A general procedure for protoplast recovery,callus and plant regeneration in plants

A portion of this work was supported by NSF grant DCB 8718314 and by the Missouri Agricultural Experiment Station. This research is a joint contribution from the Missouri Agricultural Experiment Station, Journal Series No. 10,789 and from the Massachusetts Agricultural Experiment Station, Amherst, MA 01003; Journal Paper No. MAES 2959. Ousama Zaghmout was supported by the Food for the 21st Century Program, College of Agriculture, University of Missouri, Columbia, MO.     

The ecology and electrophoretic analysis of the damselfly,Argia vivida hagen,living in a geothermal gradient

The ecology and electrophoretic properties of a damselfly, Argia vivida Hagen, inhabiting a geothermal gradient were studied. Monthly sampling of five sites revealed nymphal colonization along a 15–40°C thermal gradient; greatest densities occurred between 15–27°C. An electrophoretic analysis of proteins suggest that nymphs were adapted to a wide range of temperatures which was evidenced by differential activity of four enzyme systems (glucose-6-phosphate dehydrogenase, lactate dehydrogenase, leucine aminopeptidase, and tetrazolium oxidase). Evidence suggests the nymphs acclimated to different temperatures by altering the structure of important isozymes and expressed certain genetic features characteristic of individuals naturally found at a given temperature.Published with the approval of the Director of the University of Idaho Agriculture Experiment Station as Research Paper No. 78612.     

Influence of temperature on colonization of spring barleys by vesicular arbuscular mycorrhizal fungi

The effects of three soil temperatures on growth of spring barleys (Hordeum vulgare L.) and on their root colonization by vesicular arbuscular mycorrhizal (VAM) fungi from agricultural soils in Montana (USA) or Syria at different inoculum concentrations were tested in soil incubators in the greenhouse. The number of mycorrhizal plants as well as the proportion and intensity of roots colonized increased with higher soil temperatures. VAM fungi from Montana, primarily Glomus macrocarpum, were cold tolerant at 11°C while those from Syria, primarily G. hoi, were heat tolerant at 26°C. Inoculum potential of Montana VAM fungi was higher than Syrian VAM fungi in cool soils. Harmal, selected from Syrian barley land races, had the highest colonization by mycorrhizal fungi of the cultivars tested.Journal Series Paper: J-2532 Montana Agricultural Experiment Station.     

Identification of self-incompatibility groups in<Emphasis Type="Italic"> Hatiora</Emphasis> and<Emphasis Type="Italic"> Schlumbergera</Emphasis> (Cactaceae)

The Cactaceae, a family of about 1,800 species of succulent perennials, contains numerous species that exhibit self-incompatibility (SI). The objective of the current study was to determine the number of incompatibility groups present among diploid (2n=2x=22) cultivars of the genera Schlumbergera Lem. (Christmas cacti) and Hatiora Britton & Rose (Easter cacti). Two partial diallel crosses were performed, one with 19 cultivars of Christmas cacti [= S. truncata (Haworth) Moran and S. × buckleyi (Buckley) Tjaden] and the other with 10 cultivars of Easter cacti [= H. gaertneri (Regel) Barthlott, H. rosea (Lagerheim) Barthlott, and H. × graeseri Barthlott ex D. Hunt]. The compatibility/incompatibility status of crosses was determined by percent fruit set and presence of seed in mature fruit. None of the cultivars set fruit when selfed or crossed with a cultivar in the same incompatibility group, but fruit set ranged from 35% to 100% following compatible crosses. For the Christmas cacti, eight intra-incompatible but reciprocally compatible groups were identified, with 13 of the 19 cultivars assigned to three incompatibility groups (68%). The ten cultivars of Easter cacti yielded nine intra-incompatible but reciprocally compatible groups, with two cultivars in one incompatibility group and the other eight cultivars each assigned to a unique group. One cultivar of Christmas cactus ('Abendroth 6') was incompatible when crossed as a male with cultivars in incompatibility group 2 but was compatible in reciprocal crosses, results that suggest that this cultivar is an S-allele homozygote. The crossing relationships are consistent with a one-locus, gametophytic SI system with multiple alleles. Allozyme locus Lap-1, shown previously to be linked with the S locus (recombination frequency 7%) in Schlumbergera, exhibited insufficient allelic diversity for determining the S genotypes of the 19 cultivars of Christmas cacti. Based on the number of incompatibility groups in each diallel, at least five S-alleles occur in the 19 Christmas cacti and the 10 Easter cacti.Publication 3337 of the Massachusetts Agricultural Experiment Station. This material is based on work supported in part by the Cooperative State Research, Extension, Education Service, United States Department of Agriculture, Massachusetts Agricultural Experiment Station, under Project No. 746     

Characteristics of self-incompatibility in Schlumbergera truncata and S. x buckleyi (Cactaceae)

The influence of self-incompatibility (SI) on fruit set, seed set, and pollen tube growth was investigated in Schlumbergera truncata (Haworth) Moran and S.xbuckleyi (T. Moore) Tjaden. Four Schlumbergera clones were crossed in a complete diallel to verify the presence of SI. Fruit did not set when the clones were selfed or when two of the clones were crossed reciprocally, but all other outcrosses yielded fruit which contained 100–200 seeds each. Compatible outcrosses were characterized by large numbers of pollen tubes in the style and ovary cavity at 72 h after pollination. When pistils were selfed or incompatibly crossed, pollen tubes were inhibited in the upper third of the style and few pollen tubes reached the base of the style by 72 h after pollination. Schlumbergera exhibits several characteristics often associated with sporophytic SI systems (tricellular pollen and dry stigmas with elongate papillae), together with those commonly observed in gametophytic SI systems (stylar inhibition of incompatible pollen tubes and absence of reciprocal differences in outcrosses).Publication no. 3174 of the Massachusetts Agricultural Experiment Station     

Autolysis of high-GC isolates of Pseudomonas putrefaciens

High-GC isolates of P. putrefaciens undergo extensive autolysis after growth, resulting in a marked decrease in turbidity and the release of high-molecular-weight DNA which imparts a high viscosity to culture broths. The native DNA released is resistant to attack by the exocellular DNase activity of the culture broths. Autolysis is inhibited by a pH of 6.0 and the presence of 0.001 m Mg⁺⁺ or Ca⁺⁺, and is enhanced by elevated pH values and temperatures. This autolytic phenomenon in broth cultures readily distinguishes high- from low-GC isolates. The latter do not exhibit autolysis.Paper No. 1093, Massachusetts Agricultural Experiment Station, University of massachusetts at Amherst. This research was supported in part from Experiment Station Project No. 194 and Public Health Service Research grant FD 00153-09.     

Inheritance of the photoperiodically induced cold acclimation response in Cornus sericea L., red-osier dogwood

Summary Cold acclimation responses of latitudinal ecotypes of Cornus sericea L. (C. stolonifera Michx.) and F₁, F₂ and BC₁ hybrid progenies were measured under natural photoperiod conditions in St. Paul, MN and artificially shortened photoperiods in the glass-house. The 65 °N and 62 °N ecotypes (Alaska and Northwest Territories, respectively) were characterized by a short night length for hardiness induction, the 42 °N ecotype (Utah A and B) by a long night length for hardiness induction, while the F₁ was intermediate to the parents. Results from reciprocal crosses indicated there was no significant unilateral maternal influence on cold acclimation. Acclimation responses of the F₂ were highly variable but generally ranged between the parental extremes. However, three individuals from the 42 ° × 62 °N crosses exhibited greater cold resistance than the northern parent on two successive freezing test dates. F₂ plants were also found with less freezing resistance than the southern parent. Backcrosses to the southern parent produced progeny with acclimation patterns resembling that of the southern parent and were significantly less hardy than the F₂ in early freezing tests.Scientific Journal Series Paper No. 12,075 of the Minnesota Agricultural Experiment Station     

Penicillium urticae Bainier enumeration in soils

Summary A dilution plating method estimatedPenicillium urticae Bainier numbers in soil. This method, which used an agar layering technique and a cyclic incubation of 8 hours at room temperature (about 25°C) and 16 hours at 5°C, permitted the differential growth and sporulation in favor ofP. urticae B. over other common soil fungi.Procedures of extraction, paper chromatography, infrared analysis, and bioassay assayed for accumulated patulin. A combination of these methods routinely estimatedP. urticae B. numbers in soils and authenticated patulin production by selected isolates.Contribution from the Northern Plains Branch, Soil and Water Conservation Research Division, Agricultural Research Service, USDA, in cooperation with the Nebraska Agricultural Experiment Station, Lincoln. Published as Paper No. 2275, Journal Series, Nebraska Agricultural Experiment Station.Soil Scientist, USDA, Grand Junction, Colorado (formerly Chemist, USDA, Lincoln, Nebraska); and Microbiologist, USDA, Lincoln, Nebraska, respectively.     

Dispersal of first-instar gypsy moth larvae in relation to population quality

Summary Field studies of dispersal by first instar gypsy moth larvae indicate that almost all larvae undergo an initial dispersal episode. However, in laboratory studies large larvae (from large eggs) disperse more frequently than small larvae (from small eggs) in the presence of favored food. Large larvae may be better adapted for dispersal. When larvae encounter unacceptable food or are denied food, larvae disperse more frequently and dispersal by small larvae is nearly as frequent as dispersal by large larvae. Factors affecting egg size may contribute to shifts in dispersal patterns of gypsy moth larvae and distribution of populations.Paper No. 2041, Massachusetts Agricultural Experiment Station, University of Massachusetts at Amherst. This research supported (in part) from Experiment Station Project No. 355     

Organogenesis in pepper tissue cultures

Knowledge concerning in vitro growth and developmental responses of bell and chile peppers (Capsicum annuum L.) has been limited. Shoot and root organogenesis in cultures of seedling explants was restricted to primary cultures or those less than three months old under 12-and 16-h photoperiod at 25°C. Shoot organogenesis was extended to 5 months under continuous light at 25°C, and to 8 months under continuous light at 28.5°C. Murashige and Skoog basal media containing 0.05mg/l each of IAA and BA promoted shoot elongation and rooting of some explant sources, while 0.05-4 mg/l IAA with 10–50 mg/l BA promoted adventitious shoot bud formation. Glucose was superior to sucrose as the carbon source. Leaf discs collected from greenhouse-grown plants regenerated shoots for at least 2 months. Incubation environment, carbon source, explant source, growth regulator treatment and passage number were not independent variables as demonstrated by statistical analysis. The plant regeneration techniques described here have useful but limited applications, not extending to unorganized callus or cell suspension cultures.Journal article no. 1151 of the New Mexico Agricultural Experiment Station.     

Population quality,dispersal and numerical change in the gypsy moth,Lymantria dispar (L.)

Summary First instars from small and large gypsy moth eggs differ significantly in their head capsule width, weight, hatching time and the length of thoracic setae. Pupal weight and the developmental period of immature stages of the gypsy moth originating from small or large eggs do not differ significantly. The mean number of eggs per mass produced by females originating from small eggs is greater than that of females from large eggs although not statistically significant. Highly significant differences in mean egg size of egg masses of each type of female were also observed. The relationship between egg size and dispersal strategies are discussed.Paper No. 2229 Massachusetts Agricultural Experiment Station. University of Massachusetts at Amherst. This research supported (in part) from Experiment Station Project No. 355     

Suitability,digestibility and assimilation of various host plants of the gypsy moth Lymantria dispar L.

Summary The development and survival of gypsy moth (Lymantria dispar) larvae is strongly influenced by the host plant upon which they feed. The most rapid development and largest pupae were produced from grey birch fed larvae. Beech and maple-fed larvae produced the smallest pupae while maple-fed larvae exhibited prolonged development. White and red oak-fed larvae exhibited development and pupal weights intermediate between the above two groups. The approximate digestibility (AD) and efficiencies of conversion of food (ECD and ECI) were generally highest among grey birchfed individuals. The utilization of the relatively closely related oak species, as reflected in AD and ECD values, differed.Leaves were examined for 14 elements. The content of each element varied among host plant species and over time. For example, nitrogen levels were highest in grey-birch and dropped over time in all host plants.Lepidoptera: LymantriidaePaper No. 2277 Massachusetts Agricultural Experiment Station, University of Massachusetts at Amherst, MA U.S.A. This research supported (in part) from Experiment Station Project No. 437     

Regeneration of Gossypium hirsutum and G. barbadense from shoot apex tissues for transformation

A method of regenerating cotton plants from the shoot apical meristem of seedlings was developed for use with particle gun and Agrobacterium-mediated transformation. This method was developed to circumvent the problems of genotype restriction and chromosomal damage frequently encountered in cotton regeneration in tissue culture through somatic embryogenesis. In this procedure, the cells of the shoot meristem are targeted for transformation. Normal and fertile plants of Gossypium barbadense Pima S-6, and 19 cultivars of G. hirsutum were regenerated using this method. Shoot regeneration from these tissues was direct and relatively rapid. A MS based, hormone-free medium could be used with all the varieties tested.This project was funded by grants from Cotton Incorporated, Nisshinbo Industries, and a grant from the Texas Agricultural Experiment Station to RHS. Texas Agricultural Experiment Station Technical Article TA-25667.     

Gene dosage at the amylose-extender locus of maize: Effects on the levels of starch branching enzymes

Soluble starch branching enzymes and starch synthases from maize kernels of differing dosage of the ae locus were purified by DEAE-cellulose chromatography. A near-linear relationship between increasing dosage of the dominate amylose-extender allele (Ae) and branching enzyme IIb activity was found. In contrast, levels and properties of branching enzymes I and IIa, as well as the citrate-stimulated and primer-requiring starch synthases, remained unchanged. The near-linear increase in branching enzyme IIb activity with increasing doses of the Ae allele is consistent with the hypothesis that ae is the structural gene coding for branching enzyme IIb.Paper of the Journal Series, New Jersey Agricultural Experiment Station, Cook College, Rutgers University, New Brunswick. This work was performed as part of NJAES Projects 12442 and 12201 (NE-124), supported by the New Jersey Agricultural Experiment Station, Regional Research Funds, NSF Grant PCM 78-16127, and funds from Corn Refiners Association, Inc.     

Influence of light intensity and temperature on the locomotory and flight activity ofBrachymeria intermedia [Hym.: Chalcididae] a pupal parasitoid of the gypsy moth

The changes in activity level ofBrachymeria intermedia (Nees) were studied by exposing adults to 15.5°, 21° and 26.5°C and light intensities of about 97, 195 and 260 ft-c. Flight and locomotory movements increased as temperature and light intensity increased. Significant peaks in activity occurred during the photophase. No activity was observed during the scotophase.

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Résumé Les modifications du niveau d'activité deBrachymeria intermedia (Nees) ont été étudiées en exposant les adultes à 15,5°C, 21°C et 26,5°C et à des intensités de la lumière de 97, 195 et 260 ft.c. Les mouvements de locomotion et de vol augmentent avec l'élévation de la température et de l'intensité lumineuse. Des maxima d'activité significatifs se produisent pendant la photophase. Il n'y a pas d'activité pendant la scotophase.

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Paper No. 2181 Massachusetts Agricultural Experiment Station. University of Massachusetts at Amherst. This research supported (in part) from Experiment Station Project No. 437 and by a U. S. Department of Agriculture sponsored program entitled “The Expanded Gypsy Moth Research and Development Program (CSRS special grant No. 516-15-57)”.     

Growth of Agrostis palustris in subsurface black-layered sand induced by cyanobacteria and sulfate-reducing bacteria

The growth of Agrostis palustris was evaluated in sand columns in response to black-layer formed by the interaction of cyanobacteria in the genera Oscillatoria (isolates OS-1 and OS-2) and Nostoc (NS-1) with the sulfate-reducing bacterium Desulfovibrio desulfuricans. All plants of A. palustris transplanted into black-layered sand columns survived, and the black-layer cleared adjacent to roots as they grew down in the column. Black-layer remained in the columns below the advancing root tips. After 10 weeks of growth, numerous roots showed discontinuous reddish-brown discoloration on their surfaces. Shoot growth of A. palustris was reduced in response to all cyanobacteria and D. desulfuricans isolates alone or in combination. Root growth was unaffected by the microorganisms with the exception of stimulation by OS-1 and inhibition by D. desulfuricans. Interaction of the microbes and the formation of black-layer is discussed relative to the growth of A. palustris. Journal Paper J-14483 of the Iowa Agriculture and Home Economics Experiment Station, Ames, IA 50011. Project 2616 Journal Paper J-14483 of the Iowa Agriculture and Home Economics Experiment Station, Ames, IA 50011. Project 2616     

Somatic embryogenesis and plant regeneration from embryogenic suspension cultures of perennial ryegrass

Summary Embryogenic callus induced from mature caryopses of perennial ryegrass (Lolium perenne L.) were placed in liquid half-strength Murashige and Skoog (MS) basal medium and supplemented with 6.0 mg/l 2,4-dichlorophenoxy acetic acid (2,4-D), 3 g/l (w/v) casein hydrolysate (CH), and B5 vitamins, to initiate fast-growing highly embryogenic cell suspension cultures. Newly initiated suspension cultures contained a high level of large non-embryogenic cells (NE) with relatively few embryogenic (E) cells. Cell types were separated by discontinuous Percolls gradients or by filtering the newly initiated cultures through 31-μm nylon mesh. The growth conditions of the E cell were optimized by testing various media components including 2,4-D and sucrose, and subculture diluton ratio. Optimal shoot formation occurred after pretreatment of the embryogenic cells on solidified callus maintenance medium supplemented with 60 mg/l cefotaxime for 4 weeks prior to transfer to regeneration medium Regeneration media consisted of half-strength MS basal medium supplemented with B5 vitamins, 0.5 mg/l fluridone, and 0.5 mg/l BA. Most plants regenerated were albino with only a few green plants. Journal Paper number MAES 2959 of the Massachusetts Agricultural Experiment Station.     

A comparison of Hordeum bulbosum-mediated haploid production efficiency in barley using in vitro floret and tiller culture

Summary A high efficiency of Hordeum bulbosum-mediated haploid production in barley has been achieved using a floret culture technique in which florets pollinated with Hordeum bulbosum are cultured on modified N₆ media containing 0.5 mg/l kinetin and 1.2 mg/l2,4-D. Cultures were maintained at 25 °C with a 16 h photoperiod for 9 days before embryo rescue. In a comparison of haploid production efficiency using five F₁ hybrids from winter x winter and winter x spring barley crosses, 41.6 haploid plants/100 florets pollinated were produced using floret culture. Using detached tiller culture, 13.5 haploid plants/100 florets pollinated were produced. Higher efficiencies achieved with floret culture are attributed to the formation of larger, differentiated embryos. Such embryos lead to higher frequencies of plant regeneration. The F₁ from a winter x winter cross was inferior in haploid production compared to F₁s from winter x spring crosses. No genotype x technique interaction was observed.Oregon Agricultural Experiment Station Technical Paper No. 8653     

A method for determining the in vivo stability of Drosophila alcohol dehydrogenase (E.C.1.1.1.1.)

A rapid and accurate method of measuring the relative in vivo stability of Drosophila alcohol dehydrogenase is presented. The potential of this technique for examining posttranslational control of in vivo enzyme concentrations is discussed.This work was supported by NSF grant #DEB 7815466 to J.M.Journal Paper No. J-9977 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa. Project No. 2272.     

Chironomidae (Diptera) of Baldwin Lake,Illinois, a cooling reservoir

The chironomid fauna of Baldwin Lake, a closed-cycle cooling reservoir for a power plant, was sampled monthly to semi-monthly for one year at four stations, two in the discharge channel and two in the main basin. Qualitative samples were also taken elsewhere and with multiple-plate samplers. Twenty-four species were collected. Annual mean population density was less than 100 larvae m^–2 in the channel, 1037 M^–2 in the main basin. Three species of Tanypodinae formed over 96 percent of the larval population in the main basin: Tanypus stellatus Coquillett, Procladius bellus (Loew), and Coelotanypus concinnus (Coquillett). The principal species on the multiple-plate samplers were Dicrotendipes nervosus (Staeger), Glyptotendipes lobiferus (Say), and Parachironomus monochromus (Wulp). Temperatures of 35°C or more in the channel virtually eliminated the chironomids, whereas temperatures up to 32°C in the main basin increased the number of generations of T. stellatus from the usual two to four at Station 3 and three at Station 4. Station 3 had almost 1,000 degree-days more heat than Station 4.