Control of Ethylene Induced Permeability of Plant Cells by a Substituted Benzothiadiazole

The permeability of bulb tissues of onion (Allium cepa L.), amaryllis (Hippeastrum hybrids), tulip (Tulipa × hybrida) and daffodil (Narcissus tazetta L.) treated with Ethephon and 5-methyl-7-chloro-4-ethoxycarbonylmethoxy-2,1,3-benzothiadiazole (TH6241), to water and solutions of 1,3-dimethylurea was determined by the plasmometric methods. Ethylene treatments promoted, while those with TH6241 generally decreased the cell membrane permeability; TH6241 negated the effects of ethylene on cell permeability.     

The in vitro propagation of amaryllis (Hippeastrum spp. Hybrids)

Summary A new, rapid technique for the propagation of amaryllis (Hippeastrum spp. hybrids) by means of tissue culture is reported. Leaf bases, scapes, peduncles, inner bulb scales and ovaries were cultured successfully in vitro and plantlets were induced readily at various concentrations of growth regulators. Some plantlets also were produced in the absence of growth regulators. The most productive tissues for propagation were inverted scapes and peduncles, cultured in a modified Murashige and Skoog salt solution with added organic constituents and 1 mg per 1 (4.5μM) 2,4-dichlorophenoxyacetic acid (2,4-D) and 1 mg per 1 (4.4μM) 6-benzylaminopurine (BAP). Plantlets induced axenically also grew roots on the generalized shoot-inducing medium so that no special rooting medium was required. Although friable callus was obtained from ovary tissue cultured on a medium containing 2 mg per 1 (11μM) naphthaleneacetic acid and 4 mg per 1 (18μM) BAP, it produced shoots after 8 weeks of further subculture on the same medium. An average of 10 rooted plantlets was obtained from each scape or peduncle explant on the shoot-propagating medium. Thus, if 45 explants are obtained from each bulb, 450 rudimentary plantlets could be obtained from each mother bulb in 8 weeks of culture. This is a substantial increase over present propagation methods. This work was supported by a grant-in-aid of research, to Bruce G. Cumming, from the National Research Council of Canada.     

Seed bulb size influences the effects of exogenous brassinolide on yield and quality of Pinellia ternata

• In recent years, natural Pinellia ternata populations of have gradually been exhausted, while the cultivated yield has been limited due to lack of research and uncertain climate condition. Therefore, it is necessary to explore methods of improving yield and quality in P. ternata using brassinolide (BR) treatments and choice of a suitable seed bulb size.
• This article reports the effects of BR and two seed bulb sizes (diameter: 0.5–1.0 cm and 1.0–1.5 cm) on active and nutrient components and antioxidant activity in P. ternata. The experiment included six levels of BR (0, 0.05, 0.10, 0.50, 1.00 and 2.00 mg l^?1).
• The tuber yield of the two seed bulb sizes and bulbil yield of small seed bulbs increased 5.67%, 22.66% and 69.23% by day 105 after 0.50 mg l^?1 BR treatment, compared with the control. On day 105, only 0.05 mg l^?1 BR increased scores in principal components analysis (PCA) in tubers of small seed bulbs by 167.29%, and 0.05 and 0.50 mg l^?1 BR increased PCA score in bulbils of large seed bulbs by 145.66% and 252.97%, respectively, compared with the control. Significant BR × seed bulb size interactions were found on yield and quality of P. ternata.
• The results indicate that BR effects on yield and quality of tubers and bulbils of P. ternata are not only related to BR concentration but also to seed bulb size.

     

The effects of storage condition on starch metabolism and regeneration potentials of twin-scales and inflorescence stem explants of <Emphasis Type="Italic">Narcissus tazetta</Emphasis>

Summary The natural propagation rate of Narcissus is very slow. In vitro micropropagation of Narcissus is more efficient than conventional propagation for rapidly building up aseptic stocks of varieties, especially for the establishment of new cvs. and the production of pathogen-free stock material. In the present study, Narcissus tazetta cv. ‘Ziva’ bulbs were used as the source of mother plants. The bulbs were kept at 30°C in a dark chamber until the start of the experiments. Prior to explant preparation, the bulbs were subjected to a cold treatment at 15°C in the dark for 6 wk to break dormancy. Twin-scales and inflorescence stem discs were isolated from aseptic bulb parts and were used as the initial explants. The polar orientation affected the regeneration of the inflorescence stem. Storage duration at 30°C followed by cold treatment were found to affect starch levels, adenosine diphosphate glucose pyrophosphorylase (AGPase) activities, and regeneration potentials. Starch levels were reduced significantly during a 10 mo. storage period at 30°C in both twin-scales and inflorescence stem disc explants. Regeneration was followed by an efficient acclimatization system with 98–99% survival. More than 500 highly uniform young bulbs were propagated from one mother bulb in a 12 mo. period.     

The establishment of axenic seed and embryo cultures of endangered Hawaiian plant species: special review of disinfestation protocols

More than one third of the USA’s federally listed endangered and threatened plants are endemic to Hawaii. Because of the accelerating rate of environmental pressures resulting from habitat degradation to loss, the Hawaiian Rare Plant Program (HRPP, Lyon Arboretum, Honolulu, Hawaii) utilizes in vitro culture and seed storage to rescue, recover, and restore Hawaii’s most critically endangered native plant taxa. These in vitro methods complement existing ex situ and in situ efforts in the state. Wild-collected seeds tend to be hard to clean, and obtaining contamination-free seeds or embryos without damage or over sterilization is one of the most difficult obstacles in the successful establishment of in vitro cultures. Loss of rare and valuable propagation material occurs when seed samples succumb to contamination or mortality during the initial disinfestation stage. This study reviews pre-treatments and provides examples of three general disinfestation protocols successfully used in the HRPP to establish axenic ovule and embryo cultures of some endangered Hawaiian species. Three disinfestation protocols are described: bleach sterilization, gas sterilization, and ethanol dip and flame. Since 1998, the HRPP has been successful in establishing in vitro cultures of 135 endangered Hawaiian plant taxa representing 30 genera.     

Incidence and Control of Necrotic Leaf Mosaic Caused by Arabis Mosaic Virus in Lilium tigrinum splendens in the Netherlands

Necrotic mosaic on leaves and ring spots on bulb scales of Lilium tigrinum splendens, can be caused by arabis mosaic virus (AMV). Primarily infected bulbs can show spongy roots and large necrotic areas on creamy coloured bulb scales. Consecutive series of plants replanted for monthly periods in infested soil were mostly infected by AMV at a high rate (70%) throughout the year. Very low or undetectable numbers of Xiphinema diversicandatum, nematodes in soil dilution experiments infected lilies very efficiently (50–70 %). In general, soil disinfestation with dichloropropene, dazomet, methylbromide, and other disinfectants were variably fairly effective, particularly when yellow crocus among which couch was abundant, was previously grown for two years. The influx of AMV infected material into the soil was assumed to increase the number of AMV-carrying nematodes, and may be one cause of the failure of soil disinfestation. A survey of AMV infested soil in lily-growing regions in The Netherlands indicated its occurrence in a few fields only. Complex control measures applicable under growers' conditions, in addition to the variably effective soil disinfestation, are indicated.     

The in vitro propagation of amaryllis (Hippeastrum spp. hybrids)

A new, rapid technique for the propagation of amaryllis (Hippeastrum spp. hybrids) by means of tissue culture is reported. Leaf bases, scapes, peduncles, inner bulb scales and ovaries were cultured successfully in vitro and plantlets were induced readily at various concentrations of growth regulators. Some plantlets also were produced in the absence of growth regulators. The most productive tissues for propagation were inverted scapes and peduncles, cultured in a modified Murashige and Skoog salt solution with added organic constituents and 1 mg per 1 (4.5 micron) 2,4-dichlorophenoxyacetic acid (2,4-D) and 1 mg per 1 (4.4 micrometer) 6-benzylaminopurine (BAP). Plantlets induced axenically also grew roots on the generalized shoot-inducing medium so that no special rooting medium was required. Although friable callus was obtained from ovary tissue cultured on a medium containing 2 mg per 1 (11 micrometer) naphthaleneacetic acid and 4 mg per 1 (18 micrometer) BAP, it produced shoots after 8 weeks of further subculture on the same medium. An average of 10 rooted plantlets was obtained from each scape or peduncle explant on the shoot-propagating medium. Thus, if 45 explants are obtained from each bulb, 450 rudimentary plantlets could be obtained from each mother bulb in 8 weeks of culture. This is a substantial increase over present propagation methods.     

Promotion by Jasmonic acid of bulb formation in shoot cultures of Narcissus triandrus L.

The in vitro bulb formation by shoot clumpcultures of N. triandrus on media with jasmonicacid (JA) alone or in association with both2-isopentenyladenine (2-iP) or naphthalene acetic acid(NAA) and on medium with only NAA was studied. Themedia with JA plus 2-iP or JA plus NAA caused a highmultiplication of leaves, significantly higher on thatwith 2-iP. Leaf proliferation was low on medium withJA alone. The media containing JA promoted theformation of a bulb at the base of the leaves and thebulbs attained different sizes. On the medium with JAplus NAA, the number of bulbs that reached up to 5 mmin diameter was higher than that on the medium with JAplus 2-iP, but the highest number was formed on themedium with JA alone. The medium with NAA alone led tothe formation of few small bulbs that were elongatedinstead of roundish as those formed on media with JA.Further culture of the bulbs on a growth mediumsuitable to their enlargement increased their size butthe dimensions attained were dependent upon the mediuminitially used for bulb formation. Bulbs derived frommedium with JA alone attained the largest diameter.Almost all the bulbs grown on the growth medium wererooted. Data described here show that JA promotes in vitro bulb formation in shoot cultures of N.triandrus and suggests that JA might play animportant role in the formation and enlargement ofbulbs in Narcissus plants.     

Reduction of contamination in bulb-explant cultures ofNarcissus by a hot-water treatment of parent bulbs

After disnfestation of explants with 1% sodium hypochlorite for 30 min, 40–60% of twin-scale cultures ofNarcissus Golden Harvest remained contaminated, mainly byFusarium spp. Extension of the disinfestation time in hypochlorite from 30 min to 2h or a double disinfestation treatment did not reduce the contamination, suggesting that the fungus may be located inside the tissue. Hot-water treatment of bulbs prior to treatment with sodium hypochlorite resulted in contamination being reduced to 5%, while the regeneration was not affected. Similar results were obtained when experiments were repeated with five other cultivars ofNarcissus.     

The effect of jasmonic acid, sucrose and darkness on garlic (Allium sativum L. cv. Ptujski jesenski) bulb formation in vitro

Summary The effect of sucrose, jasmonic acid (JA) and darkness on bulb formation of garlic Allium sativum L. cv. Ptujski jesenski was studied in vitro. B5 medium supplemented with 3% sucrose, 5 μM JA and 5 μM 2-isopentenyl adenine (2iP) was used for shoot induction on garlic basal plates. For bulb induction, explants with developed shoots were transferred onto media with 3% or 8% sucrose in the presence or absence of 5 μM JA. Sucrose (8%) significantly increased the percentage of shoots which formed bulbs by 86–90%, bulb diameter and the number of bulbs per basal plate. On medium supplemented with JA, the average number of bulbs per basal plate was 11.5. Growth of explants in the dark was ineffective for stimulating bulb formation. Simultaneous use of JA and sucrose can improve garlic micropropagation via bulb formation, without intermediate callus formation.     

<Emphasis Type="Italic">In vitro</Emphasis> micropropagation and alkaloids of <Emphasis Type="Italic">Hippeastrum vittatum</Emphasis>

Different concentrations of methyl jasmonate, spermine, casein hydrolysate, or progesterone combined with 16 mg/l 2 iP + 4 mg/l naphthalene acetic acid (NAA) were investigated in order to obtain higher multiplication rate and growth of Hippeastrum vittatum bulbs in vitro on MS basal medium. The highest multiplication rate (8.2 bulbs/explant) was attained with 80 mg/l spermine, while the highest bulb fresh weight (1.23 g/bulblet) was obtained with 4 mg/l methyl jasmonate. Progesterone at 20 mg/l or casein hydrolysate at 2.0 g/l gave the highest leaf length (14.1 and 13.2 cm, respectively). So, it can be advised to use 80 mg/l spermine combined with 16 mg/l 2 iP + 4 mg/l NAA to obtain the highest number of bulbs per explant with moderate leaf length and bulb fresh weight. Chemical analysis showed alternations in the alkaloid type ratio and number of compounds in the bulbs treated with methyl jasmonate (4 mg/l).     

Central Adiposity Determines Prevalence Differences of the Metabolic Syndrome

Objective: To compare the expression of the metabolic syndrome in Spain and San Antonio, TX, two populations with major differences regarding their cardiovascular risk profile. Research Methods and Procedures: Cross‐sectional analysis of population‐based, epidemiological surveys using the metabolic syndrome definition of the National Cholesterol Education Program. In San Antonio, we limited our analysis to non‐Hispanic whites because non‐Hispanic whites are largely of European ancestry (n = 1339 in San Antonio and 2947 in Spain) Results: In men, increased central adiposity was more prevalent in San Antonio than in Spain (29.7 vs. 23.0%, p < 0.0001); in women, it was less prevalent in San Antonio than in Spain (40.2 vs. 66.4%, p < 0.0001). The metabolic syndrome followed that same pattern: more prevalent in men (28.9 vs. 20.8%, p = 0.019) and less in women from San Antonio (27.1 vs. 30.9%, p < 0.0001). In subjects with the metabolic syndrome, most women had increased central adiposity (92.6% in San Antonio and 97.5% in Spain), and most men had either increased central adiposity or blood pressure (99.2% in San Antonio and 95.0% in Spain). Discussion: Contrary to men, the metabolic syndrome is more prevalent in Spanish women than in women from San Antonio with differences that mirror differences in central adiposity. Central adiposity and blood pressure may be used to exclude the metabolic syndrome. Considering recent secular trends in obesity, we predict there will be an increase in the prevalence of the metabolic syndrome in both populations in the coming years.     

鹭兰种子无菌繁殖球根

为建立鹭兰（Habenaria radiata）种子繁殖法,对无菌培养条件下的种子发芽和幼苗生长进行了观测,并对无菌繁殖获得的球根进行了种植试验。结果表明,鹭兰种子不进行处理难以无菌培养发芽,种子通过75%乙醇10 s+1% NaClO 10 min处理,或者40 kHz超声波2.5 min+1% NaClO 10 min处理,在1/2MS培养基的发芽率可达58%以上;在1/2MS培养基中,添加GA-3 1 mg·L^-1对叶片生长有促进作用,添加IAA 1 mg·L^-1对根生长和球根形成有促进作用,球根形成率达55%;无菌繁殖的球根贮存至春季盆栽,成苗率约72%。     

Modulation of somatic embryogenesis in hypocotyl-derived cultures of geranium (Pelargonium X hortorum bailey) CV ringo rose by a bacterium

Summary The Ringo Rose cultivar of zonal geranium (Pelargonium x hortorum Bailey) has been shown to be morphogenetically unresponsive. Attempts to improve somatic embryogenesis using various seed stress treatments before germination proved ineffective. However, bacterial contamination of one of the seed-stress treatments led to infected explants that had a significant increase in frequency of high-quality somatic embryos. The co-cultivation of explants with the isolated bacterium (tentatively identified asBacillus sp.) was found to be repeatable, and potentially represents a novel way to improve morphogenesis in geranium and possibly other species.     

Flowering pathway is regulated by bulb size in Lilium longiflorum (Easter lily)

Lilium longiflorum (Easter lily) vegetative propagation occurs through production of underground bulbs containing apical and axillary meristems. In addition, sexual reproduction is achieved by flowering of elongated shoots above the bulb. It is generally accepted that L. longiflorum has an obligatory requirement for vernalisation and that long day (LD) regime hastens flowering. However, the effect of bulb size and origin, with respect to axillary or apical meristems on flowering, as well as the interactions between these meristems are largely unknown. The aim of this study was to explore the effect of bulb size, vernalisation and photoperiod on L. longiflorum flowering. To this end, we applied vernalisation and photoperiod treatments to the different bulb sizes and used a system of constant ambient temperature of 25 °C, above vernalisation spectrum, to avoid cold‐dependent floral induction during plant growth. Vernalisation and LD hasten flowering in all bulbs. Large, non‐vernalised bulbs invariably remained at a vegetative stage. However, small non‐vernalised bulbs flowered under LD conditions. These results demonstrate for the first time that cold exposure is not an obligatory prerequisite for L. longiflorum flowering, and that an alternative flowering pathway can bypass vernalisation in small bulbs. We suggest that apical dominance interactions determine the distinct flowering pathways of the apical and axillary meristems. Similar floral induction is achieved in propagated bulblets from scaling. These innovative findings in the field of geophyte floral induction represent valuable applicative knowledge for lily production.     

Spread of Exotic Cordgrasses and Hybrids (Spartina sp.) in the Tidal Marshes of San Francisco Bay, California, USA

Four species of exotic cordgrass (Spartina sp.) occur in the San Francisco estuary in addition to the California native Spartina foliosa. Our goal was to map the location and extent of all non-native Spartina in the estuary. Hybrids of S. alterniflora and S. foliosa are by far the most numerous exotic and are spreading rapidly. Radiating from sites of deliberate introduction, S. alterniflora and hybrids now cover ca. 190 ha, mainly in the South and Central Bay. Estimates of rate of aerial increase range from a constant value to an accelerating rate of increase. This could be due to the proliferation of hybrid clones capable of rapid expansion and having superior seed set and siring abilities. The total coverage of 195 ha by hybrids and other exotic cordgrass species is slightly less than 1% of the Bay's tidal mudflats and marshes. Spartina anglica has not spread beyond its original 1970s introduction site. Spartina densiflora has spread to cover over 5 ha at 3 sites in the Central Bay. Spartina patens has expanded from 2 plants in 1970 to 42 plants at one site in Suisun Bay. Spartina seed floats on the tide, giving it the potential to export this invasion throughout the San Francisco estuary, and to estuaries outside of the Golden Gate. We found isolated plants of S. alterniflora and S. densiflora in outer coast estuaries north of the Bay suggesting the likelihood for the San Francisco Bay populations to found others on the Pacific coast.     

Totipotency in Tissue Explants and Callus of Some Members of the Liliaceae, Iridaceae, and Amaryllidaceae

In vitro responses of twelve species of bulbs and conns werecompared. Plantlets could be induced directly without interveningcallus on stem tissue in nine species, on ovary tissue in fivespecies, and on leaf tissue in four species. In Gladiolus, Hyacinthus,Muscari, Ornithogahim, and Scilla plantlets were formed withoutgrowth factors added to the Murashige and Skoog medium. In Hippeastrum,Schizoslylis, Sparaxis, and Ipheion auxin was required. No plantletscould be induced directly on explants of growing tissue of Freesia,Tulipa, or Narcissus. Adventitious plantlets could be inducedon pieces of bulb or corm from ten species but such materialwas difficult to free from contamination. Callus was obtained from all species except Tulipa and Hippeastrum.Plantlets could be regenerated from callus except that of Gladiolus,Sparaxis, and Schizostylis. Differences in responses of the twelve species tend to cut acrossthe three families and no simple relation is evident betweenthe natural rate of vegetative increase and the in vitro behaviour.     

Clonal propagation,encapsulation and reintroduction of <Emphasis Type="Italic">Ipsea malabarica</Emphasis> (Reichb. f.) J. D. Hook., an endangered orchid

Summary Rapid clonal propagation and encapsulation of in vitro-formed bulbs of Ipsea malabarica (Reichb. f.) J. D. Hook., an endemic and endangered orchid of the Western Ghats of Kerala, and its reintroduction to the natural habitat were accomplished. Rhizome segments of Ipsea cultured on half-strength Murashige and Skoog (MS) medium supplemented with 6.97 μM kinetin induced the highest number of shoots, at the rate of four shoots per explant within 50 d. Transfer of the isolated shoots increased the rate of shoot multiplication to more than 10 shoots. Subsequent culture enhanced the number of shoots. No decline of shoot multiplication was observed up to the 10th subculture. Shoots developed bulbs during culture which developed into rhizomes. Sucrose at 6–8% reduced the time for the development of bulbs and rhizomes. Roots were developed from the base of the developed shoots as well as from the bulbs. Isolation and culture of bulbs also developed 5–10 shoots within 50 d. Encapsulated in vitro-formed bulbs cultured either on hormone-free halfstrength MS or 6.97 μM kinetin-supplemented medium facilitated 100% conversion. As a step to conservation in situ, 50 plantlets were reintroduced into their natural habitat, i.e. at Vellarimala (at 1300 m height) of the Western Ghats of Kerala, and flowered normally. Development of more than 40 000 plantlets starting from a single explant is possible within 250 d. This threatened endemic orchid stands to benefit greatly from the established protocol and will hopefully curtail the threat of extinction.     

Eucomis zambesiaca baker: Factors affecting in vitro bulblet induction

Eucomis species having considerable horticultural potential are used in African traditional medicine to treat various ailments. The effects of environmental and physiological parameters on the initiation and growth of bulblets using leaf explants were investigated. These included the effect of temperature (10, 15, 20, 25 and 30 °C), photoperiod (8 h light, 16 h light, continuous light and continuous dark), carbohydrates (sucrose, fructose and glucose) at different concentrations and combinations as well as various plant growth regulators; gibberellic acid (GA₃), indole-3-butyric acid (IBA), napthaleneacetic acid (NAA), N⁶-benzyladenine (BA), zeatin and others. Liquid shake and liquid static cultures versus solid cultures were investigated. Maximum number of bulblets per leaf explant was obtained at 20 °C, with an average of 3 bulbs per leaf explants and a bulblet mass of 57 mg. An 8 h light cycle produced 1.38 bulbs per leaf explant, at a mass of 42 mg. Fructose at 3% produced an average of 1.18 bulbs per leaf explant, 3.39 mm wide and weighing 56.6 mg. Of the plant growth regulators, 4.90 µM IBA was found to be the optimum treatment for bulblet induction, with an average bulb diameter of 4.36 mm and a mean bulblet mass of 79.07 mg. Liquid shake cultures exhibited poor growth while bulblet, leaf and root growth was improved in liquid static cultures. Successful micropropagation from leaf explants established that leaf explants can be used as an alternative explant source to bulbs. This protocol allows for the fast and economic mass propagation of Eucomis plants.     

Micropropagation systems for the mexican redbud (Cercis canadensis var. Mexicana L.) and other woody plants of the chihuahuan desert

Summary There are many Chihuahuan desert species that have potential as landscape plants for the arid communities of the southwestern United States [agarita, Berberis trifoliata Moric.; Mexican buckeye, Ungnadia speciosa Endl.; Texas madrone, Arbutus xalapensis var. texana (Buckl.). A. Gray]. Within these plant populations, there are superior genotypes that offer even greater interest for the landscape. However, it is difficult to clonally propagate many of these species with conventional techniques, and the seed-derived populations often do not breed true. Therefore, selection of superior genotypes in wild populations coupled with clonal propagation through tissue culture may offer an attractive option. It is relatively easy to achieve disinfestation of explants from desert plants due to a general lack of natural surface contamination by fungi and bacteria, even though interference from numerous trichomes can impede good contact with disinfesting agents. However, there is only a narrow window of time that is ideal for explant collection, because of the brief, periodic flushes of growth that characterize this unusual plant group. There may be years when, due to the harsh environment, the amount of suitable explant material is severely limited. Phenolics and exudates are also problematic in this group of plants, and acclimatization of ex vitro plantlets to the harsh desert environment is a particular challenge. For these reasons, specific adaptations and modifications were necessary to achieve success with micropropagation of desert plant species such as Mexican redbud (Cercis canadensis var. mexicana L.).