Genetic parameters for growth and wood chemical properties in <Emphasis Type="Italic">Eucalyptus urophylla</Emphasis> × <Emphasis Type="Italic">E. tereticornis</Emphasis> hybrids

Key message

Growth and wood chemical properties are important pulpwood traits. Their narrow-sense heritability ranged from 0.03 to 0.49 in Eucalyptus urophylla × E. tereticornis hybrids, indicating low to moderate levels of genetic control. Genetic correlations were mostly favorable for simultaneous improvement on growth and wood traits. Additive and non-additive genetic effects should be considered in making a hybrid breeding strategy.

Context

Eucalypt hybrids are widely planted for pulpwood production purposes. Genetic variations and correlations for growth and wood chemical traits remain to be explored in Eucalyptus interspecific hybrids.

Aims

Our objectives were to clarify the heritability of growth and wood chemical traits and determine the genetic correlations between traits and between trials in E. urophylla × E. tereticornis hybrids.

Methods

Two trials of 59 E. urophylla × E. tereticornis hybrids derived from an incomplete factorial mating design were investigated at age 10 for growth (height and diameter) and wood chemical properties (basic density, cellulose content, hemi-cellulose content, lignin content, and syringyl-to-guaiacyl ratio). Mixed linear models were used to estimate genetic parameters.

Results

Narrow-sense heritability estimates were 0.13?0.22 in growth and 0.03?0.49 in wood traits, indicating low to moderate levels of additive genetic control. Genetic correlations were mostly positively significant for growth with basic density and cellulose content but negatively significant with hemi-cellulose and lignin contents, being favourablefavorable for pulpwood breeding purpose. Type-B correlations between sites were significant for all the traits except diameter and lignin content.

Conclusion

Hybrid superiority warrants the breeding efforts. An appropriate breeding strategy should be able to capture both additive and non-additive genetic effects.

     

Comparative microsporogenesis and flower development in <Emphasis Type="Italic">Eucalyptus urophylla</Emphasis> × <Emphasis Type="Italic">E. grandis</Emphasis>

Microsporogenesis and flower development in Eucalyptus urophylla × E. grandis were examined using chromosome tableting to provide a method to predict the meiotic stages in this species. Although microsporogenesis was normal, cytokinesis during meiosis of pollen mother cells occurred simultaneously, with strong asynchronism observed in the two different lengths of stamens in a flower bud. In a single flower, the developmental period of microsporogenesis in anthers on the longer stamens was always ahead of those on the shorter stamens. Flower development was also asynchronous at different locations on a branch. Flower buds on the upper side of the branch were larger in diameter than those on the lower side. In addition, a correlation was observed between microsporogenesis development and flower bud diameter growth. The pachytene stage was first observed when the diameter of the flower buds increased to 3.0 mm, and the majority of the meiotic stages were observed when bud diameters ranged from 3.5 to 5.0 mm. This study showed that the developmental stages of microsporogenesis in Eucalyptus urophylla × E. grandis could be distinguished readily, which may be applicable to future breeding studies.     

Micropropagation of <Emphasis Type="Italic">Eucalyptus grandis</Emphasis> × <Emphasis Type="Italic">E. urophylla</Emphasis> AEC 224 clone

Genetic transformation systems require protocols that allow regenerating transgenic plants from transformed tissues. This study aimed to establish a protocol for indirect organogenesis in leaf explants of a Eucalyptus grandis  ×  E. urophylla AEC 224 clone. During callogenesis stage, several concentrations of NAA and then NAA or 2,4-D combined with TDZ were tested in JADS culture medium for 30 days, followed by subculture of the explants in the regeneration medium, containing 5.0 µM BA and 0.5 µM NAA for another 30 days. In these media, the explant oxidation rate was high (95 %). Thus, in order to reduce oxidation, different culture media were compared: WPM, MS, JADS and modified QL, followed by explant transfer onto regeneration medium. The highest percentage of regeneration and the lowest oxidation rate were achieved on WPM medium. Then, NAA and 2,4-D were tested in combination with TDZ and also TDZ and BA combined with NAA in WPM medium. The most efficient culture media in terms of shoot regeneration were WPM supplemented with 0.25 µM TDZ and 0.1 µM NAA during 30 days for callus induction and then with 5.0 µM BA and 0.5 µM NAA for another 30 days. This protocol yielded a regeneration rate of 43 %, with a low oxidation of tissues. A rooting experiment was conducted using half strength MS medium and comparing three concentrations of IBA (2.46, 4.90 and 7.35 µM). The highest rooting percentage (35 %) was obtained on medium containing 2.46 µM IBA. Once the shoots were rooted, acclimatization in a greenhouse was not challenging and plant survival reached 100 %.     

Tree and stand growth for clonal <Emphasis Type="Italic">E. urophylla</Emphasis> × <Emphasis Type="Italic">grandis</Emphasis> across a range of initial stockings in southern China

A large initial stocking × fertilizer factorial field trial established in 1993 with a Eucalyptus urophylla × grandis hybrid clone in southern Guangxi province, China, included 6 initial stocking treatments ranging from 667 to 2,222 trees ha⁻¹ and 6 fertilizer treatments. Growth data was collected at various intervals up to age 192 months and also data on outer-wood pilodyn penetration (as an indicator of wood density) and acoustic velocity through the outer-wood (as an indicator of modulus of elasticity) at the last assessment. This report examines the results for the main effect of initial stocking treatments and stocking × fertilizer interactions across ages. There were significant differences between initial stockings for both average individual tree volume and standing volume ha⁻¹ at all ages up to 144 months, but just average individual tree volume at 192 months. At ages up to 75 months total standing volume decreased as initial stocking decreased—maximum volume being obtained at 2,222 trees ha⁻¹. From 88 to 110 months the highest standing volume was obtained at 1,667 trees ha⁻¹ and then at ages 144 and 192 months, 833 trees ha⁻¹ provided the highest standing volume. Average individual tree volume at all ages generally increased as initial stocking decreased—the largest trees were obtained at 667 trees ha⁻¹. As age increased the proportional differences between tree volumes at higher and lower stockings increased—at age 27 months average individual tree volume at 667 trees ha⁻¹ was 72% greater than that at 2,222 trees ha⁻¹ and differences increased steadily with age up to 190% at 192 months. There were no significant differences between initial stockings for the properties of the outer-wood assessed by pilodyn penetration and acoustic velocity. Also, no significant interactions were found between initial stocking and fertilizer treatments for any traits at any ages implying that effects of initial stocking and fertilizer are additive for the traits assessed.     

AFLP fingerprinting of <Emphasis Type="Italic">Populus deltoides</Emphasis> and <Emphasis Type="Italic">Populus </Emphasis>×<Emphasis Type="Italic"> canadensis</Emphasis> elite accessions

Amplified fragment length polymorphism (AFLP) was used in genetic fingerprinting of 22 elite Chinese Populus deltoides and Populus × canadensis accessions. The results indicated that each of the nine AFLP primer pairs selected generated fingerprint profiles that were unique to each of the accessions. Therefore, each accession was definitively identified by any of the nine primer pairs. Furthermore, the inter-accession genetic relationships inferred based on 461 polymorphic fragments from the nine AFLP primer pairs were largely consistent with phylogenetic relationships based on morphologic traits. Bootstrap analysis showed that three AFLP primer pairs were required to obtain genetic similarity values with a maximum CV of 10% while 10 AFLP primer pairs could give a maximum CV of 5%. Thus, AFLP can readily be applied for a rapid and accurate evaluation of the degree of similarity between poplar cultivars. In this study, the number of AFLP fragments used was sufficient to establish a reliable estimate of genetic similarity among accessions, with a maximum CV of 5.12%. Therefore, the information on the genetic relationships among the poplar accessions generated in this study in connection with knowledge on agronomic traits may have an impact on poplar breeding and planting in China.     

Soil microorganisms alleviate the allelopathic effect of <Emphasis Type="Italic">Eucalyptus grandis</Emphasis> × <Emphasis Type="Italic">E</Emphasis>. <Emphasis Type="Italic">urophylla</Emphasis> leachates on <Emphasis Type="Italic">Brassica chinensis</Emphasis>

Soil microbes may be critical players in determining the allelopathic potential of some plants. Low levels of plant community biodiversity in Eucalyptus plantations have been attributed to the allelopathic potential of these tree species. The role of soil microbes in the allelopathic effect of leaf leachates of the hybrid tree Eucalyptus grandis × E. urophylla, was tested in Petri dish assays with Brassica chinensis as a receiver plant. Soils were collected from either a local garden (soil A) or a Eucalyptus plantation (soil B) and half of each soil was sterilized to remove microbes. These soils were then treated with E. grandis × E. urophylla leachates for 0–72 h. Seed germination of B. chinensis was significantly inhibited in soils treated with leaf leachates relative to untreated soils. The inhibitory effect of the leaf leachates was more pronounced in sterilized soils. Total phenolic content was obvious lower in nonsterile leachate-treated soils than in sterile soils. Biomass of B. chinensis was negatively correlated with the total phenolic content in soils. Our findings suggest that soil microbes can alleviate the allelopathic potential of Eucalyptus and thereby its negative impact on plant growth.     

Activities of some defense proteins associated with age or plant family in larch needles of <Emphasis Type="Italic">Larix olgensis</Emphasis> and <Emphasis Type="Italic">L. kaempferi</Emphasis> × <Emphasis Type="Italic">L. gmelinii</Emphasis>

Defence proteins are a kind of chemical defence compounds. They play a key role in plant restraining biotic and abiotic harm. To illuminate activities of some defense proteins associated with age or plant family of larch, the larch needles were collected from two different families of Korean larch, Larix olgensis and a hybrid larch, L. kaempferi × L. gmelinii, respectively, and then the activities of defence proteins in those were tested using a UV spectrophotometry. The results showed that the activities of protective enzymes at the16-/17- and 19-year-age groups were higher than those at the other age groups in the both larch species. While the activities of polyphenol oxidase (PPO) at the 16-/17-year-age group and phenylalanine ammonia-lyase (PAL) and protease inhibitors at the 19-year-age group were the highest among all age groups. Then, compared with L. olgensis, the hybrid larch had significant effects on the activities of protective enzymes. The effects of plant family on the activities of PAL and chymotrypsin inhibitor were significantly different, and then those on the activities of PPO were not significantly different. The effects of the two families in L. olgensis on the activities of trypsin inhibitor (TI) were significantly different, while those in the hybrid larch on the activities of TI were contrary. To conclusions showed that the different age and plant family significantly affected the activities of defence proteins in the needles of two larch species, and then enhanced the larch resistance to pests. These could play a key function in forestry tree genetic improvement and management in future.     

Differences in leaf functional traits between red and green leaves of two evergreen shrubs <Emphasis Type="Italic">Photinia</Emphasis> × <Emphasis Type="Italic">fraseri</Emphasis> and <Emphasis Type="Italic">Osmanthus fragrans</Emphasis>

Leaf functional traits are adaptations that enable plants to live under different environmental conditions. This study aims to evaluate the differences in leaf functional traits between red and green leaves of two evergreen shrubs Photinia × fraseri and Osmanthus fragrans. Specific areas of red leaves are higher than that of green leaves in both species. Thus, the material investment per unit area and per lamina of red leaves is significantly lower than that of green leaves, implying an utmost effort of red leaves to increase light capture and use efficiency because of their low leaf-chlorophyll concentration. The higher petiole length of green leaves compared with that of red leaves indicates that adult green leaves may have large fractional biomass allocation to support the lamina structures in capturing light with maximum efficiency and obtaining a high growth rate. The high range of the phenotypic plasticity of leaf size, leaf thickness, single-leaf wet and dry weights, and leaf moisture of green leaves may be beneficial in achieving efficient control of water loss and nutrient deprivation. The high range of phenotypic plasticity of leaf chlorophyll concentration of red leaves may be advantageous in increasing resource (especially light) capture and use efficiency because this leaf type is juvenile in the growth stage and has low leaf-chlorophyll concentration.     

Two novel eukaryotic translation initiation factor 5A genes from <Emphasis Type="Italic">Populus simonii</Emphasis> × <Emphasis Type="Italic">P. nigra</Emphasis> confer tolerance to abiotic stresses in <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis>

The role of plant eIF5A proteins in multiple biological processes, such as protein synthesis regulation, translation elongation, mRNA turnover, programmed cell death and stress tolerance is well known. Toward using these powerful proteins to increase stress tolerance in agricultural plants, in the present study, we cloned and characterized PsneIF5A2 and PsneIF5A4 from young poplar (P. simonii × P. nigra) leaves. The deduced amino acid sequences of PsneIF5A2 and PsneIF5A4 were 98 % similar to each other, and they are orthologs of eIF5A1 in Arabidopsis. In a subcellular localization analysis, PsneIF5A2 and PsneIF5A4 proteins were localized in the nucleus and cytoplasm. qRT-PCR analysis showed that PsneIF5A2 and PsneIF5A4 were transcribed in poplar flowers, stem, leaves, and roots. In addition, they were also induced by abiotic stresses. Transgenic yeast expressing PsneIF5A2 and PsneIF5A4 had increased salt, heavy metal, osmotic, oxidative tolerance. Our results suggest that PsneIF5A2 and PsneIF5A4 are excellent candidates for genetic engineering to improve salt and heavy metal tolerance in agricultural plants.     

The effects of phenolic acid on nitrogen metabolism in <Emphasis Type="Italic">Populus</Emphasis> × <Emphasis Type="Italic">euramericana</Emphasis> ‘Neva’

The declines in soil fertility and productivity in continuously cropped poplar plantations are related to phenolic acid accumulation in the soil. Nitrogen is a vital life element for poplar and whether the accumulation of phenolic acid could influence nitrogen metabolism in poplar and thereby hinder continuous cropping is not clear. In this study, poplar cuttings of Populus × euramericana ‘Neva’ were potted in vermiculite, and phenolic acids at three concentrations (0X, 0.5X and 1.0X) were added according to the actual content (1.0X) in the soil of a second-generation poplar plantation. Each treatment had eight replicates. We measured gas exchange parameters and the activities of key enzymes related to nitrogen metabolism in the leaves. Leaf photosynthetic parameters varied with the concentration of phenolic acids. The net photosynthetic rate (P_N) significantly decreased with increasing phenolic acid concentration, and non-stomatal factors might have been the primary limitation for P_N. The activities of nitrate reductase (NR), glutamine synthetase (GS) and glutamate synthase (GOGAT), as well as the contents of nitrate nitrogen, ammonium nitrogen, and total nitrogen in the leaves decreased with increasing phenolic acid concentration. This was significantly and positively related to P_N (P < 0.05). The low concentration of phenolic acids mainly affected the transformation process of NO₃^? to NO₂^?, while the high concentration of phenolic acids affected both processes, where NO₃^? was transferred to NO₂^? and NH₄⁺ was transferred to glutamine (Gln). Overall, phenolic acid had significant inhibitory effects on the photosynthetic productivity of Populus × euramericana ‘Neva’. This was probably due to its influence on the activities of nitrogen assimilation enzymes, which reduced the amount of amino acids that were translated into protein and enzymes. Improving the absorption and utilization of nitrogen by plants could help to overcome the problems caused by continuous cropping.     

Combined surface drip irrigation and fertigation significantly increase biomass and carbon storage in a <Emphasis Type="Italic">Populus</Emphasis> × <Emphasis Type="Italic">euramericana</Emphasis> cv. Guariento plantation

Fast-growing poplar plantations are considered of great benefit to both timber production and carbon (C) sequestration, and are increasingly planted for multiple purposes worldwide. Irrigation and fertilization are common management practices in plantations in semiarid regions. However, quantitative investigation of the integrative effect of surface drip irrigation and fertigation (SDIF) on biomass and C storage in poplar plantations remains limited. In this study, we conducted a field experiment on a fast-growing poplar cultivar (Populus × euramericana cv. Guariento) plantation to compare the combination of surface drip irrigation and fertigation in growing seasons with conventional management (control; CK). Experiments repeated over 2 years showed that SDIF significantly increased biomass and C storage in both trees and soil in the plantation compared with the CK. Tree biomass C in SDIF-treated and CK stands after the first year of the experiment (age 5) was 6.20 and 4.05 t C ha^?1, respectively, and the difference further increased, i.e., 15.18 and 8.63 t C ha^?1, respectively, after the second year of the experiment (age 6). There was 53 and 76 % higher C storage in SDIF-treated trees than in the CK trees after the first and second years of the experiment, respectively. The SDIF increased the soil C concentration, especially in the surface soil at 0- to 40-cm depth. Soil organic C at a depth of 0–60 cm under the SDIF treatment was 45.42, 50.87 and 61.32 t C ha^?1 in the 1st, 2nd and 3rd years, respectively, with annual increases of 12 and 21 % between the first and second, and second and third year, respectively. The corresponding soil organic C in the CK was 43.08, 43.57 and 47.92 t C ha^?1 in the 1st, 2nd and 3rd years; the annual increases were only 1 and 10 %, respectively. The results confirmed the significant effect of the combined management on C storage in poplar plantations, thus we suggest it can be applied in forestry management, even though it generally did not change C concentrations of tree components.     

Effects of Thidiazuron,basal medium and light quality on adventitious shoot regeneration from <Emphasis Type="Italic">in vitro</Emphasis> cultured stem of <Emphasis Type="Italic">Populus alba</Emphasis>×<Emphasis Type="Italic">P. berolinensis</Emphasis>

The effect of Thidiazuron （TDZ）, basal media and light quality on adventitious shoot regeneration from in vitro cultured stem of Populus albaxP berolinensis were determined to establish a high efficiency shoot regeneration system from stem explants of P. alba~P berolinensis. Stems ofPopulus alba~P berolinensis were collected from cultured shoots in vitro derived from dormancy buds of 3-year-old seedlings. The stem explants were cultured on MS medium containing 0.02-mg·L-1NAA （naphthaleneacetic acid）, and 0.1, 0.3, 0.5 and 1.0 mg·L-1 concentrations of TDZ to determine the effect of TDZ on shoot regeneration. Three basal media, i.e. MS, woody plant medium （WPM） and B5, were used to test their influences of different media on adventitious shoot regeneration. Green, red, blue and yellow plastic films in comparison with florescent light as control were used to observe their effects on shoot regeneration. The results showed that differ- ent concentrations of TDZ had an evident influence on shoot regeneration. Lower concentration of TDZ （0.1 mg·L-1） resulted in more ad- ventitious shoot regeneration and higher concentration of TDZ （〉0.1 mg·L-1） inhibited shoot regeneration. Among different media, MS medium exhibited a high efficiency for shoot regeneration, followed by WPM medium, while B5 medium inhibited shoot regeneration. Normal light and yellow light exhibited better effects on shoot regeneration, compared with other light.     

Effect of silvicultural treatments on growth,fertility and capsule traits in seedling seed orchards of <Emphasis Type="Italic">Eucalyptus camaldulensis</Emphasis> and <Emphasis Type="Italic">E. tereticornis</Emphasis>

The effects of nutritional treatments and the flowering promoter Paclobutrazol™ on tree growth and fertility were studied in unpedigreed seedling seed orchards (SSOs) of Eucalyptus camaldulensis and E. tereticornis at two locations in southern India. At Pudukkottai, a semi-arid site in Tamil Nadu State, five treatments were applied: (1) untreated control, (2) nitrogen (two doses of urea at 217 g N/tree), (3) nitrogen plus phosphorus (two doses of 312 g single superphosphate), potassium (two doses of 83 g muriate of potash) and trace elements (two doses of 10 g sodium borate and 25 g zinc sulphate), (4) pollarding trees at 4 m height with application of N, P, K and trace elements, (5) drenching with the flowering promoter Paclobutrazol (0.25 ml active ingredient per cm of stem girth) along with N, P, K and trace elements. At Panampalli, a higher-rainfall site in Kerala State, only treatments 1, 2 and 3 were applied. Four replicates of treatment plots sized 18 m × 18 m, with 22–32 trees per plot were set up in each orchard in randomized complete block designs. Untreated plots of E. camaldulensis and E. tereticornis at Pudukkottai displayed 12 and 21% fertile trees, respectively, in the first year of monitoring, while at Panampally the percentage of fertile trees in untreated E. camaldulensis plots was higher at 72%, and that of E. tereticornis similar at 23%. Both species attained greater stem diameter at breast height at age 9 years at the higher-rainfall location but there was no significant impact of fertilizer application on 9-year diameter at either site. Diameter was reduced by pollarding and Paclobutrazol application at Pudukkottai. Only Paclobutrazol produced a significant increase in fertility in both species for four successive years, increasing the percentage of fertile trees to 59 and 71%, respectively, for E. camaldulensis and E. tereticornis 1 year after application. The number of capsules produced per tree also increased significantly with Paclobutrazol application, although the difference was comparatively less by the fourth year. The number of capsules per umbel did not differ significantly between locations, whereas the germination rate varied between treatments although no consistent trends were observed.     

5<Emphasis Type="Italic">Α</Emphasis>-Reductase inhibitory tannin-related compounds isolated from <Emphasis Type="Italic">Shorea laeviforia</Emphasis>

Five tannin-related compounds – gallic acid, flavogallonic acid dilactone, valoneic acid dilactone, gallagyldilactone, ellagic acid – were isolated from the heartwood of Shorea laeviforia, and the inhibitory activity of each against rat liver 5-reductase was evaluated. Valoneic acid dilactone and gallagyldilactone exhibited positive inhibitory activity, but gallic acid and ellagic acid did not. Flavogallonic acid dilactone stimulated 5-reductase activity, even though this compound is structurally similar to valoneic acid dilactone. The kinetic study of valoneic acid dilactone and gallagyldilactone indicated that the inhibitory behavior of 5-reductase were not competitive against the substrate (testosterone) and were partially competitive against the cofactor (NADPH). Additionally, double inhibition analysis of valoneic acid dilactone and NADP⁺ showed synergetic inhibition. These results suggested that neither valoneic acid dilactone nor gallagyldilactone can affect the binding of testosterone but that either compound could interact with an enzyme–NADP⁺ complex to inhibit 5-reductase.     

Adventitious rooting of <Emphasis Type="Italic">Eucalyptus globulus</Emphasis> × <Emphasis Type="Italic">maidennii</Emphasis> mini-cuttings derived from mini-stumps grown in sand bed and intermittent flooding trays: a comparative study

Eucalyptus globulus Labill and hybrids thereof have low lignin content, favoring cellulose extraction, but are often recalcitrant to clonal propagation. This work analyzed biochemical and morphological changes during adventitious rooting of mini-cuttings of E. globulus × maidenni obtained from mini-stumps cultured in drip fertigated sand bed or intermittent flooding tray commercial propagation systems. Morphological (% rooting, root number and length, mean rooting time) and biochemical parameters (peroxidase activity, total phenolic content and flavonoid content) were monitored to characterize the rooting phases. All of the rooting parameters were equivalent in both systems, indicating comparable efficiency of both methods in clonal propagation. Kinetic profiles of biochemical parameters were also similar, although the activity of peroxidases was an order of magnitude higher and the phenolic content about three times lower in cuttings derived from intermittent flooding-grown mini-stumps than in those derived from sand bed-grown mini-stumps. Taken together, results suggest that rooting phases were similar in both systems: induction before day 5, formation from day 5 to 15, and elongation from day 15 to 45. These data may contribute to the development of rooting phase-specific mineral nutrient solutions to maximize clonal propagation and plant survival.     

What factors influence the stem taper of <Emphasis Type="Italic">Eucalyptus</Emphasis>: growth,environmental conditions,or genetics?

• Introduction   

Stem taper equations have been widely used for volume estimation to varied top diameter limits or for biomass calculations. However, their main drawback is that specific calibration is often necessary for each species or clone, and accounting for genetic and environmental effects is often a challenge.     

Fitness of heat and desiccation tolerant hybrid strains of <Emphasis Type="Italic">Heterorhabditis</Emphasis> <Emphasis Type="Italic">bacteriophora</Emphasis> (Rhabditidomorpha: Heterorhabditidae)

Entomopathogenic nematodes of the genus Heterorhabditis are a suitable alternative to chemical insecticides because of their high control effect against insects in cryptic environments and their environmental safety. The availability of H. bacteriophora hybrid strains with increased tolerance to environmental extremes could be a driving force for a more widespread use of nematode-based plant protection products. However, increase in heat and desiccation tolerance of hybrid strains could compromise their fitness regarding virulence, host penetration and reproductive capacity. The fitness of heat and desiccation tolerant hybrid strains was compared to the commercial strain EN 01. Only the heat tolerant strains were superior or similar in fitness to strain EN 01. The strains with increased desiccation tolerance were usually less fit, a possible result of a trade-off effect of selection for desiccation tolerance. Hybrid strains selected for enhanced tolerance after an adaptation to stress were generally better in fitness ranking compared to those for which adaptation prior to stress exposure was excluded. This might be a result of pleitropy. Host penetration and virulence was not correlated. The commercial strain had the highest reproduction per mean number of nematodes penetrating the host insect, which is a result of automatic selection of inbred lines with high reproduction potential during the commercial production process in liquid culture.     

Development of transgenic hybrid sweetgum (<Emphasis Type="Italic">Liquidambar styraciflua</Emphasis> × <Emphasis Type="Italic">L. formosana</Emphasis>) expressing γ-glutamylcysteine synthetase or mercuric reductase for phytoremediation of mercury pollution

Using Agrobacterium-mediated gene transfer, we generated transgenic hybrid sweetgum (Liquidambar styraciflua × L. formosana) overexpressing two types of genes to enhance plant remediation of mercury-contaminated soil and water: bacterial γ-glutamylcysteine synthetase gene (ECS), the first and most important enzyme in phytochelatin synthesis, or various genes encoding a mercuric ion reductase (merA9, merA18, merA77). Hybrid sweetgum proembryogenic masses (PEMs) constitutively overexpressing ECS were able to grow in the presence of 50 μM HgCl₂, which inhibited wild-type PEMs, but plantlets regenerated from the PEMs had abnormal form and did not survive for more than a few weeks following germination. In contrast, mature somatic embryos generated from PEMs constitutively overexpressing merA9 and merA18 converted to normal plantlets on germination medium containing 25 μM HgCl₂, while control embryos were killed on 25 μM Hg(II)-medium. Transgenic merA plantlets displayed enhanced resistance to Hg(II) and released Hg(0) two to three times more efficiently than the wild-type plantlets.     

Comparative data on the insecticide resistance of <Emphasis Type="Italic">Anopheles</Emphasis> <Emphasis Type="Italic">albimanus</Emphasis> in relation to agricultural practices in northern Belize,CA

Anopheles albimanus Wiedemann is one of the primary vectors of malaria in northern Belize. The primary breeding sites for this species in Central America are fresh water wetlands that are found in close proximity to agriculture and, in particular, sugarcane fields. The use of insecticides and herbicides on these crops leads to the introduction of chemicals into the surrounding wetlands. The potential for the development of insecticide resistance is, therefore, quite high. A study was undertaken to obtain comparative data on the resistance status of An. albimanus in northern Belize in relation to agriculture practices. Larval surveys were conducted at four collection points in marshes juxtaposed to different crop types: Dubloon (DB)—control site located 300 m from any agriculture; Little Belize (LB)—mixed crop agricultural with some human settlements; and Chan Chen (CC) and Buena Vista (BV)—year round sugarcane production. Time mortality curves were obtained by exposing F1 larvae to DDT 1 μg/ml, malathion 1 μg/ml, and permethrin 0.05 μg/ml. Significant differences in mortality rates from the four sites were observed only to malathion. The DB sites contained larvae that were the most susceptible overall, in congruence with its distance from crops. Larvae from LB exhibited a medium level of tolerance. Finally, the two populations collected at the edge of sugarcane fields (CC and BV) exhibited the highest tolerance to malathion. These results represent first data on the impact of agriculture on the resistance status of malaria vectors in northern Belize. The effect of malathion on An. albimanus insecticide resistance reinforces the concerns of the potential impact of agriculture usage on the efficacy of vector control program.     

Live-weight gains of Holstein × Zebu heifers grazing a <Emphasis Type="Italic">Cratylia argentea</Emphasis>/Toledo-grass (<Emphasis Type="Italic">Brachiaria brizantha</Emphasis>) association in the Mexican humid tropics

The legume Cratylia argentea associated to Brachiaria brizantha—Toledo-grass (Bb + Ca) and Toledo-grass alone (Bb) were evaluated under grazing conditions by Holstein × Zebu heifers. Three evaluation periods during three consecutive years, were performed. We measured, daily live-weight gains of heifers (DWG, g/day); biomass dry matter (BDM, kg/ha) at beginning and end of each grazing period. On plant samples, were measured percentages of crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF), lignin, in situ DM degradability (ISDMD), and voluntary biomass DM intake (VBI, g/LW^0.75) using the Cr₂O₃/in situ indigestibility technique. A completely randomized design was applied, using heifers as experimental units for DWG gains and VBI. At the beginning of the grazing, the BDM for Bb + Ca and Bb–considering the three periods-averaged, 3065 and 936 kg/ha, respectively. Crude protein content of C. argentea was around 20 %; and for Bb + Ca or Bb, ranged from 8.7 to 4.6 %. In either treatment, in situ DM degradability averaged 72.6 % (P ≥ 0.05), regardless of the botanical component type. Biomass DM intake was 83.02 (Bb + Ca) and 89.22 (Bb) g/LW^0.75. The DWG gains (g/heifer) for Bb + Ca and Bb were (per period): 829 and 574 (first); 469 and 118 (second); and 534 and 508 (third). This study showed that the Bb + Ca association was better to improve the daily gain of F1 Holstein × Zebu heifers, as compared to Bb alone. Also, C. argentea associated to a low to medium-quality grass improved the nutritional value of the diet without affecting the biomass DM intake.