting kale leaves at a mature stage of development resulted in maximum carotenoid values. Cultural management practices that increase carotenoid concentrations would be expected to improve nutritional quality for fresh markets.     

High frequency plant regeneration from microspore-derived embryos of ornamental kale (Brassica oleracea L. var. acephala)

The aim of this work was to study the effect of solid medium, developmental stage, embryonic age, cold treatment and additives to the medium on plant regeneration from microspore-derived embryos in four F₁ hybrids of ornamental kale (Brassica oleracea L. var. acephala). The results showed that all of the cultivars responded best when the embryos were cultured in solidified B5 medium with 1% agar. Optimal regeneration was gained when cotyledonary embryos were cultured for 25 days. Cold treatment significantly improved plant regeneration with a frequency of up to 79.0% under 4 °C for 2 d or 5 d. The addition of 3.0 or 5.0 mg/L silver nitrate (AgNO₃) increased the frequency of plant regeneration. In the Zhouyehongxin cultivar, the frequency of plantlet development reached 84.4%. The addition of activated charcoal reduced embryo hyperhydricity.     

Combining ability for superoxide dismutase,peroxidase and catalase enzymes in cabbage head (Brassica oleracea var. capitata L.)

Antioxidant enzymes have been touted as beneficial for enhancing the fitness, preventing disorders, and mitigating the effects of aging and senescence. Our objective was to evaluate combining ability of superoxide dismutase, peroxidase, and catalase activity in cabbage head. Head samples were frozen immediately in liquid nitrogen and placed at −80 °C for assay. Less than unity values of σ²_gca/σ²_sca ratio for all three enzymes indicated predominance of non-additive gene action. The parents CMS-GA and Red Cabbage excelled as good general combiners for all antioxidants and indicated the value and need for multiple crossing. The crosses CMS-GA × Red Cabbage, CMS-GA × C-2, 83-2 × AC-204, 83-2 × EC-490174, 83-2 × AC-1021, Pride of Asia × C-4, and Pride of Asia × AC-1019 showed significant specific combining ability, which could be exploited through heterosis breeding. The hybrid combinations with high per se performance and favorable SCA estimate and involving at least one of the parents with high GCA estimate could be useful to increase the abundance of favorable alleles for enhancing the antioxidants in cabbage head.     

Artificial cold treatment,gibberellin application and flowering response of kohlrabi (Brassica oleracea L. var. gongylodes L.)

Juvenility, cold requirement and the effect of GA₃-application on flowering have been studied for some cultivars. If present, the juvenile phase was very short. Cold treatment for 8–12 weeks at 4 °C followed by 2 weeks at 10 °C gave flowering in all cultivars. Gibberellin sprays (250 or 500 p.p.m., 3 times) after an incomplete cold treatment promoted bolting and flowering even in the slow-bolting ‘Trero’, having the highest cold requirement.     

Photosynthetic and growth responses of juvenile Chinese kale (Brassica oleracea var. alboglabra) and Caisin (Brassica rapa subsp. parachinensis) to waterlogging and water deficit

Chinese kale (Brassica oleracea var. alboglabra) and Caisin (Brassica rapa subsp. parachinensis) are leafy vegetable crops grown in south-east Asian countries where rainfall varies dramatically from excess to deficit within and between seasons. We investigated the physiological and growth responses of these plants to waterlogging and water deficit in a controlled experiment in a glasshouse. Juvenile plants were subjected to waterlogging or water deficit for 19 days in case of Chinese kale and 14 days in case of Caisin and compared with well-watered controls. Caisin tolerated waterlogging better than Chinese kale because it produced hypocotyl roots and gas spaces developed at the stem base. In Chinese kale, waterlogging reduced plant fresh weight (90%), leaf area (86%), dry weight (80%) and leaf number (38%). In contrast, waterlogging had no impact on leaf number in Caisin and reduced plant fresh and dry weights and leaf area by 60–70%. Water deficit reduced leaf area, fresh weight and dry weight of both species by more than half. Leaf number in Chinese kale was reduced by 38% but no effect occurred in Caisin. Water deficit increased the concentration of nitrogen in the leaf dry matter by more than 60% in both species and the leaf colour of water deficient plants was dark green compared with the leaf colour of well-watered plants. Soil water deficit delayed flowering of Caisin while waterlogging accelerated it. Thickening and whitening of the cuticle on the leaves of Chinese kale probably increased its ability to retain water under drought while Caisin adjusted osmotically and Chinese kale did not. Waterlogging and water deficit had strong effects on leaf gas exchange of both Brassica species. Water deficit closed the stomata in both species and this was associated with a leaf water content of 9 g g⁻¹ DW. In contrast, waterlogging reduced conductance from 1.0 to 0.1 mol H₂O m⁻² s⁻¹ in direct proportion to changes in leaf water content, which fell from 11 to 5 g g⁻¹ DW. This separation of the effects of water deficit and waterlogging on conductance was reflected in transpiration, internal CO₂ concentration and net photosynthesis. In conclusion, Chinese kale and Caisin showed rather different adaptations in response to waterlogging and water deficit. Caisin was more tolerant of waterlogging than Chinese kale and also showed evidence of tolerance of drought. There is genetic variation to waterlogging within the Brassica genus among the leafy vegetables that could be used for cultivar improvement.     

High-frequency embryogenesis,regeneration of broccoli (Brassica oleracea var. italica) and analysis of genetic stability by RAPD

High-frequency somatic embryogenesis and shoot regeneration of broccoli (Brassica oleracea var. italica) were achieved. Cotyledon and hypocotyl explants from four varieties of broccoli were cultured on MS and modified MS media (mMS, supplemented with PG-96 organic components) with different combinations of growth regulator. The effects of genotypes, different explants, growth regulator combinations, organic components and AgNO₃ on induction of calli and shoots were evaluated. The optimal media for inducting calli/shoots and roots were mMS medium containing 3% (w/v) sucrose and 0.8% (w/v) agar supplemented with NAA at 0.5 mg l⁻¹, 6-BA at 3.0 mg l⁻¹, AgNO₃ at 4.0 mg l⁻¹ and MS medium containing 3% sucrose and 0.8% (w/v) agar supplemented with NAA at 0.2 mg l⁻¹, respectively. The callus induction percentages were over 90% in all four varieties; shoot induction percentage was 92.5% and the average number of shoot per explant was 4.1 from cotyledon explant in variety Bishan. In this study, we established high-efficient embryogenesis and shoot regeneration system of broccoli and analyzed genetic stability of regenerants at DNA level using RAPD molecular marker. Out of 62 arbitrary primers screened using PCR amplification, 79 polymorphic bands were amplified from 20 primers. The results demonstrated the genetic stability of regenerants from the same variety.     

Antioxidant potency of white (Brassica oleracea L. var. capitata) and Chinese (Brassica rapa L. var. pekinensis (Lour.)) cabbage: The influence of development stage,cultivar choice and seed selection

The accumulation of total phenols (TP, Folin-Ciocalteu method) and total flavonoids (TF, colorimetric assay with AlCl₃) and the evolution of antioxidant capacity (FRAP assay, DPPH and ABTS radical scavenging assays) have been monitored in juices of Croatian white cabbage (Brassica oleracea var. capitata) cultivars Vara?dinski and Ogulinski, as well as Chinese cabbage (Brassica rapa var. pekinensis), at various developmental stages. Measurements were performed every four weeks, starting from planting to full maturity, throughout the course of eight months. In the first 8–12 weeks, the TP and TF contents as well as antioxidant capacity increased significantly in all analyzed juice samples and in most even doubled. This rapid increase was followed by a gradual decrease in values derived from all assays, over the course of 12–30 weeks, to the final values which were in all cases lower than the values measured at week 4. The results also point to significant variability in TP and TF contents and antioxidant capacity at the fully mature stage between white and Chinese cabbage juices and between juices extracted from cultivars Ogulinski and Vara?dinski.     

Dormancy and germination of Areca triandra seeds

The dormancy mechanisms of Areca triandra Roxb. Ex Buch-Ham seeds were studied by treating the intact or mechanically scarified seeds with scarification, chemical soaking and stratification. The results indicate that the seeds have exogenous and endogenous dormancy. The exogenous dormancy is imposed by the pericarp and it is the major limiting factor for germination. It can be broken by mechanical scarification, but not by chemical scarification in 98% H₂SO₄ for 30 min. Chemical treatments (soaking for 24 h in 100–200 mg/L GA₃, 0.2% KNO₃ and 0.1–0.3% NaNO₂, and for 12 h in 10% H₂O₂ or 20 min or 12 h in 15% H₂O₂) and stratifications, especially, cold stratifications for 30–120 days, broke the endogenous dormancy and significantly hastened germination of mechanically scarified seeds, although they did not increase the germination percentages.     

The culture of isolated microspores of ornamental kale (Brassica oleracea var. acephala) and the importance of genotype to embryo regeneration

The culture of isolated microspores of kale (Brassica oleracea var. acephala) was studied including the importance of genotype to embryo regeneration, medium composition chiefly the sucrose concentration and the use of colchicine, simultaneously medium renovation. It was initiated using 29 different genotypes as donor plants. Embryos were induced from six of the kale genotypes and these corresponded to the more out-bred genotypes. Embryogenesis was achieved using four different combinations of culture media: (a) microspores initially cultured in NLN medium supplemented with 13% (w/v) sucrose (NLN-13) for 48 h, followed by transfer to fresh NLN-13 medium; (b) microspores cultured for 48 h in NLN-13 medium supplemented with colchicines (50 mg/L) followed by transfer to unsupplemented NLN-13 medium; (c) microspores cultured for 48 h in NLN-16 medium supplemented with colchicines (50 mg/L) followed by transfer to unsupplemented NLN-16 medium; (d) microspores cultured for 48 h in NLN-16 medium supplemented with colchicines (50 mg/L) followed by transfer to unsupplemented NLN-13 medium. The embryos obtained from four of the genotypes developed into plantlets and these regenerated plants have been successfully transplanted to soil.     

Pollination with laser-irradiated pollens breaks cross-incompatibility between zicaitai (Brassica campestris var. purpurea) and ornamental kale (Brassica oleracea var. acephala) to produce hybrids with the aid of ovule culture

The objective of this study was to produce interspecific hybrids between an Ogura-cytoplasmic male sterile (CMS) line of zicaitai (Brassica campestris var. purupurea, 2n = 20) and cultivars of ornamental kale (Brassica oleracea var. acephala, 2n = 18) to develop a CMS system for hybrid seed production. Pollination with pollen grains of ornamental kales irradiated at a power output of 9.0 mW with a He–Ne laser for 3 min could overcome the cross-incompatibility between the species concerned. Intact hybrids could be efficiently produced from ovules cultured on Murashige and Skoog media supplemented by 0.2 mg l⁻¹ 6-benzyladenine. Chromosome number of hybrids was confirmed to be 2n = 19. Hybrids resembled ornamental kales in leaf morphology and in vernalization response. Pollens of hybrids had a sterile appearance. Moreover the hybridity of the putative hybrids was confirmed by RAPD data on a DNA fragment of 820 bp.     

Rapid seedlings regeneration from seeds and vegetative propagation with sucker and rhizome of Eremospatha macrocarpa (Mann & Wendl.) Wendl and Laccosperma secundiflorum (P. Beauv.) Kuntze

To develop efficient seedling production methods for Laccosperma secundiflorum and Eremospatha macrocarpa, a study was conducted to examine regeneration using offsets combined with several physical and chemical treatments of seeds. Offsets categorized into small, medium and large diameters, were planted in three conditions: shaded and open nursery, and greenhouse. We tested sucker from E. macrocarpa, and sucker and rhizome from L. secundiflorum. For both species, high viability percentage (ranging from 55% to 100%) were observed for small and medium suckers planted in shaded nursery and greenhouse, against less than 49% for sucker planted in open nursery. The mean seedling emergence times were estimated to 84, 77 and 75 days after planting (DAP) for small, medium and large sucker of L. secundiflorum, respectively under open nursery condition, and 76, 75, 95 DAP for small, medium and large suckers of the same species, respectively in shaded condition. Greenhouse has a significant positive effect on E. macrocarpa seedlings emergence time. For this species, the mean seedling emergence times were estimated to 43 DAP for small sucker and 76, 93 DAP for medium and large suckers. No seedling was obtained from rhizome planted in all the growing conditions tested. Concerning seed dormancy breaking, germination percentages and rates were determined for 13 treatments. The best treatments were pre-soaking unscarified seeds for 4 days in 1.01 g l⁻¹ and 0.10 g l⁻¹ KNO₃, with 79% and 68% of germination, respectively and in 3.46 × 10⁻³ g l⁻¹ GA₃ for 68% of germination. These methods are suggested to improve germination of L. secundiflorum seeds. Successful and recommended methods for E. macrocarpa are pre-soaking scarified seeds in 3.46 × 10⁻³ g l⁻¹ and 3.46 × 10⁻⁴ g l⁻¹ GA₃, 96% and 94% of germination, respectively. Dormancy, probably a combination of mechanical and chemical dormancy, is present in the two species.     

Effect of different fertilisation and irrigation practices on yield,nitrogen uptake and fertiliser use efficiency of white cabbage (Brassica oleracea var. capitata L.)

The effect of different fertilisation (i.e. broadcast application and fertigation) and irrigation practices (tank sprinkler and drip irrigation) on yield, yield quality (nitrate content), nitrogen uptake of white cabbage (Brassica oleracea var. capitata L.) and the potential for N losses was assessed on sandy-loam agricultural soil. ¹⁵N-labelled fertiliser was used as a tracer. It was found that different practices significantly affected yield, nitrate content in plants, N uptake, as well as fertiliser use efficiency. The highest yield (93 t ha⁻¹), plant N uptake (246 kg ha⁻¹), and fertiliser use efficiency (42%) were obtained under treatment with broadcast fertilisation with farmer's practice of irrigation (tank sprinkler). The N surplus after harvest was −41 kg N ha⁻¹, indicating the lowest potential for N losses. Treatment by fertigation and drip irrigation covering 100% of the crop's water requirements did not result in the highest yield as expected (72 t ha⁻¹), the N surplus after harvest was about +38 kg ha⁻¹. The lowest yield (58 t ha⁻¹), fertiliser use efficiency (30%) and hence the highest potential for N losses (N surplus after harvest +68 kg ha⁻¹) were found in treatment with broadcast fertilisation and drip irrigation covering 50% of the crop's water requirements.     

Root zone temperature,plant growth and yield of broccoli [Brassica oleracea (Plenck) var. italica] as affected by plastic film mulches

Plastic film mulches are often associated with increases in plant growth and yield of vegetable crops. Few studies, however, report on the effect of plastic film mulches on root zone temperature on broccoli. The objectives of this study were to determine the effects of colored plastic mulches on root zone temperature and broccoli plant growth and yield Broccoli (‘Packman’) plants were grown using plastic film mulch and drip irrigation. The treatments consisted of plastic film mulch (black, blue, gray on black, red, silver on black, and white on black mulches) and bare soil. Colored plastic film mulches affected root zone temperature and the accumulation of soil degree-days. Mean daily RZT, maximal daily RZT and degree day accumulation in the soil were highest in dark-colored mulches (blue, black, red, and gray) and lowest in light-colored mulches (silver and white), while minimal daily RZT was highest in silver mulch and lowest in white mulch. Silver mulch showed the smallest diurnal fluctuations in RZT among plastic mulches, with the highest RZT during the night and among the lowest in the afternoon. Colored plastic mulches also affected broccoli plant growth and yield, although the effect of mulches was more dramatic in the spring than in the fall seasons. Broccoli yield was linearly related to both the vegetative top dry weight and root dry weight of mature plants. Broccoli yield was little affected at mean RZT of <21 °C but increased with increasing mean RZTs above 21 °C up to 25 °C. Thus, broccoli plant growth and yield responded more favorably to dark-colored mulches than to light-colored mulches, suggesting that broccoli benefited from increased soil warming. No high RZT stress effects on broccoli plant growth or yield were observed in this study. Thus, this study suggests that plastic film mulches, particularly the dark-colored mulches, may be an option for spring broccoli production in areas with cool conditions early in the spring.     

In vitro shoot regeneration from immature seeds of Epimedium alpinum induced by thidiazuron and CPPU

In vitro propagation of Epimedium alpinum L. was carried out using immature seed explants. The effects of various concentrations of thidiazuron (TDZ) and 1-(2-chloro-4-pyridyl)-3-phenylurea (CPPU), on the induction of organogenic callus, were evaluated. Organogenesis occurred most efficiently when explants were transiently exposed (48 h) to 20 μM CPPU or 80 μM TDZ followed by culture on hormone-free woody-plant medium (WPM). Organogenic callus consisting of white, compact clumps of tissue proliferated slowly on hormone-free WPM. To promote adventitious shoot induction, the effects of different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzyladenine (BA) were investigated. The highest per cent shoot regeneration, 66.7% of explants, and the maximum mean number of shoots, 2.6 per explant, were obtained on WPM containing 1.1 μM 2,4-D and 22 μM BA. Shoots were rooted on hormone-free WPM and well-developed plantlets were successfully transferred to soil.     

A micropropagation system for cloning of Bambusa tulda Roxb.

The communication describes standardization of an efficient in vitro propagation and hardening procedure for obtaining plantlets from field grown culms of Bambusa tulda. Administration for 10 min of 0.05 and 0.1% mercuric chloride to explants collected in winter and summer seasons, respectively facilitated optimum culture establishment and bud break. 0.1–0.2% mercuric chloride in rainy season enhanced aseptic culture establishment but inhibited bud break due to toxicity to explants. MS liquid medium enriched with 100 μM glutamine, 0.1 μM indole-3-acetic acid and 12 μM 6-benzylaminopurine supported maximum in vitro shoot multiplication rate of two-fold. The proliferated shoots were successfully rooted on MS liquid medium supplemented with 40 μM coumarin resulting in a maximum of 98% rooting. The procedure requires 45 days cycle for the in vitro clonal propagation (15 days for shoot multiplication and 30 days for root induction) and 80 days for acclimatized plantlet production.