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.     

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.     

Weed competition in cauliflower (Brassica oleracea L. var. botrytis) in the Jordan Valley

A field experiment was carried out in 1996/1997 and repeated in 1997/1998 at the Jordan University Research Station located in the central Jordan Valley to determine the effect of weed competition on growth and yield of cauliflower Brassica oleracea var. botrytis cv. “White Cloud”. The treatments consisted of either allowing weeds to infest the crop or maintaining plots weed-free for increasing durations after transplanting. Results showed that longer periods of weed/cauliflower competition greatly reduced crop growth and head yield. Average reductions in shoot dry weight and head yield were 81% and 89%, respectively. Maintaining a weed-free crop for any period after transplanting increased cauliflower growth and head yield compared with the weed-infested control. Weed competition for 14 days after transplanting reduced cauliflower average head yield by 41%. To determine the critical period of weed competition and the influence of weed infestation on cauliflower head yield the Gompertz and logistic equations were fitted to data representing increasing duration of weed-free and weed-infested periods, respectively. Based upon an arbitrary 5% level of head yield loss, the critical period of weed competition occurred at 0–38 days after cauliflower transplanting which corresponded with the rapid increase in weed biomass. Results indicated that early weed removal is necessary to prevent yield loss.     

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.     

Production of inter-section hybrids between Primula filchnerae and P. sinensis through ovule culture

Inter-section hybrids were obtained in the reciprocal crosses between Primula filchnerae (2n = 2x = 24) of Sect. Pinnatae and P. sinensis ‘Fanfare’ (2n = 2x = 24) of Sect. Auganthus by rescuing ovules on half-strength (1/2) Murashige and Skoog's (MS) medium supplemented with 50 g l⁻¹ sucrose, 2.5 g l⁻¹ gellan gum, 0.1 mg l⁻¹ α-naphthaleneacetic acid (NAA), 0.1 mg l⁻¹ 6-benzyladenine (BA) and 50 mg l⁻¹ gibberellic acid (GA₃). In ovule culture, germination occurred with radicle elongation but no plumule was observed. The radicle kept on the initial medium showed root proliferation with callus formation. When the calluses were transferred to (1/2)MS media containing 30 g l⁻¹ sucrose and 3 g l⁻¹ gellan gum, without plant growth regulators (PGRs) or with 1 mg l⁻¹ zeatin and 0.1 mg l⁻¹ NAA, plantlets were regenerated. The plants thus obtained were confirmed to be hybrids through flow cytometry (FCM) and random amplified polymorphic DNA (RAPD) analyses. The hybrid obtained when P. filchnerae was used as the maternal parent was diploid, whereas hexaploid hybrid was obtained when using P. sinensis as the maternal parent. The hexaploid hybrid might be produced through chromosome doubling of a triploid originated from the fertilization of P. sinensis with unreduced pollen of P. filchnerae.     

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.     

Plant regeneration from in vitro cultured leaves of Lanzhou lily (Lilium davidii var. unicolor)

Lanzhou lily (Lilium davidii var. unicolor) is one of the best lilies which are edible in China but the efficient shoot regeneration system has not been developed. The purpose of the present study is to establish an efficient and reproducible protocol for induction of shoots in vitro from L. davidii var. unicolor leaves. Shoot regeneration from in vitro cultured leaves of L. davidii var. unicolor was tested on the 26 media based on NN [Nitsch, J.P., Nitsch, C., 1969. Haploid plants from pollen grains. Science 163, 85–87] basal medium, containing different concentrations of thidiazuron (TDZ) in combination with different concentrations of α-naphthaleneacetic acid (NAA). Shoot organogenesis occurred directly from the leaves without forming callus. Shoot regeneration mainly occurred from the cuts across the midvein and the base of the leaf explants. The highest frequency of regeneration (93.3%) and the largest number of shoots per leaf (3.83) were obtained on NN basal medium supplemented with 0.5 mg l⁻¹ TDZ and 1.0 mg l⁻¹ NAA. All the regenerated shoots formed complete plantlets on half-strength MS [Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant 15, 473–497] basal medium containing 0.1–0.5 mg l⁻¹ indole-3-butyric acid (IBA) with in 30 days, and 92% of the regenerated plantlets survived in the soil. This study will be useful for Agrobacterium-mediated transformation and exploitation of somaclonal variation of Lanzhou lily.