Threshold Concentrations of Nucleopolyhedrovirus in Soil to Initiate Infections in <Emphasis Type="Italic">Heliothis virescens</Emphasis> on Cotton Plants

The purpose of the research was to determine threshold concentrations of nucleopolyhedrovirus (NPV) in soil for abiotic transport to cotton plants in the field and under conducive and nonconducive conditions in the greenhouse. Under the assumption that 2% mortality would suffice to initiate foci of infection in Heliothis virescens larvae on the plants, thresholds ranged from 25 to 2,311 viral occlusion bodies (OB)/g soil in the greenhouse. Thresholds generally were smaller for rain on sandy soil and wind on clay soil than for wind on sand or rain on clay. Thresholds generally increased with height of the plant above the soil surface. In field plots, percentage mortality in bioassays of cotton plants was greatest on leaves versus other tissues, and mortality increased with soil dosage and decreased with plant height and over time. Season-long soil-NPV-transport thresholds for 2% plant-bioassay mortality of larvae ranged from 1 OB/g to 7.4 × 10⁸ OB/g soil based on the amounts of NPV applied to the soil at planting time, and they ranged from 8 OB/g to 1.2 × 10⁵ OB/g soil based on bioassays of soil samples collected concurrently with plant samples throughout the growing season. These results should contribute to NPV epizootiology, biological control, and risk assessment through better understanding of viral soil-to-plant transport.     

Exotic plant communities shift water-use timing in a shrub-steppe ecosystem

Semiarid areas in the US have realized extensive and persistent exotic plant invasions. Exotics may succeed in arid regions by extracting soil water at different times or from different depths than native plants, but little data is available to test this hypothesis. Using estimates of root mass, gravimetric soil water, soil-water potential, and stable isotope ratios in soil and plant tissues, we determined water-use patterns of exotic and native plant species in exotic- and native-dominated communities in Washington State, USA. Exotic and native communities both extracted 12 ± 2 cm of water from the top 120 cm of soil during the growing season. Exotic communities, however, shifted the timing of water use by extracting surface (0–15 cm) soil water early in the growing season (i.e., April to May) before native plants were active, and by extracting deep (0–120 cm) soil water late in the growing season (i.e., June to July) after natives had undergone seasonal senescence. We found that δ ¹⁸O values of water in exotic annuals (e.g., −11.8 ± 0.4 ‰ for Bromus tectorum L.) were similar to δ ¹⁸O values of surface soil water (e.g., −13.3 ± 1.4 ‰ at −15 cm) suggesting that transpiration by these species explained early season, surface water use in exotic communities. We also found that δ ¹⁸O values of water in taprooted exotics (e.g., −17.4 ± 0.3 ‰ for Centaurea diffusa Lam.) were similar to δ ¹⁸O values of deep soil water (e.g., −18.4 ± 0.1 ‰ at −120 cm) suggesting that transpiration by these species explained late season, deep water use. The combination of early-season, shallow water-use by exotic winter-actives and late-season, deep water-use by taprooted perennials potentially explains how exotic communities resist establishment of native species that largely extracted soil water only in the middle of the growing season (i.e., May to June). Early season irrigation or the planting of natives with established root systems may allow native plant restoration.     

Effects of AM colonization on “wild tobacco” plants grown in zinc-contaminated soil

This greenhouse study aimed to determine the effect of colonization by the arbuscular mycorrhizal (AM) fungus (Glomus intraradices Schenck & Smith) on the “wild” tobacco (Nicotiana rustica L. var. Azteca), under soil–zinc (Zn) conditions. Plants of N. rustica were grown in AM or non-AM inoculated substrate and subjected to four soil–[Zn] concentrations (0, 50, 100, and 250 mg Zn kg⁻¹ dry soil). The AM root colonization increased markedly from 14 to 81% with the increasing soil–[Zn] and the mycorrhizal structures were significantly more abundant at the highest soil–[Zn], suggesting that Zn may be involved directly or indirectly in AM root colonization. In addition, total Zn content or Zn concentrations in shoots and roots were shown to increase as soil–[Zn] increased in both AM and non-AM plants. As for the growth parameters studied, there were no significant differences between treatments despite the increase in Zn content or concentration. The AM roots subjected to the highest soil–[Zn] had a significant reduction by about 50% of total Zn content and Zn concentration compared to non-AM roots. Still, the relative extracted Zn percentage decreased dramatically as soil–[Zn] increased. Soil pH was significantly lower in non-AM than AM treatments at the highest soil–[Zn]. In summary, AM plants (particularly roots) showed lower Zn content and concentration than non-AM plants. In this regard, the AM fungi have a protective role for the host plant, thus playing an important role in soil-contaminant immobilization processes; and, therefore, are of value in phytoremediation, especially when heavy metals approach toxic levels in the soil.     

In vitro shoot-tip grafting improves recovery of cotton plants from culture

A rapid in vitro shoot-tip grafting (STG) technique was adapted to increase recovery of intact cotton plants from shoots developed in culture. Induction of root organogenesis in cotton shoots is genotype dependent and unreliable. The resulting loss of regeneration potential due to failure to form roots can vary from 30 to 80% according to genotype and represents a significant bottleneck in the overall recovery of plants from culture. If the non-rooting shoots are transgenic, the loss in regenerated plant material can be substantial. In vitro grafting of cotton shoots to seedling rootstock proved to be a simple and reliable method allowing 90–100% recovery of non-rooting shoots from culture. Success of any given graft was directly related to scion size (0.8–1.0 cm) and age (14–35 days) of the seedling rootstock. The method appeared to be genotype independent, and varietal differences between rootstock and scion did not effect the rate of plant recovery from culture. This revised version was published online in June 2006 with corrections to the Cover Date.     

Micropropagation of Hemidesmus indicus for cultivation and production of 2-hydroxy 4-methoxy benzaldehyde

Caulogenic responses of various explant types from 12-month-old plants of Hemidesmus indicus were tested. Second and third visible nodes (0.5 cm) from the apex and root segments (0.5 cm) were the most and least regenerative respectively, with the formation of 9.37 and 2.6 shoots in 4 weeks on half strength MS medium supplemented with 2.22 μM BA and 1.07 μM NAA and 4.44 μM BA and 2.69 μM NAA respectively. Caulogenic ability of the nodes decreased with increasing maturity. Shoot buds initiated upon the young nodes on day 10 developed into 7.2 cm long shoots within 4 weeks thereby making a shoot elongation phase unnecessary. Nodal explants of the in vitro raised shoots subcultured in the same medium produced 9.32 shoots of 7.1 cm length in 3–4 weeks, similar to those of the mature plant-derived nodes. Multiplication through subculture of the nodes up to 25 passages of 4 weeks each was achieved without decline. Shoot cultures were rooted in quarter salt strength MS medium containing 9.8 μM IBA and the rooted plants were hardened for establishment in pots at 96% rate. Four months after establishment, the micropropagated plants were stable and showed uniform morphological and growth characteristic. After 12 months of cultivation in the field, on an average micropropagated plant consisted of 4–5 shoots, 5–8 branches per shoot and increased root biomass (13.5 g) compared to the poor growth (single shoot and 2–3 branches) and root production (4.6 g) values obtained with plants raised from conventional rooted stem cuttings. The concentration of the root specific compound, 2-hydroxy 4-methoxy benzaldehyde per plant was 2–3 fold higher in micropropagated plants though on unit dry root biomass (0.12% per g dry wt) basis it remained the same between two sources of plants. This revised version was published online in June 2006 with corrections to the Cover Date.     

How much seed remains in the soil after a fire?

Soil seed banks that persist after a fire are important in fire-prone habitats as they minimise the risk of decline or local extinction in plants, should the fire-free interval be less than the primary juvenile periods of the species. In two common woody plant genera (Acacia and Grevillea) in southeastern Australia, we examined the size and location of the residual seed bank after fire across areas of varying seedling densities at three locations in comparison to the distribution of seeds in the soil at an unburnt site. We found viable dormant seeds remaining in the soil after fire (evidence of residual soil seed bank). A significantly lower proportion of seeds remained in the top 5 cm of soil than at 5–10 cm or 10–15 cm soil depths, independent of seedling density or plant genus. This was due to greater germination, and possibly some seed mortality, near the soil surface. Reduced germination below 5 cm was probably due to the reduced efficacy of the fire cues that break seed dormancy, a declining ability of seeds to emerge successfully from such depths, and the lower abundance of seeds in the soil at such depths. The magnitude of the residual seed bank was similar across 0–5, 5–10 and 10–15 cm soil depths in Acacia suaveolens. For two Grevillea species, most residual seeds were at 0–5 and 5–10 cm. The residual soil seed bank in the top 10 cm of soil after fire varied across sites with estimates of 0, 19 and 27% in G. speciosa and 23, 35, and 55% in A. suaveolens. At two sites, both species had similar residual seed bank sizes, while at a third, there were large differences between the species (0–55%). The observed patterns imply that the fire-related cues that break seed dormancy generally declined with soil depth. For Acacia, seed dormancy is broken by heat shock, a fire-cue that declines with soil depth. Some 250 species (approx 15% of the fire-prone flora) in the region are thought to have dormancy broken by heat shock. For Grevillea, where seed dormancy is broken by the interaction of smoke and heat shock, at two sites, we suggest three possibilities: (i) the smoke cue declined with soil depth; (ii) both heat and smoke are obligatory for breaking seed dormancy; or (iii) the cues may be independent and additive and below the zone of soil heating, only a proportion of available seeds had dormancy broken by smoke alone. At a third site (no residual seed bank detected) the smoke cue was predicted not to have declined with soil depth. Up to 900 species (just under half the fire-prone flora) in the study region are thought to have seed dormancy broken by the interaction of heat and smoke during the passage of a fire.     

Transformations and fate of sheep urine-N applied to an upland U.K. pasture at different times during the growing season

The fate of sheep urine-N applied to an upland grass sward at four dates representing widely differing environmental conditions, was followed in soil (0–20 cm) and in herbage. Urine was poured onto 1-m² plots to simulate a single urination in August 1984 (warm and dry), May (cool), July and August 1985 (cool and wet) at rates equivalent to 40–52 g N m⁻². The transformation of urine-N (61–69% urea-N) in soil over a 6–7 week period followed the same general pattern when applied at different times during the season; rapid hydrolysis of urea, the appearance of large amounts of urine-N as ammonium in soil extracts, and the appearance of nitrate about 14 days after application. The magnitude of “apparent” nitrification however differed markedly with environmental conditions, being greatest in May 1985 when a maximum of 76% of the inorganic soil N was in the form of nitrate. At all other application dates nitrate levels were relatively low. With the August 1984 application soil inorganic N returned to control levels (given water only) after 31 days but considerable amounts remained in soil for 60–90 days with the other applications. Weekly cuts to 3-cm indicated that increases in herbage dry matter and N yields in response to urine application were greatest in absolute terms after the May 1985 application and continued for at least 70 days with all applications. Relative to control plots the May application resulted in a 3-fold increase in herbage DM compared with corresponding values of 6-, 5-, and 7-fold increases with the August 1984, July and August 1985 applications. Recovery of urine-N in herbage was poor averaging only 17% of that applied at different dates, while recovery in soil extracts was incomplete. The exact routes of loss (volatilisation, leaching, denitrification or immobilisation) were not quantified but it is evident that substantial amounts of urine-N can be lost from the soil-plant system under upland conditions.     

Comparison of conditions for mycelial growth of <Emphasis Type="Italic">Lepista sordida</Emphasis> causing fairy rings on <Emphasis Type="Italic">Zoysia matrella</Emphasis> turf to those on <Emphasis Type="Italic">Agrostis palustris</Emphasis> turf

Symptoms of fairy rings caused by Lepista sordida have been reported on Zoysiagrass (Zoysia spp.) turf maintained at fairway height (2 cm), but not on bentgrass (Agrostis spp.) maintained at putting green height (0.5 cm). The mycelia of this fungus inhabit primarily the upper 0–2 cm layer of the soil extending into the thatch. To compare conditions for the mycelial growth in Z. matrella turf to those in A. palustris turf, we examined the effects of nutrients, temperature, water potential, and pH in the field as well as in the laboratory. Greater growth of the mycelia was observed in medium that included hot water extracts from soil of the 0–1 cm zone in Z. matrella turf compared to that from A. palustris. The upper soil layer in Z. matrella turf contained more organic matter from clippings than that in A. palustris. The temperature and water potential of the 0–2 cm soil zone in Z. matrella turf were also more favorable for the mycelial growth. The soil pH values of this zone in Z. matrella turf were less favorable compared to A. palustris but within the range for accelerating mycelial growth. Part of this study was presented orally at the 46th meeting of the Mycological Society of Japan in 2002     

Vertical distribution of soil nematodes in an age sequence of <Emphasis Type="Italic">Caragana microphylla</Emphasis> plantations in the Horqin Sandy Land,Northeast China

The vertical distribution of soil nematodes down to a depth of 50 cm was studied in an age sequence of 0-, 5-, 10-, and 22-year-old Caragana microphylla plantations (treatments) in the Horqin Sandy Land, Northeast China. The abundances and generic compositions of nematode fauna in five soil layers (0–10, 10–20, 20–30, 30–40, and 40–50 cm) were analyzed. 42 genera were observed in the nematode suspensions, and Acrobeles was the dominant genus in all treatments. The results showed that the total number of nematodes and the generic diversity in an age sequence of C. microphylla plantations decreased with increasing soil depth. Significant differences in the numbers of total nematodes, bacterivores (BF), plant parasites (PP), and omnivores–predators (OP) were observed between treatments and depths. BF was the most abundant trophic group in our study, followed by OP. The numbers of OP showed an obviously increasing trend with increasing age of C. microphylla plantation. The vertical distribution of the soil nematode communities was related to gradual changes in soil chemical properties, and it indicated that C. microphylla plantations have played positive roles in improving soil environmental conditions and restoring desertified ecosystems in the Horqin Sandy Land. The ecological indices selected were influenced by plantation chronosequence but not by soil depth.     

Production,storability, and regeneration of shoot tips of potato (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.) encapsulated in calcium alginate hollow beads

Summary Shoot tips, of four potato cultivars (Désirée, Genet, Tigoni, and Tomensa), 3–4 mm in size, were precultured for 2 d on Murashige and Skoog (MS) solid medium, then encapsulated in calcium alginate to produce hollow bead synthetic seed capsules averaging 0.78 cm in diameter. Regeneration and ‘regrowth’ were tested on MS solid medium and on soil in the greenhouse, respectively. The encapsulated shoot tips were stored at 4 and 10°C for up to 390 d. For all cultivars, the encapsulated shoot tips stored at both temperatures for 180 d and at 4°C,for 270 d, 100% regeneration on MS solid medium was recorded. After 360 d in storage at 4°C, 70.8% (Tigoni), 66.7% (Genet), 58.3% (Désirée), and 51.5% (Tomensa) regeneration was recorded on MS medium, reducing to 15% (Tigoni), 25% (Genet), 10% (Désirée), and 0% (Tomensa) regeneration after 390 d in storage. ‘Regrowth’ of 93–100% was recorded for non-stored encapsulated shoot tips, directly transferred on soil in the greenhouse after a 2 wk preculture on MS solid medium with an added fungicide (carbendazim) in the encapsulating gel. The ‘regrown’ shoot tips produced plants showing normal development. The results presented here demonstrate that hollow bead synthetic seed capsules are an alternative propagating method for potato seed production.     

Sowing and transplantation of Adonis ramosa (Ranunculaceae) for the establishment of a new population in a seminatural broad-leaved deciduous forest floor

Adonis ramosa (Ranunculaceae) is a plant whose risk of extinction is increasing in Japan. In this study, we show that there is a significant possibility of establishing an A. ramosa population by sowing seeds or transplanting plants in a seminatural broad-leaved deciduous forest floor. Immediately after seed collection in early June, seeds were sown either in pots that were watered periodically or in a deciduous forest floor where the soil surface was prepared. In the following spring, 30–40% of the seeds germinated. Of the one-year-old plants buried at a depth of 2 cm and two-year-old plants buried at a depth of 2 or 8 cm in the fall, 96% sprouted leaves in the following spring. However, 79% of the one-year-old plants buried at a depth of 8 cm sprouted leaves. In spring, the number of leaves, the widths of individual plants, and the heights of the two-year-old plants were greater than those of the one-year-old plants. The dry weights of one- and two-year-old plants in fall after one spring growing season increased by 1.6–2.3 times and by 2.3–5.1 times, respectively, compared to the dry weight at the time of transplanting. Also, the dry weights of the plants transplanted at a depth of 8 cm were higher than those of plants transplanted at a depth of 2 cm for both one- and two-year-old plants. The results suggest that the growth conditions at a depth of 8 cm are more favorable than those at a depth of 2 cm.     

<Emphasis Type="Italic">Organogenic callus</Emphasis> as the target for plant regeneration and transformation via <Emphasis Type="Italic">Agrobacterium</Emphasis> in soybean (<Emphasis Type="Italic">Glycine max</Emphasis> (L.) Merr.)

A regeneration and transformation system has been developed using organogenic calluses derived from soybean axillary nodes as the starting explants. Leaf-node or cotyledonary-node explants were prepared from 7 to 8-d-old seedlings. Callus was induced on medium containing either Murashige and Skoog (MS) salts or modified Finer and Nagasawa (FNL) salts and B5 vitamins with various concentrations of benzylamino purine (BA) and thidiazuron (TDZ). The combination of BA and TDZ had a synergistic effect on callus induction. Shoot differentiation from the callus occurred once the callus was transferred to medium containing a low concentration of BA. Subsequently, shoots were elongated on medium containing indole-3-acetic acid (IAA), zeatin riboside, and gibberellic acid (GA). Plant regeneration from callus occurred 90 ∼ 120 d after the callus was cultured on shoot induction medium. Both the primary callus and the proliferated callus were used as explants for Agrobacterium-mediated transformation. The calluses were inoculated with A. tumefaciens harboring a binary vector with the bar gene as the selectable marker gene and the gusINT gene for GUS expression. Usually 60–100% of the callus showed transient GUS expression 5 d after inoculation. Infected calluses were then selected on media amended with various concentrations of glufosinate. Transgenic soybean plants have been regenerated and established in the greenhouse. GUS expression was exhibited in various tissues and plant organs, including leaf, stem, and roots. Southern and T₁ plant segregation analysis of transgenic events showed that transgenes were integrated into the soybean genome with a copy number ranging from 1–5 copies.     

Drought Stress Response on Some Key Enzymes of Cowpea (<Emphasis Type="Italic">Vigna unguiculata</Emphasis> L. Walp.) Nodule Metabolism

A greenhouse experiment was carried out aiming to evaluate the response to drought stress of cowpea nodule enzymatic activities during different plant developmental stages leading to biological N₂ fixation. Stress was applied by controlling soil’s water-potential through a porous cup. Cowpea plants cv IPA 205 were grown in pots with yellow latosol soil under three different matric potential (ψ_m) treatments. Even with high evaporative demand and limited soil water availability, cowpea could not induce an extremely low leaf water potential (ψ_w). Sap ureides concentration in cowpea declined during the drought stress period. There was a decline in enzyme activity in the metabolic pathways concerned with N₂ fixation: NADH-dependent glutamate synthase (EC 1.4.1.14), glutamine synthetase (EC 6.3.1.2) and phosphoenolpyruvate carboxylase (EC 4.1.1.31). In contrast, an increase in glutamate dehydrogenase (EC 1.4.1.4) was observed as the ψ_m declined. Metabolism associated with N₂ assimilation was impaired every time that the ψ_w was reduced below −0.73 MPa as had happened in the stressed treatments. The stress applied by the porous cup was gradual and the plant recovered its turgor, avoiding permanent deleterious alterations in the cellular metabolism, even from a limited cowpea-growth ψ_m.     

High frequency plant regeneration and accumulation of tropane alkaloids in regenerated plants of Scopolia parviflora

An efficient in vitro plant regeneration system was established from callus culture of Scopolia parviflora. Callus was induced from adventitious roots on B5 medium with 0.45–9.04 μM 2,4-dichlorophenoxyacetic acid (2,4-D). In vitro plantlet regeneration was achieved on B5 medium supplemented with 44.38 μM benzyladenine (BA), 3% sucrose, and 0.38% gelrite. Plantlets were transplanted to artificial soil and grown to maturity successfully in a greenhouse. The tropane alkaloid contents in regenerated plants were analyzed using high-performance liquid chromatography (HPLC), and were found to be higher than those of adventitious roots, native growing plants, and acclimated plants. Regenerated plants from organogenic callus cultures produced a greater amount of tropane alkaloids.     

Land use intensification affects soil microbial populations, functional diversity and related suppressiveness of cucumber Fusarium wilt in China’s Yangtze River Delta

The extent of soil microbial diversity in agricultural soils is critical to the maintenance of soil health and quality. The aim of this study was to investigate the influence of land use intensification on soil microbial diversity and thus the level of soil suppressiveness of cucumber Fusarium wilt. We examined three typical microbial populations, Bacillus spp., Pseudomonas spp. and Fuasarium oxysporum, and bacterial functional diversity in soils from three different land use types in China’s Yangtze River Delta, and related those to suppressiveness of cucumber Fusarium wilt. The land use types were a traditional rice wheat (or rape) rotation land, an open field vegetable land, and a polytunnel greenhouse vegetable land that had been transformed from the above two land use types since 1995. Results generated from the field soils showed similar counts for Bacillus spp. (log 5.87–6.01 CFU g⁻¹ dw soil) among the three soils of different land use types, significantly lower counts for Pseudomonas spp. (log 5.44 CFU g⁻¹ dw soil) in the polytunnel greenhouse vegetable land whilst significantly lower counts for Fusarium oxysporum (log 3.21 CFU g⁻¹ dw soil) in the traditional rice wheat (or rape) rotation land. A significant lower dehydrogenase activity (33.56 mg TPF kg⁻¹ dw day⁻¹) was observed in the polytunnel greenhouse vegetable land. Community level physiological profiles (CLPP) of the bacterial communities in soils showed that the average well color development (AWCD) and three functional diversity indices of Shannon index (H′), Simpson index (D) and McIntosh index (U) at 96 h incubation in BIOLOG Eco Micro plates were significantly lower in the polytunnel greenhouse vegetable land than in both the traditional rice wheat (or rape) rotation land and the open field vegetable land. A further greenhouse experiment with the air-dried and sieved soils displayed significantly lower plant growth parameters of 10-old cucumber seedlings as well as significantly lower biomass and total fresh fruit yield at the end of harvesting at day 70 in the polytunnel greenhouse vegetable soil sources. The percentages of Fusarium wilt plant death were greatly increased in the polytunnel greenhouse vegetable plants, irrespective of being inoculated with or without Fusarium oxysporum f. sp. cucumerinum. Our results could provide a better understanding of the effects of land use intensification on soil microbial population and functional diversity as well as the level of soil suppressiveness of cucumber Fusarium wilt.     

The reserve of weatherable primary silicates impacts the accumulation of biogenic silicon in volcanic ash soils

Banana plantlets (Musa acuminata cv Grande Naine) cultivated in hydroponics take up silicon proportionally to the concentration of Si in the nutrient solution (0–1.66 mM Si). Here we study the Si status of banana plantlets grown under controlled greenhouse conditions on five soils developed from andesitic volcanic ash, but differing in weathering stage. The mineralogical composition of soils was inferred from X-ray diffraction, elemental analysis and selective chemical/mineralogical extractions. With increasing weathering, the content of weatherable primary minerals decreased. Conversely, clay content increased and stable secondary minerals were increasingly dominant: gibbsite, Fe oxides, allophane, halloysite and kaolinite. The contents of biogenic Si in plant and soil were governed by the reserve of weatherable primary minerals. The largest concentrations of biogenic Si in plant (6.9–7 g kg⁻¹) and soil (50–58 g kg⁻¹) occurred in the least weathered soils, where total Si content was above 225 g kg⁻¹. The lowest contents of biogenic Si in plant (2.8–4.3 g kg⁻¹) and soil (8–31 g kg⁻¹) occurred in the most weathered desilicated soils enriched with secondary oxides and clay minerals. Our data imply that soil weathering stage directly impacted the soil-to-plant transfer of silicon, and thereby the stock of biogenic Si in a soil–plant system involving a Si-accumulating plant. They further imply that soil type can influence the silicon soil–plant cycle and its hydrological output.     

Threshold distances and depths of nucleopolyhedrovirus in soil for transport to cotton plants by wind and rain

Two aspects of abiotic transport of nucleopolyhedrovirus from soil to cotton plants were examined in greenhouse experiments: the distance from the plants and depth in soil from which the virus could be transported under controlled conditions of soil type and moisture, wind, and precipitation. Transport distance and depth were tested separately under relatively conducive (precipitation/sandy soil and wind/clay soil) and non-conducive (precipitation/clay soil and wind/sandy soil) conditions, as determined in previous research. The amount of virus transported by precipitation generally decreased as distance from the plant increased, but in wind the amounts of virus transported were best described by polynomial models, with transport efficiency usually peaking at a distance of 60 cm. Depending on plant height and tissue, the farthest distances that virus was transported ranged from 30 to 60 cm by precipitation from clay soil, 60-75 cm in precipitation/sand, 60-80 cm in wind/clay, and 60-80 cm in wind/sand. In the depth experiments, transport by precipitation and wind generally decreased as the depth of virus in soil increased. The greatest depth from which NPV was transported ranged from 0 to 0.5 cm by precipitation from clay soil, 0.5-1.0 cm in precipitation/sand, 1.0-2.0 cm in wind/clay, and 0.5-1.0 cm in wind/sand. All of the experimental parameters (distance or depth, soil type, plant height, plant tissue) and all two-way interactions significantly (P<0.05) affected transport in all four experiments, except for the "soilxplant tissue" interaction in the depth/wind experiment. In all of the experiments, transport was significantly greater (P<0.05) to lower than to upper portions of plants and to leaves than to buds and squares. Transport was significantly greater from sandy soil than from clay in precipitation, and it was greater from clay than from sandy soil in wind. The results will contribute to NPV epizootiology, microbial control, and risk assessment.     

An efficient grafting system for transgenic plant recovery in cotton (Gossypium hirsutum L.)

A successful transformation program relies on the number of survival plants in soil that can be obtained. Low recovery of transgenic plants is still a key restrictive factor for transgenic cotton production. In order to utilize genetic transformation in cotton breeding program effectively, an efficient grafting system for recovering plants derived from somatic embryogenesis following Agrobacterium infection and kanamycin selection was developed. Various aspects of in vitro grafting were examined in efforts to improve the efficiency of transformant recovery. Using strong seedling rootstocks was the first important step to obtain high rate of successful grafts. Scion size >0.6 cm and seedling rootstock at age of 6–12 days were appropriate for grafting. The successful grafting ratio was higher when using hypocotyls without radicle. Shoot-tip and shoot stem with axillary bud were also suitable for in vitro grafting, which meant we could significantly improve the survival ratio of transgenic plantlets, because one plantlet has a shoot-tip but several axillary buds. Based on our data, the period from in vitro seedling rootstock germination to transplant of grafts to field usually takes one month. Over 90% successful grafting ratio could be obtained under optimal conditions, which represented a significant improvement over currently available methods for recovery of cotton plantlet from somatic embryogenesis after transformation. Ex vitro grafting could also be used for plant recovery, which gave an average of successful grafting ratio of 71.9%. However, this method was strongly affected by environmental factors.     

<Emphasis Type="Italic">In vitro</Emphasis> propagation of <Emphasis Type="Italic">Zingiber petiolatum</Emphasis> (Holttum)

Summary We describe an in vitro propagation protocol for Zingiber petiolatum (Holttum), I. Theilade, a rare species from the southern part of Thailand. Fruits were surface-sterilized and seeds germinated on Murashige and Skoog medium (MS) medium supplemented with 3% sucrose. Three-month-old seedlings were used as initial plant material for in vitro propagation. Terminal buds of the plants were inoculated on MS medium containing 6-benzylaminopurine (BA; 2.2–35.5 μM) alone or in combination with 1-naphthaleneacetic acid (0.5 μM). Eight weeks after inoculation, the cultures were transferred to MS medium without plant growth regulators for 4wk. The cultures transferred from MS medium with 17.8 μM BA revealed the highest shoot induction rate of 6.1±0.7 shoots per explant. Rooting was spontaneously achieved in MS medium without plant growth regulators. Rooted plants were successfully transplanted to soil.     

Induction of multiple shoots and plant regeneration from ‘accessory buds’ of nodal segments from field-grown mature cotton plants (Gossypium hirsutum L.)

Summary Sprouting of a single shoot was obtained from nodal segments of field-grown cotton plants (Gossypium hirsutum L. cv. DCH-32 and NHH-44) when cultured on Murashige and Skoog's (MS) basal medium devoid of plant growth regulators. Pruning of the sprouted shoot followed by re-culturing of the explant on MS basal medium supplemented with 2.22 μM benzylaminopurine triggered the dormant ‘accessory buds’ present in the node to proliferate and differentiate into multiple shoots. The shoots elongated on the same medium and rooted on MS basal medium devoid of growth regulators. Plantlets were hardened under greenhouse conditions and transferred to the field. Flowering and boll formation were observed in these plants at maturity. This technique for successful clonal propagation from mature cotton tissues should permit the rapid multiplication of plants selected based on specific phenotypic or genotypic characteristics.