Keragaman Genetik Klon Lokal Kopi Robusta Asal Temanggung Berdasarkan Marka SSR

<em>Temanggung is one of the centers of Robusta coffee production in Central Java, with a variety of potential local coffee clones. The exploration found that several numbers of local Robusta coffee clones had the potential to have high productivity and were resistant to pests/diseases. However, their level of genetic similarity to the superior clones that have been released has not been clearly identified. This study aimed to investigate the genetic variability of Temanggung local Robusta coffee clones based on SSR markers. The study was conducted at the Molecular Laboratory of the Integrated Laboratory, Indonesian Industrial and Beverage Crops Research Institute from February to December 2018. A total of 29 local coffee clones derived from Temanggung along with two control coffee clones (BP 42 and BP 358) were used. All the 14 SSR markers used in the present study were polymorphic and could cluster those local coffee clones into 5 major groups at a genetic similarity coefficient of 0.57. Four local coffee clones (Putih Daun Lebar, Lokal, Tugusari Hijau and Tugusari Kuning) were in the same group with control clones in group I. One local clone (Tugusari Hijau) had a genetic similarity with the control clone BP 358 at 0.91. Meanwhile, the other 25 local clones were in different groups from the control clones. These local clones, which showed genetic distance far from the control clones, can be selected as candidates for local superior clones in coffee breeding program.</em>

Productivity and Biomass Properties of Poplar Clones Managed in Short-Rotation Culture as a Potential Fuelwood Source in Georgia

Georgian forests are very valuable natural resources, but due to the lack of affordable alternatives to firewood, people are forced to use forest resources illegally and unsustainably. The aim of this study was to determine the productivity and biomass properties of four poplar clones from Aigeiros and Tacamahaca and one control clone, considering their wood and bark characteristics and their proportion in the stems. Short-rotation woody crops with these clones represent a potential source of commercial fuelwood production in Georgia as an alternative to natural forests. These tree characteristics were evaluated after three years of growth. The survival of the clones was generally high. No significant differences in biomass production (dry matter, DM) were found among the four clones tested (DM of approximately 4 Mg ha−1 yr−1), while the control clone achieved significantly lower values for DM. The biomass specific density was exceptionally high, at 481–588 kg m−3, which was a result of the high proportion of bark mass in the stem (23.3–37.7%), with a density almost twice that of wood. On the other hand, the tested clones had a very high ash content in the biomass (2.6–4.5%), which negatively affected their energy potential expressed as a lower heating value (17,642–17,849 J g−1). Our preliminary results indicated that both the quantity and quality of biomass are important factors to justify the investment in an intensive poplar culture. The four clones should be further considered for commercial biomass production and tested at different sites in Georgia to evaluate the genotype-by-environment interactions and identify the site conditions required to justify such an investment.

The Epidemic Spread of Seiridium cardinale on Leyland Cypress Severely Limits Its Use in the Mediterranean

Leyland cypress (× Hesperotropsis leylandii) is a fast-growing conifer used in most temperate regions as an ornamental tree for hedges and screens, and is one of the most commercially important trees in Europe. In recent years, severe diebacks and mortality due to cypress canker have been observed on Leyland cypress plantations in Southern Europe. This study was conducted to evaluate (i) the spread and impact of cypress canker caused by Seiridium cardinale in plantations of a sample area of 1,250 km2 in central Italy, (ii) the response of the most commonly grown Leyland cypress varieties to artificial inoculation with to S. cardinal, and (iii) the pathogenicity of S. cardinale isolates obtained from Leyland cypress. Of the 1,411 surveyed trees, 11.4% had been killed by cypress canker and 43.9% of the living trees were affected by the disease. The number of diseased or dead trees and the percentage of cankered trunks was significantly correlated with the mean trunk diameter of the plantations. Six months after inoculation, the size of developed cankers was significantly different among the inoculated Leyland cypress cultivars but all of them showed markedly larger cankers than the C. sempervirens canker-resistant control clone. All of the tested S. cardinale isolates obtained from Leyland cypress also caused cankers on Cupressus sempervirens when inoculated as conidial suspensions or mycelia. Leyland cypress is highly prone to contract cypress canker in the Mediterranean due to its high susceptibility to S. cardinale infections, low genetic variability among the grown cultivars, and cracks which form on fast-growing trunks, favoring entry of the fungus into the inner bark and the occurrence of infections.

Vegetative Propagation of American Elm (Ulmus Americana) Varieties from Softwood Cuttings

Softwood cuttings of American elm varieties ‘Jefferson’, ‘New Harmony’, ‘Princeton’, ‘R18-2’, ‘Valley Forge’, and a tissue-cultured non-transformed control clone (BP-NT) were rooted using three different treatments to determine which method would be most suitable for small-scale propagation. The treatments included aeroponic chambers, an intermittent-mist bench in a greenhouse, and Grodan rootplugs soaked in a nutrient solution. The rootplug treatment had the highest percentage of rooted shoots (44%) followed by the intermittent-mist bench treatment (20%) and lastly by the aeroponics chambers (10%). The rooted cuttings from the rootplug treatment also looked substantially healthier and had more fresh growth four weeks after potting than the other two treatments. The Grodan rootplug treatment is recommended, but additional testing can be useful to improve the overall rooting percentage.

Cytotoxicity Is Not Required for the Anti-Leukemia Efficacy of Human H-Y-Specific CD4+ T Lymphocytes In Vivo.

Abstract 3553 Poster Board III-490 Introduction Minor histocompatibility antigens (mHag) play a major role in the graft-versus-leukemia (GvL) effect following HLA-matched allogeneic hemopoietic cell transplantation (allo-HCT). Clinically, the GvL effect coincides with the emergence of mHag-specific CD8+ cytotoxic T lymphocytes (CTL). Experimentally, targeting a single mHag with human CD8+ CTL has a major anti-leukemia effect in NOD/scid mice. Altogether, these observations suggest that mHag-specific cytotoxicity by CD8+ T cells is an important component of the GvL effect. In contrast, little is known on the contribution of mHag-specific CD4+ T cells. Female-to-male allo-HCT is characterized by a low rate of leukemia relapse, indicating that H-Y-encoded mHag are potent leukemia-regression antigens. Earlier, we described a DRB3*0301-restricted H-Y mHag epitope inducing CD4+ helper T-cell responses in H-Y-mismatched HLA-matched allo-HCT. Aim: The aim of this study is to elucidate the role of mHag-specific human CD4+ T lymphocytes on the GvL effect. Methods The ALL-CM leukemia cell line, derived from a male (i.e. H-Y+) HLA-A0201+, DRB30301+ patient, reproducibly engrafts in NOD/scid mice after administration of 10×106 cells. Both an HLA-A0201-restricted H-Y-specific CD8+ CTL clone and the DRB30301-restricted H-Y-specific CD4+ helper T-cell clone that we earlier described were used to investigate the anti-leukemia efficacy of CD8+ and CD4+ T cells in NOD/scid mice. Results In vitro, the CD8+ H-Y specific CTL clone was highly cytotoxic against the ALL-CM leukemia. The H-Y specific CD4+ helper T-cell clone did not lyse the leukemia, but produced IFN-γ upon recognition. Infusion of the H-Y-specific CD8+ CTL clone (25×106 cells/mouse) 3 days after ALL-CM leukemia challenge significantly delayed leukemia progression by 3 weeks compared to a CMV-specific CD8+ CTL control clone (p<0,001). Despite no measurable in vitro cytotoxicity, the H-Y-specific CD4+ helper T-cell clone (25×106 cells/mouse) delayed leukemia progression by 2 weeks compared to a leukemia non-reactive HLA-DR1-specific CD4+ helper T-cell control clone (p<0,001). In vitro co-incubation of the H-Y-specific CD4+ helper T-cell clone did not influence leukemia proliferation but induced up-regulation of MHC-class I and II, CD80, CD86 and CD40. In vitro, pre-incubation of leukemia cells with the H-Y-specific CD4+ helper T-cell clone irradiated did not improve the in vivo anti-leukemia efficacy of the H-Y-specific CD8+ CTL clone. Co-infusion of the H-Y specific CD4+ helper T-cell clone did not augment the in vivo persistence of the H-Y-specific CD8+ CTL T-cell clone. Nevertheless, the co-infusion resulted in a delay in leukemia progression of approximately 5 weeks, suggesting an additive, non overlapping anti-leukemia mechanism. Conclusions Minor Hag-specific human CD4+ T lymphocytes may contribute to the GvL effect through a direct, non cytotoxic mechanism, which could be additive to that of CD8+ CTL. The nature of this non cytotoxic GvL effect is currently under investigation. A.B. and L.H. equally contributed to this study. Disclosures: No relevant conflicts of interest to declare.

Performance of new Hevea clones from IAC 400 series

The Hevea breeding program of Instituto Agronômico de Campinas (IAC) has completed clonal evaluation on the following series: IAC 100, IAC 200 and IAC 300. The performance of 22 clones of Hevea brasiliensis (Willd. ex Adr. de Juss.) Muell.-Arg., evolved at IAC, over a period of eleven years was evaluated in the Western Central part of the São Paulo State, Brazil. Among these 22 new clones, six were intraspecific hybrid clones (IAC 400, IAC 404, IAC 405, IAC 406, IAC 410, IAC 412) and the remaining are primary those resulted from selected ortets within half-sib progenies. An old popular clone RRIM 600, of Malaysian origin, was used as the control. The trial was laid out in a randomized block design with three replications. Yield performance over a period of four years, mean girth at the 11th year, girth increment before tapping and on tapping, thermal property of natural rubber produced, bark thickness, number of latex vessel rows in seven year virgin bark, percentage incidence of tapping panel dryness, wind damage and diseases like leaf and panel anthracnose have been observed. Sixty one percent of the clones were superior in relation to the control for yield. The clone IAC 400 recorded the highest yield (97.40 g tree-1 tap-1) over four years of tapping, followed by IAC 411 (78.87 tree-1 tap-1), whereas the control clone RRIM 600 recorded 50.86 g tree-1 tap-1. All selected clones were vigorous in growth. Girth increment of these clones was average to above average. Except for IAC 423, other clones had thick virgin bark at opening ranging from 4.84 mm for IAC 401 to 6.38 mm for IAC 416. The natural rubbers from IAC clones have shown good thermal stability up to 300ºC and no differences in the thermal behavior among rubber from clones of the IAC series and the clone RRIM 600 were found in inert atmosphere.

The Role of EHZF in Erythroid Differentiation of K562 Cells.

Early hematopoietic zinc finger protein (EHZF), the human homolog to mouse Evi3, is a transcription factor with 30 zinc fingers and a highly conserved N-terminal FOG repression motif. The EHZF mRNA expression is abundant in early hematopoietic progenitors and declines during differentiation. In most of the leukemic cells from acute myelogenous leukemia patients, significant levels of EHZF mRNA are detected. These findings suggest that EHZF play an important role in hematopoietic differentiation. In the present study we investigated whether siRNA-mediated depletion of EHZF affected the erythroid differentiation of K562 cells. Three kind of EHZF siRNA were designed and their expression vectors were constructed. After transient expression of these siRNAs by electroporation, hemoglobinization of K562 cells were analyzed by Benzidine staining. In all of these siRNAs introduced cells, the levels of hemoglobinizations were abundant compared to control cells, which suggested that EHZF might influence the erythroid differentiation of K562 cells. To further analyze the effects of EHZF siRNA in erythroid differentiation, we established K562 clones in which EHZF was depleted by siRNA. EHZF siRNA expression vectors were introduced into K562 cells by electroporation and the expression of EHZF mRNA were examined by RT-PCR in G418 resistant clones. The depressions of EHZF mRNA in the clones were confirmed by Northern blotting. We isolated several EHZF depleted clones in each kind of EHZF siRNA introduced cells. Most of the EHZF depleted clones showed marked hemoglobinization by Benzidine staining compared to control clones, and surface expressions of glycophorin A were also increased in these clones. These results confirmed the relevant role of EHZF in erythroid differentiation of K562 cells. Comparison of mRNA expressions between an EHZF depleted clone and a control clone using DNA array showed increase expressions of hemoglobin alpha, beta, delta mRNA in EHZF depleted clone compared to control clone. Several transcription factors which are involved in erythroid differentiation also showed the difference mRNA levels between two clones. Studies are underway to elucidate the mechanisms underlying the role of EHZF in erythroid differentiation, the relationship to other transcription factors (GATA-1, GFi-1B, FOG-1 etc) which involved in erythroid differentiation.

Evaluation of Some Processing Characteristics of Cultivated Guava Clones

An evaluation of the processing characteristics of the fruit of cultivated guava clones was conducted in a two-stage study. The first stage consisted in pre-screening the cultivated clones using the percent soluble solids, pH, and color of the fruit pulp as criteria for selecting and combining the clones into representative samples. Eleven composite clonal samples were compared against a wild-fruit control. Pulp and nectar yields were determined and chemical analyses performed on each of the samples. Viscosity was used as a criterion for diluting the fruit pulps to nectar consistency. The canned nectar samples were appraised using a 9-point hedonic scale for scoring purposes. Nine of the 11 samples examined showed better stability during storage 85° F. (29.4° C.) than the control. Composite sample number 10 received the highest score and changed less in the 9-month storage period than other samples under study, also produced the highest pulp yield. In the second stage of the study, an individual evaluation was conducted on each of 13 new guava clones and a wild-fruit control. Pulp and nectar yields and chemical analyses were performed on each fruit sample. Pulp dilutions for nectar preparation were based on solid content rather than on viscosity. Samples were evaluated organoleptically using a 9-point hedonic scale. The keeping quality of seven of the cultivated guava clones appeared comparable to that of the wild fruit. Three of the samples, i.e., clones 57-9-114 (Rico 18), 57-2-142 (Rico 19), and 57-2-51 (Rico 20), were rated higher than the control. Clone number 57-2-142 (Rico 19) scored highest in the quality evaluation of 9-month storage at 85° F. (29.4° C). This clone also produced the second highest nectar yield among the 13 clones under evaluation and was superior to the wild-fruit control.