Commercial Clones Research Articles

Effects of Planting Density of Rubber Tree Clone (RRIM 2020 Clone and RRIM 2025 Clone) Wood to Particleboard Properties

The depletion of log resources encourages research into alternative ways to sustain the wood supply. Therefore, the 4-year-old Rubber Research Institute of Malaysia (RRIM) clones series, RRIM 2020 and RRIM 2025, were chosen as potential raw materials for particleboard in this study. The purpose of this study was to assess the effects of planting density and rubber tree clones on the mechanical and physical properties of single-layer particleboard. The planting densities used were low, moderate-low, moderate-high, and high, representing 500, 1000, 1500, and 2000 trees/ha, respectively. Prior to manufacturing, the RRIM 2000 series clone trees were harvested, cut, chipped, flaked, and screened. The mechanical and physical properties were evaluated in accordance with the Japanese Industrial Standard (JIS A 5908-2003). The findings revealed that both planting density and clone had a signifi- cant impact on the mechanical and physical properties of particleboard with a thickness of 10 mm and a density of 700 kg/m3 . RRIM 2020 specimens with low planting density had superior modulus of elasticity (MOE), modulus of rupture (MOR), and internal bonding (IB) values of 2415, 19, and 1.7 MPa, respectively. Furthermore, moderate-low planting density demonstrated the lowest thickness swelling (TS) and water absorption (WA) values and was comparable to control particleboard from commercial clone Prang Besar (PB), PB260. In terms of rubber clones, RRIM 2020 particleboard met the minimum requirements of the JIS standard for mechanical properties and outperformed RRIM 2025. This study recommended a low planting density of 500 trees/ha and the RRIM 2020 clone as a suitable raw material for particleboard manufacturing with a ten percent urea formaldehyde resin content.

Rootstock-Mediated Genetic Variance in Cadmium Uptake by Juvenile Cacao (Theobroma cacao L.) Genotypes, and Its Effect on Growth and Physiology.

Grafting typically offers a shortcut to breed tree orchards throughout a multidimensional space of traits. Despite an overwhelming spectrum of rootstock-mediated effects on scion traits observed across several species, the exact nature and mechanisms underlying the rootstock-mediated effects on scion traits in cacao (Theobroma cacao L.) plants often remain overlooked. Therefore, we aimed to explicitly quantify rootstock-mediated genetic contributions in recombinant juvenile cacao plants across target traits, specifically cadmium (Cd) uptake, and its correlation with growth and physiological traits. Content of chloroplast pigments, fluorescence of chlorophyll a, leaf gas exchange, nutrient uptake, and plant biomass were examined across ungrafted saplings and target rootstock × scion combinations in soils with contrasting levels of Cd. This panel considered a total of 320 progenies from open-pollinated half-sib families and reciprocal full-sib progenies (derived from controlled crosses between the reference genotypes IMC67 and PA121). Both family types were used as rootstocks in grafts with two commercial clones (ICS95 and CCN51) commonly grown in Colombia. A pedigree-based best linear unbiased prediction (A-BLUP) mixed model was implemented to quantify rootstock-mediated narrow-sense heritability (h2) for target traits. A Cd effect measured on rootstocks before grafting was observed in plant biomass, nutrient uptake, and content of chloroplast pigments. After grafting, damage to the Photosystem II (PSII) was also evident in some rootstock × scion combinations. Differences in the specific combining ability for Cd uptake were mostly detected in ungrafted rootstocks, or 2 months after grafting with the clonal CCN51 scion. Moderate rootstock effects (h2> 0.1) were detected before grafting for five growth traits, four nutrient uptake properties, and chlorophylls and carotenoids content (h2 = 0.19, 95% CI 0.05–0.61, r = 0.7). Such rootstock effects faded (h2< 0.1) when rootstock genotypes were examined in soils without Cd, or 4 months after grafting. These results suggest a pervasive genetic conflict between the rootstock and the scion genotypes, involving the triple rootstock × scion × soil interaction when it refers to Cd and nutrient uptake, early growth, and photosynthetic process in juvenile cacao plants. Overall, deepening on these findings will harness early breeding schemes of cacao rootstock genotypes compatible with commercial clonal scions and adapted to soils enriched with toxic levels of Cd.

Assessment of drought effects on survival and growth dynamics in eucalypt commercial forestry using remote sensing photogrammetry. A showcase in Mato Grosso, Brazil

The plantation of fast-growing tree species plays a crucial role in supplying forest products to growing economies while relieving pressure on native forests reducing the overexploitation of native ecosystems. Grown in more than 90 countries, Eucalyptus spp. are fast-growing tree species driving economic production comprising pulp, wood, or thermal energy. Climate change is increasing the frequency and severity of extreme drought episodes: the 2019 drought in Mato Grosso, Brazil, was the second driest episode ever recorded. Forest inventory data comprising thousands of individual tree measurements collected in consecutive surveys - years 2019, 2020, 2021 – were modelled with mixed-effects models to identify significant factors influencing tree mortality after drought. Time-series of growth dynamics of the four eucalypt commercial clones were tested. For the landscape assessment, surveys using unmanned aerial vehicle (UAV) supported a high-resolution image mosaicking to derive digital surface models and the vegetation indexes driving segmentation and classification methods using ground plot-level observations as training data. The impact of drought was measured in terms of mortality, growth dynamics and the financial losses computed when valuing the 1400-ha plantation along the 3-year series. The modelling results showed high-stocking plantations suffered more from extreme water-stress conditions. The interaction between stocking and clone was analysed. On average, the 2019 drought triggered an estimate loss of $3500 per hectare. The estimation of live and dead conditions using UAV-based outcomes reached an overall accuracy of 89% and 0.7 in the Cohen’s kappa used to map forest health wall-to-wall. Our findings advocate for a reduction of tree planting density and enhanced clone selection to turn eucalypt plantations more resilient to extreme water-stress. Combining UAV-based remote sensing technology, ground truth data and management expertise in eucalypt management eases the operationalization of data-driven solutions towards more resilient silvicultural guidelines and increased efficiency of forest plantations.

Salix spp. Bark Hot Water Extracts Show Antiviral, Antibacterial, and Antioxidant Activities-The Bioactive Properties of 16 Clones.

Earlier studies have shown that the bark of Salix L. species (Salicaceae family) is rich in extractives, such as diverse bioactive phenolic compounds. However, we lack knowledge on the bioactive properties of the bark of willow species and clones adapted to the harsh climate conditions of the cool temperate zone. Therefore, the present study aimed to obtain information on the functional profiles of northern willow clones for the use of value-added bioactive solutions. Of the 16 willow clones studied here, 12 were examples of widely distributed native Finnish willow species, including dark-leaved willow (S. myrsinifolia Salisb.) and tea-leaved willow (S. phylicifolia L.) (3 + 4 clones, respectively) and their natural and artificial hybrids (3 + 2 clones, respectively). The four remaining clones were commercial willow varieties from the Swedish willow breeding program. Hot water extraction of bark under mild conditions was carried out. Bioactivity assays were used to screen antiviral, antibacterial, antifungal, yeasticidal, and antioxidant activities, as well as the total phenolic content of the extracts. Additionally, we introduce a fast and less labor-intensive steam-debarking method for Salix spp. feedstocks. Clonal variation was observed in the antioxidant properties of the bark extracts of the 16 Salix spp. clones. High antiviral activity against a non-enveloped enterovirus, coxsackievirus A9, was found, with no marked differences in efficacy between the native clones. All the clones also showed antibacterial activity against Staphylococcus aureus and Escherichia coli, whereas no antifungal (Aspergillus brasiliensis) or yeasticidal (Candida albicans) efficacy was detected. When grouping the clone extract results into Salix myrsinifolia, Salix phylicifolia, native hybrid, artificial hybrid, and commercial clones, there was a significant difference in the activities between S. phylicifolia clone extracts and commercial clone extracts in the favor of S. phylicifolia in the antibacterial and antioxidant tests. In some antioxidant tests, S. phylicifolia clone extracts were also significantly more active than artificial clone extracts. Additionally, S. myrsinifolia clone extracts showed significantly higher activities in some antioxidant tests than commercial clone extracts and artificial clone extracts. Nevertheless, the bark extracts of native Finnish willow clones showed high bioactivity. The obtained knowledge paves the way towards developing high value-added biochemicals and other functional solutions based on willow biorefinery approaches.

Development of Astatine-211 (211At)-Based Anti-CD123 Radioimmunotherapy for Acute Leukemias and Other CD123+ Hematologic Malignancies

Background: Radioimmunotherapy (RIT) has long been pursued to improve outcomes in acute leukemia. Of current interest are alpha-particle emitting radionuclides as they deliver a very large amount of radiation over just a few cell diameters, enabling efficient and selective target cell kill. So far, alpha-emitters including astatine-211 (211At) have been primarily explored with monoclonal antibodies (mAbs) targeting CD45 or CD33 but their broad display on non-malignant target-expressing cells can lead to marked “on-target, off tumor cell” toxicities. To overcome this limitation, we developed a novel form of 211At-based RIT targeting CD123. CD123 is displayed widely on acute leukemia cells, including underlying leukemic stem cells, but is expressed only on a discrete subset of normal hematopoietic cells and is virtually absent on non-blood cells.Methods: We immunized BALB/c mice with peptides consisting of the extracellular domain of human CD123 to generate anti-CD123 mAbs. Flow cytometry-based assays with human acute leukemia cell lines were used to characterize binding of hybridoma supernatants and mAbs to CD123. mAbs were conjugated with isothiocyantophenethyl-ureido-closo-decaborate(2-) (B10), a boron cage molecule for subsequent astatination, and were then labeled with 211At. In vivo leukemia cell targeting (“biodistribution”) and efficacy studies were conducted in immunodeficient NOD-Rag1 null IL2rɣ null/J (NRG) mice xenografted with MOLM-13 cells, a CD123+ human acute myeloid leukemia cell line.Results: Based on initial hybridoma screening studies, we selected 4 mAbs (10C4, 5G4, 11F11, and 1H8) for further characterization. Phenotyping studies with CD123+ and CD123- human acute leukemia cell lines (including CD123+ cell lines in which CD123 was deleted via CRISPR/Cas9) confirmed specific binding of all mAbs to human CD123 (binding intensity: 10C4>5G4=11F11=1H8), with 10C4 yielding a higher median fluorescence intensity than the widely used commercial anti-CD123 mAb clones, 7G3 and 6H6 (Figure 1). In vitro internalization with a panel of human acute leukemia cell lines studies demonstrated uptake of all mAbs by CD123+ target cells with a kinetic slower than that for anti-CD33 antibodies (typically, 30-50% of the anti-CD123 mAb internalized over 2-4 hours). All 4 anti-CD123 mAbs could be conjugated to B10 and subsequently labeled with 211At. Unlike a non-binding 211At-labeled control mAb, 211At-labeled anti-CD123 mAbs showed uptake at MOLM-13 flank tumors in NRG mice carrying MOLM-13 xenografts. After additional leukemia cell targeting studies to optimize the dosing of 10C4, we conducted proof-of-concept efficacy studies in NRG mice injected intravenously with luciferase-transduced MOLM-13 cells (disseminated leukemia model). Animals were either untreated or treated with 10 µCi, 20 µCi, or 40 µCi of 211At-labeled 10C4-B10 mAb (9-11 animals/group). This was followed by the infusion of bone marrow cells from donor mice as stem cell support 3 days later. As shown in Figure 2 and Figure 3, 211At-10C4-B10 led to a dose dependent decrease in tumor burden. Further, the treatment significantly prolonged survival compared to untreated animals (median survival: 49 days [40 µCi of 211At] vs. 31 days [10 µCi of 211At] vs. 21 days [Ctrl]; P<0.0001 for Ctrl vs. 10 µCi, P<0.004 for 10 µCi vs. 40 µCi), demonstrating potent in vivo anti-leukemia efficacy of a single dose of 211At-CD123 RIT.Conclusion: Our data support the further development of 211At-CD123 RIT for the treatment of patients with acute leukemia and other CD123+ hematologic malignancies. [Display omitted] DisclosuresGreen: Bristol Myers Squibb: Membership on an entity's Board of Directors or advisory committees, Patents & Royalties, Research Funding; Cellectar Biosciences: Research Funding; GSK: Membership on an entity's Board of Directors or advisory committees; JANSSEN Biotech: Membership on an entity's Board of Directors or advisory committees, Research Funding; Juno Therapeutics: Patents & Royalties, Research Funding; Legend Biotech: Consultancy; Neoleukin Therapeutics: Membership on an entity's Board of Directors or advisory committees; Seattle Genetics: Membership on an entity's Board of Directors or advisory committees, Research Funding; SpringWorks Therapeutics: Research Funding. Walter: Kite: Consultancy; Janssen: Consultancy; Genentech: Consultancy; BMS: Consultancy; Astellas: Consultancy; Agios: Consultancy; Amphivena: Consultancy, Other: ownership interests; Selvita: Research Funding; Pfizer: Consultancy, Research Funding; Jazz: Research Funding; Macrogenics: Consultancy, Research Funding; Immunogen: Research Funding; Celgene: Consultancy, Research Funding; Aptevo: Consultancy, Research Funding; Amgen: Research Funding.

Artificial light improves productivity of mini-cuttings in a clonal minigarden of Eucalyptus benthamii and E. dunnii

Eucalypts are economically important worldwide, being largely propagated by clonal means, that is, frequently achieved by mini-cuttings. Regional climatic conditions, however, interfere with this strategy. Southern Brazil, especially in higher altitudes, has a considerable number of cloudy and cold days annually. It is a challenge for clonal propagation, even for cold-tolerant species such as Eucalyptus benthami and E. dunnii. The supplementation with artificial light has proved to be effective for improving growth in agricultural crops, but little work has been done with trees. Here we conducted a set of experiments for producing mini-cuttings of two commercial clones of E. benthamii and one clone of E. dunnii under the supplementation with artificial light, using red/far-red (660:730 nm) and red/blue (660:450 nm) light emitting diodes (LED), in comparison to a control (sunlight only). The supplementation with red/far-red and red/blue light increased the production of mini-cuttings in colder months, but more consistently positive impacts were observed with red/far-red lamps. The shoots produced under red/far-red light were more elongated and implicated in higher leaf canopy coverage. Overall, this work is a first step aimed at improving clonal propagation systems of E. benthamii and E. dunnii through mini-cuttings supplemented with artificial light.

Prospecting Endophytic Bacteria Endowed With Plant Growth Promoting Potential Isolated From Camellia sinensis.

Endophytes are well-acknowledged inoculants to promote plant growth, and extensive research has been done in different plants. However, there is a lacuna about the endophytes associated with tea clones and their benefit to promote plant growth. The present study focuses on isolating and characterizing the beneficial endophytic bacteria (EnB) prevalent in commercially important tea clones cultivated in North Eastern India as plant growth promoters. Diversity of culturable EnB microbiome, in vitro traits for plant growth promotion (PGP), and applicability of potent isolates as bioinoculant for in vivo PGP abilities have been assessed in the present study. A total of 106 EnB identified as members of phyla Proteobacteria, Firmicutes, and Actinobacteria were related to 22 different genera and six major clusters. Regarding PGP traits, the percentage of isolates positive for the production of indole acetic acid, phosphate solubilization, nitrogen fixation siderophore, ammonia, and 1-aminocyclopropane-1-carboxylic acid deaminase production were 86.8, 28.3, 78.3, 30.2, 95.3, and 87.7, respectively. In total, 34.0, 52.8, and 17.0% of EnB showed notable production of hydrolytic enzymes like cellulase, protease, and amylase, respectively. Additionally, based on the bonitur score, the top two isolates K96 identified as Stenotrophomonas sp. and M45 identified as Pseudomonas sp. were evaluated for biofilm formation, motility, and in vivo plant growth promoting activity. Results suggested strong biofilm formation and motility in K96 and M45 which may attribute to the colonization of the strains in the plants. Further in vivo plant growth promotion experiment suggested sturdy efficacy of the K96 and M45 as plant growth promoters in nursery condition in commercial tea clones Tocklai vegetative (TV) TV22 and TV26. Thus, this study emphasizes the opportunity of commercialization of the selected isolates for sustainable development of tea and other crops.

Cacao (Theobroma cacao L.) Response to Water Stress: Physiological Characterization and Antioxidant Gene Expression Profiling in Commercial Clones.

The increase in events associated with drought constraints plant growth and crop performance. Cacao (Theobroma cacao L.) is sensitive to water deficit stress (DS), which limits productivity. The aim of this research was to characterise the response of seven (CCN51, FEAR5, ICS1, ICS60, ICS95, EET8, and TSH565) commercially important cacao clones to severe and temporal water deficit stress. Ten-month-old cacao trees were submitted to two treatments: well-watered and water-stressed until the leaf water potential (Ψleaf) reached values between −3.0 and −3.5 MPa. The effects of hydric stress on water relations, gas exchange, photochemical activity, membrane integrity and oxidative stress-related gene expression were evaluated. All clones showed decreases in Ψleaf, but TSH565 had a higher capacity to maintain water homeostasis in leaves. An initial response phase consisted of stomatal closure, a general mechanism to limit water loss: as a consequence, the photosynthetic rate dropped by approximately 98% on average. In some clones, the photosynthetic rate reached negative values at the maximum stress level, evidencing photorespiration and was confirmed by increased intracellular CO2. A second and photosynthetically limited phase was characterized by a drop in PSII quantum efficiency, which affected all clones. On average, all clones were able to recover after 4 days of rewatering. Water deficit triggered oxidative stress at the early phase, as evidenced by the upregulation of oxidative stress markers and genes encoding ROS scavenging enzymes. The effects of water deficit stress on energy metabolism were deduced given the upregulation of fermentative enzyme-coding genes. Altogether, our results suggest that the EET8 clone was the highest performing under water deficit while the ICS-60 clone was more susceptible to water stress. Importantly, the activation of the antioxidant system and PSII repair mechanism seem to play key roles in the observed differences in tolerance to water deficit stress among clones.

Technological potential of fibers from 20 Hevea brasiliensis clones for use as pulp, paper, and composite materials

Based on the use of Hevea brasiliensis in latex production, the species has a consolidated role in the Brazilian economy. However, at the end of the production cycle, which lasts from 25 to 30 years, resulting wood, in general, has no added value and is normally used for firewood, without further exploring its technological properties. Seeking to introduce this species into the pulp and paper industry, we aimed to determine the fiber quality of 20 commercial clones. Wood samples were collected from planted trees (11 to 12 years old) in the municipality of Selvíria - MS. We calculated wood quality indexes of H. brasiliensis for cellulose and paper, including Flexibility coefficient, Wall proportion, Runkel ratio, Slenderness ratio (Aspect ratio), and Luce's Shape Factor. The fiber quality indexes that best indicate the potential use of wood from Hevea brasiliensis clones for cellulose and paper production were Wall Fraction, Runkel Ratio and Luce's Shape Factor, indicating that H. brasiliensis fibers, if used for proper industrial purposes, will give origin of a rigid paper with greater bonding contact on the surface. The clones that presented better results for fiber quality indexes were IAC 311, IAC 41, IAN 873, IAC 326, IAC 40 and RRIM 725. Enabling the use of wood after the latex exploratory cycle for pulp and paper will serve as a basis for genetic improvement of these clones and expansion of forest plantations for this purpose.

Abstract 1671: Evaluation of total PD-1 expression using multi-color flow cytometry in metastatic non-small Cell lung cancer patients treated with multi-neoantigen vector (ADXS-503) alone and in combination of pembrolizumab to assess T-cell &amp; T-cell memory subsets

Abstract Precision for Medicine (Precision) developed and qualified two 17 color flow immunophenotyping assays to be used as pharmacodynamic biomarkers for Advaxis clinical studies in patients with Metastatic Non-Small Cell Lung Cancer treated with ADXS-503 alone and in combination with Pembrolizumab (Pembro). ADXS-503 (A503) is an off-the-shelf, attenuated Listeria monocytogenes (Lm)-based immunotherapy bioengineered to elicit potent T-cell responses against 22 tumor antigens commonly found in NSCLC. Pembrolizumab (Pembro) is a programmed death receptor-1 (PD-1)-blocking antibody approved for the treatment of advanced lung cancer. A503 and Pembro have complementary mechanisms of immune activation and reversal of immune tolerance. Here, we qualified two multi-color flow cytometry assays to quantify total PD-1 expression in cryopreserved peripheral blood mononuclear cells (PBMCs) from individuals that were treated either with A503 only or with A503 in combination with Pembro. The detection of free PD-1 and Pembro-bound PD-1 was achieved by co-staining a partially competing αPD-1 antibody (clone PD1.3.1.3) with a biotinylated αHu-IgG4 antibody. The robustness of the assay was demonstrated using a nine-point half-log serial dilution of Pembro, where the highest concentration was 10µg/mL and the lowest concentration was 0.001µg/mL, including a no drug control. The assay conditions were optimized for sensitivity, optimal signal:noise ratio, detection of free and drug bound receptor by titrating and testing various commercial αPD-1 antibody clones and tertiary reagents to detect biotinylated αHu-IgG4. The Pembro bound receptor was detected using a biotinylated αHu-IgG4 antibody, while the free receptors were quantified using a commercial αPD-1 antibody. The assay was able to quantify free and drug bound PD-1 in the intended immune cell types without compromising the staining of other cell surface and intra-nuclear markers. Majority of the evaluable patients, 6 out of 8, had increased counts of NK, CD4+ and CD8+ T-cells, including TCM, TEM and memory stem cells after the administration of ADXS-503 ± Pembro. PD1 expression on circulating CD4+, CD8+ and NK T-cells was also increased while PD-L1 expression was elevated in on-therapy tumor biopsies in some of these patients. Measuring total PD-1 in T-cells can be more challenging in patients on Pembrolizumab therapy as no known commercial non-competing αPD-1 antibody clones are available. This novel assay will facilitate the evaluation of total PD-1 expression as a pharmacodynamic biomarker in T-cells when PD-1 blockade is being used. These results also support that combination of ADXS-503 with PD-1 blockade could lead to enhancement of efficacy of anti-tumor immunotherapy. Citation Format: Venkat Mohanram, Natalya Belkina, Angelina R. Bisconte, Jonathan W. Goldman, Gregory J. Gerstner, Missak Haigentz, Thomas Stinchcombe, Balazs Halmos, Surya Vangala, Victor Kabala, Dinesh Simkhada, Cristiane Metran, Darren Davis, Megan Parsi, Andres A. Gutierrez, Deborah Phippard, Suresh S. Ramalingam. Evaluation of total PD-1 expression using multi-color flow cytometry in metastatic non-small Cell lung cancer patients treated with multi-neoantigen vector (ADXS-503) alone and in combination of pembrolizumab to assess T-cell &amp; T-cell memory subsets [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1671.

Abstract 2678: Tumoral CXCR4 and CXCL12 but not CXCR7 nor PD-L1 expression predicts disease free survival (DFS) and cancer specific survival (CSS) in resected pancreatic adenocarcinoma patients

Abstract Introduction: Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive tumor types and the 4th leading cause of tumor-related mortality in developed nations. PDAC is characterized by a highly fibrotic stroma that can physically exclude cytotoxic T cells from the vicinity of tumor cells. Targeting the axis CXCR4-CXCL12 improved T cell access to PDAC Tumor Microenvironment (TME) making tumors more susceptible to anti-PD-1/anti PD-L1 therapy. With the intent to evaluate the role of the entire CXCR4-CXCL12-CXCR7 axis in a context of PD-1/PD-L1 expression, 80 primary pancreatic cancers were evaluated in cancer cells and TME. Methods: 80 primary PDAC samples from patients undergone surgery collected at Ospedale P. Pederzoli Peschiera del Garda (Verona, Italy) were included in the study. Immunohistochemistry was conducted using primary antibodies: Anti-CXCR7/RDC-1 clone 11G8; Anti-CXCL12 Clone #79018; two commercial anti-CXCR4 clones #44716 and the UMB-2; two commercial anti-PD-L1, clone E1L3N and 22C3. Results: CXCR4 was expressed in 53 out of 73 tumors (72.6%). CXCL12 was predominantly identified into the membrane of cancer cells in 16 out of 75 (21.3%) tumors. CXCR7 staining was detectable in 44 out of 76 (57.8%) tumors. PD-L1 identified into the membrane of cancer cells in 35 out of 70 (50%) tumors. Patients with high CXCR4 tumor expression experienced worse outcome (DFS: 11.77 vs 22.85 months p=0.0071; CSS: 21.83 vs 35 months p&amp;gt;0.05, not significant) and predicted short DFS in R0 patients subgroup (n=48) (DFS: 11.20 months vs 33.40 months p=0.0140). CXCL12 expression predicted DFS (p=0.0002) and CSS (p=0.0046). Of note CXCL12 predicted short DFS (p=0.0015) and CSS (p=0.0213) in R0 patient's subgroup (n=48). Although stromal expression of CXCR4, CXCL12 and CXCR7 were not prognostic, CXCR4 positive inflammatory cells and tumoral CXCL12 significantly and directly correlated with the vascular invasion. Neither tumoral nor stromal expression of PD-L1 was prognostic but PD-L1 interestingly stained the perineural invasion and tumor-infiltrating inflammatory cells of body or tail tumors. In multivariate analysis, tumoral CXCR4 expression (p=0.0043), perineural invasion (p=0.0030) and AJCC lymph node status (p=0.0016) were independent prognostic factors for DFS. Tumoral CXCL12 expression (p=0.01467), AJCC Stage (p=0.0004) and perineural infiltration (p=0.03164) were independent prognostic factors for CSS. Conclusion: Tumoral CXCR4 and CXCL12 were prognostic factors at the univariate and multivariate analysis as for DFS and CSS in PDAC. Stromal PD-L1 stained perineural infiltration and stromal CXCR4 characterized inflammatory cells in the vascular invasion. Thus tumoral and stromal characterization for CXCR4 and CXCL12 in PDAC is worth to identify prognostic categories and to orient therapeutic approach. Citation Format: Crescenzo D'Alterio, Alessandro Giardino, Giosuè Scogmaliglio, Fabiana Tatangelo, Giovanni Butturini, Luigi Portella, Giuseppe Guardascione, Isabella Frigerio, Stefano Gobbo, Gerardo Botti, Stefania Scala. Tumoral CXCR4 and CXCL12 but not CXCR7 nor PD-L1 expression predicts disease free survival (DFS) and cancer specific survival (CSS) in resected pancreatic adenocarcinoma patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2678.

Biomass yield, dry matter partitioning and physiology of commercial and Erianthus introgressed sugarcane clones under contrasting water regimes

The productivity of sugarcane is limited by several biotic and abiotic stresses, of which drought is an important environmental stress, causing substantial yield losses especially in tropical parts of the world. The present day sugarcane cultivars are derivatives of interspecific hybrids involving Saccharum officinarum and S. spontaneum and the drought tolerance is governed by S. sponaneum. Of late considerable attention has been given to utilize Erianthus Spp, the related wild genera of Saccharum, shows remarkable levels of drought tolerance under water limited condition. In this study, experiments were conducted to compare the influence of drought on biomass, yield and physiology of commercial and Erianthus introgressed clones. The results showed that the cane yield was reduced 37.98% in commercial clones compared to 24.67% in Erianthus introgressed clones under drought. The highest stress tolerance index (STI) of 1.256 was recorded in the clone CYM 08–922 which is a backcross hybrid involving Erianthus arundinaceus. Significant differences were observed for leaf area index, relative water content, lipid peroxidation and epicuticular wax content among the genotypes and between treatments. Sugar yield was most affected by drought (38.86%) followed by cane yield (33.30%), NMC (26.79%) and single cane weight (22.49%). Cane yield showed a positive and significant correlation with dry biomass under both irrigated (r = 0.303) and drought situations (r = 0.404) and among the physiological parameters, only RWC showed significant and positive correlation with cane yield (r = 0.307). The leaf dry matter was 14.55% higher under drought indicating influence of drought on translocation of dry matter from leaves to canes. Among clones CoM 0265, Co 06022, CYM 08-922, CYM 09-1369 and GU 12-31 showed greater ability to partition dry matter in to canes under drought than the other clones and had higher stress tolerant index. Cane height, number of millable stalks and RWC% showed positive and significant association with cane yield under both drought and irrigated condition and could be important parameters in screening clones under sugarcane improvement programmes.

Sexual compatibility in cacao clones drives arrangements in the field leading to high yield

Sexual self-incompatibility is a very salient trait of woody plants, including cacao (Theobroma cacao L.). In this species, most commercial clones are self-incompatible and, therefore, productivity depends on pollen flow for fruit formation due to the combinatorial ability of each clone to accept or reject pollen tubes in the embryo sac. To determine the combinatorial ability of commercial cacao clones, artificial (manual) pollinations were performed between cacao clones of the same and between groups for three years. In total, 46 cacao clones of five geographical groups were evaluated: ‘FHIA’ from Honduras, ‘ICS’ from Trinidad and Tobago, ‘UF’ from Costa Rica, ‘CAUCASIA’ from Colombia, and ‘EET’ from Ecuador. The results showed that, except for ‘CAUCASIA’, there is a high level of inter-compatibility between groups. Cacao clones exceed the established threshold of 30% in Fruit Set Success when used as a female or a male donor. As expected, low self-compatibility rates were found among the clones studied. We propose the use of sexual inter-compatibility information for the design of planting arrangements to maximize cacao yields.

Competitive ability among Eucalyptus spp. commercial clones in Mato Grosso do Sul state

In general, genetic improvement programs for Eucalyptus seek to increase productivity. Diallelic methods are commonly used in crossbreeding designs, but few studies use this methodology to assess competitive ability. The aim of this study was to estimate the competitive capacity and effects of intra- and inter-genotypic in Eucalyptus spp. clones and identify combinations that support increased productivity. To do this, 12 commercial clones were evaluated in a competitive trial located in Três Lagoas, Mato Grosso do Sul State, Brazil, established in a randomized block design, with three replicates and five plants per plot. Data for the mean annual increment (MAI) obtained at five years of age were assessed using two statistical approaches. Groups were classified according to specific (ability of the two clones to compete each other) and general competitive ability (average ability of a clone to compete with others). There were differences in the genotypes' ability to suffer or exercise competition. The results obtained showed the best combinations resulted in up to 10% increase in productivity. That combined use of diallelic analysis with the statistical approach which allows for the assessment of the effects of intra- and inter-clonal competition, may be useful for Eucalyptus spp. improvement programs, especially in terms of identifying the best combinations for planting when the objective is to obtain gains in crop productivity.

Establishment of Regional Phytoremediation Buffer Systems for Ecological Restoration in the Great Lakes Basin, USA. II. New Clones Show Exceptional Promise

Poplar tree improvement strategies are needed to enhance ecosystem services’ provisioning and achieve phytoremediation objectives. We evaluated the establishment potential of new poplar clones developed at the University of Minnesota Duluth, Natural Resources Research Institute (NRRI) from sixteen phytoremediation buffer systems (phyto buffers) (buffer groups: 2017 × 6; 2018 × 5; 2019 × 5) throughout the Lake Superior and Lake Michigan watersheds. We divided clones into Experimental (testing stage genotypes) and Common (commercial and/or research genotypes) clone groups and compared them with each other and each NRRI clone (NRRI group) at the phyto buffers. We tested for differences in clone groups, phyto buffers, and their interactions for survival, health, height, diameter, and volume from ages one to four years. First-year survival was 97.1%, with 95.5%, 96.2%, and 99.6% for the 2017, 2018, and 2019 buffer groups, respectively. All trees had optimal health. Fourth-year mean annual increment of 2017 buffer group trees ranged from 2.66 ± 0.18 to 3.65 ± 0.17 Mg ha−1 yr−1. NRRI clones ‘99038022’ and ‘9732-31’ exhibited exceptional survival and growth across eleven and ten phyto buffers, respectively, for all years. These approaches advance poplar tree improvement efforts throughout the region, continent, and world, with methods informing clonal selection for multiple end-uses, including phytotechnologies.

Relative performance of coppice versus seedlings of 16 eucalypt taxa over two rotations in northern coastal Zululand, South Africa

From the 1980s, Eucalyptus grandis × E. camaldulensis (GC) and E. grandis × E. urophylla (GU) were planted as alternatives to E. grandis for improved productivity in the Zululand coastal regions of South Africa. Although these hybrid combinations provided a degree of drought and/or disease tolerance together with the ability to regenerate the stand via the management of coppice growth, there was the need to test better-suited species for the drier, northern parts of coastal Zululand. Species, provenances and inter-specific hybrid clones of the genera Eucalyptus, Corymbia, Acacia and Casuarina were tested as alternatives to GC and GU in four site × species trials over one rotation (1996 to 2006). Although E. longirostrata, Corymbia henryi, C. citriodora subsp. variegata and E. argophloia showed promise as alternatives to GC and GU, their coppicing ability remained unknown. To determine this, one of the trials (Palm Ridge) was re-implemented and managed for regeneration via coppice over a second rotation (2R). Coppicing ability, growth and susceptibility to pests and disease data were compared for the 16 species at Palm Ridge over the two successive rotations (1R — planted = 8.9 years; 2R — coppiced = 8.2 years). Except for E. argophloia and one E. camaldulensis provenance, all treatments had equivalent, or improved stem stocking in 2R relative to 1R (trial mean: 1R = 1 174 stems ha-1; 2R = 1 288 stems ha−1). The two commercial hybrid clones had the highest volumes in both the 1R (GU = 174 m3 ha−1; GC 163 m3 ha−1) and 2R (GU = 149 m3 ha−1; GC 124 m3 ha−1) rotations. The mean annual volume increments (MAI in m3 ha−1 y−1) were not significantly different between rotations (1R = 10.74 m3 ha−1 y−1; 2R = 9.95 m3 ha−1 y−1), nor were there any significant rotation × treatment interactions indicating that the treatments performed in a similar manner over both rotations. Although there were differences among provenances within taxa, these were not as great as the differences amongst the taxa tested: GC/GC > C. citriodora subsp. variegata > E. logirostrata > C. henryi, with E. tereticornis, E. camaldulensis, E. argophloia and E. watsoniana not performing well over both rotations. Despite the higher rainfall recorded over 2R (837 mm y−1) relative to 1R (765 mm y−1), the incidence of pests and diseases that was not recorded in 1R most likely contributed to the reduced growth of the coppiced trees. Most notable of these were Leptocybe invasa on the GC, E. tereticornis and E. camaldulensis treatments, Botryosphaeria spp. on all but the Corymbia treatments, kino on the Corymbia treatments, and Coniothyrium spp on the GU treatment. Relative to the two commercial hybrid clones (GC and GU), E. longirostrata, and C. henryi were the most promising of the alternative species tested over both rotations (planted and coppiced), with C. citriodora subsp. variegata also showing promise. Although all treatments were affected by pest(s) or disease(s), some of the species showed resistance and provenances within species tolerance, indicating the potential for selection for a reduction in biotic risk.

Investigating the wood production and adaptability potential of Corymbia hybrid progeny in northern coastal Zululand, South Africa

During August 2013, two ‘new’ eucalypt taxa × site interaction trials were established in northern coastal Zululand. The main purpose of these trials was to investigate the commercial forestry potential of a range of Corymbia inter-specific hybrids for the region. Two sites contrasting in mean annual precipitation (850 mm and 1 155 mm respectively) and productivity potential, namely Mfezi (Mtubatuba) and Flatcrown (Kwambonambi), were selected for these trials. The 28 Corymbia hybrid seedlots comprised controlled crosses between C. torelliana and C. citriodora subsp citriodora, C. citriodora subsp variegata and C. henryi. Trees were assessed annually for the presence of pests and diseases and at mid-rotation (48 months) comprehensively measured for growth. By mid-rotation, none of the Corymbia hybrid families showed susceptibility to the suite of insect pests plaguing many of the commercial Eucalyptus hybrid clones in the region. Furthermore, no noteworthy disease infection symptoms were observed in any of the Corymbia or Eucalyptus species and hybrids in the trials. At the drier site of the two (Mfezi), at 24 months following 2 successive dry (<700 mm rainfall) years, trees of some of the Corymbia hybrid families manifested drought stress symptoms in the form of mild to moderate kino exudations from stems and branches. However, by mid-rotation, most of the Corymbia hybrid families in the trial showed excellent tree health, survival and stem volume relative to that of the commercial hybrid controls. At both sites, the seven top-ranking treatments for volume were Corymbia hybrid families. Based on the overall excellent growth performances of several of the Corymbia hybrid families in the two trials, more intensive investigations, for example in the areas of wood properties, vegetative propagation and clonal performance testing, would appear well-warranted.

Variation of growth and phenology traits in poplars planted in clonal trials in Northern Europe\u2014implications for breeding

The increased demand for wood to replace oil-based products with renewable products has lifted focus to the Baltic Sea region where the environment is favorable for woody biomass growth. The aim of this study was to estimate broad-sense heritabilities and genotype-by-environment (G×E) interactions in growth and phenology traits in six climatically different regions in Sweden and the Baltics. We tested the hypothesis that both bud burst and bud set have a significant effect on the early growth of selected poplar clones in Northern Europe. Provenance hybrids of Populus trichocarpa adapted to the Northern European climate were compared to reference clones with adaptation to the Central European climate. The volume index of stemwood was under low to medium genetic control with heritabilities from 0.22 to 0.75. Heritabilities for phenology traits varied between 0.31 and 0.91. Locally chosen elite clones were identified. G×E interactions were analyzed using pairwise comparisons of the trials. Three different breeding zones for poplars between the latitudes of 55° N and 60° N in the Baltic Sea Region were outlined. The studied provenance hybrids with origin from North America offer a great possibility to broaden the area with commercial poplar plantations in Northern Europe and further improve the collection of commercial clones to match local climates. We conclude that phenology is an important selection criterion after growth.

Low-density PD-1 expression on resting human natural killer cells is functional and upregulated after transplantation

Expression of programmed cell death protein 1 (PD-1) on natural killer (NK) cells has been difficult to analyze on human NK cells. By testing commercial clones and novel anti-PD-1 reagents, we found expression of functional PD-1 on resting human NK cells in healthy individuals and reconstituting NK cells early after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Peripheral blood samples from healthy individuals and transplant recipients were stained for PD-1 expression using the commercial anti-PD-1 clone PD1.3.1.3, fluorescein isothiocyanate (FITC)–labeled pembrolizumab, or an FITC-labeled single-chain variable fragment (scFv) reagent made from pembrolizumab. These reagents identified low yet consistent basal PD-1 expression on resting NK cells, a finding verified by finding lower PD-1 transcripts in sorted NK cells compared with those in resting or activated T cells. An increase in PD-1 expression was identified on paired resting NK cells after allo-HSCT. Blockade of PD-1 on resting NK cells from healthy donors with pembrolizumab did not enhance NK function against programmed death-ligand 1 (PD-L1)–expressing tumor lines, but blocking with its scFv derivative resulted in a twofold increase in NK cell degranulation and up to a fourfold increase in cytokine production. In support of this mechanism, PD-L1 overexpression of K562 targets suppressed NK cell function. Interleukin-15 (IL-15) activity was potent and could not be further enhanced by PD-1 blockade. A similar increase in function was observed with scFv PD-1 blockade on resting blood NK cells after allo-HSCT. We identify the functional importance of the PD-1/PD-L1 axis on human NK cells in which blockade or activation to overcome inhibition will enhance NK cell–mediated antitumor control.

Bark characterization of a commercial Eucalyptus urophylla hybrid clone in view of its potential use as a biorefinery raw material

The bark of an Eucalyptus urophylla hybrid clone used by the charcoal industry was studied regarding anatomy and chemical composition, including the characterization of its polar extracts, in view of its potential use. The bark is characterized by a dominant homogeneous phloem with little ray expansion and alterations in the non-conducting phloem and by a rhytidome with a large portion of dead phloem tissue between the periderms. The cell proportion was axial parenchyma 37.9%, fibers 26.8%, radial parenchyma 19.4%, expanded cells 12.2%, and sieve tubes elements (3.7%). The chemical composition was ash 2.99%, total extractives 19.49% (92% polar extractives), suberin 0.57%, total lignin 25.09%, and polysaccharides 39.90%. The carbohydrates were composed mainly of glucose and xylose (83.7% and 9.7% of all monosaccharides, respectively). Polar (ethanol-water) extracts had a high phenolic content of 324.4 mg of gallic acid equivalents/g (GAE/g) of extract, flavonoids 215.0 mg of catechin equivalents/g (CE/g) extract, and tannins 325.5 mg of catechin equivalents/g (CE/g) extract. The antioxidant activity was low, corresponding to 310.1 mg Trolox equivalents/g extract and an IC50 of 10.59. In view of an integrated potential use, the bark of this E. urophylla hybrid clone is interesting as a source of tannins and a subsequent use for glucose-based applications, while the high-fiber content also allows considering it as a fiber source.