Responses of non-primed or primed seeds of ‘Marketmore 76’ cucumber (Cucumis sativus L.) slurry coated with Trichoderma species to planting in growth media infested with Pythium aphanidermatum

Aqueous slurries of commercial preparations of Trichoderma harzianum Rifai strain KRL-AG2 G41 (Th), T. virens Strain G-41 (Tv), or their combination (ThTv, at half rates each of the single application rate) were applied to ‘Marktetmore 76’ cucumber seeds (Cucumis sativus L.) that were non-primed or primed for 3 days at 25 °C either osmotically (−2.5 MPa from 0.337 molal Ca(NO₃)₂) or osmomatrically (−1.0 MPa from 0.135 molal Ca(NO₃)₂ plus −1.5 MPa from 50% water in exfoliated grade 5 vermiculite). Slurries were applied to seeds (1 mg per seed) either before or after priming. Seeds were sown in soilless, peat-based media with or without inoculation with Pythium aphanidermatum (Pa). Protection against damping-off caused by high pressures of Pa (16% emergence in non-coated, non-primed seeds) was increased by slurry coating Th on non-primed (76.4% emergence) or on osmotically primed seeds, with coating either before or after priming having no effect on efficacy (average 62.6% emergence). Slurry coating Th on osmomatrically primed seeds failed to increase final emergence percentage (FEP). Colony forming units per three seeds (CFU, all 10³) was 2.8 for non-primed seeds, and 3.2 and 2.6, respectively, when osmotically and osmomatrically primed seeds were coated after priming. In a second study with lower disease pressure (58.1 FEP from non-coated, non-primed seeds), slurry coating of non-primed or osmotically primed seeds with Th, Tv or ThTv reduced percentage damping-off and increased FEP. The combination coating eliminated damping-off only in non-primed seeds, and tended to reduce percentage damping-off in osmotically or osmomatrically primed seeds compared to coating with Th or Tv alone. In a third study using only non-primed seeds, slurry coatings with mefenoxam fungicide, Th, Tv, or ThTv decreased total damping-off to 2.6%, 7.4%, 2.0%, and 0%, respectively, from the 30.1% occurring in non-coated seeds. Th, Tv or ThTv applied to growth media at the same rate as the seed coating (1 mg per seed) were generally as effective as the seed coatings, and only the ThTv growth medium application eliminated damping-off. A fourth experiment revealed that Th, Tv or ThTv remained viable on non-primed seeds for up to 4 weeks (the longest storage duration) at 21 or 4 °C, but 21 °C storage resulted in faster seed germination by week 3 and higher CFU per three seeds by week 4. In summary, coating of non-primed seeds with Th, Tv or ThTv was more effective than coating primed seeds in reducing percentage damping-off. While priming treatments generally led to faster seedling emergence and greater seedling shoot fresh weight than was achieved with non-primed seeds, only for non-primed seeds was damping-off eliminated by the ThTv seed coating or growth medium application.     

Biocontrol of mildew with Bacillus subtilis in bitter gourd (Momordica charantia L.) seeds during germination

The bitter gourd seed has a thick, hard seed coat. Mildew often occurs during germination and causes uneven and low rates of seed germination. However, the problems caused by mildew can be overcome by treating seeds with hot water, by soaking in water, or by using microorganisms. Seeds of the ‘Ching Pi’ bitter gourd were treated in a water bath at 60 °C for 10 min and then soaked in tap water at 25 °C for 24 h. The resulting germination percentage was 86.7%, and the resulting percentage of mildewed seeds was 10%. The biocontrol potential of three commercially available Bacillus subtilis solutions was examined. For seeds primed with Huodijun B. subtilis solution, the germination percentage was 73.3% and the mildewed percentage 6%. In dual cultures, the antibiotic content in the Huodijun B. subtilis solution was significantly greater than in Yunghsing and Huolibao, the other B. subtilis solutions examined. B. subtilis effectively inhibited the growth of Rhizoctonia solani mycelium and caused abnormal mycelial growth.     

Ethylene influences development and flowering of Ptilotus spp. in vitro and ex vitro

The genus Ptilotus has immense potential for ornamental horticulture but its commercial development has been hindered by propagation limitations. Poor seed quality and germination are reported. Cutting propagation is limited by cutting supply as the juvenile phase of Ptilotus is short. Micropropagation has been used in an attempt to overcome these difficulties but explants become floral in vitro and this causes plantlets to elongate. Ethephon has been used to control flowering of stock plants of many ornamental species. This study investigated the effect of ethephon applied to young (3-week-old, deflasked from tissue culture) and mature (1-year-old) Ptilotus plants in a greenhouse. A system of applying gaseous ethylene at 0, 100, 200 and 300 mg l⁻¹ to the headspace of in vitro plantlets in glass jars was developed and the response of in vitro plantlets to ethylene studied. One-year-old Ptilotus plants were treated with 500 mg l⁻¹ ethephon 2 days before pruning or 1 or 2 weeks after pruning. Ethephon application 2 days before pruning decreased the number of inflorescences and increased the number of shoots (compared to the control) but was phytotoxic. Ethephon applications of 150 or 300 mg l⁻¹ applied weekly or fortnightly to 3-week-old plants deflasked from tissue culture reduced plant height and number of inflorescences and at low concentrations increased the number of new shoots. A fortnightly application at 150 mg l⁻¹ is recommended. Previous reports on the effects of ethylene on inflorescence production on plantlets in vitro are limited. Our study showed that exposure of in vitro plantlets of P. nobilis to ethylene gas at 100 mg l⁻¹ for 1 h significantly increased the number of shoots and plant height but this did not occur for plantlets of P. spicatus. Plantlets of P. spicatus exposed to transient ethylene at 200 and 300 mg l⁻¹ showed significantly greater rooting (52.4%) than the control (13.6%).     

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.     

Regeneration and characterization of plants derived from leaf in vitro culture of two sweet orange (Citrus sinensis (L.) Osbeck) cultivars

In the current work attempts were made to investigate culture of leaf explants derived from in vitro seedlings of two sweet orange (Citrus sinensis (L.) Osbeck) cultivars, Bingtangcheng and Valencia. Effects of several factors, including culture medium, lighting condition, explant age and genotype on regeneration response were examined based on three parameters, percentage of explants producing shoots, mean number of shoots per explant and shoot forming capacity. Culture of the explants on shoot-inducing media (SIM) composed of MT salts supplemented with different growth regulators gave rise to disparate shoot regeneration, in which SIM₁ (MT + 0.5 mg L⁻¹ BA + 0.5 mg L⁻¹ Kinetin + 0.1 mg L⁻¹ NAA + 3% sucrose + 0.8% agar, pH 5.8) was shown to be the most effective medium for direct induction of shoots from leaf explants. Highly significant difference in the response of shoot bud regeneration was noted between the two cultivars, with Bingtangcheng being more responsive than Valencia. Culture of explants from fully developed leaves led to better shoot regeneration capacity in comparison to undeveloped ones. However, the two lighting conditions used herein did not cause significant difference in shoot regeneration. Phenotypic observation and randomly amplified polymorphic DNA (RAPD) analysis confirmed that all the regenerated plants from both genotypes were genetically identical to their donor plants, suggesting absence of detectable genetic variation in the regenerated plants. The data presented here demonstrated that direct initiation of plants from leaf explants has been successfully accomplished. To our knowledge, this is the first report on direct regeneration of shoots from leaf explants in Citrus, which will provide an alternative source for citrus genetic manipulation in the future.     

Development of seedless and Mal Secco tolerant mutant lemons through budwood irradiation

Mal secco (caused by Phoma tracheiphila (Petri) Kantsch. and Gik.) is the most destructive fungal disease of lemon plantations worldwide and seedless lemons would be preferred by most consumers. Five dosage levels, 0, 3, 5, 7, and 9 kiloradian (krad), of cobalt (⁶⁰Co) gamma irradiation were applied to budstick of ‘Kutdiken’ lemon (Citrus limon (L.) Burm. f.) clone KT-2A. Mutations were stabilized in three vegetative generations. Three hundred fifty-eight and 478 M1V3 (mutation one and vegetation three) plants were evaluated for seed number and mal secco tolerance in the field and the greenhouse, respectively. LD₅₀ was around 5 krad gamma irradiation for ‘Kutdiken’ lemon. The seed number varied from 0 to 34 per fruit. The level of mal secco tolerance also varied significantly among the plants from 1.0 (no symptom) to 4.3 (high level of disease occurance). The stable seedless and mal secco tolerant plants were obtained from 5 and 7 krad irradiation: the three mutants from 5 krad irradiation gave more lemon-like fruits, while 7 krad irradiation caused altered tree morphology and early maturation of fruits. This study shows considerable potential for lemon cultivar improvement aiming to obtain seedless and mal secco tolerant lemons.     

Factors affecting haploid induction through in vitro gynogenesis in summer squash (Cucurbita pepo L.)

Haploid production using in vitro ovule cultures has long been recognized as an important tool to produce haploid and homozygous double-haploid plants for genetic studies and plant breeding programs. In the present study, four experiments were carried out to study the influence of genotype, position of female flowers on plant stem, temperature and sucrose concentration on the in vitro gynogenesis induction of squash. (1) Ovules of 12 genotypes were excised from female flowers, 1 day before anthesis, and cultured onto MS medium containing 3% sucrose and 1 mg l⁻¹ from each of kinetin and 2,4-D (2,4- dichlorophenoxy acetic acid). Differences in response among genotypes were demonstrated. Raad F₁ showed the highest percentage of responding ovules and number of plantlets per dish with 48.8% and 15 plants, respectively. The results revealed that genotype is a key factor influencing the in vitro gynogenesis in squash. (2) Ovules were excised from first, second and third female flower of two hybrids (Giad and Raad) and cultured onto the mentioned above medium. The highest percentage of responding ovules and number of plantlets per dish were obtained from ovules excised from the second female flower on the plant stem. (3) Effect of temperature (4 and 32 °C) for 0, 4, 7 and 12 days on the ovule culture of Queen F₁ was studied. Ovules incubated at 4 or 32 °C for 4 days produced a better embryogenic response. (4) Three sucrose concentrations (30, 60 and 90 g l⁻¹) were tested with the ovule cultures of the local cultivar (Eskandrani). Differences among sucrose concentrations were statistically significant and ovules cultured on the MS medium containing 30 g l⁻¹ produced the best result. MS medium containing 90 g l⁻¹ did not produce gynogenic ovules.     

Breaking the intergeneric hybridization barrier in Carica papaya and Vasconcellea cauliflora

The present investigation was undertaken to develop PRSV (Papaya ringspot virus) resistant hybrids through intergeneric hybridization. Intergeneric hybridization was done involving nine Carica papaya cultivars as female and Vasconcellea cauliflora as male. To break the intergeneric hybridization barrier, various nutrient combinations were used. Among the combinations used, sucrose 5%, sucrose 5% + boron 0.5% and sucrose 5% + CaCl₂ 0.5% improved the fruit set and seed set percentage. A total number of 1197 flowers were pollinated and 308 fruits were obtained. On extraction, 721 seeds were obtained from CO 7, Pusa Nanha and CP 50. Out of 721 F₀ seeds (crossed seeds) sown, 419 seeds germinated and artificial screening for PRSV was carried out 27 days after sap inoculation. Out of 29 F₁ hybrid plants from CO 7 x V. cauliflora cross, only six plants namely CO 7V1 to CO 7V6 were found free from PRSV symptoms. Similarly, out of 55 F₁ hybrids from cross involving Pusa Nanha x V. cauliflora only 23 plants namely PNV1 to PNV23 were found free from the symptoms and 70 plants namely CPV1 to CPV70 out of 335 plants of CP50 x V. cauliflora cross were found free from PRSV symptoms. Among the crosses, Pusa Nanha x V. cauliflora had higher yield under PRSV infected conditions, however, total soluble solids and total sugars were found lesser than the CO 7 x V. cauliflora cross. The hybridity of the progenies were confirmed by using ISSR (Inter Simple Sequence Repeats) primers by the amplification of DNA from progenies and their parents. ISSR primers UBC 856, UBC807 and ISSR primer combinations UBC 856-817, UBC 810-817, UBC 861-817, UBC 856-810, UBC 861-810 and UBC 856-817 clearly amplified specific bands of the male parent, which were present in F₁ progenies, but it was absent in female parents.     

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.     

An improved protocol for adventitious shoot regeneration and plant formation in Psoralea corylifolia L.

An effective protocol was developed for in vitro regeneration of Psoralea corylifolia through enriched cotton moistened-liquid (CML) and solid culture systems. Prolific adventitious shoot buds were achieved from hypocotyl explants of 2-week-old cultures on enriched CML Phillips and Collins (L2) medium supplemented with different concentrations and combinations of thidiazuron (TDZ), benzyladenine (BA), kinetin (KIN), naphthalene acetic acid (NAA), indole-3-acetic acid (IAA) and bavistin (BVN). Combination of 2 μM TDZ, 0.5 μM BA, 100 mg l⁻¹ BVN and 2 μM NAA produced a greater number of adventitious shoots per explant (93.5) when transferred to half-strength enriched solid L2 medium. Regenerated shoots (40–50 mm in length) were exposed simultaneously for rooting as well as hardening in moistened (1/8-L2 basal salt solution with 5 μM IBA and 100 mg l⁻¹ BVN) soil mixture and vermiculite (3:1, v/v). The plants were subsequently established in the field. The survival percentage differed with seasonal variations.     

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.     

Taxonomic revision of Dendrobium moniliforme complex (Orchidaceae)

Taxonomic revision of Dendrobium moniliforme complex is presented. D. moniliforme complex is characterized by the even slim stems, bracts with brownish zone, semi-spherical anther cap and the hairy disc of lip. Dendrobium tosaense, Dendrobium officinale and Dendrobium guangxiense were excluded by having membranous bracts lacking brownish zone, anther cap conical and bifid. Two species are recognized in this complex, i.e., D. moniliforme and Dendrobium wilsonii. D. wilsonii differs from D. moniliforme by having elliptic leaves about 1.3–2 cm wide, dorsal sepal 3.0–4.0 cm long, 0.6–0.9 cm wide, petals elliptic to oblong, 3.0–4.0 cm long, 1.0–1.5 cm wide, lip elliptic to ovate–lanceolate, 2.6–3 cm long, 1.2–1.5 cm wide.     

Both anti-oxidation and lipid-carbohydrate conversion enhancements are involved in priming-improved emergence of Echinacea purpurea seeds that differ in size

This study evaluated the effects of priming on emergence responses of purple coneflower (Echinacea purpurea L. Moench) seeds. The seeds that differ in seed size were either primed with moistened vermiculite (solid matrix priming) or primed in non-aerated −0.5 MPa polyethylene glycol 6000 solution at 25 °C for 6 days (osmopriming), followed by air-drying to their initial moisture level. The tetrazolium staining tests indicated that both large and small seeds were biochemically viable. No notable difference in germination percentage was found between large and small seeds. However, extensive cavity was visible in portions of small seeds in comparison with large seeds. Large seeds accumulated more antioxidants and had greater activities of anti-oxidative enzymes than small seeds. They also had greater isocitrate lyase and malate synthase activities than small seeds. As a result, large seeds had higher emergence percentage and faster emergence speed as compared to that of small seeds. Both solid matrix priming and osmopriming increased emergence percentage and shortened mean emergence time of purple coneflower seeds by increasing the activities of anti-oxidative enzymes and decreasing the levels of malondialdehyde and total peroxide accumulation. Moreover, priming also enhanced the anti-oxidative activities of treated seeds. The activities of isocitrate lyase and malate synthase were also increased in primed seeds. The enhanced anti-oxidation and lipid-carbohydrate conversion activities might explain in part why primed purple coneflower seeds emerged better than non-primed seeds.     

The genotypic effects on the chemical composition and antioxidant activity of sea buckthorn (Hippophae rhamnoides L.) berries grown in Turkey

In this study, chemical composition of berries of 10 sea buckthorn (Hippophae rhamnoides L.) genotypes in Turkey was investigated. The total phenolic content of the berries ranged from 21.31 mg gallic acid equivalents (GAE) per g dry weight basis to 55.38 mg GAE per g. The highest antioxidant activity was 93.54% (similar to the standard BHT at 200 mg/L) and the lowest was 80.38%. There was no correlation (R = 0.688) between the total phenolic content and the antioxidant activity. The major fatty acids in berries were palmitoleic acid (35.48%), followed by palmitic acid (28.13%), oleic acid (22.89%) and linoleic acid (3.96%). Total soluble solid content of sea buckthorn genotypes varied from 10.15 to 14.80%, titratable acidity varied from 2.64 to 4.54%, the pH varied from 2.63 to 2.98 and Vitamin C varied from 19 to 121 mg/100 mL. The average content of minerals in the sea buckthorn berries of different genotypes was 20,800 ppm N, 7100 ppm P, 7260 ppm K, 1960 ppm Ca, 1465 ppm Mg, 32 ppm Zn, 24 ppm Cu, 22 ppm Mn and 7 ppm Fe.     

Callus induction and plant regeneration from embryonic axes of Kosteletzkya virginica

Kosteletzkya virginica, a perennial dicot halophytic species of the Malvaceae, is native to American salt marsh. It was introduced into China as a potential species to improve coastal wetlands and to develop ecologically sound saline agriculture. K. virginica adapts excellently to the tidal-flat habitats in China's east coast, with multiple eco-benefits; in particular, its seed oil could be used to produce biodiesel. The purpose of this study was thus to develop a standardized protocol to induce a high frequency of callus and subsequent plantlet regeneration system for a K. virginica breeding program with the final objective of applying transgenic techniques to improve seed oil yield. The embryonic axes of K. virginica were used as explants for callus induction, shoot induction from the callus and then adventitious root induction from the shoots on nine culture media with different hormone combinations. The best results were achieved on the following media: (1) 93.94% callus induction on MS medium supplemented with 1.0 mg L⁻¹ indole-3-acetic acid (IAA), 0.3 mg L⁻¹ kinetin, 30 g L⁻¹ sucrose and 8 g L⁻¹ agar; (2) 65.83% shoot induction on 1/2MS medium supplemented with 0.1 mg L⁻¹ IAA, 0.5 mg L⁻¹ zeatin, 30 g L⁻¹ sucrose and 8 g L⁻¹ agar; (3) 96.67% rooting on MS medium containing 30 g L⁻¹ sucrose and 8 g L⁻¹ agar. The survival rate of plantlets by organogenic regeneration was 85% after being transplanted into potting soil in flowerpots and placed in the greenhouse. This experiment indicates that we established successful callus induction and plant regeneration protocols for K. virginica.     

The influence of desiccation and rehydration on the survival of polyembryonic seed of Citrus suhuiensis cv. limau madu

A study was carried out to investigate the influence of desiccation and freezing followed by various presowing rehydration procedures on the desiccation sensitivity of the seed of Citrus suhuiensis cv. limau madu. The freshly harvested seeds of limau madu were desiccated under a broad range of relative humidity (RH) to various equilibrium water contents (g H₂O g⁻¹ dw). The desiccated and desiccated–frozen seeds were either directly sown under germination conditions or subjected to presowing rehydration procedures: seed preheating, prehumidification and osmoconditioning. The hydrated and desiccated seeds were sown in controlled germination conditions and the survival was evaluated 4–6 weeks after sowing. The results showed that desiccation progressively reduced the percentage of normal seedling of the seeds of limau madu and the viability is almost lost at water contents below 0.08 g H₂O g⁻¹ dw. The estimated desiccation sensitivity was substantially high (WC₅₀ = 0.143 g H₂O g⁻¹ dw) when the desiccated seeds were rapidly rehydrated (uncontrolled rehydration). In contrast, seed prehumidification, preheating and osmoconditioning (controlled rehydration procedures) markedly enhanced normal seedling percentages decreasing the estimated values of WC₅₀ (between 0.08 and 0.127 g H₂O g⁻¹ dw). While the rapidly rehydrated desiccated–frozen seeds were almost killed at water content of 0.15 g H₂O g⁻¹ dw, prehumidification and preheating have noticeably increased percentage of frozen seeds survival at the same water content. However, at water content of 0.21 g H₂O g⁻¹ dw preheating significantly (P < 0.05) increased percentage of normal seedling of the frozen seeds. Seed desiccation markedly reduced the percentages of germinated seeds with multiple seedlings. Seed controlled rehydration remarkably increased the survival of polyembryos. The beneficial effect of seed controlled rehydration on the survival of the desiccated seeds was pronounced at medium water contents (0.08–0.25 g H₂O g⁻¹ dw).     

High frequency multiplication of Phalaenopsis gigantea using trimmed bases protocorms technique

This study was conducted to determine the effects of coconut water (CW) and activated charcoal (AC) on multiplication of Phalaenopsis gigantea protocorms. The protocorms used for this study were obtained by germinating seeds in vitro. Protocorms with trimmed and untrimmed bases were cultured on XER basal medium containing 0, 10, 15 or 20% (v/v) CW; and 0, 1, 2 or 2.5 g AC l⁻¹. Trimmed protocorms exhibited the highest percentage of proliferation on a medium containing 15% (v/v) CW and 2.5 g AC l⁻¹ (56.82 ± 38.86%) with an average of 4.24 ± 2.89 protocorms formed per protocorm. Untrimmed protocorms cultured on a medium containing 20% (v/v) CW without AC produced the highest percentage of new protocorms (6.93 ± 6.28%) with an average of 0.72 ± 0.57 per protocorm. When CW was added to a medium singly, 10% (v/v) CW induced a higher degree of proliferation on trimmed protocorms (5.68 ± 10.14%) with an average 0.50 ± 0.84 new protocorms per protocorm. Untrimmed protocorms proliferate to a much lower extent (2.57 ± 2.74%) with an average of 0.72 ± 0.57 protocorms per protocorm when cultured on a similar medium. A high concentration of CW enhanced proliferation on untrimmed protocorms, but increased mortality of trimmed protocorms. The addition of CW with AC to media increased protocorm proliferation and survival of both trimmed and untrimmed protocorms. When cultured on all media, trimmed protocorms produced a higher number of new protocorms (an average 0.5–7.0) as compared to untrimmed protocorms (0.3–1.9). Comparative studies showed that trimmed protocorms produced up to 10 times more new protocorms than untrimmed ones. Altogether this study showed that trimmed protocorms cultured on a medium containing CW and AC can be used for high-frequency multiplication of P. gigantea seedlings.     

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.     

Interspecific variations in seed germination of Corylopsis

This study was initiated to investigate the differences in germination percentages and rates between Corylopsis coreana Uyeki and Corylopsis sinensis var. calvescens Rehder & E.H. Wilson following a warm stratification (WS) and cold stratification (CS), and to study the effect of different WS temperatures interacting with different durations of CS. Warm stratification at 10 °C, 15 °C, 20 °C, and 25 °C was given for 1 month (1 M 10 °C, 15 °C, 20 °C, and 25 °C WS) followed by 0 M, 1 M, 2 M, and 3 M of CS at 5 °C (0 M, 1 M, 2 M, 3 M CS) and seeds were germinated in an air conditioned greenhouse maintained at 18.5 °C/18 °C. On average, less than 1% of C. coreana seeds germinated when sown without any WS and CS or with 1 M 15 °C, 20 °C, and 25 °C WS without CS treatment. However, 26% C. coreana seeds germinated after 1 M 10 °C WS without any CS treatment. Germination was not affected by WS temperatures when followed by 2 M 5 °C CS. It is concluded that C. coreana exhibited low seed germination at 10 °C and that this temperature could be considered the upper limit of CS for C. coreana. Only 2 M CS was required for more than 90% seeds to germinate. However, C. sinensis var. calvescens required longer than 3 M CS for more than 29% seeds to germinate. This clearly shows that there is an interspecific variation in optimum dormancy-breaking requirements.     

Response of gladiolus (Gladiolus spp) plants after exposure corms to chitosan and hot water treatments

The state of Morelos, Mexico has gradually become an important producer of gladiolus. Some preconditioning treatments of corms are empirically done causing uneven emergence and low quality of flowers. In this investigation, before planting, gladiolus corms var. ‘Blanca Borrego’ were dipped in chitosan (chitosan reagent and commercial chitosan Biorend^®), in hot water at various temperatures and in treatments combined with Biorend^® at 1.5% and hot water. Results indicated that the most influenced variables were corm germination, number of flowers per spike, number of cormlets and vase life. Overall, the commercial product Biorend^® at 1.5% accelerated corm emergence in approximately 4 days, the number of flowers increased by 2–7 and the vase life extended for 3 days. The number of cormlets was also duplicated. Corms dipped in the commercial chitosan Biorend^® at 1.5% at different intervals of time were not greatly affected except for the emergence and number of cormlets. However, for this experiment there were significant effects on the number of leaves and flowers because of the interactions between chitosan and the immersion time. The temperature of 55 °C affected plant development because emergence was delayed by 6 days; and there were less number of leaves, flowers and cormlets. On the other hand, the incidence of Fusarium oxysporum in naturally infected corms was 0% at temperatures of 55 °C and 50 °C. Immersion times (0, 10, 15 and 20 min) in hot water at 50 °C did not show significant effects on plant development and vase life. Corms dipped in Biorend^® at 1.5% and hot water at 50 °C accelerated their emergence for about 1–7 days, the number of flowers increased by two, extended the storage life for 1–3 days and increased the number of cormlets. The integration of these two treatments -Biorend^® and hot water- might be a good option for increasing the gladiolus plant quality and vase life.