Fruit set,seed development and embryo germination in interploid crosses of citrus

Interploid crosses of diploid Kinnow mandarin, Succari sweet orange and Sweet lime with tetraploid Kinnow were made reciprocally for the production of triploid plants. Low fruit set and occurrence of underdeveloped or empty seeds was common in all interploid crosses. The hybrid fruits harvested after 12–14 weeks or 7 months after pollination produced many underdeveloped and few developed seeds; however, tetraploid Kinnow as seed parent yielded more developed seeds per fruit. The under developed seeds were devoid of endosperm. Nucellar embryony was higher in the diploid than in the tetraploid strain of Kinnow. Immature embryos of seeds harvested 12–14 weeks after pollination were cultured in vitro on MS media supplemented with adenine sulphate (20 mg l⁻¹) and malt extract (500 mg l⁻¹). Maximum germination (75%) of hybrid embryos from underdeveloped seeds was obtained in 4× × 2× cross of Kinnow strains. The germinated plantlets were transferred into pots and noted 59–89.3% survival rate in various crosses.     

Chitosan specificity for the in vitro seed germination of two Dendrobium orchids (Asparagales: Orchidaceae)

The effects of different types of chitosan on seed germination and protocorm development were determined for two orchid species, Dendrobium bigibbum var. compactum and Dendrobium formosum. Six chitosan types derived from polymer or oligomer chitosan each with 70, 80 or 90% levels of deacetylation (P70, P80, P90, O70, O80 and O90, respectively), were evaluated as direct medium supplements at 0, 10, 20, 40 or 80 mg/L in modified VW medium by following seed germination and protocorm growth for 12 weeks. Chitosan of all six tested types and four concentrations were found to significantly enhance the proportion of D. formosum seeds that germinated, when compared to these germinated without chitosan. In contrast, chitosan caused no enhanced germination rate was noted for D. bigibbum var. compactum with all tested chitosans and doses tested. However, almost all types of chitosan at 10 mg/L, except O90, were able to significantly improve the growth of D. bigibbum var. compactum protocorms, whilst 10 or 20 mg/L of P70 chitosan was the best formula to enhance the growth of D. formosum protocorms. It is concluded that chitosan responses in seed germination and protocorm development were somewhat species and developmental stage dependent. Therefore, the appropriate chitosan application for each plant species should be evaluated first before use.     

In vitro embryo germination and proliferation of pistachio (Pistacia vera L.)

In vitro development of isolated embryos and axillary bud proliferation were studied in Pistacia vera L. Different regulator-free nutrient media were compared to determine the effects of the mineral solution, agar and sucrose concentrations on seedling development from mature embryos. Nutrient-rich MS [Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tabacco tissue cultures. Physiol. Plant. 15, 473–479] and DKW [Driver, J.A., Kuniyuki, A.M., 1984. In vitro propagation of Paradox walnut rootstock. HortScience 19, 507–509] mineral solutions were more efficient for the development of aerial parts than nutrient-poor KN [Knop, W., 1884. Bereitung einer concentrierten nährstofflosung für pflanzen. Landwersuhssat 30, 292–294] and WT [Withe, P.R., 1936. Plant tissue cultures. Bot. Rev. 2, 419–437] solutions. Reducing the agar concentration enhanced fresh matter production and balanced seedling development. When tested at different concentrations, sucrose was found to orient mature embryo development, with the best results obtained at concentrations of 2–4%, whereas high concentrations (6 and 12%) mainly inhibited elongation of the aerial parts. Plantlets obtained previously from in vitro cultured embryos were propagated by axillary budding. High bud proliferation (six shoots per explant) was achieved when using 17.8 μM benzyladenine (BA) combined with 0.5 μM indole-3-butyric acid (IBA). The addition of 2.9 μM gibberellic acid (GA₃) to the propagation medium did not improve axillary shoot yields and on average, media with GA₃ produced 2.3–2.6 elongated stems per cultured explant. Shoots were rooted in vitro in half-strength MS medium containing 12.3 μM IBA.     

Effects of seed maturity,seed storage and pre-germination treatments on seed germination of cleome (Cleome gynandra L.)

The influence of seed maturity, seed storage and germination pre-treatments on seed germination of cleome (Cleome gynandra L.) were investigated. Seed maturation studies showed that capsules harvested at 18 days after anthesis possessed the highest dry weight with 19.2% moisture and 1% germination. Development of fresh-ungerminated seed was observed with increasing maturity of fruit, suggesting that cleome exhibited forms of seed dormancy. Storing mature seed at 15 °C and at room temperature for 5 months showed that seed dormancy was broken after 3 months under both storage regimes. When mature seeds were subjected to different treatments including various levels of GA₃, KNO₃, leaching, pre-chilling, soaking and pre-heating at different temperatures, it was found that pre-heating at 40 °C for period of 1–5 days was the most effective method in breaking dormancy in cleome.     

In vitro asymbiotic seed germination and seedling growth of Ansellia africana Lindl.

The effects of sodium hypochlorite treatment and growth medium on in vitro seed germination and seedling growth of the leopard orchid (Ansellia africana Lindl.) were investigated. Forty minutes treatment of seeds with 1.75% sodium hypochlorite stimulated germination (70.6%) and seedling development when measured at Stage 5 (emergence of first leaf) on P668 medium after 8 weeks of culture. The growth medium played a significant role in determining the germination response of A. africana seeds. Dark pretreatment of seed cultures significantly enhanced the germination percentage and the growth of rhizoids on the protocorms. Leaf growth in terms of length was substantially enhanced on P668 medium. It was significantly reduced in modified Knudson C medium after 16 weeks of culture. Seedlings developed on P668 medium showed a significantly better growth performance in relation to leaf length, leaf number, root length, fresh and dry weights per plant in vermiculite after 12 weeks of ex vitro growth in a mist house with 90% relative humidity. Further studies developing a suitable method for in vitro symbiotic germination could assist in reintroduction of this threatened orchid species into the wild.     

The effect of seed treatment on the germination and early growth of Euterpe edulis (family Palmae)

The germination rate of palm seeds can be improved by pre-treatment of the seed. Several methods proved effective for Euterpe edulis seed, but sulphuric acid immersion and hydrogen peroxide leaching treatments both decreased emergence of germinated seedlings and restricted plant growth up to 6 months after germination.The best overall results in germination and subsequent seedling development were from seed leached in water for 72 hours at 30°C. This treatment can be related to the natural conditions encountered by the seed in the highly leached soils of a tropical rain forest.     

Seed dormancy and germination of Michelia yunnanensis (Magnoliaceae)

Michelia yunnanensis Franch. is a Chinese endemic ornamental shrub with potential for greater utilization as a landscape and medicinal plant if propagation was less difficult. Seed development and breaking of seed dormancy were investigated to improve propagation of M. yunnanensis. No fresh seeds germinated when tested at the time of dispersal. Newly matured seeds of M. yunnanensis contained differentiated linear underdeveloped embryos that were physiologically dormant. The embryo/seed length ratio of M. yunnanensis was 0.15. Warm stratification did not break seed dormancy. Dormancy was broken by cold stratification at 4 °C but not by flowing water or nitrate. Embryos developed grew inside seeds during cold stratification at 4 °C. In newly harvested dormant seeds, embryos were 0.94 mm long and increased in length 139% before radicle emergence (germination). GA₃ substituted for cold stratification to break dormancy in seeds of M. yunnanensis incubated at 25 °C or 20/25 °C. Mature M. yunnanensis seeds exhibited intermediate complex morphophysiological dormancy. Optimal germination of non-dormant seed in terms of both germination percentage and rate occurred at 20/25 °C.     

Gibberellin-like stimulation of celery (Apium graveolens L.) seed germination by N-substituted phthalimides

Thermodormancy of celery seeds incubated in the dark at 22°C was alleviated by two N-substituted phthalimides to the same extent as that normally achieved with a mixture of the gibberellins A₄ and A₇. As previously demonstrated with GA₄₊₇, the highly dormant cultivar ‘Lathom Blanching’ responded more to phthalimides in the presence of cytokinin, whereas ‘Florida 683’ required no cytokinin to elicit the maximum response to phthalimide treatment. Seed-soak treatments with either N-substituted phthalimide improved both the rate of germination and the percentage seedling emergence of both cultivars sown in compost under glasshouse conditions.     

平邑甜茶(Malus hupehensis)胚囊种类与发生特性

应用石蜡切片技术对结果期平邑甜茶的胚囊发生与形成过程进行了详细观察,以确定胚囊种类和发生特性。结果表明,平邑甜茶胚珠倒生,大孢子母细胞在盛花期前3～5d的花蕾期能减数分裂形成四分体并由合点端的大孢子发育成有性胚囊;到盛花期,多数有性胚囊败育,少数发育为八核胚囊。平邑甜茶的无性胚囊原始细胞始于盛花期前后靠近合点端的珠心细胞,并很快发育为单核无性胚囊,盛花后5～7d的坐果初期形成无性的八核胚囊。无孢子生殖胚囊可以单个发育,也可以多个发生而形成复合胚囊;其原始细胞发生的持续时间为10d左右。     

Asymbiotic seed germination,mass propagation and seedling development of Vanda coerulea Griff ex.Lindl. (Blue Vanda): An in vitro protocol for an endangered orchid

The application of modern biotechnology for conservation of any endangered species requires an efficient in vitro regeneration protocol. In this study a reliable protocol was developed for in vitro seed germination, protocorm multiplication and subsequent plantlet regeneration of Vanda coerulea, an endangered orchid species. Among the four basal media evaluated for asymbiotic seed germination, Phytamax was found to be the best followed by Murashige and Skoog (MS). Phytamax was also found good for protocorm development. For protocorm like body (PLB) regeneration, protocorms were then further cultured on Phytamax media fortified with different phytohormones either individually or in combinations. The frequency of protocorm like body (PLB) regeneration significantly relied on kinds and concentrations of plant growth regulators used. A combination of 1-naphthaleneacetic acid (NAA) (5.36 μM) and 6-benzyle amino purine (BAP) (3.80 μM) was found to be suitable for maximum PLB regeneration. Healthy plantlets were induced from PLBs when cultured on same basal medium supplemented with activated charcoal (AC – 3.0 g/l). Plantlets with well developed leaves and roots were transplanted to pots filled with a mixture of charcoal, brick pieces and sphagnum moss and transferred to the greenhouse. This protocol will enable mass propagation and conservation of this exquisite orchid.     

Symbiotic seed germination of Grammatophyllum speciosum Blume and Dendrobium draconis Rchb. f., native orchids of Thailand

In vitro symbiotic seed germination is an important tool not only for the study of orchid-fungus specificity but also for the production of mycobiont-infected healthy seedlings that could be valuable for both horticultural and conservation purposes. The current study compared effectiveness of eight putative orchid mycorrhizal fungi obtained from mature orchids in the genera Paphiopedilum, Cymbidium and Dendrobium, in promoting in vitro seed germination and protocorm development of Grammatophyllum speciosum Blume and Dendrobium draconis Rchb. f., native Thai orchids. The developmental stages of seeds and protocorms cultured on Murashige and Skoog (MS) medium, oat meal agar (OMA), or OMA inoculated with one of the eight fungal isolates were evaluated weekly. Two isolates of Epulorhiza repens (Bernard) Moore (=anamorphic species of Tulasnella calospora (Boud.) Juel), Da-KP-0-1 and Pv-PC-1-1, were found to be the most effective fungi in promoting protocorm development of G. speciosum. At week 13, protocorms co-cultured with either one of these two fungal isolates, on the average, were significantly more advanced than those sown on OMA. Protocorms co-cultured with isolate Pv-PC-1-1 were also significantly more advanced than those cultured on MS medium. For D. draconis seed germination, three fungal isolates of different anamorphic species of Tulasnella, C1-DT-TC-1, Pv-PC-1-1, and C3-DT-TC-2, were found to be the most effective fungi in promoting protocorm development. However, none of these fungal isolates outperformed MS medium. Additionally, the compatibility between the fungal isolates tested and the two orchid species was discussed.     

Effects of light,macronutrients, and sucrose on germination and development of the endangered fern Adiantum reniforme var. sinense (Adiantaceae)

Effects of light, macronutrients strength of Murashige and Skoog (MS) (1962), and sucrose in the culture medium on spore germination and gametophyte development of the endangered fern Adiantum reniforme var. sinense were investigated. The presence of light was found to be essential for both spore germination and gametophyte growth. Moreover, a medium consisting of 1/4 MS with 15 g/l sucrose was optimal for spore germination and early gametophyte development; whereas, MS medium with 30 g/l sucrose was optimal for further gametophyte development. Increasing amounts of sucrose (45–60 g/l) in the medium delayed gametophyte growth and development. Additionally, sporophyte formation and early growth of gametophytes in a medium consisting of clay and peat (v/v = 1:2) was higher than those in a medium consisting of pure river sand. These findings indicated that requirements for nutrients for spore germination and early gametophyte development of A. reniforme var. sinese were relatively low, but these increased with further gametophyte development, formation and growth of sporophytes.     

Effect of seed age,stratification, and soaking on germination of wild asparagus (Asparagus acutifolius L.)

Wild asparagus (Asparagus acutifolius L.) is a widespread species found in all the Mediterranean areas. The spears are highly valued by consumers and owing to its frugality, this species is a feasible new crop with high income potential, especially for Mediterranean marginal areas. Currently, the cultivation of this species is limited because of its low and erratic seed germination that makes difficult the production of seedlings for plant propagation. In this research, non-after-ripened (1 month-old) and after-ripened seeds (dry stored at room temperature for 13 months) were exposed for 30 days in the dark to three moist stratification treatments: cold (5 °C), warm (23 °C) or no stratification; subsequently they were soaked for 12 h in warm water (35 °C) or not soaked. The effect of these pre-germination treatments on three germination parameters (germination percentage, time to 50% of final germination – T₅₀ – and germination pattern) was studied, as well as some possible seed dormancy forms involved therein. The 1-year dry storage period proved to be effective in after-ripened seeds by enhancing seed sensitivity to the subsequent pre-germination treatments. After-ripened seeds exhibited higher and more rapid germination compared to non-after-ripened seeds. Soaking, cold or warm moist stratification had similar single effect on non-after-ripened seeds (27% germination). With after-ripened seeds, only soaking or warm stratification were effective (47% germination) when singularly applied, while cold stratification did not improve germination. By combining stratification and soaking treatments, a higher germination for both non-after-ripened and after-ripened seed-lots was achieved. The highest germination was obtained when after-ripened seeds were stratified and soaked (76%), without any significant difference between cold or warm stratification. Single or combined application of moist stratification (regardless of the temperature used) and soaking resulted always in a faster germination compared to that of no-treated seeds and especially with after-ripened seeds (T₅₀ = 6 days). A non-deep type 1 physiological dormancy can be hypothesized for the seeds of this species. Low stratification temperature induce secondary dormancy in after-ripened seeds that can be removed by soaking them at 35 °C for 12 h.     

In vitro germination and seedling development of cryopreserved Dendrobium hybrid mature seeds

In vitro germination and seedling development from Dendrobium Swartz. hybrid ‘Sena Red’, ‘Mini WRL’, ‘Jaquelyn Thomas’, and ‘BFC Pink’ seeds cryopreserved through vitrification (PVS2) were evaluated. Germination percentages after cryopreservation (LN) were variable among different controls and treatments, despite the initial high seed viability for all hybrids. Seeds exposed to PVS2 at ice temperature from 1 to 3 h prior to LN exhibited significantly higher germination than seeds exposed to PVS2 at room temperature for the same time periods. No significant differences in germination percentages were observed for time exposure to PVS2 at 1, 2 or 3 h for ‘Sena Red’, ‘Mini WRL’, and ‘BFC Pink’. Seeds of ‘Jaquelyn Thomas’ exposed to 1 h prior to LN showed higher germination percentage than for exposure to 2 or 3 h. The combination of a pre-cooling treatment (ice) with a dehydration treatment (PVS2) for a period of time of 1–3 h was essential to allow proper germination of cryopreserved seeds. Although variability in seed germination among different hybrids and treatments was observed, germination was above 50% of the controls and all germinated seeds developed into normal seedlings with healthy shoot and root formation. No abnormalities, nutritional deficiencies, or diseases were observed in developed seedlings and no significant differences were observed for seedling growth and development from germinated seeds among the different hybrids. Seedlings transplanted to pots acclimatized well and developed into normal plants within 6–8 months in greenhouse. Transplanted seedlings exhibited 100% survival for all hybrids.     

Influence of fruit ripeness at the time of seed extraction on pepper (Capsicum annuum) seed germination

The influence of fruit ripeness (half ripe, fully ripe, overripe) at the time of seed extraction on seed germination behaviour, at 25 °C and 13 °C, was studied in two Spanish pepper cultivars for canning. Seeds from half ripe fruits had a poorer behaviour than those taken from fully ripe fruits, especially at 13 °C. Room ripening and overripening of half ripe fruits improved the germination behaviour of the seeds although this behaviour was poorer than that of seeds extracted from fully ripe fruits that were allowed to overripen. Overripening of fully ripe picked fruits slightly improved germination behaviour.     

Cold storage of oil rose (Rosa damascena Mill.) flowers

Oil rose flowers were stored at 0 °C in four different packaging materials [plastic box + stretch film (PB + SF), Xtend^®, Smartbag^® and polyethylene (PE)] for 60 days. During storage, weight loss, O₂ and CO₂ concentrations in the packages, petal color and sensorial attributes were investigated besides essential oil content and composition. Storage duration and packages had significant (p < 0.01) effects on weight loss. At the end of storage, the lowest weight loss was in PE package (1.696%) whereas the highest weight loss was in Xtend^® (10.081%). The essential oil content was significantly (p < 0.01) affected by storage duration and packages. In addition, the essential oil contents obtained from all packages for a storage period of 10 days and the essential oil contents obtained from unstored (control) petals were included in the same group. At the end of storage, the essential oil contents decreased by 91.3, 57.7, 80.0 and 64.3% in PB + SF, Xtend^®, Smartbag^® and PE packages, respectively as compared to control. In addition, storage duration and package types significantly (p < 0.01) affected petal color, O₂ and CO₂ concentrations in the packages and sensorial scores. The concentration of citronellol, a main component of rose oil, increased in all packages during storage of 10 days in comparison to the control group while it varied in other storage durations and package types. However, nerol and geraniol were lower than the control group during storage while concentrations of nonadecane, heneicosane and eicosane were higher. In conclusion, loss of oil yield and quality, due to various reasons and particularly due to fermentation in oil rose from the harvest of petals to their distillation, can be minimized with storage of petals in all package types for up to 10 days.     

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.     

Light acclimation in rose (Rosa hybrida cv. Grand Gala) leaves after pruning: Effects on chlorophyll a fluorescence,nitrate reductase,ammonium and carbohydrates

Changes in light intensity have a particular effect on the photosynthetic apparatus. Most of the studies on light acclimation in higher plants have focused on the effects of fixed light intensity. Few works deal on sudden changes in light intensity due to pruning for long periods of time. Pruning the bent shoots in roses (cv. Grand Gala) can modify the light interception in internal leaves and change their photosynthetic reactions. Before pruning, internal leaves were acclimated to low light intensity and after pruning, internal and external leaves received the same light intensity. The aim of this work was to find out how the photosynthetic light reaction, measured by chlorophyll (Chl) a fluorescence in internal and external leaves of arched shoot could be modulated by light changes. Other parameters described as light dependent were also measured: nitrate reductase (NR) activity, NH₄⁺, sucrose and starch levels. Internal and external leaves in rose have shown a high plasticity, rapid and dynamical acclimation, in response to changes in incident sunlight produced by pruning, that can be explained by Chl a fluorescence parameters. The modified NR activity, NH₄⁺, sucrose and starch levels were difficult to associated with light intensity changes, and their modulation could be the response to long-time light acclimation.     

African spider flower (Cleome gynandra L./Gynandropsis gynandra (L.) Briq.) as a red spider mite (Tetranychus urticae Koch) repellent in cut-flower rose (Rosa hybrida L.) cultivation

Companion planting of Cleome gynandra, of Kenyan origin, in beds of cut-flower roses reduces significantly red spider mite (Tetranychus urticae Koch) infestation without any detrimental effect on productivity or flower quality. The level of reduction is dependent upon the density of the C. gynandra plants with 15 plants in a 1.8 m² bed (8.3 plants m²) being the most effective, planted either around the bed perimeter or within the rows of roses. The relatively high density of C. gynandra plants required may limit the direct application of this technology in export-focused, greenhouse rose production yet may be of significant value as a supplement to other mite-control strategies. The potential benefits of such companion planting for growers of field roses and those involved in some domestic markets are also evident. Research into the nature and extraction of the active, volatile mite-repellant components of C. gynandra is indicated.